Analysis of HLA-DQA1 in Japanese patients with type 1 diabetes mellitus, using DNA-PCR-RFLP typing. DNA-polymerase chain reaction(PCR)-RFLP(restriction fragment length polymorphism) typing of the DQA1 gene was performed on 39 patients with type 1 diabetes and 30 controls. Analysis of the frequency of the subtypes of DQA1 alleles, using the DNA-PCR-RFLP typing technique, showed that the DQA1*0301 subtype was most strongly associated with the disease (97.4% vs 56.7%, R.R. = 19.8, pc less than 0.00005). These results indicated that DQA1*0301 may determine the disease susceptibility in the Japanese. In addition, we analyzed the frequency of the subtypes of DQA1 genes in the ten patients carrying the DRW8-DQW8 haplotype, and found that at least eight of them (80%) were classified as having DQA1*0301. The DRW8-DQW8-DQA1*0301 may be one of the susceptible haplotypes among the Japanese.


Ontogenic expression of somatostatin-messenger RNA in the intestinal tract of neonatal rats. Ontogenic expression of somatostatin (SRIF) -messenger RNA (mRNA) in the gastrointestinal tract was examined in neonatal rats aged from 1 day preterm to 60 days postpartum in comparison with that in the hypothalamus. SRIF-mRNA in the hypothalamus was already expressed in prenatal rats and its developmental change was relatively small. In contrast, a unique pattern of SRIF-mRNA expression was seen in the different intestinal regions, gastric antrum, duodenum, jejunum and colon. In the duodenum, SRIF-mRNA level was low at birth, markedly increased during the postnatal 3 days and declined to the previous level by day 21. Jejunal SRIF-mRNA was found in neonates but progressively decreased in a similar way to duodenum. On the contrary, gastric SRIF-mRNA level, which was low during early development, rose rapidly to a peak on day 21 and gradually declined to an adult level. In the colon age-related change was not conspicuous, remaining at a low level. These results indicate that (1) expression of SRIF gene in the intestinal tract is regulated by local factor(s) as well as developmental stage, and (2) shift of SRIF-mRNA pattern occurs during weaning from the duodenum-dominant infantile pattern to the gastric-dominant adult pattern.


[The characteristics of the anxiolytic action of benzodiazepine and non-benzodiazepine tranquilizers in different tests of anxiety]. In experiments on rats the presence or absence was studied of the phenomenon of potentiation of anxiolytic action, estimated by the time of the animal stay in the light part of the chamber in tests of avoidance of "the lighted ground" or "menacing situation" at combined application of the pairs of substances of benzodiasepine and non-benzodiasepine series (elenium, indoter, campiron and campironin). Spectra of their neurochemical activity were determined in experiments on neurones of isolated spinal ganglia of rats with intracellular biopotentials records. It has been established that GABA-potentiating action of indoter and elenium, dopamine-negative action of campiron and campironin are significant in their anxiolytic action in the states of alarm of aversive genesis of different modalities. Serotoninmimetic effect of non-diasepine tranquilizers is of functional significance in the test of avoidance of "the lighted ground", but not of the "menacing situation". It is suggested that differences of neurochemical mechanisms of anxiolytic action of the tested tranquilizers testify to different neurochemical profiles of the neuronal "matrices" of the studied states of alarm.


Hypnotherapy for reflex sympathetic dystrophy. Reflex sympathetic dystrophy (RSD) is an unusual, debilitating, chronic pain syndrome thought to be the result of a continuous excessive discharge of regional sympathetic nerves. Supportive and stress-reduction psychotherapies are commonly recommended as adjunctive treatments. Biofeedback is a more direct symptomatic treatment. Although hypnotherapy is effective in altering sympathetic reflex and pain responses, there are no reports of its use for the treatment of RSD. This article reviews some promising results of hypnotherapy with three RSD sufferers. I discuss the role of hypnotherapy as a supportive adjunct to medical treatment. I also explore the possible role of hypnotherapy as a complementary treatment.


Incidence of liver toxicity associated with the use of flutamide in prostate cancer patients. The incidence of flutamide-related liver toxicity was studied in 1,091 consecutive patients treated for stage C or D prostate cancer with the antiandrogen flutamide and the luteinizing hormone-releasing factor (LHRH) agonist [D-Trp6, des-Gly-NH2(10)] LHRH ethylamide. Liver function tests, namely measurement of serum aspartate amino-transferase (AST) and alanine aminotransferase (ALT), total bilirubin, alkaline phosphatase, gamma-glutamyl transpeptidase (gamma-GT), and prothrombin and thromboplastin times, were performed at 4, 8, and 12 weeks and every 3 months thereafter. Clinical signs and symptoms of liver dysfunction were also sought. The causal link between the antiandrogen used and liver injury was assessed on the basis of the temporal relationship with the use of the drug in the absence of other possible causes and, in two patients, through rechallenge of the putative causative drug after a period of normalization of liver function. An increase in AST and ALT at fourfold or more above upper normal limits was observed in only four patients (0.36%). Total serum bilirubin and alkaline phosphatase were elevated in only one patient at 126 mmol/L and 640 IU/L, respectively. Among the four patients, only two developed clinical manifestations of liver disease (0.18%). Biopsy was performed in one patient, and the histopathologic findings showed a mixed pattern of cytotoxic and cholestatic changes. All clinical and biologic manifestations of liver toxicity rapidly disappeared upon discontinuation of flutamide alone. No sequelae were observed in the long-term follow-up at 18, 22, 31, and 62 months. The 1,087 remaining patients experienced no or mild (less than fourfold upper normal limit) and transient elevation in aminotransferase serum levels during the first 6 months of treatment, with normalization at later time intervals. Despite the fact that the cases reported so far, along with our large series, indicate that the incidence of flutamide-induced liver toxicity is very low, we recommend serial blood aminotransferase measurements at 2 and 4 weeks of treatment in order to detect early signs of possible flutamide-induced hepatic injury, thus avoiding the low potential risk of clinically significant liver toxicity.


The paradox of beta-adrenergic blockade for the management of congestive heart failure. To review the current data regarding the use of beta-adrenergic blockers for the treatment of congestive heart failure. Relevant studies published between 1975 and 1991 were reviewed. Key data from each study were extracted. The significance of conclusions reached by each author(s) was identified. beta-adrenergic blockade, although still considered an investigational therapy for the treatment of congestive heart failure, has been proven in several studies to improve ventricular function, including myocardial contractility and relaxation. In addition, since beta-blockade up-regulates myocardial beta-receptors, the myocardium becomes more responsive to graded doses of beta-agonists. Speculation regarding the possible mechanisms of these effects is presented. In addition, since beta-blockers have been shown to reduce neurohormonal activation, they may have a beneficial effect on survival. Although small pilot studies or subgroup analysis of larger studies suggest beta-blockade therapy improves survival in heart failure, this has yet to be proven. Large prospective trials are warranted to study this issue. As current data suggest, beta-blockers improve ventricular function and reduce neurohormonal activation in heart failure. beta-blockers should be considered as adjunctive therapy in patients with congestive heart failure. In addition, future studies are warranted to better elucidate their effects on ventricular function and survival.


Reassessment of the role of theophylline in the current therapy for nocturnal asthma. Symptoms of asthma commonly increase at night for a variety of reasons. While different options are available for the management of nocturnal asthma, theophylline has maintained a prominent role in treating this problem, and it is widely promoted as a first-line agent. Very recent reports stress the use of anti-inflammatory agents as the drugs of choice for the long-term treatment of asthma. Using the key words "asthma," "theophylline," "bronchodilators," "adrenal cortex hormones," "beclomethasone," "triamcinolone acetonide," and "disodium cromoglycate," the MEDLINE files were searched from 1982 to the present. Articles dating before 1982 were accessed from cross-reference of the more recent articles. Sustained-release theophylline preparations are effective in decreasing the rate of exacerbations of nocturnal asthma symptoms, but theophylline has risks of major toxicity and serum concentrations must be monitored. Inhaled corticosteroids are also useful in controlling nocturnal asthma, and they reduce inflammation, which is the basic problem in asthmatics. Inhaled anticholinergics and oral controlled-release albuterol are also helpful in reducing symptoms of nocturnal asthma. A suggested approach to managing nocturnal asthma includes corticosteroids inhaled through a spacer device at optimum doses, adding inhaled albuterol or oral sustained-release albuterol and then ipratropium with a spacer. If control is not maintained with this regimen, sustained-release theophylline may add benefit to inhaled steroid therapy in reducing nighttime asthma symptoms. Long-acting inhaled beta 2 agonists show promise and could be the adjunctive treatment of choice when they become available in the United States.


The synthesis and antineoplastic activity of 2'-deoxy-nucleoside-cyanoboranes in murine and human culture cells. The guanine, inosine, adenine and cytidine deoxyriboside cyanoboranes proved to be cytotoxic and possess in vivo antineoplastic activity against murine and human single cells and cultured cells derived from solid tumor lines. The agents preferentially inhibited DNA synthesis in Tmolt3 leukemic cells. The enzyme sites of drug inhibition of two active derivatives were DNA polymerase-alpha and de novo purine synthesis at the regulatory sites, PRPP amido transferase and IMP dehydrogenase. Moderate inhibition by the agents of TDP kinase and ribonucleoside reductase activities also occurred. Kinetic studies showed that IMP dehydrogenase activity was inhibited the earliest of all of the enzymes affected by the drugs. The d(ATP) pools were also reduced by drug treatment. DNA strand scission was evident after 24 hr incubation at 100 microM of drug. The 14C-cytidine-cyanoborane drug was rapidly taken up over 6 hr and most effective uptake was shown by rapidly dividing cells. The drug was bound to DNA, RNA and protein in Tmolt3 leukemic cells.


Prediction of drug resistance in human tumors using immunohistochemical techniques. An in vitro chemosensitivity testing procedure based on the immunodetection of protein products which reflect the expression of various tumor resistance mechanisms is proposed. This protocol may be of predictive value in choosing drugs which should be selected for chemotherapy, and avoiding drugs which might be expected to be ineffective due to the enhanced expression of specific resistance factors.


HER-2/neu amplification and overexpression in primary human breast cancer is associated with early metastasis. The etiology of human breast cancer is poorly understood and no specific marker of transformation has been identified. Amplification of HER-2/neu, as reported in a comprehensive study by Slamon et al, was found to be the most powerful predictor of disease-free and overall survival after the status of the axillary lymph nodes. Our study examines the HER-2/neu oncogene in 61 primary human breast cancers at both the DNA level (by Southern blotting) and the protein level (by immunohistochemical methods). Of the 61 tumors analyzed in our study, 17 (28%) had amplification of HER-2/neu. There was no significant correlation of HER-2/neu amplification with age, tumor diameter or hormone receptor status; however, amplification and overexpression of HER-2/neu was significantly correlated with the status of the axillary lymph nodes (P = 0.02). Of 16 patients with amplification of HER-2/neu, 14 (88%) had positive regional nodes. One of the two node negative cases with amplified HER-2/neu had bone marrow micrometastasis. Overall, 16 out of 17 (94%) tumors of the patients having amplified HER-2/neu had metastatic disease at the time of diagnosis. In summary, HER-2/neu amplification is associated with early tumor dissemination in primary human breast cancer and may be a marker of poor prognosis.


