Peripheral blood stem cell transplantation: impact on procedure load and workload in an apheresis unit.

Peripheral blood stem cells (PBSC) reinfusion appears to hasten hematologic reconstitution following myeloablative therapy. While procurement of PBSC adds apheresis procedures, rapid engraftment could decrease the demand for platelet transfusions. To determine the impact of PBSC collection on workload in our apheresis unit, we studied 3 consecutive groups of patients with metastatic breast cancer given comparable high-dose chemotherapy and autologous bone marrow transplant, with or without PBSC or granulocyte-colony stimulating factor (G-CSF). Forty-one transplants were performed with bone marrow cells only: 31 patients (Group A) did not receive G-CSF, while the following 10 patients (group B) received daily G-CSF until neutrophil engraftment. Bone marrow cells and PBSC were used for the most recent 11 transplants (group C), followed by daily G-CSF until engraftment. PBSC were mobilized with cyclophosphamide (4 g/m2) and etoposide (1 g/m2), followed by G-CSF, 8 micrograms/kg/day. PBSC collection was carried out on a Fenwal CS3000+ cell collector, using modified procedure 1, to obtain a minimum of 5 x 10(8) mononuclear cells/kg. The times to neutrophil count over 500/microL, platelet count over 20,000/microL, and discharge from the hospital after transplant were significantly shorter for patients in group C (medians of 8, 8, and 21 days, respectively) compared to group A (medians of 14, 14, and 29 days; P = 0.001) or group B (medians of 11, 24, and 32 days; P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


[Efficacy and tolerability of apraclonidine (ALO 2145, 1%) in the prevention of early ocular hypertonia, after trabeculoretraction using argon laser and posterior capsulotomy using Nd:YAG laser].

ALO 2145 (Apraclonidine Chlorhydrate)*, is a new hypotensive agent; it acts as a selective alpha-2-adrenergic agonist to produce a marked reduction in intraocular pressure with minimal effect on the cardiovascular system. 38 patients undergoing ocular laser surgery were allocated to treatment with either ALO 2145 1% (n = 20), or to placebo (n = 18), in a double-masked fashion. One drop of study medication was instilled into the operative eye one hour before laser surgery, and one drop immediately after the laser surgery. ALO 2145 treated eyes exhibited a significantly lower mean intraocular pressure increase in the immediate postoperative phase, compared with placebo treated eyes. ALO 2145 also significantly reduced the postoperative incidence of intraocular pressure spikes, defined as intraocular pressure increases of more than 10 mmHg over baseline (17% of cases for active treatment vs 60% of cases for placebo). ALO 2145 1% was shown to be effective and well tolerated in the dosage regimen employed in this study.


[Medico-economic assessment of neuroleptics in schizophrenia. Amisulpride versus haloperidol].

The aim of this study is to assess the economic impact of neuroleptic strategies in the long-term treatment of schizophrenic patients. In this respect a new neuroleptic strategy (amisulpride) was compared to a reference drug (haloperidol) using a cost minimization method. Clinical, demographic and economic (direct medical costs) data were obtained retrospectively from patients' charts. Patients (n = 160) were randomly selected according to diagnosis (schizophrenia, DSM III-R), treatment (outpatient, amisulpride or haloperidol) and follow up period (at least 6 months). The health insurance point of view was selected for the economic analysis. We found a significant reduction of the annual number of days of relapse when patients were treated with amisulpride compared to haloperidol. This reduction was associated with a significant reduction of direct costs mainly related to shorter length of hospitalization. This result was only partly explained by demographic and clinical variables such as the severity of the disease. The differences remained significant when populations were matched. This finding illustrates the validity of the concept of efficiency in psychiatry.


Pathogenesis of unstable angina with 0- or 1-vessel disease. Important role of coronary artery spasm.

In order to examine the possible role of coronary artery spasm in the pathogenesis of unstable angina, provocative testing for coronary spasm was performed in 43 patients with unstable angina who had 0- or 1-vessel disease. Coronary spasm was induced in 20 (65%) of 31 patients by hyperventilation testing (ST increases in 18, ST decreases in 2). Anginal attacks with either ST-segment elevation or ST-segment depression in patients without a significant organic stenosis were induced in 23 (55%) of 42 patients during treadmill exercise testing. Coronary artery spasm, showing severe (> or = 90%) vasoconstriction with angina and/or ischemic electrocardiographic ST-segment deviation, was also documented angiographically in 42 (98%) of 43 patients following intracoronary injection of acetylcholine. We conclude that dynamic coronary obstruction plays an important role in the genesis of attacks in patients with unstable angina who had 0- or 1-vessel organic coronary artery disease.


[Excitatory amino-acids, a new class of neurotransmitters. Pharmacology and functional properties].

The pharmacology of excitatory amino acids (EAA) like glutamate or aspartate, has defined three main types of receptors: NMDA, quisqualate (now named AMPA) and kaïnate receptors, associated to cationic channels. The NMDA receptor, the best characterized, is a macromolecular complex with multiple specific sites: the agonist binding site (glutamate, aspartate, NMDA); the glycine site and polyamine site mediating allosteric regulations; the site located inside the channel for activity-dependent antagonists (phencyclidine, MK-801). This channel, permeable to calcium, is blocked by magnesium in a voltage-dependent manner. The structural complexity of the NMDA receptor suggests the existence of subtle regulations, but also offers many targets for pharmacological drugs. The calcium influx induced by NMDA receptor stimulation may account for the diversity of its functional properties. First, NMDA receptors modulate neuronal plasticity during the development and even long after. Indeed, NMDA receptor can induce long term potentiation (LTP; an experimental model of synaptic facilitation) and are involved in learning and memory. On the other hand, when over-stimulated, they induce neurotoxicity. The death of the cell occurs after several hours, during which NMDA antagonists can prevent irreversible damages. EAA systems are distributed in the whole brain, interacting with numerous other neurotransmitters, but particularly concentrated in the cortico-striatal and cortico-cortical fibers and in the hippocampus. Several neuro-psychiatric disorders could be related to a glutamatergic dysfunction: acute neuronal lesions (stroke, viral disease like AIDS) and epilepsy; but also chronic neurodegenerative disorders (Alzheimer's dementia, Huntington and Parkinson diseases). A glutamatergic hypothesis of schizophrenia arose from the phencyclidine model of psychosis, arguing for an imbalance between glutamate and dopamine. The therapeutic perspectives of glutamatergic substances in these diseases will be discussed.


Effects of denopamine with or without diltiazem on the ischemic heart of anesthetized dogs.

Effects of denopamine with or without diltiazem on the ischemic heart were investigated in anesthetized open-chest dogs. Partial occlusion of the left circumflex coronary artery (LCX) produced significant decreases in LCX flow and regional myocardial segment shortening rate (%SS) in the LCX-perfused area, and a significant increase in left ventricular enddiastolic pressure (LVEDP). Heart rate (HR) and mean aortic pressure (mAoP) were not altered, but aortic flow (AoF), positive first derivative of left ventricular pressure ((+)LVdP/dt), stroke volume (SV), stroke work index (SWI) and double product showed a tendency to decrease. Total peripheral vascular resistance (TPR) tended to increase. During coronary stenosis, saline infusion (vehicle group) did not change any parameter, but diltiazem infusion (diltiazem group) decreased HR, mAoP, TPR and double product and increased SV and SWI. Under these conditions, denopamine infusion produced increases in HR, mAoP, AoF, (+)LVdP/dt and double product and decreases in LVEDP and TPR in both groups. %SS in the left anterior descending coronary artery-perfused area was increased, but %SS in the LCX-perfused area was slightly decreased in both groups. SV and SWI were decreased by denopamine infusion in the vehicle group, while they were increased in the diltiazem group. Differences in changes in SV and SWI between the groups were statistically significant. Results suggest that combined treatment of denopamine and diltiazem may exert an advantage in alleviation of heart failure due to coronary stenosis.


Correlation between cell-adherent activity and surface structure in Porphyromonas gingivalis.

The cell-adherent ability of 6 strains of Porphyromonas gingivalis (381, ATCC 33277, SU63, KD1, W50 and W83) was compared by using radiolabeled bacterial cells and human gingival fibroblasts (Gin 1), human periodontal ligament fibroblasts (HPLF) and human epithelial cells (Ca9-22) that had been grown on collagen beads. The cell-adherent activity of these organisms varied among strains; P. gingivalis strains 381, ATCC 33277 and SU63 bound to the target cells at a range of 14% to 72%, but the other 3 strains (KD1, W50 and W83) were scarcely bound (0.6% to 3.5%). On the other hand, whole bacterial cells and culture supernatants of all strains showed distinct hemagglutinating activity. The 3 strains showing high cell-adherent activity were hydrophobic and the other strains showing less activity were relatively hydrophilic. Furthermore, a number of peritrichous fimbriae were found on the surface of P. gingivalis strains 381, ATCC 33277 and SU63, which showed high adherent activity, whereas, fimbriae on the other 3 strains showing low adherent ability were barely apparent. Therefore, it was assumed that the cell-adherent activity of P. gingivalis was related to the hydrophobicity of the cell surface, which was related to the number of fimbriae.


FimC, a chaperone-like periplasmic protein of Escherichia coli involved in biogenesis of type 1 fimbriae.

The product of the fimC gene of Escherichia coli K12 is required for the biogenesis of type 1 fimbriae. Mutations within the fimC gene abolish fimbrial synthesis. The FimC protein was found to be processed and the mature version was located in the periplasm. Unlike similar fimbrial systems, the major type 1 fimbriae structural protein FimA was found to be significantly resistant to proteolytic degradation when present in the periplasm in a fimC- host background. The fimC gene was sequenced, and the deduced primary structure of the FimC protein was compared to other similar known proteins involved in the biogenesis of various fimbriae.


[Arrhythmogenic effects of sultopride chlorhydrate: clinical and cellular electrophysiological correlation].

This study was designed following the first documented case of torsades de pointes induced by sultopride hydrochloride, a substituted benzamide neuroleptic drug. The patient, a 48 year-old woman with no known cardiovascular disease, had been treated for several years with this drug. She was admitted for severe bronchospasm requiring artificial ventilation. Twenty-one hours after her admission, she developed several episodes of torsades de pointes, which were successfully treated with magnesium sulphate. At that time, the QT interval was 500 ms for a heart rate of 108 b.min-1 (QTc of 668 ms, and theoretical QTc 370 ms). On the fourth day, QTc was 548 ms and theoretical QTc 370 ms. The sultopride was stopped on the fifth day. Two days later, QTc was 397 ms. Six months later, there was no recurrence. Several cases of TdP or sudden death have been reported in patients receiving neuroleptic drugs. The effects of sultopride hydrochloride were therefore tested on isolated ferret Purkinje fibres, using the microelectrode technique. Three concentrations of the drug (D1, D2, D3) were tested, as well as normal Tyrode solution. Maximum diastolic potentials (Vmax) were -88.37 +/- 0.89 mV (control), -89.08 +/- 1.20 mV (D1), -90.00 +/- 1.06 mV (D2), and -90.14 +/- 1.20 mV (D3). Vmax was not affected by sultopride during pacing at 1,000 ms of cycle length. The duration of the action potential increased with the drug concentration. There was no early after-depolarisation (EAD) during control, and 7 out of 9 fibers had EAD and 3 out of 9 triggered activity in D3. The solvent (benzyl alcohol) did not modify the action potential.(ABSTRACT TRUNCATED AT 250 WORDS)


Characteristics of haemolytic Escherichia coli with particular reference to production of cytotoxic necrotizing factor type 1 (CNF1).

