Responses of enterocyte microvilli in experimental diabetes to insulin and an aldose reductase inhibitor (ponalrestat).

The small intestine of 12-week-old streptozotocin-diabetic rats was examined by light and transmission electron microscopy in order to study the effects of alternative treatments on microvillous morphology. Four groups were examined: untreated diabetic rats, insulin-treated diabetics and rats treated with an aldose reductase inhibitor (ponalrestat) given with and without insulin. Numbers and dimensions of microvilli at the apex of columnar absorptive epithelial cells (enterocytes) were estimated using stereological principles. Values were obtained for the organ as a whole as well as for different sites along its length. In the untreated diabetic intestine, the mean (standard error of mean) number of microvilli was 4.5 (0.8) x 10(12) with a total surface area of 1.9 (0.50) m2. On average, the microvilli were 1.1 (0.08) microns long, 104 (3.8) nm in diameter and packed on the villous surface at a density of 3400 (50) per 100 microns 2. Their length at least varied with intestinal location. Significant effects of insulin therapy were detected. In contrast, the study failed to find any significant effect of aldose reductase inhibition on any variable except microvillous packing density.


Biochemical and immunohistochemical studies on overgrown gingival tissues associated with mannosidosis.

The gingival tissues of a male patient suffering from mannosidosis and presenting with gingival overgrowth have been studied. Routine histological assessment highlighted the presence of highly enlarged and vacuolated lymphocytes. The morphology of the connective tissues, fibroblasts and epithelium appeared normal. Immunohistochemical staining of the tissues for chondroitin sulfate proteoglycan demonstrated a normal distribution of this component throughout the connective tissues and intense staining associated with the vacuolated lymphocytes. In vitro studies indicated that fibroblasts isolated from the overgrown tissue did not differ from age and sex matched control fibroblasts with respect to proliferation, protein and proteoglycan synthesis. Taken together, these findings imply that the gingvial overgrowth noted in this patient was not due to a defect in the resident fibroblasts but rather reflected a secondary response to the tissues to impaired host defence mechanisms.


[The prospects and problems of drug-induced anesthesia].

The problem of pain has a major medico-social significance and necessitates detailed investigation both from the position of etiology and pathogenesis and new therapy methods. Among many methods of pain control drug-induced anesthesia is of great importance. The multiplicity of pain requires optimal use of analgetics and their combination with other agents strictly in coordination with individual needs of the patient. Special anesthesia systems are to be designed for this purpose.


Advantages of achiral h.p.l.c. as a preparative step for chiral analysis in biological samples and its use in toxicokinetic studies.

1. Achiral reverse-phase h.p.l.c. with semi-automated post-column fraction collection and solid-phase sample reconcentration, has been applied as the purification procedure during the enantiomeric quantification of two widely differing experimental drugs; an HMG-CoA reductase inhibitor (I) and an alpha 2-adrenoceptor antagonist (II). 2. The robust and specific achiral methodologies were available prior to the need for chiral analyses and recovery of drug from the fractions provided clean samples from a variety of biological matrices, without the need to develop compatible achiral/chiral mobile phases. 3. Compared with direct chiral chromatography of plasma extracts, this approach decreased the potential for metabolites and endogenous components to interfere or impair the performance of the chiral stationary phase. 4. The availability of quantitative data from achiral analysis of samples negated the need for internal standardization of the chiral analyses, helped confirm assay specificity and provided potential to determine enantiomeric ratios where only one isomer could be accurately measured. 5. Routine enantiomeric analyses were successfully carried out on samples taken from animals dosed orally with the racemic drugs, providing important data on the possible levels of exposure to individual enantiomers during toxicity testing.


Effects of adrenaline, dopamine, serotonin, and different cholinergic agents on smooth muscle preparations from the ansa proximalis coli in cattle: studies in vitro.

The effects of adrenaline, dopamine, serotonin, and different cholinergic agents on isolated muscle strips from the ansa proximalis coli in cattle were studied. Adrenaline (5 x 10(-5) mol/l) evoked a relaxing effect (p < 0.05) on longitudinal and circular muscle strips, mediated via beta- and alpha 1-receptors respectively. High concentration of dopamine (5 x 10(-4) mol/l) caused a non-significant inhibitory effect on longitudinal smooth muscles, which was probably mediated by beta- and not by dopamine-receptors. Serotonin (5 x 10(-5) mol/l) and cisapride (75 x 10(-6) mol/l) had no effect. Carbachol (2 or 10 x 10(-6) mol/l) and bethanechol (5 or 10 x 10(-5) mol/l) caused a dose-dependent (p < 0.05) contraction of both smooth muscle types. This excitatory effect was inhibited (p < 0.05) by metoclopramide (1 x 10(-4) mol/l), as well as the muscarinic agents RS-86 (5 x 10(-5) mol/l) and CGP-37,218 (37.5 or 75 x 10(-6) mol/l).