The genetics of multiple endocrine neoplasia syndromes. The familial nature of multiple endocrine neoplasia syndromes (FMEN) has been recognized for some time but little is known about their cause. Recent application of new molecular techniques has mapped FMEN syndromes to specific chromosomes and provided evidence for the mechanism through which neoplasia occurs. In this review, we describe the techniques that led to recent advances in our understanding of FMEN and we discuss the evidence supporting proposed molecular mechanisms of neoplasia.


Further characterization of two distinct adriamycin-resistant sublines from LoVo human colon carcinoma cells. We previously reported that two multidrug resistant sublines, AdR1.2 and SRA1.2, derived from LoVo human colon carcinoma cells, apparently expressed different resistance phenotypes including differential expression of p-glycoprotein (Pgp). Here, we further examined and compared other potential resistance mechanisms between AdR1.2 and SRA1.2 resistant cells. Our results showed that the Pgp-mediated AdR1.2 cells possessed an activated drug efflux pump and decreased nucleus binding of Adriamycin, while the non-Pgp-mediated SRA1.2 cells only held the second feature. Verapamil, however, partially reversed resistance in both sublines. Although glutathione-s-transferase was overexpressed in AdR1.2 but not in SRA1.2, both sublines had lower susceptibilities to drug-induced DNA strand breaks and greater capacities to repair such damage than did LoVo cells. These data suggest that, despite the differences in multidrug resistance phenotypes, the features of decreased susceptibility to DNA damage and enhanced DNA repair capacities may represent the common mechanisms responsible for drug resistance in both Pgp- and non-Pgp-mediated multidrug resistant cells.


Isolation and characterization of putative intrinsic multidrug resistant Chinese hamster ovary cells by fluorescence activated cell sorting. Most multidrug resistant cell lines reported in the literature were established by long-term continuous exposure of cells to stepwise increasing concentrations of antitumor drugs. However, these resistant cell lines may not be relevant to the majority of clinically resistant cells. In this study, we described the establishment of doxorubicin (Dox)-resistant Chinese hamster ovary cells by repeated flow cytometric cell sorting using the intrinsic fluorescence of Dox. In each sorting, the 15% least fluorescent cells were fractionated, grown to mass culture and sorted again. Results from a total of nine sorting cycles showed that the intracellular levels of Dox in the sorted cells were inversely proportional to the number of sorting cycles. The levels of P-glycoprotein mRNA in the sorted cells were increased, but reached a plateau of 2-3 fold after the fifth sorting cycle. The sorted cells exhibited a moderate but stable multidrug-resistant phenotype. Because the procedure involved minimal exposure of cells to the drug, the isolated cells are most likely related to naturally occurring (intrinsic) MDR cells.


A single 24h contact time with adriamycin provokes the emergence of resistant cells expressing the Gp 170 protein. We investigate here, in A549 cells, the influence of a single short (1h) or long (24h) adriamycin (ADR) contact time on the long-term cytotoxicity of the drug and on the emergence of resistant cells. In contrast to a 1 h ADR contact, a 24 h treatment provokes the emergence of resistant cells overexpressing the Gp170 protein and this overexpression is maintained for at least three months without any drug selection pressure.


Anticancer role of dendritic cells (DC) in human and experimental cancers--a review. The bone marrow-derived dendritic cells (DL/LC) are antigen-presenting accessory cells functioning as part of the immune system. In addition, DC/LC in epithelial tissues may have the capacity to be involved in cellular interactions which may have regulatory functions. Such properties can also be noted when LC/DC interact with cancer cells in tumors. The present review summarizes reports which suggest that the outcome of a primary tumor in patients depends on the presence or absence of DC/LC in the tumor. The evidence showing that the presence of DC/LC in primary tumors indicates that a good prognosis may be reached are presented and discussed. Based on these observations and the ability of immunomodulators to enhance the activity of DC/LC and the ability of these cells to enter into tumors, it is suggested that the molecular basis of DC/LC activity against primary tumors cells should be investigated. It is possible that activation of DC/LC, thereby enhancing their ability to enter primary tumors, and the abrogation of the ability of DC/LC-resistant tumors to destroy or prevent DC/LC from entering the tumor, could be developed as an effective anti-cancer approach.


Expression of a human multidrug resistance gene in human ovarian carcinoma cell lines. To investigate the possible role of the multidrug resistance phenotype to chemoresistance in human ovarian carcinoma, we have analyzed human multidrug resistance gene (mdr 1) expression in 8 human ovarian adenocarcinoma cell lines. An increase in P-glycoprotein level specific to multidrug-resistant tumor cells was not apparently associated with the increase in resistance to vincristine (VCR) or doxorubicin (Adriamycin). Mdr 1 transcripts (4.5 kilobases) were observed in the RNA preparation obtained from only one cell line (SHIN-3) that showed the highest resistance to both drugs in vitro and in vivo. No cell lines showed mdr 1 DNA amplification. These results suggest that the insensitivity of human ovarian carcinoma to chemotherapy could be partly explained by the expression of mdr 1.


Cell proliferation of transitional cell bladder cancer determined by PCNA/cyclin immunostaining. A histopathological description. The fraction of proliferating cells in 178 transitional cell carcinomas (TCCs) was determined by proliferating cell nuclear antigen (PCNA/cyclin) immunostaining. The fraction of positive nuclei ranged between 0% and 100%. In WHO grade 1 tumours only occasional cells were positive for PCNA/cyclin, whereas nearly all of the nuclei in WHO grade 3 tumours were positive for PCNA/cyclin (p less than 0.0001). Nodular tumours showed a higher growth fraction than papillary tumours (p less than 0.0001). Superficial Ta-T1 tumours showed a significantly lower fraction on nuclei positive for PCNA/cyclin than muscle invasive tumours (p less than 0.0001). Tumours with pelvic lymph node metastasis also had a high growth fraction (p less than 0.0001). The growth fraction as determined by PCNA/cyclin immunostaining was significantly related to aneuploidy (p less than 0.0001, S phase fraction (p less than 0.0001) and mitotic frequency (p less than 0.0001). The results show that cell proliferation in the context of histopathology can be assessed by PCNA/cyclin immunostaining in TCC. The results suggest that PCNA/cyclin immunostaining has prognostic value in TCC and consequently is clearly a subject for further studies as a prognostic variable.


Drug-induced lysosomal disorders in laboratory animals: new substances acting on lysosomes. Several substances with lysosomotropic activity were investigated in toxicological studies. AR-L 115 BS (sulmazol, a cardiotonic agent) was tested on beagle dogs; HX-CH 44 BS (a beta-blocker) and SX-AB 1316 SE (an antithrombotic agent) were tested on rats, and AF-CX 1325 XX (an antiepileptic agent) was tested on both rats and beagle dogs. All organ systems were examined morphologically by light and/or electron microscopy. When an increase in the number of lysosomes occurred this was confirmed by the pigment scheme according to Krutsay (1971) as well as by the detection of acid phosphatase and compared with earlier histochemical results. At higher dosages, all substances caused very marked proliferation of lysosomes in the liver and/or kidneys. HX-CH 44 BS also caused such proliferation in striated muscles and in the lungs. A brown discolouration of the kidneys was found with sulmazol and AF-CX 1325 XX. This finding corresponded to the microscopically detectable occurrence of numerous lipofuscin granules. The reticulum cells in the lymph nodes of dogs were also affected by AF-CX 1325 XX. It is concluded that the proliferation of lysosomes in various organs after administration of the above-mentioned substances is due to an excess of substance. The increased substance in the body is then stored in the lysosomes. With HX-CH 44 BS, lysosomal autodigestion of mitochondria in the skeletal musculature and in the alveolar macrophages of the lungs was found. The selective lysosomal incorporation of mitochondria has not been described up to now and in our opinion, this constitutes a special feature. The results otherwise largely correspond to those already described in the literature. Systemic phospholipidosis such as occurs with some other substances was not detectable. The incorporation of the substance causes several types of lysosomal inclusion. Uptake of the substance in lysosomes either leads to overt autodigestion of organelles such as mitochondria (HX-CH 44 BS) or peroxisomes or to residual lysosomes of dense structure which histochemically resemble lipofuscin. SX-AB 1316 SE serves as an example of a substance which is stored directly by lysosomes in crystalline form. Above all, in the liver the substance is taken up not only by the sinusoidal stellate cells but also by hepatocytes.


Cost of beta-adrenergic receptor blocking agents for ocular hypertension. We compared the cost per drop and cost per year of therapy for five commercially available ocular beta-blockers (Timoptic, Betagan, Betoptic, Betoptic S, and Optipranolol) based on retail prices in Philadelphia, Pa; New Orleans, La; and San Francisco, Calif. Variations in bottle volumes and drop size combined to yield up to a 48% differential in number of drops per bottle. Average retail price variations were 28% within brands (between pharmacies), 29% between brands, and 29% between regions. Yearly costs were consistently less with larger bottles. Factoring drops-per-bottle, cost of brand, and cost by pharmacy, the cost of a year's supply of beta-blocker in 5-mL bottles in the least expensive region (New Orleans) ranged from $52.25 for Optipranolol to $278.91 for Betagan (534% of the price for Optipranolol). The marked differential in yearly cost among antiglaucoma medications should perhaps be a factor in the recommendations made by physicians and health-care providers.


Microscopic polyarteritis presenting with chest infections and acute appendicitis. We describe a 38-year-old male with antineutrophil cytoplasmic auto-antibody (ANCA) positive microscopic polyarteritis who presented with recurrent chest infections, lung haemorrhage, renal insufficiency and acute appendicitis. Appendectomy was followed by resolution of abdominal symptoms and the surgical specimen revealed vasculitis of the serosal vessels. A renal biopsy was performed because of impaired renal function and this revealed focal necrotising glomerulonephritis with absence of immune deposits. Chest infections were treated with antibiotics resulting in partial clinical response, but pulmonary symptoms relapsed and a complete resolution was achieved only after plasma exchange and the administration of cyclophosphamide. Our observation emphasises the protean manifestations of microscopic polyarteritis and the relationship between ANCA and disease activity.


Animal models for immunodeficiency diseases. Immunodeficient animals continue to provide insights into the development and function of the immune system. These animals also serve as models for the propagation of normal and malignant human cells and as hosts for infectious agents, including HIV. New developments in basic and applied research using immunodeficient animals were discussed at a recent workshop.