A total of 1,106 Escherichia coli strains isolated in Spain between 1986 and 1991 from extraintestinal infections and faeces of healthy controls were examined for production of alpha-haemolysin (Hly). Among strains causing urinary tract infections, sepsis and other extraintestinal infections, Hly production was detected in 51% (P < 0.001), 32% (P < 0.001) and 18% (P < 0.02), respectively. In contrast, only 9% of faecal isolates from healthy individuals synthesized Hly. The 356 haemolytic E. coli strains characterized in this study belonged to 28 different serogroups. However, 284 (80%) were of one of eight serogroups (02, 04, 06, 08, 018, 022, 075 and 083); 40% and 31% of haemolytic strains expressed P fimbriae and mannose-resistant haemagglutination (MRHA) type III, respectively. We have found that haemolytic isolates of E. coli may clearly be divided into two categories on the basis of the ability to produce cytotoxic necrotizing factor type 1 (CNF1). The serogroups and adhesins determined in Hly+CNF1+ strains were generally different from those found in Hly+CNF1- strains. Thus, serogroups 02, 06 and 075 were associated with haemolytic E. coli producing CNF1+, whereas serogroups 01, 08, 018, 028 and 086 were established more frequently among Hly+CNF1- strains. While expression of P fimbriae was more frequently detected in Hly+CNF1- strains (70 versus 29%, P < 0.001), MRHA type III was usually identified in Hly+CNF1+ E. coli (42 versus 1%, P < 0.001). Furthermore, the sonic extracts of Hly+CNF1+ strains caused necrosis in rabbit skin (96 versus 25%, P < 0.001) and death in intraperitoneally injected mice (73 versus 11%, P < 0.001) more frequently than sonic extracts of Hly+CNF1- strains.


Evidence for catecholaminergic control of alpha-melanotropin (alpha-MSH) content in hypothalamic areas.

A possible catecholaminergic regulation of hypothalamic alpha-melanocyte-stimulating hormone (alpha-MSH) has been investigated in male rats by an in vivo approach. The hormone was measured by radioimmunoassay in three hypothalamic regions: medial basal hypothalamus, preoptic hypothalamic area and dorsolateral hypothalamus. The tyrosine hydroxylase inhibitor alpha-methyl-para-tyrosine (300mg/kg) increased the hypothalamic alpha-MSH content in medial basal hypothalamus and preoptic hypothalamic area when it was measured at 22:00 h. Diethyldithiocarbamate (600mg/kg), which inhibits dopamine beta-hydroxylase, as well as 2-3-dichloromethylbenzylamide (25mg/kg), which acts on the phenylethanolamine-NCH3 transferase also increased the alpha-MSH content in the above mentioned discrete areas. The alpha-adrenoceptor antagonist phenoxybenzamine (15mg/kg), as well as the alpha 1-adrenoceptor antagonist prazosin (1.0mg/kg), also increased the hypothalamic alpha-MSH content in medial basal hypothalamus and preoptic hypothalamic area. None of these agents modified alpha-MSH content in dorsolateral hypothalamus. Haloperidol (1.2mg/kg), a dopaminergic receptor antagonist, propranolol (6.0mg/kg) and yohimbine (10mg/kg) (non selective beta- and alpha 2-adrenergic antagonist drugs respectively) had no effect on the alpha-MSH in any of the hypothalamic areas studied. These results indicate that the catecholaminergic system is involved in the control of proopiomelanocortin derived hypothalamic alpha-MSH through an alpha 1-adrenoreceptor. The data suggest that the control mechanism in the two alpha-MSH hypothalamic pools are different.


[Febrile convulsions: should some drugs be contraindicated?].

Should certain drugs be contraindicated in children who have had febrile seizures or who present a risk of convulsions? There are no publications dealing specifically with this problem. However, many drugs can induce convulsions and may be dangerous if they are associated with another determining factor (e.g. fever). Camphor known to be toxic and its use must be avoided in young children. Other terpenes given to children with colds may be convulsant if they are used for prolonged treatment or associated with other convulsant drugs (sympathomimetics, piperazine derivatives, antihistamines, etc.). On the basis of a retrospective study of 23 cases of febrile convulsion among 343 cases of infantile convulsion reported to the Poison Control Center and the Pharmacovigilance Center of Marseille between 1973 and 1991, we propose that camphor and sympathomimetics be avoided and that potential convulsant drugs and their association be used with caution. A prospective study is underway to determine responsibility of certain drugs in the occurrence of recurrence of febrile convulsions.


Molecular analysis for the myotonic dystrophy mutation in neuromuscular disorders.

A variable expansion of an unstable CTG repeat has been identified as the causal mutation for myotonic dystrophy. Standard molecular genetic techniques can now supplement traditional assessment protocols in a variety of clinical neurological situations where diagnostic uncertainty prevailed. Southern analysis using DNA probes which identify the expanded sequence, supplemented by direct PCR analysis for repeat number, provides a specific sensitive diagnostic test for myotonic dystrophy.


[Tightly linked DNA probe for presymptomatic diagnosis and carrier detection of Wilson disease].

Haplotype analysis of the polymorphic loci, D13S26 and retinoblastoma (RB) gene which were closely linked to the gene responsible for Wilson disease (WD), was carried out to predict the presymptomatic stage or to detect carrier status in phenotypically normal sibs in 9 Chinese families with WD syndrome. By analysis of D13S26/HphI and RB/XbaI sites, 72% parents in these families were haplotypically heterozygote and therefore informative for linkage study. In 9 phenotypically normal sibs in these families, presymptomatic status was predicted with 99.2% confidence in 1 and excluded in 4. In the other 4 cases, 2 were unpredictable and 2 were at least heterozygote and had 50% chance of being WD homozygote, depending on which chromosome they have got from their fathers.


[Comparison study on distribution of two types of hemorrhagic fever with renal syndrome (HFRS)].

IFAT-positive serum samples were typed by HI and divided into Appodemus-type (type A) and Rattus-type (type R). The results showed that in Pingdu and Jimo counties, the patients were mainly of type A, whereas in Junan and in Tengzhou counties, the patients were mainly of type R. The age distribution showed that 96.0% of the type A patients belonged to age groups between 15-54 years, and no patients were younger than 15 years, or older than 65 years. The age distribution in type R patients, however, showed 76.5% of them were 15-54 years of age. Male/Female ratio was 3.3:1 in type A patients and 1.5:1 in type R patients. 80% of the type A patients were farmers. There were more housewives, students and children in type R than in type A patients. Type A patients usually occurred during Oct. and Dec. with only one peak in type A patients and two peaks in type R patients.


Immunologic characteristics of fibrillary glomerulonephritis.

IgG and IgA immune complexes, mononuclear phagocytic system function, interleukin-2 (IL-2) production by peripheral blood lymphocytes (PBL), serum-soluble IL-2 receptors, tumor necrosis factor, beta 2-microglobulin and IL-1 beta, HLA-DNA polymorphisms, immuno-isoelectrofocusing, phenotype of PBL, lymphocyte cytotoxicity, activation of lymphokine-activated killer cells and natural killer cell activity were evaluated in 8 patients with tubular/fibrillary glomerulonephritis (GN). No common serologic, immunologic or immunogenetic features suggestive of plasma cell dyscrasias were found. No elements to state whether these GNs represent a new entity or just atypical forms of known GN were found.


Partial coexistence of NADPH-diaphorase and somatostatin in the rat hypothalamic paraventricular nucleus.

Coexistence of NADPH-diaphorase (ND) activity and somatostatin (SRIF) immunoreactivity was studied in the paraventricular nucleus (PVN) of the rat hypothalamus by successive incubations of the same sections. ND was found in all PVN subdivisions, mainly in the magnocellular ones. SRIF was practically restricted to the parvicellular periventricular subdivision. Contrary to other brain regions where a wide SRIF-ND coexistence has been observed, the periventricular parvicellular subdivision was the only place of the PVN where some neurons colocalize both markers. The combination of the immunocytochemical and the histochemical labelings allows a further permanent and easy-to-perform parcellation of periventricular PVN neurons.


The non-NMDA antagonist CNQX prevents release of amino acids into the rat spinal cord dorsal horn evoked by sciatic nerve stimulation.

Basal extracellular concentrations of 9 amino acids (AAs: aspartate, Asp; glutamate, Glu; asparagine, Asn; serine, Ser; glycine, Gly; threonine, Thr; alanine, Ala; taurine, Tau; and glutamine, Gln) were determined in the spinal cord dorsal horn of anesthetized rats using microdialysis and HPLC techniques. The concentrations of all measured AAs but Gln increased significantly (P < 0.05) during sciatic nerve stimulation at C-fiber strength. The concentration of Tau remained elevated following stimulation, while the other AAs returned to prestimulation values. Addition of the specific non-NMDA antagonist, CNQX, to the perfusing solution prevented the nerve stimulation-evoked AA release. Since the measured increases in extracellular AA concentrations are probably mainly due to activation of interneurons, these results suggest that blockade of non-NMDA receptors prevented activation of interneurons in the dorsal horn and support a major role of non-NMDA receptors at the first synapse of primary afferent fibers in the dorsal horn. Complete block of AA release and decreased basal levels of Glu after infusion of TTX into the dorsal horn also implies increased neuronal activity as the main source of higher AA levels during nerve stimulation.


A double-label analysis demonstrating the non-coexistence of tyrosine hydroxylase-like and GABA-like immunoreactivities in amacrine cells of the larval tiger salamander retina.

Previous studies have localized tyrosine hydroxylase, the rate-limiting enzyme for the production of dopamine, and gamma-aminobutyric acid (GABA) to amacrine cell populations in the larval tiger salamander retina. Double-label immunocytochemistry was used to examine if tyrosine hydroxylase-like and GABA-like immunoreactivities colocalize in tiger salamander amacrine cells. A total of 2,162 tyrosine hydroxylase-like immunoreactive amacrine cells were observed in double-labelled sections. None of these cells were observed to express GABA-like immunoreactivity. Therefore, the present study demonstrates that dopamine and GABA are localized to distinct neuronal populations in the larval tiger salamander retina.


Proliferating cell nuclear antigen (PCNA) in atypical and malignant meningiomas.