[Approach to the optimal therapy of the psycho-autonomic syndrome].

To potentiate the psycho- and autonomic activity of phenazepam and to decrease its side effects, cranio-cervical electrophoresis of the drug is suggested. The method is realized with the aid of a special electrode helmet which permits localizing electrodes in the frontal and occipital areas and at different levels of the lateral surfaces of the neck. Phenazepam in the amount of 1 ml of 3% solution was administered from the cathode. 194 patients with the psychovegetative syndrome in the clinical structure of neurosis or neurosis-like condition were treated. The dynamics of the psycho-vegetative status was verified with the aid of psychometric and vegetative techniques. Cerebral hemodynamics was investigated by rheoencephalography. The data obtained demonstrate that the treatment suggested improves the patients' functional status, decreases reactive anxiety, raises the tempo of sensomotor reactions and attention activity, ameliorates short-term memory, exerts a sympatholytic action, and makes cerebral hemodynamics return to normal. The method produces fewer side effects as compared to the enteral intake of phenazepam.


[Use of intravascular laser irradiation of blood in the treatment of drug-resistant forms of schizophrenia].

Overall 62 schizophrenic patients resistant to all forms of treatment were examined for the clinical efficacy of the treatment method on the basis of intravascular laser blood irradiation. A detailed description of the method, equipment, optimal doses, and the degree of effect in different psychopathological syndromes are provided. The best results were obtained in cases where the depressive paranoid symptomatology was predominant. The authors give recommendations for practical use of laser therapy under the conditions of the psychiatric hospital, discuss its action mechanism.


[Use of plasmapheresis in the complex treatment of patients with endogenous psychoses].

The results of the use of plasmapheresis in multimodality treatment of patients with endogenous psychoses are provided for the first time. The use of plasmapheresis was shown to be highly effective in overcoming resistance to psychotropic agents as well as in the treatment of lingering and chronic extrapyramidal neuroleptic disorders. The beneficial effect on the mental and neurologic status of the patients is coupled with pronounced immunocorrective action of the procedure.


[Various indicators of the hormonal profile in patients with neuroses after psychopharmacotherapy and functional test with sinacten].

Studies of adenohypophyseal hormones, cortisol, T3, T4, thyroxine-binding globulin, testosterone, progesterone and estradiol in 90 neurotic men, treated or not with amitriptyline and benzodiazepines, allowed a conclusion that in the majority of them, endocrine functions were activated. 11 healthy men and 11 male patients underwent the three-day test with sinacten-depot (1 mg intramuscularly). The functional test data were compared over time by the methods of nonparametric statistics. The patients manifested a decrease of reserve potentialities of the pituitary-adrenal system, redistribution of the fractions of thyroid hormones, gonadotropins, the progesterone/estradiol ratio, a rise of prolactin and testosterone secretion. The drugs were found to improve the functioning of the pituitary-adrenal system. However, they did not contribute much to the positive changes in the pituitary-thyroid and other systems.


Effects of l-stepholidine on tyrosine hydroxylase activity in rat corpus striatum.

l-Stepholidine (SPD) 2.5 and haloperidol (Hal) 1.0 mg.kg-1 ip increased rat striatal L-3,4-dihydroxyphenylalanine (DOPA) and 3,4-dihydroxyphenylacetic acid (DOPAC) accumulation induced by NSD 1015 (50 mg.kg-1, ip), a decarboxylase inhibitor. SPD (2.5 mg.kg-1, ip) did not alter but apomorphine (2.0 mg.kg-1, ip) decreased the dopamine (DA) content elevated by gamma-butyrolactone (GBL, 750 mg.kg-1, ip) in the rat striatum. Ip injection of either SPD 5.0 or Hal 2.5 mg.kg-1 after NSD 1015 50 mg.kg-1 or NSD 1015 plus GBL 750 mg.kg-1 also augmented tyrosine hydroxylase (TH) activity in the rat striatum. These results suggest that SPD produces an antagonistic effect on presynaptic DA receptors.


Comparison of effects of tetrahydropalmatine enantiomers on firing activity of dopamine neurons in substantia nigra pars compacta.