Inhibition of synthesis of endothelium-derived nitric oxide in conscious dogs. Hemodynamic, renal, and hormonal effects. The role of endothelium-derived nitric oxide (EDRF/NO) for control of systemic and regional vascular resistances and for regulation of neurohumoral systems was investigated by studying the effects of the inhibitor of EDRF/NO-synthesis NG-nitro-L-arginine (L-NNA; 5 mg/kg) in six conscious dogs. L-NNA increased mean arterial pressure by an increase in total peripheral resistance, increased renal vascular, and total pulmonary resistances and reflexly decreased heart rate and cardiac output. Renal plasma flow, urine flow, and urinary sodium excretion were reduced, glomerular filtration rate was not affected. These changes were reversed by additional treatment with L-arginine (150 mg/kg). Plasma concentrations of renin, norepinephrine, vasopressin, and atrial natriuretic peptide were not changed by L-NNA. Our conclusions were that basal release of EDRF/NO plays an important physiologic role for control of systemic and regional vascular resistances, thereby controlling blood pressure, organ blood flow, and function. Neurohumoral systems are not affected by the inhibition of EDRF/NO synthesis and do not contribute to the observed vasoconstriction.


Heart rate changes evoked by hypoxia in the anaesthetized, artificially ventilated cat. Reflex changes in heart rate evoked by hypoxia were investigated in cats anesthetized with chloralose and ventilated by positive pressure during administration of vecuronium or gallamine. In five cats receiving vecuronium and with aortic pressure stabilized, systemic hypoxia (arterial O2 pressure (Pa, O2) 34.9 mmHg) reduced heart rate by 55.8 +/- 7.5 beats min-1 (mean +/- S.E.M.). After administration of atropine, hypoxia (Pa, O2 32.1 mmHg) increased heart rate by 28.2 +/- 3.4 beats min-1. After subsequent bilateral ablation of carotid sinus and vagus nerves, hypoxia (Pa, O2 31.9 mmHg) increased heart rate by 7.1 +/- 1.8 beats min-1. The cardiac accelerator response to hypoxia was further examined in groups of cats treated with gallamine and atropine. In four vagotomized cats, local perfusion of both carotid sinuses with hypoxic blood (Pa, O2 37.7 mmHg) increased heart rate by 15.5 +/- 2.3 beats min-1. In the same cats, systemic hypoxia (Pa, O2 38.3 mmHg) increased heart rate by 16.4 +/- 2.3 beats min-1. The heart rate increment in cats which had undergone either bilateral adrenalectomy or cardiac sympathectomy was similar to the increment in unoperated cats. The increment was significantly less in cats which had both adrenal glands and cardiac sympathetic nerves ablated. It is concluded that stimulation of the carotid bodies in the cat excites both parasympathetic and sympathetic cardiac nerves simultaneously.


Acute disseminated encephalomyelitis after Japanese B encephalitis vaccination. A 6-year-old girl (Patient 1) and a 5-year-old boy (Patient 2) with acute disseminated encephalomyelitis after Japanese B encephalitis vaccination are reported. Drowsiness, paresthesias, and gait disturbance were observed at 14 days (Patient 1) and 17 days (Patient 2) after the vaccination; however, transient impairment of visual acuity was only found in Patient 1. Laboratory examinations revealed slow theta waves on electroencephalography and elevated myelin basic protein in the cerebrospinal fluid in both patients. The most striking feature on magnetic resonance imaging was the combination of white matter lesions and abnormal intensity signals of the thalamus. The administration of oral prednisolone (2 mg/kg/day) markedly improved the clinical findings and abnormal magnetic resonance imaging findings. A similar magnetic resonance imaging finding of abnormal intensity of the thalamus with deep white matter lesions has been reported in patients with Japanese B encephalitis; therefore, thalamic lesions may be related to the naturally occurring encephalitis.


Gamma-glutamyltransferase, aspartate and alanine aminotransferases and their ratio, mean cell volume and urinary dolichol in pregnant alcohol abusers. To study the activities of serum gamma-glutamyltransferase (GGT), aspartate and alanine aminotransferases (AST, ALT), and their ratio (AST/ALT), mean erythrocyte cell volume (MCV) and urinary dolichol output in alcohol-abusing pregnant women, and compare the results to those of abstinent pregnant women. Prospective descriptive study. Special outpatient clinic for pregnant problem-drinkers in the department of Obstetrics and Gynaecology, Helsinki University Central Hospital, Finland. 25 pregnant women referred to the special clinic at between 12 and 24 weeks gestation, they consumed at least 150 g of ethanol weekly, and a control group of 20 abstinent pregnant women matched for age, parity and smoking habits. The women were encouraged to visit the clinic at 2-4 week intervals. At each visit blood and urine samples were obtained, and the women were interviewed on their alcohol consumption during the previous weeks and encouraged to abstain in the future. Neonatal condition, fetal alcohol effects (FAE) in the newborn. Serum activities of GGT, AST and ALT, the AST/ALT ratio, the MCV, and the urinary concentration of dolichol in alcohol-abusing women with either healthy or FAE infants, compared with those of abstinent women with healthy infants. Of the 25 alcohol-abusing women 13 gave birth to infants with FAE and 12 to healthy infants. All the women in the control group gave birth to healthy infants. GGT, AST and ALT activities were increased in all alcohol-abusing women, regardless whether the infant had FAE or not. GGT was the best of these markers, GGT activities above the 95th normal centile were found in 33% of the samples from all alcohol-abusing women. The AST/ALT ratio, MCV and urinary dolichol concentration were poor indicators of abusive drinking. Maternal alcohol abuse is difficult to assess by laboratory tests. Of the commonly used and easily available tests, GGT proved to be the best in our study.


Plasma concentrations of motilin and somatostatin are increased in late pregnancy and postpartum. To examine plasma levels of motilin and somatostatin throughout pregnancy. Prospective observational study. University Hospital, Norway. Eight healthy pregnant women (aged 24-38 years) six of them primigravidae and eight healthy non-pregnant women of similar age with ovulatory menstrual cycles. In the pregnant women blood samples were obtained at 4-week intervals from 8 weeks gestation throughout pregnancy and again at 5 days and 28 days postpartum. In the non-pregnant controls blood samples were obtained on cycle days 4, 7, 10, 13, 14, 15, 18, 21 and 24. Plasma levels of motilin and somatostatin. Plasma concentrations of both motilin and somatostatin rose continuously during pregnancy, and motilin levels increased still further to a peak of 165.1 (SE 35.8) pmol/l at 5 days postpartum. Plasma motilin levels were significantly higher during the third trimester and at 5 days postpartum compared with non-pregnant controls (P less than 0.0001). The highest plasma somatostatin levels were found at 40 weeks gestation and at 5 days postpartum (mean 32.1 SE 1.1 pmol/l). Somatostatin levels were significantly higher during the second and third trimester and the postpartum period compared with levels in the follicular phase of the non-pregnant controls (P less than 0.0001). Circulating levels of motilin cannot play a major role in the relaxation of the gut in pregnancy, but somatostatin may play a part in regulating motility.


Initiation and beyond: molecular determinants of gene regulation. Mechanisms of Transcription Control, A Jacques Monod Conference, sponsored by the Centre National de la Recherche Scientifique, Roscoff, France, September 30-October 4, 1991. The study of the mechanisms of transcriptional control continues to be an exciting area of research. The characterization of the constituents of the initiation complex and their interactions are leading to a greater understanding of gene regulation. The findings presented at this meeting emphasized the need to understand these interactions in three-dimensional space to effectively account for the observed regulation of the initiation of transcription.


Developmental and neurologic correlates of treatment response in schizophrenia. Reliable predictors of outcome in schizophrenia remain elusive, and assessment of unidimensional variables is unlikely to provide new information. We examined developmental, neurologic and psychosocial variables together to assess their correlation with several separate aspects of outcome in male schizophrenic patients (N = 31) treated with neuroleptics for a minimum of six months. Outcome measures evaluating social performance were significantly inter-correlated, but these measures did not correlate significantly with "positive" symptom measures. Persistent positive symptoms were predicted by post-natal neurologic impairment. Persistent negative symptoms and social dysfunction were predicted by psychosocial dysfunction during the developmental years. Poor early treatment response significantly correlated with persistent positive symptoms and psychosocial dysfunction. Impairment on neurobehavioral testing correlated significantly with post-natal neurologic impairment and with persistent positive symptoms.


The use of carbamazepine in the treatment of schizophrenic and schizoaffective psychoses: a review. This article reviews current literature on the clinical efficacy of carbamazepine (CBZ) administration in schizophrenic and schizoaffective psychoses. With respect to the use of CBZ in cases of aggression, overactivity and other behavioral dyscontrol syndromes, only a few, mainly open, studies have been conducted. Attention to the efficacy of CBZ in schizophrenia and related psychoses was rather late in developing, with most of the studies done since 1981. Although the results of the different controlled and uncontrolled experiments are very difficult to compare, the results generally indicate beneficial effects--particularly if CBZ is used as an adjunct to neuroleptic medication. Suggestions for future research strategies to maximize the usefulness of CBZ in schizophrenia and related disorders are given.


Cerebrospinal fluid concentrations of neuropeptide Y in depressed patients and in controls. Concentrations of Neuropeptide Y-like (NPY-LI) immunoreactivity in cerebrospinal fluid (CSF) were measured in a group of depressed patients (n = 24) and compared with that of control subjects (n = 12). CSF-NPY-LI was significantly reduced in the group of non-endogenously depressed patients when classified according to Newcastle Rating Scale for Depression--1971 (N-II). No significant correlation was found in the control or depressed groups between lumbar concentrations of NPY-LI and a number of other neurotransmitters.


Neurotoxicity related to lithium and neuroleptic combinations? A retrospective review. The question of toxic interactions resulting from combinations of lithium and neuroleptic drugs is largely based on anecdotal reports. We replicated the methods of Miller and Menninger (1987) who reported that 27% of manic patients on treatment with lithium and neuroleptics developed toxicity. We found no cases of neurotoxicity as defined in the earlier report. Pharmacologic mechanisms and differences in the clinical findings of the two studies are discussed.


Neuroleptic malignant syndrome: a preventive program. Neuroleptic Malignant Syndrome is a very serious side effect of antipsychotic medications. The paper describes a preventative program which was instituted in an inpatient unit of a provincial psychiatric hospital. There have been no mortalities from Neuroleptic Malignant Syndrome (NMS) during this program. A suspicion of NMS by clinical evaluation and laboratory tests resulted in prompt management. The recommended management plan is described.


Sequence and expression of rat ICAM-1.

Sequence and expression of rat ICAM-1. We have isolated cDNA clones-coding for rat intercellular adhesion molecule-1 (RICAM-1) from a cDNA library constructed from rat Ax cells stimulated with IL-1 beta using the mouse ICAM-1 cDNA as a hybridization probe. The RICAM-1 sequence shows 79.1% homology with mouse ICAM-1 and 55.6% homology with human ICAM-1 at the nucleic acid level. In order to examine the expression of RICAM-1 on Chinese hamster ovary (CHO) cells, we constructed the vector, pSV-RICAM1-neo, containing the SV40 promoter. Flowcytometric analysis showed that CHO-K1 cells transfected with pSV-RICAM1-neo expressed high amounts of RICAM-1 on their surfaces.