Because it is not easy to determine the tumor status of meningiomas by current diagnostic procedures, we investigated these tumors immunohistochemically using the monoclonal antibody PC 10. This antibody recognizes a fixation- and processing-resistant epitope of the proliferating cell nuclear antigen (PCNA), which is a 36-KD nuclear antigen associated with the cell cycle. We studied paraffin-embedded and formalin-fixed tissue specimens of a group of 21 atypical/malignant meningiomas together with 18 benign meningiomas. PCNA staining results were compared with the mean number of silver-stained nucleolar organizer region-associated proteins (AgNORs), tumor grading, and mitotic indices of these tumors. The percentage of PCNA-positive cells was found to range between 0.1% and 40%, irrespective of the tumor grade. When all tumors were collectively considered, no positive correlation was found between PCNA scores and histologic grading and only a weak one between PCNA score and mitotic index. A higher correlation was seen between AgNOR counts and tumor grading. Our results suggest that PCNA labeling and histologic grading seem to be independent parameters. The correlations found between AgNOR counts and tumor grading should be substantiated in further series.


Histochemical study of dipeptidylpeptidase IV in the nasal cavity organs of laboratory rodents and man.

The localization of dipeptidylpeptidase IV (DPP IV) activity was studied at light microscope level in the mucosa, glands and capillary endothelium of the human foetuses and rat, mouse and guinea pig in some prenatal and postnatal terms. Alkaline phosphatase reaction was used for complete histochemical demonstration of capillary endothelium. The two enzymes were mapped by Lojda's methods (12). The study brings knowledge of the localization of DPP IV activity in the nasal cavity structures and gives a different time survey of the onset of DPP IV activity during ontogenesis. Considering this activity we can presume that this membrane-bound protease DPP IV may participate in the metabolism of neuropeptides in the nasal cavity and play some role in immunological disturbances of patients with rhinitis.


Alfentanil combined with vecuronium or pancuronium for use in eye surgery.

The usefulness of alfentanil for eye surgery, when combined with either vecuronium or pancuronium, was evaluated in 44 fit anticholinergized patients. Following thiopentone, four min after the administration of the relaxant (0.1 mg/kg), and two min after the administration of alfentanil (15 micrograms/kg), a sixth of the patients bucked on introducing the endotracheal tube. The anesthesia for cataract extraction was maintained using alfentanil-relaxant-N2O in 18 patients. In 16 of them, a low dose alfentanil regimen, occasionally supplemented with thiopentone, was satisfactory, and the recovery of alertness was rapid. In 26 patients, alfentanil, 25 micrograms/kg, supplemented with isoflurane, efficiently obtunded the hemodynamic response after traction of extrinsic eye muscles at the commencement of major surgery. The patients receiving vecuronium presented with lower heart rates, than did those receiving pancuronium (P < 0.05). During major operations, nodal rhythm was more frequent in the vecuronium group, than in the pancuronium group (P < 0.0001).


Polyarteritis nodosa related to hepatitis B virus. A retrospective study of 66 patients.

In an attempt to establish the characteristics, circumstances leading to infection and development of polyarteritis nodosa (PN) related to hepatitis B virus (HBV), prognostic factors and outcome, and to define the most effective treatment, 66 patients observed between 1972 and 1989 were analyzed. Hepatitis was clinically present in 19/66 patients before PN. In most cases, PN occurred less than 6 months after infection. Clinical manifestations of PN were comparable to those observed in patients without HBV infection except for orchitis which was present in 13.6% and for pulmonary signs which were absent. Transaminases were normal in 38 cases for SGOT and 31 for SGPT and twice the normal range or more in the other cases. Antineutrophil cytoplasmic antibodies (ANCA) were tested in 22 patients and present in 2 (9%). Twenty-eight patients were treated with prednisone +/- oral cyclophosphamide +/- plasma exchanges. Thirty-eight patients were given a short-term treatment with prednisone followed by the association of vidarabine, 15 mg/kg bw/d for one week and 7.5 mg/kg bw/d for 2 weeks, and plasma exchanges: 14 sessions during the 3 weeks of vidarabine infusion, then tapered until stopping treatment after 2 to 3 months depending upon the clinical results obtained. The mean duration of follow-up was 50.3 +/- 46.1 months. At the end of follow-up, 13 of the 28 patients (46.4%) treated with steroids +/- cyclophosphamide +/- plasma exchanges died and 7/38 (18.4%) of those treated with vidarabine and plasma exchanges (p < 0.001) died. HBe/anti-HBe seroconversion was observed in 2 patients treated with prednisone +/- cyclophosphamide +/- plasma exchanges who were alive at the time of final analysis and in 16 patients receiving the other regimen. The outcome of patients treated with a few days of prednisone, vidarabine and plasma exchange was good and, therefore, we propose this protocol as the first viable treatment for polyarteritis nodosa related to HBV, surpassing the conventional treatment with steroids and cyclophosphamide, which stimulates viral replication.


[Long-term outcome of a hospital series of patients with atrio-ventricular accessory pathway].

Ninety five patients with a mean age of 39 +/- 19 years, 82 of whom were symptomatic, having an accessory atrioventricular bidirectional conduction pathway (WPW syndrome: 77; "concealed": 18) were followed up for an average of 7.3 +/- 2.6 years. The objectives were to analyse: the incidence and causes of death and the possible predictive factors of death due to the WPW syndrome--the influence of medical treatment and type of medication on survival and symptoms. Of the 8 cardiac deaths, 6 seemed to be related to the WPW syndrome, a prevalence of 7.8% and an annual incidence of 1.1/1000. The main risk factors which were identified were: age 62 +/- 8 years versus 37 +/- 15 years in survivors; p < 0.02--associated organic heart disease, especially ischaemic heart disease (5/6)--the description of severe symptoms, in particular recurrent syncope--documented malignant spontaneous or induced arrhythmias (5/6)--anterograde AV conduction with an effective refractory period < or = 230 msec in 4, though it was only 270 msec in the other 2 patients, indicating that this parameter is not specific--amiodarone (6/6) did not prevent the fatal outcome in this particular group of patients. In the "benign" forms, only betablocker drugs could significantly reduce the frequency and severity of symptoms, especially when compared with Class I or IC antiarrhythmics. These results suggest that the indications of radical treatment should be widened in high risk patients, especially when elderly and with associated coronary artery disease. They also suggest that the role of betablocker drugs should be reevaluated in the so-called "benign" symptomatic forms.


[Which anti-ischemic treatment can be prescribed during and after the acute phase of myocardial infarction?].

After myocardial infarction, calcium channel blockers are the most prescribed anti-ischemic drugs followed by nitrate derivatives and beta blockers. In order to assess whether this attitude is justified by published data on their efficacy, a meta-analysis of trials of anti-ischemic drugs in myocardial infarction was performed. The early mortality was 13.3% in the group treated by IV nitrates in the acute phase of myocardial infarction and 17.2% in control groups, reducing the risk by a quarter (95% confidence interval of the odds ratio (CI): 0.55-0.95). When all nitrate derivative trials were grouped together, the reduction in the risk of death of 21% was significant (from 15% to 11.8%) (CI: 0.59-0.94). Although oral nitrate derivatives introduced during the acute phase and continued for several weeks induced a non-significant reduction in mortality of 16%, when given intravenously, the benefits on early and longer term mortality were unquestionable. The mortality was 9.8% in the groups treated by calcium channel blockers and 9.3% in control groups (NS); the recurrent infarct rate was 4.8% and 5.4% respectively (NS). In this family of drugs, there was no product which distinguished itself from the others with regard to beneficial or adverse effects. The early mortality decreased from 9.2% to 8.2% in the groups treated by oral beta-blockade--a risk reduction of 10% (NS) and from 4.2% to 3.7% with intravenous beta-blockers--a risk reduction of 12% (p = 0.03). Late mortality decreased from 9.4% to 7.6%, a reduction of 20% (p < 0.00001) in long term trials.2+ contraindication of betablockers in patients without cardiac failure.


Effect of neutrophils on the growth of human pre-S1-reactive T-cells in vitro.

Granulocyte factor (GF) derives from the specific granules of polymorphonuclear neutrophils. GF possesses an immunoregulatory activity and augments the immune response to antigens in vitro and in vivo. The Pre-S1 sensitive T-cells reactive to Pre-S1 protein in vitro were developed from peripheral blood mononuclear cells of hepatitis B convalescent by in vitro Pre-S1 protein stimulation. The GF involvement in the activation and development of the Pre-S1 specific regulatory T-cells in vitro was studied.


Catecholamine modulation of magnesium plasma levels in the rat.

Epinephrine (0.3 microgram/min) or isoproterenol (0.2 microgram/min) were infused for 40 min in rats to determine the role of catecholamines in the short-term control of plasma magnesium levels. Phentolamine (0.2 and 1 mg/kg) or propranolol (5 mg/kg), injected i.p. 10 min following the beginning of the infusion with catecholamines, were used to block alpha- or beta-adrenoceptors. Epinephrine alone elevated the systolic while decreasing the diastolic blood pressure and initially increased heart rate by 8% to 455 +/- 25 beats/min. Plasma potassium declined by 24% but magnesium levels remained constant. Following 0.2 mg/kg of phentolamine, systolic and diastolic blood pressure decreased by 40% and 55% to 95 +/- 11 and 39 +/- 3 mmHg, respectively. Potassium levels were further reduced by 8%. Administration of this dose of phentolamine increased magnesium levels by 8%. Magnesium levels were elevated by 19% to 2.38 +/- 0.20 mEq/l after 1 mg/kg of phentolamine, possibly due to excessive hemodynamic derangements. Propranolol reversed the hypokalemic and hemodynamic effects of the epinephrine infusion without altering plasma magnesium. Similarly, propranolol abolished the tachycardia and hypokalemia elicited by isoproterenol without affecting magnesium levels. These results suggest that catecholamines play no major role in the short-term control of plasma magnesium in the rat.


Quality of life after open heart surgery 16-18 May 1991.

This is a review of papers presented at the international symposium on 'Quality of Life after Open Heart Surgery' comprising the experience in more than 20,000 patients. Early identification and operation of premorbid personalities, psychological counselling before and after the operation, explanation of true risks, an educational programme in redefinition of family roles, stressing the importance of returning to normal activity in education, employment and society, financial support through health insurance to cover the high cost of open heart surgery, and formulating socioeconomic policies which encourage returning to work yield improvement in all patient groups. Specific to valve replacement is patient participation in deciding the kind of prosthesis and improvement of anticoagulation strategies. Coronary bypass patients benefit broadly through a comprehensive rehabilitation programme. For congenital heart disease, emphasis was laid on the need to educate parents about the disease, the importance of open discussion between parents, the patient and the paediatric cardiologist, and the function of a parents' self-help group. For heart transplantation the endeavour must be to increase donor availability and to develop better immunosuppression schedules. Lastly, general awareness of patients' abilities will facilitate their social integration and improve their quality of life.