Alpha 2-adrenoceptors in the brain of suicide victims: increased receptor density associated with major depression. To examine directly in the brain the status of the alpha 2-adrenoceptor in major depression, the specific binding of the agonists [3H]clonidine and [3H]UK 14304 was quantitated in various brain regions of suicide victims with a retrospective diagnosis of depression or other psychiatric disorders. In depressed suicides, the binding capacity of [3H]clonidine was found to be increased in the hypothalamus (Bmax 35%-55% greater), and to a lesser extent in the frontal cortex, as compared with that in matched controls, schizophrenic suicides, or suicides with various diagnosis. The binding capacity of [3H]UK 14304 also was found increased in the frontal cortex (Bmax 30% greater), and to a lesser extent in the hypothalamus, of depressed suicides. In other brain regions such as the amygdala, hippocampus, and cerebellum there also was a tendency for an increased receptor density associated with suicide. Moreover, in the frontal cortex of suicides, the potency of norepinephrine in displacing the binding of the antagonist [3H]idazoxan also was found increased (Ki decreased eight-fold). The results indicate that the density and affinity of alpha 2A-adrenoceptors in the high-affinity state are increased in the brain of depressed suicides.


Paranoid schizophrenia may be caused by dopamine hyperactivity of CA1 hippocampus. Explicit consolidation of memory, or fixation of declarative belief, appears to be physically represented in changes of synaptic conductances of neurons in the parietal-temporal-occipital association cortex (PTO) of the mammalian forebrain. This fixation of belief in PTO is postulated to be critically dependent on a diffuse reinforcement signal via the inferior temporal cortex (ITC) ultimately caused by an increased output of the CA1 pyramidal cells of hippocampus. Analogous to the reinforcing mechanisms of other forebrain systems, this updating of the connection weights of the neural nets in PTO by the output of the critical neurons in CA1 is directly related to concentrations of dopamine (DA). We propose that the delusions (i.e., unreasonable beliefs) of paranoid schizophrenia are caused by a hyperactivity of the same DA-sensitive CA1 neurons that are responsible for the fixation of normal beliefs. The dramatic reduction in delusions with administration of neuroleptics, as DA D2 blockers, in schizophrenics may thus be explained by their acting to ameliorate the hyperactivity of these CA1 DA D2 receptors.


A new homeobox-leucine zipper gene from Arabidopsis thaliana. We have isolated a homeobox-containing gene from Arabidopsis thaliana using a degenerate oligonucleotide probe corresponding to the most conserved region of the homeodomain. This strategy has been used previously to isolate homeobox-containing genes from Caenorhabditis, and recently from A. thaliana. The Arabidopsis genes have an unusual structure in that they have a leucine zipper motif adjacent to the carboxy terminal region of the homeo domain, a feature not found in homeobox-containing genes isolated from animals. We report the isolation and primary structure of a new member of this Arabidopsis homeobox-leucine zipper gene family. This new member has the homeodomain and leucine-zipper motif similar to the two genes previously identified, but differs from these genes in the part corresponding to the carboxy terminus of the polypeptide, as well as in size and isoelectric point of the protein.


Detection of a single base substitution of the gene for prothrombin Tokushima. The application of PCR-SSCP for the genetic and molecular analysis of dysprothrombinemia. The genetic and molecular basis of a mutant prothrombin of 'prothrombin Tokushima' was studied by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and PCR-restriction fragment length polymorphism (PCR-RFLP) analyses. The abnormal gene was detected by altered migration by PCR-SSCP and by the loss of an MspI site by PCR-RFLP. The gene for prothrombin Tokushima was shown to be inherited from the mother of the proband. Sequencing analysis using PCR-amplified genomic DNA clarified a substitution of thymine (T) for cytosine (C) at position 9,490, changing arginine (Arg) to tryptophan (Trp) at position 418 of the polypeptide chain. This point mutation is assumed to be the molecular basis of prothrombin Tokushima, firstly, because of the absence of distinct changes in Southern blot analysis of the proband's DNA (using a full-length human prothrombin cDNA as a probe), secondly, because it has the same molecular weight as the abnormal gene product, and, thirdly, because of the absence of other amino acid abnormalities in the proteolytic peptide-fragments. It is concluded that PCR-SSCP and PCR-RFLP were useful for detecting the abnormal gene and for directly diagnosing the carrier status of dysprothrombinemia. This is the first report of gene analysis of dysprothrombinemia.


cDNA clones encoding Arabidopsis thaliana and Zea mays mitochondrial chaperonin HSP60 and gene expression during seed germination and heat shock. Mitochondria contain a nuclear-encoded heat shock protein, HSP60, which functions as a chaperonin in the post-translational assembly of multimeric proteins encoded by both nuclear and mitochondrial genes. We have isolated and sequenced full-length complementary DNAs coding for this mitochondrial chaperonin in Arabidopsis thaliana and Zea mays. Southern-blot analysis indicates the presence of a single hsp60 gene in the genome of A. thaliana. There is a high degree of homology at the predicted amino acid levels (43 to 60%) between plant HSP60s and their homologues in prokaryotes and other eukaryotes which indicates that these proteins must have similar evolutionarily conserved functions in all organisms. Northern- and western-blot analyses indicate that the expression of the hsp60 gene is developmentally regulated during seed germination. It is also heat-inducible. Developmental regulation of the (beta-subunit of F1-ATPase, an enzyme complex that is involved in the cyanide-sensitive mitochondrial electron transport system, indicates that imbibed embryos undergo rapid mitochondrial biogenesis through the early stages of germination. Based on the functional role of HSP60 in macromolecular assembly, these data collectively suggest that the presence of higher levels of HSP60 is necessary during active mitochondrial biogenesis, when the need for this protein is greatest in assisting the rapid assembly of the oligomeric protein structures.


An in vivo alpha-2 assay reversal of opioid-induced muscular rigidity and neuroleptic-induced ptosis. A method is described to detect selective alpha-2 adrenergic agonists in vivo. Palpebral ptosis is induced in rats by the neuroleptic agent haloperidol (Hal), or by tetrabenazine (TBZ) methanesulfonate. Twenty minutes later, test compounds are injected, and ptosis is scored. In a separate test, muscular rigidity is induced by the opioid, fentanyl, and subsequently, test compounds are assessed for their ability to reverse muscular rigidity. Results indicate that only alpha-2 agonists reliably reverse neuroleptic-induced and TBZ-induced ptosis, as well as opioid-induced rigidity. An alpha-1 antagonist reversed only rigidity, whereas, alpha-2 antagonists and beta-agonists were generally ineffective in all tests. Therefore, the ability to reverse neuroleptic and TBZ-induced ptosis along with the ability to reverse opioid-induced muscular rigidity is a characteristic unique to alpha-2 agonists.


Synthesis and neurotoxicological evaluation of putative metabolites of the serotonergic neurotoxin 2-(methylamino)-1-[3,4-(methylenedioxy)phenyl] propane [(methylenedioxy)methamphetamine]. Theoretical considerations and recent experimental data have prompted an investigation of the neurotoxicological properties of the 6-hydroxydopamine analogue 2-(methylamino)-1-(2,4,5-trihydroxyphenyl)propane (5) and its possible precursor 1-[2-hydroxy-4,5-(methylenedioxy)phenyl]-2- (methylamino)propane (4), potential metabolites of the serotonergic neurotoxin MDMA. Systemic, intracerebroventricular, and intraparenchymal (intrastriatal and intracortical) administration of 4 led to no detectable alterations of hippocampal or cortical serotonin or striatal dopamine levels in the rat under conditions that caused significant biogenic amine depletions by established neurotoxins. By contrast, intraparenchymal administration of 5 caused profound depletions of dopamine and serotonin, with the former being more severely depleted than the latter. Although not conclusive, these data suggest a possible role for 5 in the mediation of MDMA's neurotoxic actions.


A reliable method for the use of oligonucleotides as probes in blot-hybridization experiments. We have developed a ligation and specific-primer radiolabeling method that allows the use of oligonucleotides as probes in blot-hybridization experiments. The major advantage of the protocol is that standard hybridization and washing conditions may be used and yield high signals and low background. The observed increase in the stability and intensity of the hybridization signals appears to result from both increased length and specific radioactivity of the hybridization probe.


Mapping of ornithine aminotransferase gene sequences to mouse chromosomes 7, X, and 3. Ornithine aminotransferase (OAT), a mitochondrial matrix enzyme, is deficient in patients with gyrate atrophy of the choroid and retina. In human, the OAT structural gene maps to Chromosome (Chr) 10q26 and several OAT-related sequences, some of which are known to be processed pseudogenes, which map to Xp11.3-11.21. Here, we report chromosomal localization in the mouse of the OAT gene and related sequences. Genomic DNA blot analysis of a well-characterized panel of Chinese hamster x mouse somatic cell hybrids using a human OAT probe revealed two murine loci, one on mouse Chr 7 and the other on Chr X. In addition, segregation of restriction fragment length polymorphisms (RFLPs) detected by the OAT probe in recombinant inbred (RI) strains detected a third locus on Chr 3 and positioned the X locus near Cf-8 and Rsvp. Progeny of an intersubspecific backcross were used to map the Chr 7 locus between Tyr and Int-2, near Cyp2e-1.


Neurophysiologic basis of acupuncture.

Neurophysiologic basis of acupuncture. Research has shown that the benefits of acupuncture are truth, not fiction. Acupuncture effects are due to local effects, stimulation of neuroendocrine systems, and modulation of the body's electromagnetic energy. The exact mechanisms activated depend on point selection, type of stimulation, and probably time of day. The effectiveness of the treatment will depend on the disease entity treated and the skill and knowledge of the acupuncturist. Knowledge should include both the fundamental principles of Chinese medicine and the more recent scientific understanding of acupuncture's physiologic basis.


Fatal neutropenic enterocolitis associated with sulphasalazine therapy for rheumatoid arthritis. Neutropenia is a recognized complication of sulphasalazine therapy in patients with rheumatoid arthritis. It is usually mild, transient and rarely associated with serious sequelae. We describe a patient with rheumatoid arthritis who developed fatal neutropenic enterocolitis complicated by tracheo-oesophageal fistula following treatment with sulphasalazine.


Pulsed dye laser lithotripsy--the Toa Payoh Hospital experience. We report our experience with pulsed dye laser lithotripsy in the treatment of 100 ureteric stones in 95 patients over a 14-month period from July 1989 to September 1990. The overall rate of successful stone fragmentation was 97%. There was a low incidence of minor complications--mild haematuria, ureteric colic and urinary tract infection; ureteric perforation occurred in only 3 patients, all of whom were successfully treated conservatively. Pulsed dye laser lithotripsy is a safe and effective mode of treatment for ureteric stones. Current indications for laser fragmentation of stones are ureteric stones, impacted pelviureteric junction stones and Steinstrasse.