Report from a National Cancer Institute (USA) workshop on quality of life assessment in cancer clinical trials.

To promote the inclusion of quality of life (QOL) end-points in clinical research on cancer, the National Cancer Institute (USA) sponsored a workshop on QOL assessment in cancer clinical trials in July, 1990. Experts in clinical trials and QOL research formed four working groups to identify current areas of cancer treatment in which QOL end-points are most important; to discuss methodologic problems in QOL assessment; to address common problems in implementing clinical studies with QOL end-points; and to consider statistical issues in design, implementation, and data analysis. Recommendations made by the working groups are summarized in this paper.


Gene targeting for somatic cell manipulation: rapid analysis of reduced chromosome hybrids by Alu-PCR fingerprinting and chromosome painting.

The techniques of reverse genetics rely heavily on parasexual methods for manipulating the human genome. However, the application of somatic cell genetics is severely limited by the availability of suitable endogenous selectable markers in the genome. We have addressed this problem by targeting a universally selectable marker into a predetermined region of the genome, using a stringent selection for homologous recombination. Correct gene targeting to human chromosome 7q11 was screened for by Southern blotting and confirmed by fluorescent in situ hybridization. Reduced chromosome 7 hybrids were generated by chromosome mediated gene transfer and selection for the neo gene. The resultant transgenomes were characterized by a combination of L1 fingerprinting, locus specific marker analysis, Alu-PCR and chromosome 'painting'. Alu-PCR and L1 'fingerprints' are complementary and mutually consistent. Chromosome 'painting' reflects and extends the results obtained for specific marker co-transfer. Thus Alu-PCR 'fingerprinting' and 'painting' combine to rapidly provide an accurate picture of transgenome content and complexity. Gene targeting, chromosome tagging and subsequent isolation can be applied to any region of the genome for which a molecular probe is available.


[Pharmacologic defense of the brain in radiation injury: some arguments].

Based on an analysis of the latest data on the problem of pharmacologic defense, the pro and contra scientific arguments of using pharmacochemical countermeasures to radiation damages of the brain are discussed in succession. A comparative characteristic of positive and negative effects of countermeasures possessing various neurotropic properties is presented. A general scheme of searching for the means of pharmacologic correction of radiocerebral syndrome involving the following stages: pathophysiologic analysis of cerebral disorders; selection of behaviorally significant (psychophysiologic) criteria; search for pharmacochemical correction means; experimental animals studies with extrapolating their results to a man; radiocerebral syndrome simulation (pharmacologic, physical and chemical); field tests; decision-making (medico-biological and formalized approaches) is presented. "Pyramid of complexity" of radiocerebral effects extrapolation from the animals to a man to suit an hierarchy of the biological structures is presented. The effectiveness of conditionally-quantitative relationships of the drugs of prolonged and short-term effect as applied to various radiation syndromes (erythropoietic, gastrointestinal and cerebral) is discussed.


De novo mutation in the COL4A5 gene converting glycine 325 to glutamic acid in Alport syndrome.

Southern blot analysis of the COL4A5 gene in a 6 year old Italian Alport patient (proband VIZ) showed the loss of an MspI site that was present in the mother and control DNAs. PCR amplification and DNA sequencing revealed a single G-->A nucleotide change. The mutation results in substitution of a glutamic acid for a glycine residue at position 325 in the triple helical region of the alpha 5(IV) chain.


Two hot spots of recombination in the DMD gene correlate with the deletion prone regions.

Genetic mapping has indicated that meiotic recombination occurs about 4 time more frequently in the dystrophin gene than expected on the basis of its length. To detect where recombinations occur within the gene, we have studied the CEPH families panel using highly polymorphic microsatellite markers located at the ends of the gene or flanking the major deletion hot spot in intron 44. We found a major hot spot of recombination between markers STR44 and STR50(1), i.e., between exons 44 and 51. Within this hot spot, a peak of recombination was located in the large intron 44. A second minor recombination prone region was found between DXS 206, (XJ, in the large intron 7) and the 5' end of the DMD gene. The distribution of the recombination events in the gene of healthy individuals was very similar to that of deletion breakpoints in DMD/BMD patients, suggesting that the two phenomenon may share a common mechanism. These results should also improve efficiency and accuracy of linkage analysis applied to carrier detection and prenatal diagnosis. In particular, if markers located at the very 3' end of the gene are not informative, the highly polymorphic ones located between exons 50 and 60 can be used instead of presently available extragenic markers, with a very low risk of diagnostic error due to recombination.


Mapping of the formin gene and exclusion as a candidate gene for the autosomal recessive form of limb-girdle muscular dystrophy.

Limb-Girdle Muscular Dystrophy (LGMD) is a myopathy with clinical and transmission heterogeneity. The recessive form, LGMD2, has been recently mapped by linkage analysis to 15q. As an attempt to identify the gene involved in this pathology, we tested as candidate gene the LD locus, called LD for limb deformity. This gene has recently been identified and mapped to chromosome 15q13-q14. It is homologous to the murine formin gene which is localized to mouse chromosome 2. Mutations in this murine gene have been shown to cause limb deformity and kidney defect. YAC clones containing the LD gene were isolated and utilised to confirm the cytogenetic localisation. Internal DNA polymorphisms of the LD locus were analyzed in LGMD2 and CEPH families. The LD gene was mapped between the alpha cardiac actin gene and the D15S24 locus. Crossovers between the LGMD2 and the LD loci excluded the LD gene as a candidate for LGMD2.


Analysis of exon 7 of the human phenylalanine hydroxylase gene: a mutation hot spot?

Complete sequence analysis of 194 human phenylalanine hydroxylase genes from PKU patients originating from West Germany and Bulgaria revealed 13 different mutations within exon 7 of the gene. Four of these mutations (T238P: ACT-->CCT; L242F:CTC-->TTC; R252G:CGG-->GGG; and 1043 delta 11: nt 1043-nt 1053 deleted) have so far not been described in the literature. Including these new mutations at least 21 different gene lesions and one sequence polymorphism exist for exon 7. Despite this large number unbiased calculation of the mutation frequency/exon size ratio does not provide conclusive evidence that exon 7 is a hot spot for disease causing mutations. Extensive screening during our experiments also failed to demonstrate the existence of excessive polymorphism in this part of the gene. It might therefore be speculated that the functional importance of the highly conserved exon 7 sequence accounts for the clustering of observed mutations which result in clinically manifest PKU. In addition we report our experience in regard to the resolution capacity of denaturing gradient gel electrophoresis (DGGE), a nonradioactive technique for the rapid screening of unknown mutations in exon 7.


Modulation of lipoprotein lipase activity in the rat by the beta 2-adrenergic agonist clenbuterol.

This study evaluated the effects of beta 2-adrenoceptor stimulation on some determinants of triglyceride metabolism. Male Sprague-Dawley rats were injected twice daily with clenbuterol (30 micrograms.kg-1) for 7 days, or with an equivalent volume of vehicle. Serum triglycerides, hepatic triglyceride secretion rate, and lipoprotein lipase activity in white and brown adipose tissues as well as in red vastus lateralis muscle and heart were evaluated in the fasting state and following a fat-free, high-sucrose meal, 3 h after the last agonist injection. In rats killed in the fasting and postprandial states, clenbuterol reduced the mass of white adipose tissue (-25 and -12%, respectively; p < 0.02), whereas it increased the mass of vastus lateralis muscle (+11 and +7%; p < 0.002) and heart (+13 and %; p < 0.0001). In vehicle-injected animals, the fasting state was associated with lower lipoprotein lipase activity in white and brown adipose tissues, and higher enzyme activity in vastus lateralis and heart, compared with the postprandial state. Postprandially, treatment with clenbuterol reduced lipoprotein lipase activity in white adipose (-24%), whereas it increased enzyme activity in brown adipose (+107%) as well as in vastus lateralis (+35%). In fasted animals, no significant variation of enzyme activity in these tissues was observed following clenbuterol treatment, whereas in the heart, a decrease of lipoprotein lipase activity was observed (-22%). Clenbuterol lowered serum triglycerides significantly (-23%), but not their rate of secretion, whereas the agonist decreased the insulin to glucagon ratio only in the postprandial state.(ABSTRACT TRUNCATED AT 250 WORDS)


Regression of left ventricular hypertrophy--are there differences between antihypertensive agents?

Echocardiographically determined left ventricular mass (LVM) is currently considered to be the most powerful risk indicator for cardiovascular disease, yielding prognostic information beyond that provided by the evaluation of traditional cardiovascular risk factors, high blood pressure included. It has been considered logical to try to obtain regression of cardiac hypertrophy, even though the risk-reducing implications of such a measure remain to be fully established. Experimental and clinical studies have shown that some classes of antihypertensive compounds are less effective than others in causing reversal of left ventricular hypertrophy (LVH) in spite of being similarly efficacious in lowering blood pressure. In order to extract the maximum amount of information from clinical studies, a meta-analysis was performed. This analysis included 109 treatment studies, each conformed to strict present rules. Only studies with pharmacological antihypertensive therapy and echocardiographically determined LVM were included. An analysis of the effect of the four first-hand antihypertensive treatment principles, adjusted for differences between studies with ANCOVA, showed that the ACE inhibitors, beta-blockers and calcium antagonists all reduce LVM by reversing wall hypertrophy and that the effect is most pronounced with ACE inhibitors. Diuretics reduce LVM mainly by a reduction in left ventricular diameter. If the difference in ability to reverse LVH, between ACE inhibitors and beta-blockers/diuretics would correspond to a difference in prognosis, then the outcome of antihypertensive therapy might be expected to improve. This hypothesis is currently under investigation.


Distribution of glutamate-like and glutamine-like immunoreactivities in the rat organ of Corti: a light microscopic and semiquantitative electron microscopic analysis with a note on the localization of aspartate.

The light- and electron microscopic localization of glutamate and glutamine in the rat organ of Corti was studied by means of antisera raised against the respective amino acids coupled to carrier proteins. The light microscopic analysis was performed in semithin sections treated according to the peroxidase-antiperoxidase procedure. The two amino acids were visualized in the same ultrathin sections by use of postembedding immunocytochemistry with two different gold particle sizes. The distribution of aspartate-like immunoreactivity was also recorded, but only at the light microscopic level. In the hair cells, the level of glutamate-like immunoreactivity was higher than that in supporting cells but lower than that in the presumed glutamatergic terminals of cerebellar parallel and mossy fibres. The latter types of terminal were sampled from ultrathin sections that had been incubated under the same conditions as the cochlear sections. Within the hair cells, gold particles signalling glutamate were enriched on mitochondria but not on clusters of synaptic vesicles. Glutamine-like immunoreactivity was present in hair cells as well as supporting cells. The glutamate/glutamine ratio, expressed as the ratio between the respective gold particle densities, was considerably lower for hair cells compared with the cerebellar excitatory terminals. No consistent difference was found between outer and inner hair cells in relation to the levels and subcellular distribution of glutamate and glutamine immunoreactivities. Aspartate-like immunoreactivity was accumulated in outer hair cells, with some labelling also of border cells and Böttcher cells. While the present study confirmed the presence of glutamate in hair cells and demonstrated that these cells are also endowed with the important glutamate precursor glutamine, it revealed notable differences between hair cells and presumed glutamatergic terminals in the CNS. These could reflect differences in the synthesis and compartmentation of transmitter glutamate. Methodological factors could also contribute. Alternatively, the differences could be interpreted to suggest that the hair cell transmitter is not glutamate, but a similar compound. Aspartate could be a candidate in the case of the outer hair cells.


Molecular cloning and cDNA sequence analysis of coho salmon stanniocalcin.

Stanniocalcin (STC) (formerly known as both teleocalcin and hypocalcin) is an anti-hypercalcemic, glycoprotein hormone that is produced by the corpuscles of Stannius (CS), endocrine glands that are confined to bony fishes. The hormone has a unique amino acid sequence and exists as a disulfide-linked homodimer in the native state. In previous studies, we have described the purification and characterization of two salmon STCs, and examined the regulation of hormone secretion in response to calcium using both in vitro and in vivo model systems. This report describes the molecular cloning and cDNA sequence analysis of a coho salmon STC messenger RNA (mRNA) from a salmon CS lambda gt10 cDNA library. The STC mRNA in salmon is approximately 2 kilobases in length and encodes a primary translation product of 256 amino acids. The first 33 residues comprise the prepro region of the hormone, whereas the remaining 223 residues make up the mature form of the hormone. One N-linked, glycosylation consensus sequence was identified in the protein coding region as well as an odd number of half cysteine residues, the latter of which would allow for interchain bonding or dimerization of monomeric subunits. In addition, three sites were identified in the mature protein core of STC (two dibasic, one tribasic) that may be acted upon by endopeptidases to produce truncated forms of the hormone. In support of this latter possibility, Western blot analysis revealed multiple molecular weight forms of sTC within salmon glands.


Muscarinic involvement in the regulation of gonadotropin-releasing hormone in the cyclic rat.

The release of gonadotropin-releasing hormone (GnRH) from the median eminence (ME) in cyclic rats was stimulated to a significant extent by the selective muscarinic antagonists 11[(2)(diethylamino)methyl][-1-piperidinyl]-acetyl-5, 11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepine-6-one (AF-DX-116) and methoctramine, and to a lesser extent also by other ligands selectively antagonistic to m1 and m3 receptors. Such stimulation was estrous-cycle-dependent and was not achieved by muscarinic agonists. We suggest that the effect is induced via the m4 receptor subtype. Attempts to block the muscarinic-antagonist-induced stimulation of GnRH release with a variety of drugs were successful only in the presence of prazosin, an antagonist to alpha 1-adrenergic receptors. One possible explanation for this muscarinically mediated stimulation of GnRH release is that it results from cross-talk between the muscarinic and the alpha 1-adrenergic receptors, i.e., muscarinic agonists might inhibit the release induced by alpha 1-agonists, and muscarinic antagonists, by cancelling this inhibitory effect, might thus allow the endogenous alpha 1-agent, norepinephrine, to induce the release of GnRH.


[Clinical assessment of a group of children with juvenile rheumatoid arthritis receiving long-term treatment with oral gold salt].

The authors assess the efficacy of gold salt treatment for juvenile rheumatoid arthritis. The study was carried out on 16 children suffering from mono-pauciarticular, polyarticular and systemic arthritis. Treatment consisted of the administration of auranofin alone in a group of 8 children and auranofin associated to corticosteroids in a second group of 8 children. A marked improvement in clinical conditions was observed with slight transitory side effects at follow-up after 12 and 24 months.


Reaction patterns of tumor infiltrating lymphocytes in different renal cell carcinomas and oncocytomas.

1. Only clear cell and chromophilic carcinomas of the kidney exhibit a considerable lymphocytic infiltration which is compatible with some immunological responsiveness. Chromophobic carcinomas and benign oncocytomas seem to be immunologically reactive. This reflects the different antigen spectrum and histogenesis of these tumors (Störkel and Jacobi, 1989). Clear cell and chromophilic carcinomas are derived from the proximal tubule and chromophobic carcinomas and oncocytomas from the collecting duct. 2. The tumor periphery seems to be the place of greatest immunological importance, as basic requirements of a sufficient lymphocyte/tumor cell interaction can only be expected there. If these data are taken into account for a therapeutical approach with biological immune modifiers the size of the tumor (tumor burden) and proliferation index must be considered too. This might be a likely explanation for the positive effect of inhaled interleukin-2 on lung metastasis in renal cell cancers. Harvesting of tumor infiltrating lymphocytes for therapeutical purposes should take these findings into account. 3. In spite of dense lymphocytic infiltration only 3% of the tumor infiltrating lymphocytes exhibit the activation marker CD 25. There seems to be a sufficient T cell locomotion in renal cell carcinomas but an insufficient T-cell activation. Whether this fact is induced by lacking cytokine stimulation of involved lymphocytes or by still unknown mediators of the tumor cells is not yet known and needs further investigation. 4. Clear cell carcinomas exhibit most adhesion molecules and the highest amount of infiltrating cytotoxic T-cells and natural killer cells.(ABSTRACT TRUNCATED AT 250 WORDS)


[Need of risk reevaluation in morphine dependence in pain patients].

It is commonly recognized than opioids analgesics have an major place in the treatment of pain. In spite of guidelines, opioids drugs remain underutilized in chronic cancer pain and acute severe pain. Among the possible factors, involved in the insufficient use of opioids drugs, is the fear (opiophoby) of physicians, nurses, patients and family to induce or to maintain an addiction. This review examines the potential of iatrogenic addiction. We will examined the place of morphine-like drugs in the treatment of severe acute pain and chronic cancer pain, the definition of dependency in pain patients, the assessment of the dependency potential in patients treated for pain. Available studies indicate that iatrogenic addiction is quite scarce and that the risk for a major tolerance is very small. Further studies will be necessary, since opioids analgesics may also be useful in some non-cancer chronic pain.


The potential ocular phototoxicity of antidepressant drugs.

Light Therapy is a new treatment for patients with Seasonal Affective Disorder (SAD), a depressive state occurring during the winter as a result of decreased sunlight. The treatment involves placing the patient in front of a light box (2-10,000 lux) for approximately 30 min to 1 hour per day during the winter months. Although there have been no reports of damage to the eye from this treatment with light alone there is increased risk in light damage to the lens and retina if these depressed patients are being treated with antidepressant/neuroleptic drugs concurrently with their light therapy. As we have been previously reported certain drugs, having absorptions longer than 295 nm can act as photosensitizers resulting in enhanced light damage to the eye. Using a screening method developed by Roberts, we examined the potential phototoxicity of a variety of antidepressant and neuroleptic drugs.


[Clinical and experimental study of qing wen oral liquid in the treatment of viral infectious fever].

110 cases of viral fever patients receiving Qing Wen oral liquid were observed. The total effective rate was 94.5% in comparing to 86.5% in the control group, P < 0.01. The shortening of time concerning both the beginning on the declining of fever and normalization of body temperature were obvious in comparison with the control. The remedy was also effective in improving symptoms and signs, alleviating renal failure, improving microcirculation and providing bi-directional regulation to the immune system, thus the progression of the disease was controlled. Animal experiments showed that Qing Wen oral liquid could protect the rabbits with hemorrhagic fever, delay the incubation period and the peak of fever, lower the febrile index and PGE content, improve the hemorheology and enhance the cell-mediated immunity in CSF.


Molecular dissection of the Prader-Willi/Angelman syndrome region (15q11-13) by YAC cloning and FISH analysis.

Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are distinct mental retardation disorders associated with deletions of proximal 15q (q11-q13) of different parental origin. Yeast artificial chromosome (YAC) clones were isolated for 9 previously mapped DNA probes from this region, and for one newly derived marker, LS6-1 (D15S113). A YAC contig of 1-1.5 Mb encompassing four markers (ML34, IR4-3R, PW71, and TD189-1) was constructed. Multi-color fluorescence in situ hybridization (FISH) analysis of interphase nuclei was combined with YAC contig information to provide the following order of markers: cen-IR39-ML34-IR4-3R-PW71-TD189-1-LS6++ +-1-TD3-21-GABRB3-IR10-1-CMW1-tel. FISH analysis was performed on 8 cases of PWS and 3 cases of AS, including 5 patients with normal karyotypes. All eleven patients were deleted for YACs in the interval from IR4-3R to GABRB3. On the proximal side of the deletion interval, 10/10 breakpoints fell within a single ML34 YAC of 370 kb. On the distal side, 8/9 breakpoints fell within a single IR10-1 YAC of 200 kb. These results indicate a striking consistency in the location of the proximal and distal breakpoints in PWS and AS patients. FISH analysis on a previously reported case of familial AS confirmed a submicroscopic deletion including YACs corresponding to LS6-1, TD3-21 and GABRB3 and supports the separation of the PWS and AS critical regions. Since these three YACs do not overlap each other, the minimum size of the AS critical region is > or = 650 kb.


Mutations in the medium chain acyl-CoA dehydrogenase (MCAD) gene.

Medium chain acyl-CoA dehydrogenase (MCAD) catalyzes the first reaction of the beta-oxidation cycle for 4-10-carbon fatty acids. MCAD deficiency is one of the most frequent inborn metabolic disorders in populations of northwestern European origin. In the compilation of data from a worldwide study of 172 unrelated patients each representing an independent pedigree, a total of 8 different mutations have been identified. Among them, a single prevalent mutation, 985A-->G, was found in 90% of 344 variant alleles. 985A-->G causes glutamate substitution for lysine-304 in the mature MCAD subunit, which causes impairment of tetramer assembly and instability of the protein. Three of 7 rarer mutations have been identified in a few unrelated patients, while the remaining 4 have each been found in only a single pedigree. In addition to tabulating the mutations, the acyl-CoA dehydrogenase gene family, the structure of the MCAD gene and the evolution of 985A-->G mutation are briefly discussed.


Antinuclear antibody profile in Italian patients with connective tissue diseases.

In the present work we report data on the specificity of antinuclear antibodies (ANA) in a large series of Italian patients suffering from a broad spectrum of connective tissue diseases (CTD), by using a series of homogeneous and validated techniques. The present study confirms, on the one hand, generally accepted concepts, i.e. that certain autoantibodies are strictly associated to certain disease states (such as anti-PCNA and anti-Sm in systemic lupus erythematosus, Jo 1 in polymyositis, and ACA and Scl-70 in scleroderma); the presence of 'marker' antibodies is, however, restricted to a relative minority of CTD patients. The application of a new methodological approach that considers the entire profile of ANA can greatly augment their diagnostic relevance and may provide useful indications for their interpretation, allowing us to establish for the first time the diagnostic usefulness not only of marker autoantibodies but also of certain associations between non-marker autoantibodies. Finally, the application of a more appropriate and powerful statistical tool (multiple correspondence analysis) has further emphasized the clear relationship existing between antibody specificities and certain disease states.