Therapy of an animal model of human gastric cancer using a combination of anti-erbB-2 monoclonal antibodies. Amplification and/or overexpression of the erbB-2 gene have been demonstrated in 20-30% of adenocarcinomas of the breast, ovary, lung, and stomach and are associated with aggressive clinical course and poor prognosis. Interference with erbB-2 function by the use of monoclonal antibodies is a promising approach to the treatment of these diseases. In this study we demonstrate that a combination of two anti-erbB-2-specific antibodies inhibited the growth of human gastric tumor cells in vitro. This combination antibody therapy also inhibited the growth of human tumor cell lines growing as xenografts in nude mice and was able to dramatically reduce established tumors. This is the first reported observation of tumor regression induced by anti-erbB-2 monoclonal antibodies. Treatment was not curative in that tumors regrew after 6 weeks. Treatment with either single antibody alone did not inhibit cell growth or tumor formation. Pulse chase and tyrosine kinase activity experiments were used to investigate the activity of the erbB-2 gene product (gp185erbB-2). The formation of complexes by two antibodies was found to interfere with receptor function and mimic some properties of a typical receptor ligand. Selective interference of the erbB-2 receptor by combination antibody therapy may be advantageous for the treatment of human cancers.


p53 mutations are associated with 17p allelic loss in grade II and grade III astrocytoma. Loss of genetic material on the short arm of chromosome 17 is observed in approximately 40% of human astrocytomas (WHO grades II and III) and in approximately 30% of cases of glioblastoma multiforme (WHO grade IV). Previous studies of glioblastoma multiforme have shown that the p53 gene, located on the short arm of chromosome 17, is frequently mutated in these glioblastomas. To explore whether lower-grade astrocytomas are also associated with corresponding mutations of the p53 gene, we have investigated a series of 22 human astrocytomas of WHO grades II and III both for loss of heterozygosity on chromosome 17p and for p53 mutations. Mutations in the conserved regions of the p53 gene were identified by single strand conformation polymorphism analysis of exons 5, 6, 7, and 8 and were verified by direct DNA sequencing of the polymerase chain reaction products. p53 mutations were observed in 3 of 8 grade II astrocytomas and 4 of 14 grade II astrocytomas. In all 22 tumors, allelic loss of the short arm of chromosome 17 was investigated by restriction fragment length polymorphism analysis. One-half of the grade II astrocytomas (4 of 8) and grade III astrocytomas (7 of 14) exhibited allelic loss on chromosome 17p. Mutations in the p53 gene were exclusively observed in tumors with allelic loss on 17p. Our results show that p53 mutations are not restricted to glioblastoma multiforme and may be important in the tumorigenesis of lower-grade astrocytomas and that p53 mutations in lower-grade astrocytomas are associated with loss of chromosome 17p. These findings are consistent with a recessive mechanism of action of p53 in WHO grade II and III astrocytoma tumorigenesis.


[Study on natural infection, biting and transovarial transmission of epidemic haemorrhagic fever virus in Leptotrombidium (L.) scutellare]. In order to clarify the significance of Leptotrombidium (L.) scutellare in transmitting EHF, from Oct. to Nov. 1988, lungs of rodents captured in endemic areas of EHF in Shangxi Province were taken for detecting EHF antigen by IFAT, Twenty-six out of 459 lungs were positive (5.7%). Six strains of EHFV were isolated from L. (L.) scutellare collected from the rodents. In Oct. 1989, 28 Apodemus captured from areas without reported cases of EHF were placed on grassland in endemic areas, free L. (L.) scutellare were lured in all mice and 3 strains of EHFV were isolated. The above results demonstrate that L. (L.) scutellare can naturally be infected by EHFV and can be transmitted via bites. It is also suggested that this species of mites could transmit the disease transovarially. These results further indicate that L. (L.) scutellare can serve as a transmitting vector of EHF.


Three-dimensional structure of the beta subunit of E. coli DNA polymerase III holoenzyme: a sliding DNA clamp. The crystal structure of the beta subunit (processivity factor) of DNA polymerase III holoenzyme has been determined at 2.5 A resolution. A dimer of the beta subunit (M(r) = 2 x 40.6 kd, 2 x 366 amino acid residues) forms a ring-shaped structure lined by 12 alpha helices that can encircle duplex DNA. The structure is highly symmetrical, with each monomer containing three domains of identical topology. The charge distribution and orientation of the helices indicate that the molecule functions by forming a tight clamp that can slide on DNA, as shown biochemically. A potential structural relationship is suggested between the beta subunit and proliferating cell nuclear antigen (PCNA, the eukaryotic polymerase delta [and epsilon] processivity factor), and the gene 45 protein of the bacteriophage T4 DNA polymerase.


Neu differentiation factor: a transmembrane glycoprotein containing an EGF domain and an immunoglobulin homology unit. We recently reported that a 44 kd glycoprotein secreted by transformed fibroblasts stimulates tyrosine phosphorylation of the product of the neu proto-oncogene and induces differentiation of mammary tumor cells to milk-producing, growth-arrested cells. A partial amino acid sequence of the protein, termed Neu differentiation factor (NDF), enabled cloning of the corresponding complementary DNA. The deduced structure of the precursor of NDF indicated that it is a transmembrane protein whose extracellular portion contains an EGF-like domain that probably functions as a receptor recognition site. In addition, the ectodomain contains one immunoglobulin homology unit. Despite the lack of a recognizable hydrophobic signal peptide at the N-terminus, a recombinant NDF, like the natural molecule, is released into the medium of transfected COS-7 cells in a biologically active form. Northern blot analysis indicated the existence of several NDF transcripts, the major ones being 1.8, 2.6, and 6.7 kb in size. Transformation by the ras oncogene dramatically elevated the expression of NDF in fibroblasts.


Doxazosin: a new alpha 1-adrenergic antagonist. The physicochemical properties, pharmacology, pharmacokinetics, cardiovascular and metabolic effects, adverse effects, dosage, and administration of doxazosin are described, and comparative clinical studies of doxazosin therapy in patients with mild to moderate hypertension are reviewed. Doxazosin mesylate, an alpha 1-adrenoceptor antagonist, is rapidly absorbed after oral administration and undergoes extensive hepatic metabolism. The drug decreases blood pressure by reducing peripheral resistance. Maximum hypotensive effects occur four to eight hours after the dose. Doxazosin favorably affects serum lipids by increasing concentrations of high-density lipoprotein (HDL) cholesterol, increasing the HDL:total cholesterol ratio, and decreasing concentrations of low-density lipoprotein cholesterol, total cholesterol, and triglycerides. In comparative clinical trials, doxazosin lowered standing and supine systolic and diastolic blood pressures as effectively as other alpha-adrenoceptor antagonists, beta-adrenoceptor antagonists, diuretics, angiotensin-converting-enzyme inhibitors, and calcium-channel-blocking agents. The most frequently reported adverse effects are dizziness, headache, nausea, lethargy, and fatigue. Doxazosin may be used either alone or in combination with a beta-adrenoceptor inhibitor or a diuretic. Orthostatic hypotension after the first dose occurs infrequently and may be minimized by initiating therapy at a dosage of 1 mg/day. The dosage may be increased at two-week intervals as needed, and blood pressure should be closely monitored. Doxazosin has blood-pressure-lowering effects comparable to those of other alpha 1-adrenoceptor inhibitors and to those of antihypertensives in other drug classes.


Species specificity and developmental patterns of expression of the beta amyloid precursor protein (APP) gene in brain, liver and choroid plexus in birds. 1. Human APP cDNA hybridized to a 3.5 kb mRNA in liver and brain RNA from chickens, pigeons, quail and ducks as well as in RNA from choroid plexus of chicken and quail. In contrast to all other species hitherto examined a 1.6 kb mRNA hybridizing to APP cDNA was found in abundant amounts in RNA from chicken and quail livers. 2. In the chicken, before hatching, the levels of APP mRNA in total RNA from liver and choroid plexus were higher than those in RNA from liver and choroid plexus of adults. However, RNA from the rest of the brain of chicken embryos contained less APP mRNA than RNA from brain of adults. 3. In the chicken, between 10 and 40 days after hatching, APP mRNA levels in RNA from liver were higher than adult levels, APP mRNA levels in RNA from choroid plexus were similar to adult levels and APP mRNA levels in RNA from the rest of brain were below the adult levels.


Linkage map of eight human chromosome 11q markers, including DRD2, spanning 60 cM. We have constructed a linkage map of eight RFLP markers located on chromosome 11q in the region of the dopamine D2 receptor gene (DRD2) recognized by probe hD2G1. Abnormalities in dopaminergic neurotransmission mediated by this receptor have been implicated in several psychiatric disorders. The map was generated using six large reference families (from 294 to 419 individuals per locus), which are largely independent of the CEPH families, primarily using the LINKMAP and ILINK programs of the LINKAGE package of Lathrop and Lalouel. The most likely order and recombination frequencies are: [sequence: see text] The relative order of D11S84-STMY, DRD2-D11S29, and D11S146-INT2 could not be resolved reliably. There were no significant sex differences in recombination frequency. We introduce here a version of LINKMAP adapted to run under distributed parallel processing (LINDA-LINKMAP). Using pairwise analyses, we have also placed D11S421 proximal to this group.


Unusual distribution of Zfy and Zfx sequences on the sex chromosomes of the wood lemming, a species exhibiting XY sex reversal. Sex reversal occurs naturally in the wood lemming (Myopus schisticolor) due to the presence in populations of this species of a variant (mutated) X chromosome, designated X*. Thus, X*Y animals develop into females, whereas XY animals develop into normal males. Chromosome mapping by in situ hybridization of DNA sequences homologous to the human ZFY gene localized the wood lemming Zfx sequences to region p12----p11 on both the wild-type X and the mutated X* chromosomes, at or proximal to a presumed breakpoint (Xp12) involved in the generation of the X* chromosome from the normal X, and Zfy sequences along the entire short arm of the Y chromosome. Differences between Zfx and Zfx* were readily detected by Southern blot analysis. However, both the Zfx and Zfx* genes expressed similarly sized transcripts in all adult somatic tissues investigated. Although the precise molecular difference between the Zfx and Zfx* genes is still unknown, their chromosomal location suggests that either Zfx or some other closely linked gene(s) on the X chromosome may be a major X-linked sex-determining gene, Tdx, which in the X* chromosome fails to interact properly with the Y-linked testis-determining gene, Tdy, thus causing X*Y embryos to develop into females. At least 15 copies of wood lemming Zfy sequences are distributed along the short arm of the Y chromosome. Northern hybridization analyses of adult tissues and somatic cell lines indicated that these Zfy repeats were transcriptionally inactive. Normally, 3-kb Zfy (ZFY) transcripts are readily detected in mouse and human testes, especially in the germ cells. It has therefore been postulated that expression of the Zfy (ZFY) gene may be important for spermatogenesis. Whether the lack of sufficient Zfy transcripts in the testis of the adult wood lemming has any impact on spermatogenesis in this species is still to be elucidated by further studies.


Analysis of sex-chromosome aneuploidy in interspecific backcross progeny between the laboratory mouse strain C57BL/6 and Mus spretus. Sex-chromosomal aneuploidy was identified in four female progeny of 200 interspecific backcrosses between laboratory mice (C57BL/6Ros) and Mus spretus. The progeny included two 39,XO monosomy mice resulting from a backcross with M. spretus, as well as a 41,XXX trisomic mouse and a 40,XX/41,XXX mosaic mouse resulting from two separate backcrosses with C57BL/6 mice. The parental origin and meiotic stage of the aneuploidies was determined for each of the mice using a series of markers that identified allelic differences in the parental X-chromosome genes present in the hybrid female. Two of the probes identified differences in repeated elements between the M. spretus and laboratory mouse X chromosomes, whereas the remaining sites involved restriction fragment length differences of single-copy genes detectable by Southern analysis. These markers indicated that the aneuploidies were most likely of maternal origin and that the trisomy resulted from a nondisjunction at the second meiotic division. In contrast, the mosaic female could have originated either from a trisomic embryo that had lost a single X in a portion of its cells or from a mitotic nondisjunction during early embryogenesis that resulted in XXX and XO daughter cells, with subsequent loss of the XO cells.