Monoallelic expression of the human H19 gene.

Monoallelic expression of several genes has been observed in mice in which transcripts from parental homologues are distinguishable, but this phenomenon has not been demonstrated in humans. One monoallelically expressed murine gene, H19, encodes an abundant fetal RNA. We have found restriction site polymorphisms in the human H19 gene, located on chromosome 11p15, and examined the representation of these polymorphisms in cDNAs from fetal organs. Expression of H19 is largely or exclusively from a single allele; a similar analysis of the WT1 gene, on 11p13, shows biallelic expression. In the context of previous studies of 11p15 allelic losses in human embryonal tumours, our findings support the possibility of single-step inactivation of monoallelically expressed growth-regulating genes in human oncogenesis.


Linkage of the Indiana kindred of Gerstmann-Sträussler-Scheinker disease to the prion protein gene.

The Indiana kindred variant of Gerstmann-Sträussler-Scheinker disease has amyloid plaques that contain prion protein (PrP), but is atypical because neurofibrillary tangles like those of Alzheimer disease are present. To map the position of the disease causing gene, we used three markers for linkage analyses. A missense mutation at codon 198 of the PrP gene (PRNP) is found in all definitely affected individuals and yields a maximum lod score of 6.37 (theta = 0). The disease also is concordant with the two other PRNP-region markers. These results demonstrate tight linkage of the disease-causing gene to PRNP and support the hypothesis that the codon 198 mutation is the cause of IK-GSS. Our studies also suggest that methionine/valine heterozygotes at PRNP codon 129 have a later age of onset of the disease than codon 129 valine/valine homozygotes.


Mutant prion proteins in Gerstmann-Sträussler-Scheinker disease with neurofibrillary tangles.

Two families with Gerstmann-Sträussler-Scheinker disease (GSS) are atypical in possessing neocortical neurofibrillary tangles (NFTs), which are few or absent in other kindreds with GSS, in addition to amyloid plaques that react with prion protein (PrP) antibodies and protease-resistant PrP accumulation in the brain. A leucine substitution at PrP codon 102 has been genetically linked to GSS in some families. We examined the PrP gene in these families. A serine for phenylalanine substitution was found at codon 198 in the Indiana patients; arginine for glutamine substitution at codon 217 in the Swedish patients. These mutations in PrP are the first to be associated with the appearance of both PrP amyloid plaques and neocortical NFTs in GSS patients.


Congenital adrenal hyperplasia due to point mutations in the type II 3 beta-hydroxysteroid dehydrogenase gene.

Classical 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase (3 beta-HSD) deficiency is an autosomal recessive form of congenital adrenal hyperplasia characterized by a severe impairment of steroid biosynthesis in both the adrenals and the gonads. We describe the nucleotide sequence of the two highly homologous genes encoding 3 beta-HSD isoenzymes in three classic 3 beta-HSD deficient patients belonging to two apparently unrelated pedigrees. No mutation was detected in the type I 3 beta-HSD gene, which is mainly expressed in the placenta and peripheral tissues. Both nonsense and frameshift mutations, however, were found in the type II 3 beta-HSD gene, which is the predominant 3 beta-HSD gene expressed in the adrenals and gonads, thus providing the first elucidation of the molecular basis of this disorder.


Cosegregation of blood pressure with angiotensin converting enzyme and atrial natriuretic peptide receptor genes using Dahl salt-sensitive rats.

We have evaluated the genes for angiotensin converting enzyme (ACE) and guanylyl cyclase A/atrial natriuretic peptide receptor (GCA) for genetic effects on blood pressure response to high salt diet. In F2 rats derived from Milan normotensive and Dahl salt-hypertension sensitive (S) rats, both ACE and GCA cosegregated with blood pressure, and rats that were homozygous for the S allele at both the ACE and GCA loci had inordinately high blood pressure. In F2 derived from Wistar Kyoto (WKY) and S rats, GCA revealed positive cosegregation with blood pressure, but ACE did not. We conclude that certain alleles at the GCA and ACE loci (or at loci closely linked to them) have a significant genetic impact on blood pressure response to high salt in specific rat strains.


The human pseudoautosomal GM-CSF receptor alpha subunit gene is autosomal in mouse.

The gene encoding the granulocyte macrophage colony stimulating factor receptor alpha subunit (CSF2RA) has previously been mapped to the pseudoautosomal region of the human sex chromosomes. In contrast, we report that the murine locus, Csf2ra, maps to an autosome in the laboratory mouse. By in situ hybridization and genetic mapping, Csf2ra maps at telomeric band D2 of mouse chromosome 19. This first instance of a pseudoautosomal locus in human being autosomal in mouse, indicates incomplete conservation between the human and mouse X chromosomes and suggests that the genetic content of the pseudoautosomal region may differ between species of eutherian mammals due to chromosomal rearrangements.


C1 inhibitor hinge region mutations produce dysfunction by different mechanisms.

Heterozygosity for a mutant dysfunctional C1 inhibitor protein, a member of the serine proteinase inhibitor (serpin) superfamily, results in type II hereditary angioneurotic oedema. We identified a "hinge" region mutation in C1 inhibitor with a Val to Glu replacement at P14 Val-432. Recombinant C1 inhibitors P10 Ala-->Thr and P14Val-->Glu did not form stable complexes with fluid phase C1s or kallikrein. The P14 Val-->Glu mutant, however, was cleaved to a 96K form by C1s, while the P10 Ala-->Thr mutant was not. The recombinant P10 mutant also did not complex with C1s, kallikrein or beta-factor Xlla-Sepharose. The two mutations, therefore, result in dysfunction by different mechanisms: in one (P14 Val-->Glu), the inhibitor is converted to a substrate, while in the other (P10 Ala-->Thr), interaction with target protease is blocked.


Heterogeneity of glutathione S-transferase enzyme and gene expression in ovarian carcinoma.

Expression of the three major cytosolic classes of glutathione S-transferases (GST; Pi, Alpha and Mu) was examined by 2D gel analysis and Western blotting of biopsies from 26 patients diagnosed with ovarian carcinoma. In contrast to other tissues, at least one 'constitutive' subunit from each of the three major cytosolic GST classes was expressed. In most cases, pi appeared to be the major form present, although levels of alpha and mu subunit expression were approximately equal to pi in some patients. There was no detectable effect of prior chemotherapy on enzyme activity. Mean transferase activity for primary carcinoma was 79.9 +/- 11.9 (mean +/- SEM; nmol min-1 mg-1), with three pair-matched normal tissues showing minor decreases in transferase activity. One sample, in which a 32% increase in tumour enzyme activity was noted, was from a patient with primary disease and was associated with marked overexpression of a relatively basic form of alpha which was absent from the matching normal tissue, but present in 20% of all tumours examined. RFLP analysis of genomic tumour DNA using a human mu class cDNA probe indicated that at least two of the three mu forms (the 'constitutive' form and one other) observed in ovarian tissue were allelic variants, as a one-to-one correlation was observed between the presence of two Hind III fragments at 13.1 and 2.2 kb and expression of a second, more basic, variable form. This latter form was positively identified as the mu class subunit mu based on Southern analysis and was seen to be present in 40% of the samples examined. However, in the absence of mu expression, at least one other mu class subunit probably corresponding to GST psi, was seen to be present. Thus, at least in ovarian tissues, absence of the mu subunit does not necessarily imply a lack of ability to metabolize mu substrates, as psi has similar catalytic activity. A third mu subunit, probably corresponding to GST phi based on its relatively acidic pI, was also noted in 72% of samples examined, but has unknown substrate specificity. Increased expression of both alpha and mu forms may be of relevance to disease diagnosis and drug response.


Eye movement desensitization versus image confrontation: a single-session crossover study of 58 phobic subjects.

Eye movement desensitization (EMD) and a control procedure, image confrontation (IC) were compared in a group of 58 phobics, 31 of them arachnophobes. Subjects confronted disturbing images in a single-session crossover trial. Anxiety levels were recorded on the SUD Scale. Whenever practicable, SUDs to feared objects were also recorded. EMD and IC were equally effective in reducing anxiety levels. After 1 month, during which subjects were encouraged to use IC daily, improvement was maintained. Since exposure to the disturbing image is common to both methods it must be presumed to be the basis of change when EMD is used in cases of phobia.


Use of mobile phones in the behavioral treatment of driving phobias.

Results of two case studies are presented to illustrate the use of mobile phones with in vivo exposure treatment of refractory driving phobias. Number of miles driven and subjective ratings of anxiety were recorded during a baseline phase and 8 weeks of treatment involving a total of 24 driving practices. One subject's use of a mobile phone increased the number of miles driven alone, but the second subject made little progress and regressed following removal of the phone. These two cases and our experience with other patients suggest that mobile phones can benefit many individuals whose therapeutic progress is impeded by a fear of driving alone, but that phones are counterproductive for certain patients. The potential benefits and disadvantages of using mobile phones are discussed.


Post-traumatic stress disorder due to devastating burns overcome by a single session of eye movement desensitization.

This article reports on the effective use of a single session of eye movement desensitization (EMD) in the treatment of an exceptionally severe case of post-traumatic stress disorder (PTSD). The patient was the survivor of burns that left him with massive scarring, total deafness, bilateral amputations of the upper extremities above the elbow, severe contractures, and severely damaged feet and ankles. He had endured 8 years of intense suffering from symptoms of post-traumatic stress disorder.


Changes in somatostatin-like immunoreactivity in lungs from perinatal guinea pigs and the effects of somatostatin-14 on lung liquid production.