Stimulant effects of 3,4-methylenedioxymethamphetamine in the nucleus accumbens of rat. This study examined the behavioral effects in rats of intracerebral administration of S(+)-3,4-methylenedioxymethamphetamine (S-MDMA) using an automated holeboard and open-field apparatus. Administration of S-MDMA into the nucleus accumbens septi produced locomotor hyperactivity. Although the stimulant effects of S-MDMA administered systemically are antagonized by fluoxetine pretreatment, the activating effects of S-MDMA administered into the nucleus accumbens were not antagonized by fluoxetine. A similar increase in locomotor activity was observed after S-amphetamine administration into the nucleus accumbens. In contrast, the selective 5-HT-releasing drug and S-MDMA congener N-methyl-1-(1,3-benzodioxol-5-yl)-2-butanamine (MBDB) produces MDMA-like locomotor hyperactivity when administered systemically but did not alter locomotor activity when injected into the nucleus accumbens. These data indicate that S-MDMA actions in the nucleus accumbens are pharmacologically distinct from the primary effects of systemically administered S-MDMA. Behavioral effects of S-MDMA in the nucleus accumbens may result from the catecholamine-releasing properties that S-MDMA shares with S-amphetamine and not via the 5-HT-releasing properties that it shares with MBDB.


Production of endogenous nitric oxide and activation of soluble guanylate cyclase are required for N-methyl-D-aspartate-produced facilitation of the nociceptive tail-flick reflex. The intrathecal (i.t.) administration of either N-methyl-D-aspartate (NMDA, 10 fmol to 10 pmol) or L-arginine (1 pmol to 10 nmol), but not D-arginine (1 pmol to 10 nmol), produced a rapid, transient, dose-dependent facilitation (maximal response of 30.9 +/- 6.0% and 33.7 +/- 1.5%, respectively) of the nociceptive tail-flick reflex (ED50 = 47.8 +/- 15.4 fmol and 11.4 +/- 2.7 pmol, respectively). Maximal NMDA-produced facilitation of the tail-flick reflex (1 pmol i.t.) was completely abolished by prior treatment (10 min prior) with either N omega-nitro-L-arginine methyl ester (L-NAME, 10 nmol i.t.), methylene blue (10 nmol i.t.) or DL-5-aminophosphonovaleric acid (AP5, 100 pmol i.t.). NMDA-produced facilitation was completely recovered 40 min after L-NAME, 50 min after methylene blue and 30 min after AP5. L-NAME, methylene blue or AP5 did not significantly alter baseline tail-flick latency. These results suggest that NMDA-produced facilitation of a thermal nociceptive reflex is mediated through activation of an NMDA receptor that results in an increase in endogenous nitric oxide and activation of soluble guanylate cyclase in lumbar spinal cord.


Dynorphin-degrading cysteine protease is highly specific for paired arginine residues. The cleavage of dynorphin and three analogs containing paired basic residues by several proteases was investigated. The cysteine protease of neuroblastoma cells cleaved only the bond between Arg-Arg residues. Submandibular arginyl-endopeptidase, however, cleaved bonds between both Arg-Arg and Arg-Lys residues, and pancreatic trypsin at the carboxyl sides of both arginine and lysine residues. This shows that the cysteine protease is highly specific for paired arginine residues.


Effect of retinoic acid on liver transglutaminase activity and carbon tetrachloride-induced liver damage in mice. Transglutaminase (TGase) activity in the cytosol fraction of the mouse liver increased following intraperitoneal injection of retinoic acid. Retinoic acid inhibited the carbon tetrachloride-induced increase in serum alanine transaminase activity. These findings suggest that TGase is involved in the effect of retinoic acid on carbon tetrachloride-induced liver damage.


The influence of chronic nicotine treatment on stress-induced gastric ulceration and emptying rate in rats. Ten-day treatment with nicotine (5, 25 or 50 micrograms/ml drinking water) dose-dependently intensified gastric ulceration induced by cold-restraint, and emptying rate. Stomach contractions produced by graded doses of bethanechol i.v. were elevated further by nicotine treatment. It is suggested that chronic nicotine administration produces hypersensitivity of the gastric muscarinic receptors; stomach hypermotility contributes to the ulcer-worsening action of the alkaloid.


CD11b-bearing mononuclear leucocytes and IgA levels in the staging of human immunodeficiency virus infection. Certain immunological parameters (i.e. low CD4+ T cell numbers, high serum soluble CD8) have been described as prognostic factors for the progression of human immunodeficiency virus (HIV) infection to later clinical stages. In the present study we have found in one hundred HIV-infected Spanish patients (81% drug abusers, 7% homosexuals, 6% heterosexuals, and 6% other or unknown risk groups) that CD11b+ peripheral blood mononuclear cells are increased in those with persistent lymphadenopathy as compared to other clinical stages (asymptomatic, AIDS-related complex and AIDS). Serum IgA was significantly increased in AIDS patients, and in patients at any other clinical stage who had concomitant infections (mainly mycobacterial and fungal). CD11b (an integrin with complement receptor functions) may thus be of clinical interest for the staging of HIV-infected patients, and reflect stage-selective immunological changes in mononuclear cell biology during HIV infection. High IgA on the other hand, would be a marker of concomitant infection as well as of disease progression. The results concern mostly drug addicts (the main risk group in Spain), but may apply to the other risk groups because no significant differences were detected between drug addicts (n = 81) and non-drug addicts (n = 19) for the studied variables (p greater than 0.05).


[Natriuretic peptide family].

[Natriuretic peptide family]. The natriuretic peptide system consists of at least three endogenous ligands: atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP), and three receptors, ANP-A receptor (guanylate cyclase A), ANP-B receptor (guanylate cyclase B) and clearance receptor (C receptor). ANP, the prototype of natriuretic peptides, is mainly produced in the atrium and secreted into the circulation as a cardiac hormone. ANP is also produced in the ventricle and in the central nervous system. BNP, first isolated from the porcine brain, has a marked divergence in its molecular size and sequence among species. In humans and rats, the major site of production of BNP is the ventricle of the heart. BNP is also secreted into the circulation as a cardiac hormone. The plasma BNP level in normal subjects is approximately one sixths of the plasma ANP level; however, the plasma BNP level markedly increases in heart failure, renal failure and hypertension and the augmentation of the BNP secretion is much larger than that of the ANP secretion. In addition, clearance of BNP from the circulation is slower than that of ANP. Furthermore, BNP is secreted more urgently than ANP in acute heart failure. CNP distributes mainly in the central nervous system and pituitary gland. No significant amount of CNP is detectable in the heart and plasma. Thus, CNP is a local regulator rather than a cardiac hormone. Three natriuretic receptors have ligand selectivity.(ABSTRACT TRUNCATED AT 250 WORDS)


Purification and properties of factor XIII from human placenta. 1. FXIII was isolated and purified over 4000 fold from human placenta to apparent electrophoretic homogeneity by a new procedure including ethanol precipitation. DEAE-Cellulose, molecular sieving on Sephacryl S-300 and Phenyl-Sepharose chromatography. 2. Its pI was about 5.1. Under appropriate conditions, the incubation of FXIII in the presence of thrombin did not lead to inactivation cut in the polypeptidic chain. 3. FXIII was also activated by CaCl2 and, in a lesser extent, by other divalent cations like SrCl2, BaCl2 or MgCl2. 4. The binding of calcium to FXIII exhibited a negative cooperativity. 5. The activity-pH curve of the calcium-activated enzyme did not appear very different from that of the thrombin-activated enzyme.


[Orthotopic liver transplantation for metastases of bronchial carcinoid tumor]. We report the case of a 42 year old man in whom orthotopic liver transplantation was performed successfully for unresectable hepatic metastases of a bronchial carcinoid tumor. Prior to transplantation, somatostatin therapy, pulmonary lobectomy, and systemic chemotherapy (streptozotocin and fluorouracil) were performed. After 9 months there were no signs of clinical or biological recurrence. Orthotopic liver transplantation might be indicated for unresectable and limited liver metastases of neuroendocrine tumor.


Molecular genetics of the Drosophila melanogaster ovo locus, a gene required for sex determination of germline cells. The Drosophila melanogaster ovo gene is required for survival and differentiation of female germline cells, apparently playing a role in germline sex determination. We recovered 60 kb of genomic DNA from its genetic location at 4E1,2 on the X chromosome. A transcription unit coding for an apparently female-specific germline-dependent 5-kb poly(A)+ RNA size class is located substantially in a 7-kb region, within which three DNA-detectable lesions for mutations that inactivate the ovo function are located at two sites approximately 4 kb apart. The breakpoint of a deficiency that removes the neighboring lethal complementation group shavenbaby (svb) but leaves the ovo function intact maps approximately 5 kb to the molecular left of the leftmost ovo mutant site. A class of mutations that inactivates both the svb function and the ovo function affects genomic DNA between the two ovo sites. Sequences required for the two genetic functions are partly overlapping. In spite of this overlap, P element-mediated gene transfer of a 10-kb genomic DNA segment containing the 5-kb poly(A)+ RNA transcription unit rescues the female sterility phenotypes of ovo mutations, but not the svb lethality.


Genetic analysis of the additional sex combs locus of Drosophila melanogaster. Additional sex combs (Asx) is a member of the Polycomb group of genes, which are thought to be required for maintenance of chromatin structure. To better understand the function of Asx, we have isolated nine new alleles, each of which acts like a gain of function mutation. Asx is required for normal determination of segment identity. AsxP1 shows an unusual phenotype in that anterior and posterior homeotic transformations are seen in the same individuals, suggesting that AsxP1 might upset chromatin structure in a way that makes both activation and repression of homeotic genes more difficult. Analysis of embryonic and adult phenotypes of Asx alleles suggests that Asx is required zygotically for determination of segment number and polarity. The expression pattern of even-skipped is altered in Asx mutant embryos, suggesting that Asx is required for normal expression of this gene. We have transposon-tagged the Asx gene, and can thus begin molecular analysis of its function.


Independence of VNTR alleles defined as fixed bins. An analysis is presented of data collected by the Federal Bureau of Investigation at six unlinked variable number of tandem repeats (VNTR) loci for the United States population. Databases have been constructed of VNTR profiles of Caucasians, Blacks and Hispanics from Florida, Texas and California. There was very little evidence for correlations between lengths for pairs of VNTR fragments, within or between loci. When the fragment lengths were amalgamated into discrete bins, there was also little evidence for disequilibrium over all genotypes, within or between loci, for the Caucasian database, although some disequilibrium was found for the Black and Hispanic databases. No disequilibrium was found for the Caucasian or Black databases when tests were confined to heterozygous individuals. In cases of global disequilibrium, local tests can be applied to specific genotypes. The results suggest that, at the bin level, frequencies of VNTR profiles can generally be estimated as the products of the frequencies of the constituent elements. This overcomes the problem of estimating population frequencies when any particular profile does not exist in the database. There is some evidence for different frequencies, at the individual bin level, between geographic samples within each of the Caucasian, Black and Hispanic databases, and considerable evidence for differences between the three databases. These differences are less evident for the frequencies of four-locus profiles.


Risk estimation in familial adenomatous polyposis using DNA probes linked to the familial adenomatous polyposis gene. The familial adenomatous polyposis gene has recently been assigned to the long arm of chromosome five through linkage to several 5q DNA probes. These probes can now be used to trace inheritance of the disease gene in affected families. In this study, DNA samples from 152 members of 10 Australian familial adenomatous polyposis families have been examined for restriction fragment length polymorphisms detected by DNA probes C11P11, ECB27, and YN5.48. Linkage analysis confirmed linkage between the familial adenomatous polyposis gene and each probe with a maximum combined LOD score of 2.82 for C11P11, 2.90 for ECB27 and 5.49 for YN5.48 all at a recombination fraction of zero. Risk estimates were determined for the 51 at risk individuals in these families based on their restriction fragment length polymorphism data alone or in addition by including the effect of age dependent penetrance. Thirty two of those at risk (63%) could be assigned specific high (greater than or equal to 95%) or low (less than or equal to 5%) risks of developing familial adenomatous polyposis on the basis of their probe results. When the effect of age dependent penetrance was included, 26 (51%) fell at the extremes of risk (greater than or equal to 99% or less than or equal to 1%). Such estimates provide a sound basis for planning sigmoidoscopic screening of at risk family members and will thus facilitate surveillance in familial adenomatous polyposis families.


GM and KM allotypes and GM RFLP allogenotypes in Micronesians from Nauru. Immunoglobulin allotypes of the GM and KM systems were determined in a sample of Micronesian subjects from Nauru. Four GM haplotypes were identified in the sample: GM*1,3 23 5, 10,11,13,14, GM*1,17 23' 21, GM*1,3 23' 5,10,11,13,14, and GM*1,2,17 23' 21, although the last of these may have been introduced by non-Micronesian admixture. The frequency of the KM*1 allele is 0.115 +/- 0.033, which is slightly lower than reported in Micronesians from the Caroline Islands. RFLPs generated by the enzymes Taq I and Pvu II and detected by a Hu gamma 4 probe were related to GM phenotypes. The haplotypes GM*1,3 +/- 23 5,10,11,13,14 were strongly associated with a Taq I 5.0-kb band. The presence and absence of the allotype G2M 23 were marked by a Pvu II 7.0 + 2.0 kb pair and a Pvu II 9.0-kb fragment, respectively. GM*1,17 23' 21 was strongly associated with a Pvu II 5.0 + 2.7 kb pair. The different relationships between GM haplotypes and Hu gamma 4 RFLPs in Micronesians and Caucasians indicate that a universal GM allogenotyping procedure cannot yet be developed; instead, population-specific procedures are necessitated by differences in GM allotype arrangements between populations.


Glucocorticoid receptor numbers in the brain and liver of the obese Zucker rat. Scatchard analysis of ligand binding activity has been used to investigate type I and type II glucocorticoid receptor subtypes in the hippocampus, hypothalamus and liver of lean and genetically obese fa/fa rats. Despite normal levels of corticosterone in the hypothalamus and hippocampus of fa/fa rats, maximum binding to both type I and type II receptors was increased in the hippocampus and hypothalamus. The KD value of type I receptors was normal in fa/fa rats (lean 0.13 nM, obese 0.15 nM) whereas the KD for corticosterone binding to type II receptors was increased (lean 0.48nM, obese 1.02nM). Adrenalectomy increased Bmax of type II receptors in lean rats in a time dependent manner but had no effect on binding to receptors of obese rats after the initial clearance of endogenous bound ligand. KD values were not altered by adrenalectomy in either genotype. There were no differences in binding of corticosterone to hepatic receptors of lean and obese rats seven days after adrenalectomy. The data suggests that glucocorticoid binding to type I and type II receptors is abnormally regulated in the brains of fa/fa rats.


Dexmedetomidine-induced ocular hypotension in rabbits with normal or elevated intraocular pressures. This study covered the ocular hypotensive effects of the stereoisomers of the alpha 2-adrenoceptor agonist medetomidine. The dextro-isomer, dexmedetomidine, is known from pharmacologic experiments to be a specific, potent, and selective full agonist at alpha 2-adrenoceptors, whereas the levo-enantiomer seems to be almost inactive. Thus, the levo-isomer (0.5 mg/ml, 25 microliters) had no significant effect on intraocular pressure. After unilateral topical administration, dexmedetomidine (0.5 mg/ml, 25 microliters) lowered intraocular pressure bilaterally in normal rabbits and in rabbits with intraocular pressure elevated after laser irradiation of the pigmented trabecular band of the anterior chamber angle. In the treated (ipsilateral) eye of normal rabbits, a maximum decrease of 4.6 +/- 0.6 mmHg was observed at 2 hr post treatment. In the contralateral eye, the maximum decrease was 4.1 +/- 0.5 mmHg at 1 hr after treatment. In rabbits with laser-induced elevation of intraocular pressure, the maximum decrease in treated hypertensive eyes was 13.5 +/- 0.3 mmHg 1 hr after dexmedetomidine administration. These results indicate that the selective alpha 2-adrenoceptor agonist, dexmedetomidine, is a potent and effective drug for decreasing intraocular pressure in rabbits.


The assessment of cellular proliferation by immunohistochemistry: a review of currently available methods and their applications. Immunohistochemical methods using antibodies to cell cycle-related antigens may be used as a means of assessing various aspects of proliferation in tissue, and have the important advantage of preserving the spatial orientation of proliferating cells in histological sections. Currently, the most widely available antibodies for this purpose are antibodies to bromodeoxyuridine (BrdU), Ki67 and antibodies to proliferating cell nuclear antigen (PCNA). BrdU is a thymidine analogue incorporated during the S phase of the cell cycle, which can be introduced by 'in vitro' incubation, and monoclonal antibodies are available to display its localization. Ki67 demonstrates a nuclear antigen expressed in all phases of the cell cycle, except G0 and early G1, but can only be applied to frozen tissue. PCNA is a nuclear antigen which is essential for DNA synthesis, two commercially available antibodies to PCNA work in paraffin-embedded tissue, but may have different staining characteristics under different conditions of fixation. The main advantages and disadvantages of these different techniques are discussed, together with their main applications to date.


Immunogold labelling is a quantitative method as demonstrated by studies on aminopeptidase N in microvillar membrane vesicles. Microvillar membrane vesicle preparations with varying content of aminopeptidase N were prepared from enterocytes of the pig small intestine. Postembedding immunogold labelling of aminopeptidase N was performed on these glutaraldehyde/paraformaldehyde-fixed, osmium tetroxide-treated and Epon-embedded microvillar membrane vesicles. The number of gold particles per micrometre microvillar membrane (labelling intensity) was calculated and compared to the corresponding enzymatic activity. A very close relationship was found between labelling intensity and aminopeptidase N activity, demonstrating that postembedding immunogold labelling can be used quantitatively.


The ultrastructural immunolocalization of gamma-glutamyltranspeptidase in rat lung: correlation with the histochemical demonstration of enzyme activity. gamma-Glutamyltranspeptidase (gamma-GT) was localized in slices of rat lung, at the ultrastructural level, by pre-embedding immunogold labelling. Antiserum was raised against the protein purified from rat kidney. The enzyme was found to be concentrated on the lumenal surface of the non-ciliated ("Clara") cells of the bronchiolar epithelium and, to a lesser degree, on the surface of type II alveolar pneumocytes. This immunological localization was consistent with the distribution of reaction product, in both slices and resin sections incubated to demonstrate gamma-GT activity. gamma-GT is probably involved in the utilization of reduced glutathione (GSH) present in the fluid lining the airway epithelium.


Retail cut yields of Rambouillet wether lambs fed the beta-adrenergic agonist L644,969. Twenty Rambouillet wether lambs were given ad libitum access to a diet with (BAA, n = 10) or without (control, n = 10) 1 ppm of the beta-adrenergic agonist L644,969. Lambs were fed to a constant slaughter weight end point of 54.5 kg. Carcasses were fabricated to yield bone-in and boneless cuts that were trimmed progressively to 1.27, .64, .32, and .00 cm of s.c. fat remaining. Addition of BAA did not affect growth traits. Actual and adjusted fat thickness, body wall thickness, and percentage of kidney-pelvic fat did not differ between control and BAA lambs. However, BAA increased longissimus muscle area, longissimus muscle depth, and leg score while decreasing USDA yield grade. The BAA increased carcass conformation scores and decreased flank lean color scores. No other carcass quality measurements were affected by BAA. Addition of BAA did not affect overall carcass yields of bone-in retail cuts. However, BAA increased overall carcass yields of boneless retail cuts regardless of fat trim level. The BAA increased bone-in leg yield. Yield of boneless sirloin, bone-in loin and boneless loin were not affected by BAA. For these cuts, the percentage change from the control was highly dependent on fat trim level. There was no difference in short-cut, shank-off, semiboneless leg yield between control and BAA. Addition of BAA did not affect yield of bone-in rack regardless of fat trim level. However, BAA greatly increased yield of boneless ribeye. The BAA did not affect yield of bone-in or boneless shoulder.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of dietary level of lysine and of level and source of protein on feed intake, growth performance, and plasma amino acid pattern in the finishing pig. The effects of dietary level of lysine and of level and source of CP on voluntary feed intake, growth performance, plasma free amino acids, and carcass characteristics were investigated in a study involving 60 female and 60 castrated male Large White finishing pigs (from 42 to 101 kg live weight) with ad libitum access to feed. Six treatments were compared according to a 2 x 3 factorial plan, with two levels of lysine (.55 and .65% selected below the recommended levels for both sexes) and three types of CP (N x 6.25) supply: a 13% CP diet based on wheat, peanut meal and soybean meal; a 15.6% CP diet providing the same amino acid pattern as that of the basal diet; and a 15.2% CP diet containing the same levels of essential amino acids as the 13% CP diet, with the addition of glutamic acid as a source of nonessential amino acids. By maintaining a constant amino acid pattern separate changes in dietary lysine and CP levels resulted in a relative independency of their effects on feed intake, growth performance, and body composition. Muscle gain increased with supplementary lysine, with a lower response at the lower CP level (13%). At the same level of lysine (.55 or .65%), increasing protein content from 13 to 15.6% did not affect feed intake, but growth rate was lower and feed/gain was increased, partly because of an additional energy cost resulting from catabolism of excess protein.(ABSTRACT TRUNCATED AT 250 WORDS)


The role of autologous blood stem cells in support of high-dose therapy for multiple myeloma. During the last few years, high-dose therapy with hemopoietic stem cell support has become a well-admitted therapeutic option for young patients with MM. The role of allogeneic or autologous graft and of blood rather than bone marrow as the source of hemopoietic stem cells must be further investigated. Autologous PBSC transplantation has, however, both practical and theoretic advantages over allogeneic and autologous BMT: (1) It can be applied to most patients, especially if blood stem cells are collected early in the course of therapy. (2) It usually induces relatively rapid hematologic reconstitution. (3) In comparison with autologous BMT, it appears to minimize the hazard of the reinfusion of malignant cells.