Somatostatin-like immunoreactivity was measured by radioimmunoassay with a monoclonal antibody in lungs from perinatal guinea pigs (62 +/- 2 days of gestation). Fetuses delivered by Caesarean section and dissected before breathing showed 4748 +/- 758 pg/lung (n = 25). Fetuses allowed to breathe (neonates) showed marked increases in activity: 7629 +/- 1355 pg/lung (n = 12) after breathing 30 seconds, and 10729 +/- 1064 pg/lung (n = 6) after breathing 3 minutes (2.3-fold increase, P < 0.005). Values then declined (5203 +/- 1050 pg/lung (n = 9) at 30 minutes; 1458 +/- 105 pg/lung (n = 4) at 60 minutes). Changes were similar in pg/g wet tissue. HPLC characterized the immunoreactive peptides as somatostatin-14 (SS-14) and somatostatin-28 (SS-28) in both fetuses and neonates (n = 11). SS-28 made up only 13.7 +/- 1.7% of the activity; this percentage did not change with breathing. The effects of synthetic SS-14 on lung liquid production were investigated in in vitro lungs from 42 fetal guinea pigs. All 21 preparations immersed in 10(-5)-10(-7) M SS-14 during the middle hour of 3 h incubations reduced production, often approaching zero after treatment (rates, ml/kg body weight per h, succeeding hours: 10(-5) M (n = 9), 3.09 +/- 0.68, 0.93 +/- 0.39, -0.05 +/- 0.60 (fall significant during and after treatment, P < 0.025-0.005); 10(-6) M (n = 6), 3.06 +/- 0.68, 1.29 +/- 0.58, 0.36 +/- 0.38 (P < 0.05-0.005); 10(-7) M (n = 6), 1.96 +/- 0.66, 1.11 +/- 0.34, 0.64 +/- 0.28 (P < 0.05-0.025).(ABSTRACT TRUNCATED AT 250 WORDS)


The cellular immunology of bovine paratuberculosis: the predominant response is mediated by cytotoxic gamma/delta T lymphocytes which prevent CD4+ activity.

Peripheral blood T-cell subsets were obtained from an experimentally sensitized bovine and from nine bovines naturally infected with Mycobacterium paratuberculosis and tested for their ability to respond to antigen. It was determined that following antigen challenge, proliferative responses followed a biphasic pattern: an initial CD4+ proliferative response, a period of anergy, and a final response governed by gamma/delta T lymphocytes. The anergic phase was characterized by a dramatic drop in peripheral blood CD4+ cells; the nature of the non-responsiveness could not be determined. The anergic phase was followed by increased proliferative responses of non-MHC restricted gamma/delta T lymphocytes. Although CD4+ cells had the ability to proliferate in response to M. paratuberculosis antigens in the absence of gamma/delta T cells, antigen-primed CD4+ lymphocytes failed to incorporate [3H]-thymidine in the presence of gamma/delta T cells and M. paratuberculosis antigen. It was concluded that M. paratuberculosis-specific gamma/delta T lymphocytes have immunoregulatory function and exhibit cytotoxic activity against antigen-primed CD4+ helper cells. The data suggest that the inability of effector cell populations to prevent intracellular proliferation of M. paratuberculosis may be a result of cytotoxic killing of the T helper lymphocyte population required for macrophage activation.


Role of T cells, TNF alpha and IFN gamma in recall of immunity to oral challenge with virulent salmonellae in mice vaccinated with live attenuated aro- Salmonella vaccines.

The SL3261 Salmonella typhimurium aroA live vaccine strain confers solid protection against oral challenge with virulent salmonellae, immunity persisting long after the vaccine has been cleared from the tissues. BALB/c mice immunized with SL3261 and later subjected to in vivo depletion of both CD4+ and CD8+ T cells had impaired recall of immunity to oral challenge with the virulent S. typhimurium C5, with increased mortality and higher bacterial loads in the reticuloendothelial system (RES). Selective depletion of CD4+ cells alone significantly impaired resistance both 8 and 14 weeks after vaccination as determined by estimation of bacterial numbers in organ homogenates. Depletion of CD8+ cells alone had less effect on immunity when performed at 8 weeks than at 14 weeks after immunization. Administration of anti-IFN gamma or anti-TNF alpha antibodies also impaired recall of immunity, exacerbating a secondary infection in vaccinated mice. Challenge of T cell-depleted immune mice with virulent salmonellae caused hepatosplenomegaly with minute grossly visible focal lesions, and a marked increase in the number and severity of necrotic foci in spleen, liver and lymph nodes. A widespread mononuclear cell infiltrate was present. The histopathology in anti-IFN gamma-treated mice was qualitatively similar to that seen in T-cell depleted mice. In contrast, in the anti-TNF alpha-treated mice splenomegaly was much less than in T cell-depleted mice. Granulomas were absent, no mononuclear infiltration was observed and there was severe necrosis; the lesions appeared similar to or worse than those seen in naïve mice. Surprisingly, IFN gamma was detectable in sera of both controls and T cell-depleted mice on day 8 of the secondary infection, as well as in sera of anti-TNF alpha-treated mice on day 6 of infection. The results indicate that T cells, IFN gamma and TNF alpha are all important in the specific recall of immunity to virulent salmonellae conferred by immunization with live vaccines, with the effect of T cell and IFN gamma depletion (marked macrophage infiltration) being qualitatively very different from that of TNF alpha neutralization (no mononuclear infiltrate or granuloma formation).


[The activity of cholestatic enzymes in viral hepatitis B].

The examination of 5-cholestasis indicating enzymes was carried out in patients with viral B hepatitis. The changes in activity of such enzymes: alkaline phosphatase, its liver isoenzyme, gamma glutamyltranspeptidase, 5'nucleotidase, leucine aminopeptidase and alanine aminopeptidase were estimated as for being useful in discovering the states of cholestasis. Already in the first week of observation all examined enzymes showed the highest level of activity, only in the case of FZ and FW, however the difference between the patients with and without cholestasis were markedly static. In the following weeks the differences in average activities between group with and without cholestasis were present in the case of 5'N and LAP. The activity of GGTP and AAP were high in both groups of patients. And so alkaline phosphatase is the enzyme which discovers the states of cholestasis in viral B hepatitis quickly and markedly.


[Menadione induction of Ca2+ efflux from the mitochondria and of mediator secretion from the nerve endings].

It is shown that menadion increases the rate of Ca2+ efflux from mitochondria which decreases in the presence of ditiotreitol. Menadion at concentration of 50 microM induces Ca2+ efflux from mitochondria and its effect on synaptic transmission is discussed.


[The involvement of D-1 and D-2 dopamine receptors in the effects of neuroleptics and the pathogenesis of schizophrenia].

The authors presents the literature concerned with the role of D-1 and D-2 dopamine receptors in the pathogenesis of schizophrenia and the effects of neuroleptics. The author concludes that substances which stimulate the D-1 receptors may decrease symptoms of schizophrenia and restrain the formation of tardive dyskinesia, whilst blocking the D-1 receptors may act anti-psychotically without causing tardive dyskinesia. However, they cannot be used with people because of their considerable toxicity. The introduction of new generation of neuroleptics.


[Sigma receptors: the key to therapeutic action or the side-effects of neuroleptics?].

The authors discuss psychotropic effects of phencyclidine (PCP) in the context of neurochemical mechanisms of schizophrenia. They concentrate on sigma receptors and PCP receptors and tie their activity with the dopaminergic system. The authors describe neuroleptic effects on sigma receptors and the therapeutic consequences. They include that the question whether the interaction of neuroleptics with sigma receptors results in therapeutic effects or rather in undesirable symptoms will decide about the future direction of treatment of schizophrenia.


Cell proliferation in rat colon measured with bromodeoxyuridine, proliferating cell nuclear antigen, and [3H]thymidine.

Epithelial cell proliferation was studied in the normal colonic mucosa of 5-week-old Sprague-Dawley rats, comparing [3H]thymidine incorporation (group 1) with two newer proliferation markers, bromodeoxyuridine (group 2) and proliferating cell nuclear antigen (group 3). Microautoradiography (group 1) or immunoperoxidase assays (groups 2 and 3) were carried out. Cells were counted for positive reaction and position along 50 colonic crypt columns/animal. No significant differences were found in number or distribution of labeled epithelial cells in proliferative compartments in crypt columns of normal colonic mucosa; labeled cells were mainly in the lower 60% of colonic crypts. Thus, in this model, bromodeoxyuridine and proliferating cell nuclear antigens were comparable to [3H]thymidine as reliable markers of proliferating epithelial cells in rat colon.


[The specific nature of vagus influences on the heart rhythm under the action of humoral regulators].

Effects of taurin, somatostatin, serotonin and noradrenaline upon the vagal chronotropic action and its components were studied in anesthetised cats. The data obtained suggests the existence of a specific transmitter link for vagal tonic and synchronizing effects upon cardiac rhythm.


Frequency and type of dental traumas in mandibular body and condyle fractures.

Dental injuries in association with 207 mandibular fractures were evaluated from patient files and radiographs. It was shown that 32% of the dentulous patients with condylar fractures had dental injuries, approximately 3.7 traumatised teeth per patient, most of which were dental hard tissue injuries and situated typically in the maxillary molar region. Dental injuries were diagnosed in 30% in association with mandibular corpus fractures, approximately 3.6 traumatised teeth per patient, and injuries were more often dental luxations in the anterior part of the mandible. It was found that 143 teeth were involved in the line of mandibular corpus fracture of 105 patients (1.4 per patient). More than half of the fracture lines were diagnosed as passing the periodontal ligament.


[The participation of the small intestine in the inhibition of gastric secretion induced by H2-receptor blockade].

Resection of the proximal portion of small intestine impaired the inhibition of spontaneous gastric secretion induced by the hystodile H2-receptors blockade in rats. The data obtained suggests participation of enteral mechanisms in the inhibition of gastric secretion.


Histamine skin sensitivity in chronic schizophrenic smokers not responding favourably to neuroleptics.

Histamine, the neurotransmitter involved in both immune and allergic responses to stress, has been reported to affect acute and chronic schizophrenic patients differently in histamine skin sensitivity tests. The authors tested for histamine skin sensitivity patients who had been hospitalized with diagnosed schizophrenia for many years who had not responded favourably to conventional therapy and who had been consistently exposed to active and/or passive smoking. These patients were found to be markedly sensitive to the histamine skin test, whereas a control group of non-hospitalized, non-smoking, non-allergic, non-medicated, 'healthy' volunteers, of similar age and sex, were not sensitive to the histamine intracutaneous test. These findings suggest a possible screening indicator in respect of which patients with a diagnosis of schizophrenia will respond favourably or not favourably to neuroleptics.


[The surgical treatment of malignant tumors of the head of the pancreas and of the periampullary area].

Under analysis are results of pancreatoduodenal resections in 71 patients. In patients with mechanical jaundice lymph drainage with sorption and return of lymph into the vascular bed was used during the preoperative preparation. Occlusion of the duct system of the pancreas stump with glue KL-3 was performed on 62 patients, in 9 patients choledochocholecysto-anastomosis was used in order to improve the quality of the biliodigestive opening. A cryoprocedure on the nonremoved portion of the tumor was performed in 3 cases on the posterolateral surface of the portal vein. These measures allowed the postoperative lethality to be reduced up to 11.2%.


[The effect of epidermal growth factor on the expression of erb B2/neu oncogene in mouse embryo fibroblast cells].