Expression of proliferating cell nuclear antigen during the cell cycle of human diploid fibroblasts. Proliferating cell nuclear antigen mRNA levels were determined in human diploid fibroblasts as they progressed through the cell cycle. PCNA message levels were low at G0, gradually increased following entrance into G1, peaked at G1/S, and declined during S phase. PCNA mRNA was determined to have a half life of 12 hours when cells were blocked at the G1/S interface. PCNA protein levels increased two- to three-fold as cells moved from G0 to S phase.


Pharmacological analysis of positive chronotropic and inotropic responses to etilefrine in isolated dog heart preparations. Positive chronotropic and inotropic responses to etilefrine (alpha-[(ethylamino)methyl]-m-hydroxybenzyl alcohol), an orally active cardiovascular agent, were investigated in isolated dog right atrial and left ventricular preparations. Intravenous administration of etilefrine to a support dog increased heart rate and mean systemic blood pressure, and increased sinus rate and atrial contractile force in the isolated right atrium perfused with blood from the support dog. Etilefrine injected intra-arterially to isolated atria and ventricles induced dose-dependent positive chronotropic and inotropic effects. Etilefrine was about 100 times less potent than isoproterenol. The effects of etilefrine in isolated atria were significantly inhibited by treatment with atenolol, but were not significantly inhibited by ICI 118,551. The effects of etilefrine were partially inhibited by imipramine. These results indicate that etilefrine is a highly selective beta-1 adrenoceptor agonist and suggest a moderate catecholamine-releasing activity by tyramine-like action in the blood-perfused dog heart.


Renal alpha 2-adrenoceptors in New Zealand genetically hypertensive rats. 1. Renal alpha 1- and alpha 2-adrenoceptors were characterized in the New Zealand strain of genetically hypertensive (GH) rat and the Otago random-bred albino normotensive (NT) control rat at 4 and 12 weeks of age with [3H]-prazosin and [3H]-rauwolscine. 2. At 4 weeks of age, the density of alpha-adrenoceptors in NT and GH rats was similar to both the alpha 1- (193 +/- 11 vs 163 +/- 14 fmol mg-1 protein) and alpha 2- (347 +/- 34 vs 319 +/- 41 fmol mg-1 protein) adrenoceptor. At 12 weeks of age, GH rats had a greater density of renal alpha 1- (152 +/- 27 vs 238 +/- 17 fmol mg-1 protein) and alpha 2- (175 +/- 42 vs 350 +/- 23 fmol mg-1 protein) adrenoceptors compared to the NT rats. 3. Pre-incubation of kidneys from GH rats (12 weeks of age) with 1 and 10 microM clonidine decreased the density of receptors identified by unlabelled clonidine displacement of [3H]-rauwolscine to 77% and 56% of control. Pre-incubation with adrenaline, 2,6 dimethylclonidine or phenylephrine failed to alter binding. 4. Pre-incubation of kidneys from NT rats (12 weeks) or young GH rats (4 weeks) with 10 microM clonidine failed to alter displacement of [3H]-rauwolscine by unlabelled clonidine. 5. These studies demonstrate that in another strain of hypertensive rat, the GH rat, alpha 2-adrenoceptor density is increased as compared to the normotensive control at 12 but not 4 weeks of age.(ABSTRACT TRUNCATED AT 250 WORDS)


Activation of alpha 1- and alpha 2-adrenoceptors inhibits noradrenaline release in rabbit renal arteries: effects of pertussis toxin and N-ethylmaleimide. 1. The effects of selective alpha 1- and alpha 2-adrenoceptor agonists and antagonists on the stimulation-induced (S-I) outflow of radioactivity at 2 Hz were investigated in superfused rabbit renal arteries incubated with [3H]-noradrenaline. 2. The alpha 1-adrenoceptor agonist methoxamine (10 microM) inhibited S-I outflow of radioactivity and this effect was abolished by the alpha 1-adrenoceptor antagonist prazosin (0.1 microM) but not by the alpha 2-adrenoceptor antagonist rauwolscine (1 microM). Neither the prostaglandin synthesis inhibitor indomethacin (10 microM) nor the adenosine receptor antagonist 8-phenyl-theophylline (1 microM) prevented the inhibitory effect of methoxamine. 3. The alpha 2-adrenoceptor agonists clonidine (0.1 microM) and UK 14304 (0.1 microM) both inhibited S-I outflow of radioactivity. The inhibitory effect of clonidine was blocked by rauwolscine but not by prazosin. The inhibitory effect of UK 14304 was markedly reduced by rauwolscine. 4. Prazosin (0.1 microM) alone did not enhance the S-I outflow of radioactivity at 2 Hz and slightly enhanced S-I outflow at 4 Hz. Rauwolscine (1 microM) alone markedly enhanced S-I outflow of radioactivity at 2 and 4 Hz. 5. Pretreatment of the arteries with pertussis toxin (1 microgram ml-1) did not significantly alter the inhibitory effects of methoxamine or UK 14304 or the potentiation by rauwolscine. However, pretreatment of the arteries with a higher concentration of pertussis toxin (5 micrograms ml-1) prevented the inhibitory effect of methoxamine but still did not affect the responses to UK 14304 and rauwolscine. 6. Pretreatment of the arteries with N-ethylmaleimide (NEM, 10 microM) for 30 min did not alter the inhibitory effect of methoxamine but markedly attenuated the inhibitory effect of UK 14304 and the facilitatory effect of rauwolscine. 7. The results suggest that both alpha 1- and alpha 2-adrenoceptors take part in the modulation of noradrenaline release from sympathetic nerves in rabbit renal arteries. Alpha 1-adrenoceptor mediated inhibition may be coupled to G-proteins which are pertussis toxin sensitive and alpha 2-adrenoceptor mediated inhibition to G-proteins which are NEM-sensitive.


Reversal of multidrug resistance by B859-35, a metabolite of B859-35, niguldipine, verapamil and nitrendipine. It has been shown previously that verapamil and other calcium antagonists and calmodulin inhibitors can reverse multidrug resistance. We compared the potency of the dihydropyridine derivatives (4R)-3-[3-(4,4-diphenyl-1-piperadinyl)-propyl]-5-methyl-1,4-dihydr o-2,6- dimethyl-4-(3-nitrophenyl)-pyridine-3,5-dicarboxylate-hydrochloride (B859-35), a metabolite of B859-35, niguldipine and (R)-nitrendipine to that of (RS)-verapamil in reversing multidrug resistance. The accumulation of the fluorescent dye rhodamine 123, which is transported by the P-glycoprotein, was determined by a flow cytometer. Multidrug-resistant human HeLa KB-8-5 and Walker rat carcinoma cells were incubated in the presence and in absence of the drugs indicated above. We found that 0.1 microM B859-35 increases the accumulation of rhodamine 123 in multidrug-resistant KB-8-5 and Walker cells more effectively than 1 microM (RS)-verapamil. In sensitive KB-3-1 cells addition of the drugs had no significant influence on the accumulation of rhodamine 123. IN KB-8-5 cells, 10 nM Adriamycin caused a reduction of cell growth to 85% compared to untreated controls (= 100%). If 1 microM B859-35, B859-35 metabolite, niguldipine, verapamil or (R)-nitrendipine was added to 10 nM Adriamycin, growth reduction compared with untreated controls increased to 12%, 11%, 23%, 63%, and 82% respectively. The effect of 0.1 microM B859-35 was a reduction in proliferation to 38%, that of 0.1 microM verapamil to 72%. These data illustrate that B859-35, a compound with antitumor activity in several tumors, is at least ten times more potent than racemic verapamil in reversing multidrug resistance.


Expression and release of phosphatidylinositol anchored cell surface molecules by a cell line derived from sensory neurons. Early postnatal mouse dorsal root ganglion neurons were found to express several glycosylphosphatidylinositol-anchored (GPI) molecules from the immunoglobulin superfamily (neural cell adhesion molecule 120 kD isoform, F3, Thy1) whose expression is developmentally regulated. A hybrid cell line (ND26), made by fusing postmitotic rat dorsal root ganglion (DRG) neurons with the mouse neuroblastoma N18Tg2, could be induced to differentiate by manipulating the composition of the culture medium and expressed similar GPI molecules to DRG neurons. We used this model system to investigate the metabolism of GPI-anchored molecules. We found that neural cell adhesion molecule 120 Kd isoform expression decreased upon differentiation, whereas the level of F3 and Thy1 increased, suggesting a role in neurite outgrowth processes. The ratio of molecules cleavable by exogenous phosphatidylinositol phospholipase C (PI-PLC) was similar for all the GPI-anchored molecules, which could mean that cell-specific modifications of the basic anchoring structure determine the level of potentially releasable molecules. Measurements of spontaneous release indicated that this reflected the overall level of expression of these molecules by the ND26 cell line. Finally, we observed an effect of dibutyryl cAMP on the level of expression of F3 and Thy1 but not of N-CAM. However, we could not detect any significant effect of nerve growth factor (NGF) either on the level of expression or on the amount of spontaneously released molecules.


Acetyl-CoA carboxylase in Reuber hepatoma cells: variation in enzyme activity, insulin regulation, and cellular lipid content. Reuber hepatoma cells are useful cultured lines for the study of insulin action, lipid and lipoprotein metabolism, and the regulation of acetyl-CoA carboxylase (ACC), the rate-limiting enzyme of fatty acid biosynthesis. During investigations in different clonal lines of these cells, we have uncovered marked intercellular variability in the activity, enzyme content, and insulin regulation of ACC paralleled by differences in cellular neutral lipid (triglyceride) content. Two contrasting clonal lines, Fao and H356A-1, have been studied in detail. Several features distinguish these two lines, including differences in ACC activity and enzyme kinetics, the content of the two major hepatic ACC isozymes (Mr 280,000 and 265,000 Da) and their heteroisozymic complex, the extent of ACC phosphorylation, and the ability of ACC to be activated on stimulation by insulin and insulinomimetic agonists. As studied by Nile Red staining and fluorescence-activated cell sorting, these two lines also display marked differences in neutral lipid content, which correlates with both basal levels of ACC activity and inhibition of ACC by the fatty acid analog, 5-(tetradecyloxy)-2-furoic acid (TOFA). These results emphasize the importance of characterization of any particular clonal line of Reuber cells for studies of enzyme regulation, substrate metabolism, and hormone action. With respect to ACC, studies in contrasting clonal lines of Reuber cells could provide valuable clues to understanding both the complex mechanisms of intracellular ACC regulation in the absence and presence of hormones and its regulatory role(s) in overall hepatic lipid metabolism.