The erb B2/neu oncogene encodes a protein which sequence is closely similar to the epidermal growth factor receptor (EGFR). We have previously found that EGF can induce the expression of erb B1/EGFR gene in normal and 3H-TdR transformed C3H/10T1/2CL8 mouse embryo fibroblast cells i.e. NC3H10 and TC 3H10 respectively, but we do not know whether the neu oncogene expression can be induced by EGF. In this study, the effect of EGF on NC3H10 and TC3H10 has been observed by Northern blot analysis. The result indicated that EGF had a obvious induction effect on neu oncogene expression in these cells. Thus, the expression of both erbB 1/EGFR gene and erbB 2/neu oncogene can be induced by EGF. This result may provide a novel clue to the molecular mechanism of EGF action in cell nucleus.


Health aspects of the use of beta-2 adrenergic drugs in animal production.

In the zootechnical field, there is a strong need to correlate analytical results with biological effects of beta-2 adrenergic agonist drugs on animal health, food processing and human toxicology, taking into account the peculiarity of their administration (long-term treatments with doses tenfold as high as the therapeutical ones). The opportunity to use ELISA tests to readily detect illegal treatments, suspected on the basis of clinical and inspective data, can allow appropriate preventive medicine action by monitoring the food chain during its early steps (in living animals). Sanitary implications will not be limited only to serious clinical signs confirmed by analyses, but should also lead to educational programmes intended for farmers, showing them the main obvious risks in animal production associated with beta-2 agonist side-effects.


Improvement of nitrogen supply for L-threonine production by a recombinant strain of Serratia marcescens.

Serratia marcescens T-2000 was previously reported to be an L-threonine-producing strain that harbors the recombinant plasmid carrying the mutant-type threonine operon. This strain produced 55 g of L-threonine/L of the medium containing urea as a nitrogen source after 72 h of cultivation. In the urea-containing medium, transitory stop of the growth was observed during the early period of cultivation when the entire amount of ammonium ion formed from urea via heat decomposition disappeared in the medium. This indicated that the shortage of ammonium supply in cells might delay both the cell growth and the L-threonine production. The use of ammonia water as a nitrogen source for L-threonine production was therefore studied, because microbial cells generally assimilate this source more readily than urea. When ammonia water was automatically fed to the medium so as to maintain the pH of the medium at around 7, the growth was accelerated, and the L-threonine production reached a maximum of 65 g/L at 48 h. Under these conditions, sucrose, a carbon source, was continuously fed to the medium, resulting in the production of 100 g of L-threonine/L at 96 h. Thus, the L-threonine production of the recombinant L-threonine-producing strain could be increased by devising the method for supply of a nitrogen source.


HIV infected patients: correlation of cutaneous disease with degree of immunosuppression.

The AIDS epidemic has become a world-wide health problem since the disease was identified ten years ago. An HIV-dermatology outpatient clinic was established at the Albion Street Centre in Sydney to manage patients with cutaneous manifestations of AIDS. This study reviews the spectrum of skin disorders seen, and attempts to correlate skin disease with the degree of immunosuppression as reflected by CD4 or T helper lymphocyte counts, whereas previous reports have attempted to correlate skin disease with clinical signs and symptoms. Brief summaries of HIV disease and the cutaneous manifestations of AIDS are given.


Effects of ifenprodil on the N-methyl-D-aspartate receptor ionophore complex in rat brain.

The effects of a cerebral anti-ischemic drug ifenprodil on the receptor ionophore complex of an N-methyl-D-aspartate (NMDA)-sensitive subclass of central excitatory amino acid receptors were examined using [3H](+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10- imine (MK-801) binding in rat brain synaptic membrane preparations as a biochemical measure. The binding in membrane preparations not extensively washed was markedly inhibited not only by competitive NMDA antagonists such as (+/-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic, D-2-amino-5-phosphonovaleric and D-2-amino-7-phosphonoheptanoic acids, but also by competitive antagonists at the strychnine-insensitive glycine (Gly) site including 7-chlorokynurenic acid and 6,7-dichloroquinoxaline-2,3-dione. Among several proposed ligands for alpha-adrenergic receptors tested, ifenprodil most potently inhibited the binding in these membrane preparations due to a decrease in the density of the binding sites without significantly affecting the affinity. Ifenprodil also inhibited the binding of [3H]N-[1-(2-thienyl)cyclohexyl]piperidine as well as of [3H]MK-801 to open NMDA channels in a concentration-dependent manner at concentrations above 10 nM in membrane preparations extensively washed but not treated by a detergent, with a Hill coefficient of less than unity. Further treatment of extensively washed membrane preparations with a low concentration of Triton X-100 resulted in an almost complete abolition of [3H]MK-801 binding, and the binding was restored to the level found in membrane preparations not extensively washed following the addition of both L-glutamic acid (Glu) and Gly. Ifenprodil was effective in inhibiting [3H]MK-801 binding via reducing both initial association and dissociation rates in Triton-treated membrane preparations, irrespective of the presence of Glu and Gly added. The binding in Triton-treated membrane preparations was additionally potentiated by the polyamine spermidine in a concentration-dependent manner at concentrations above 10 microM in the presence of both Glu and Gly at maximally effective concentrations. Ifenprodil invariably diminished the abilities of these three stimulants to potentiate [3H]MK-801 binding at concentrations over 1 microM in a manner that the maximal responses each were reduced. These results suggest that ifenprodil does not interfere with the NMDA receptor complex as a specific isosteric antagonist at the polyamine domain in contrast to the prevailing view.


Circadian and seasonal rhythms of 5-HT receptor subtypes, membrane anisotropy and 5-HT release in hippocampus and cortex of the rat.

Specific serotonin binding (5-HT1, 5-HT1A, and 5-HT2 subtypes) and membrane anisotropy were measured at 2 h intervals over a 24 h period in the hippocampus and cortex of Wistar WU rats, housed under a 12 h light-dark cycle, with lights on at 07.00. All experiments were performed both in March and December. In the hippocampus significant circadian rhythms could be ascertained for 5-HT1 binding sites in March and December while for 5-HT1A (subtype of 5-HT1) binding sites the circadian rhythm was only significant in March. The membrane anisotropy also showed significant variations only in March. Circadian rhythms were also found in the cortex for 5-HT1 (December) and 5-HT2 (March and December) binding sites as well as for the membrane anisotropy (December). A correlation was found between membrane anisotropy and 5-HT1 and 5-HT2 binding sites in hippocampus and cortex, respectively. A circadian rhythmicity was also observed for serotonin release as measured by in vivo voltammetry in both brain areas. The results obtained on the diurnal variations of serotonin receptor subtypes and serotonin release and the probable inverse relationship of these two parameters may be relevant in understanding the coupling of pre- and postsynaptic activity.


Structure and expression of human and rat D2 dopamine receptor genes.

D2 dopamine receptor may be related with the pathogenesis of Parkinson's disease and schizophrenia. Furthermore, the antipsychotic drugs have high affinity for D2 dopamine receptor. We carried out the cloning of the genomic DNA for human D2 dopamine receptor and clarified the structure of this gene. Our isolated gene spans about 15 kbp and consists of seven exons interrupted by six introns. However, putative first exon was not yet identified. Spot blot hybridization analysis of cell sorter fractionated human chromosomal DNA with D2 receptor genomic DNA revealed the localization of this gene in the chromosome 11 fraction. We analyzed human genomic DNA by Southern blot hybridization with D2 dopamine receptor genomic DNA as a probe, but so far we could not find RFLP. Northern blot analyses of brain RNA of several animals and rat brain RNA after various treatments were carried out. Developmental changes of D2 dopamine receptor mRNA were observed in the rat brains.


The amino acid substrate of bovine tyrosine hydroxylase.

Tyrosine hydroxylase catalyzes the tetrahydropterin-dependent hydroxylation of tyrosine to form 3,4-dihydroxyphenylalanine. Several nonphysiological aromatic amino acids have been examined as inhibitors and substrates for bovine adrenal tyrosine hydroxylase. The Ki values for para-substituted phenylalanines increase as the size of the substituent increases. For each A2 increase in surface area of the substituent, the free energy of binding becomes 50 cal more positive. Replacement of the phenyl ring with a pyridyl ring decreases the affinity about one order of magnitude. A number of these aromatic amino acids are also substrates for the enzyme. The KM values again increase in size with increasing size of the substituent, but the Vmax value is independent of the reactivity of the amino acid. The effect of size on binding is consistent with a tight interaction between the para position region of the substrate and the enzyme. The lack of a change in the Vmax value is consistent with the rate-limiting step in catalysis by bovine tyrosine hydroxylase being formation of the hydroxylating intermediate rather than hydroxylation of the amino acid. These results will be useful in designing mechanism-based inhibitors of catecholamine biosynthesis and establish that the mechanisms of rat and bovine tyrosine hydroxylase do not differ significantly.


Adhesion molecules on the plasma membrane of epidermal cells. IV. Immunolocalization of the intercellular adhesion molecule-1 (ICAM-1, CD54) on the cell surface of a small subpopulation of keratinocytes freshly isolated from normal human epidermis.

The intercellular adhesion molecule-1 (ICAM-1) is a cell membrane glycoprotein displaying a pivvtal role in cell-cell interactions in the immune system, and is a ligand for LFA-1, which is expressed on leukocytes. ICAM-1 is expressed in different cell types, including epithelial cells in a number of organs; the universal feature on all these cells is ICAM-1 induction from very low ICAM-1 constitutive levels on unstimulated resting cells to very high ICAM-1 levels triggered by mediators released at sites of inflammation. Therefore, since a strong expression of ICAM-1 on keratinocyte (KC) surface was recently demonstrated in various inflammatory skin lesions, in this investigation we asked whether very low ICAM-1 levels might be present on the plasma membrane of unstimulated KC in normal skin. Crude epidermal cell suspensions, freshly isolated from normal human skin, were immunolabeled by anti-ICAM-1 monoclonal antibody and stained by two highly sensitive ultrastructural detection systems, namely, the immunogold (5-nm-sized particles) method and the immunogold-silver-enhancement method. The quantitative analysis of 1000 KC scrutinized under the electron microscope revealed that 17.2% KC were ICAM-1-positive, although a density per KC section (midplane) of merely 18.92 +/- 13.02 5 nm-sized particles was scored (n = 100), indicating that the amounts of ICAM-1 moieties on this KC subset are presumably low. The ICAM-1 expression on a subset of KC in normal skin might account for the trafficking to and from normal epidermis of LFA-1-positive cells, including migrating Langerhans cells and occasional leukocytes.


Localization and characterization of aminopeptidase P in bovine adrenal medulla.

Aminopeptidase P (EC 3.4.11.9) is demonstrated for the first time in the cytosolic fraction of chromaffin cells of the bovine adrenal medulla. The enzyme is inhibited by metal chelators and by sulfhydryl-reactive agents, which suggests that both a tightly bound metal ion and a cysteine residue are necessary for enzymatic activity. Aminopeptidase P might be important for the modulation of the biological activity of neuropeptides. Its occurrence in the adrenal chromaffin cells provides a useful tool for studying the function of this unique proline-specific peptidase in neuropeptide processing and secretion.