Extracellular single unit recording techniques were used to elucidate the effects of enantiomers of tetrahydropalmatine (THP) on the firing activity of dopamine (DA) neurons in substantia nigra pars compacta (SNC). (-)-THP rapidly reversed the apomorphine (Apo)-induced inhibition of the SNC DA cell firing activity (ED50 = 0.77, 0.52-1.14, mg.kg-1), while much larger doses of (+)-THP were required to reverse the Apo-induced inhibition (ED50 = 23, 15.2-34.7, mg.kg-1) and the maximal reversal caused by (+)-THP was 79 +/- 9% of the basal firing rate. In paralyzed rats, (-)-THP (0.5-16 mg.kg-1) significantly increased the spontaneous firing rate of SNC DA neurons dose-dependently, while (+)-THP did not until the dose reached 16 mg.kg-1. Pretreatment with (-)-THP 4 mg.kg-1 attenuated Apo-induced inhibition of SNC DA cell firing rate, while (+)-THP 32 mg.kg-1 revealed a similar potency to block the Apo-induced inhibition. In addition, (+)-THP did not potentiate the effect caused by d-amphetamine (Amp) as some behavioral experiments have shown, but large dose of (+)-THP (32 mg.kg-1) blocked the Amp-induced inhibition of SNC DA cell firing activity as (-)-THP (4 mg.kg-1) did. These results suggest that the interaction between D2 receptors and THP enantiomers has stereoselectivity and that (-)-THP is a D2 antagonist while (+)-THP seems to be not.


A 10-year follow-up study of tardive dyskinesia.

In a 10-year follow-up study of 44 patients with tardive dyskinesia (TD), the majority (22 or 50%) had no change in their TD severity, 9 (20%) had an improvement and 13 (30%) had a worsening of their TD. Little difference was noted in those patients whose medication was decreased (n = 12) and those whose medication remained unchanged (n = 32). Of the women, 26% showed improvement as compared with 11% of the men. Also, patients whose TD improved had lower present neuroleptic dose than those whose TD worsened. These two factors should be studied in larger patient cohorts.


The effects of alcohol and benzodiazepines on the severity of ski accidents.

Urine samples from 402 victims of ski accidents were analyzed for the presence of benzodiazepines (BZD) and alcohol. Eighty-one (20%) samples were positive for alcohol; BZD were detected in 34 (8.5%) cases. Ten of the samples (2.5%) were found to be positive for both alcohol and BZD. Subjects who were positive for either alcohol or BZD or both were older than the other persons examined. The prevalence of alcohol was significantly higher among male accident victims. BZD intake could be demonstrated to have a significant influence on the severity of injuries. Besides an increased awareness of the need for skier education regarding drug use, heightened attention of medical caregivers is warranted to inform their patients about potential accident hazards in sport activities when BZD are prescribed.


Regulation of calcitonin gene expression by hypocalcemia, hypercalcemia, and vitamin D in the rat.

High calcium leads to the secretion of calcitonin, and the administration of 1,25-dihydroxyvitamin D3 leads to a decreased transcription of the calcitonin gene. We now report the effect of chronic hypercalcemia, hypocalcemia, and vitamin D deficiency on calcitonin gene expression in vivo in the rat. Hypercalcemia was created by calcium infusions for 6 h, a high-calcium diet given to weanling rats for 3 weeks, and the transplantation of the Walker carcinosarcoma 256 cell line. Despite serum calcium as high as 22 mg/dl, there was no difference in calcitonin mRNA levels among these rats. The control genes studied, actin and somatostatin, which is specific for C cells in the thyroparathyroid tissue, also did not differ among the different groups of rats. Injected 1,25-(OH)2D3 decreased calcitonin mRNA levels at 6 h, as previously reported. Hypocalcemia, created by feeding diets deficient in calcium and vitamin D to weanling rats for 3 weeks, had no effect on calcitonin mRNA levels, in contrast to the large increases in PTH mRNA levels. These results demonstrate that calcitonin gene expression in vivo in the rat is regulated by administered 1,25-(OH)2D3 but not by changes in serum calcium.


Electrophysiological characterization of histamine receptor subtypes in sheep cardiac Purkinje fibers.

The histamine-receptor-subtype-mediated effects on action potentials of electrically driven and spontaneously active isolated sheep cardiac Purkinje fibers were investigated using H1- and H2-selective agonists and antagonists. In electrically stimulated Purkinje fibers, histamine (3 mumol/l) increased the action potential plateau height, decreased the action potential duration measured at a repolarization level of -60 mV and enhanced the pacemaker activity. These effects were abolished by the H2-selective antagonist cimetidine (30 mumol/l), but were not impaired by the H1-selective antagonist dimetindene (0.3 mumol/l). In spontaneously active Purkinje fibers, histamine (10 mumol/l) increased the spontaneous rate by 24%, the slope of diastolic depolarization by 45% and shortened the duration of the diastole by 32% of the respective control measurements. These effects were blocked by 30 mumol/l cimetidine, but remained unchanged in the presence of 0.3 mumol/l dimetindene. Concentration-response curves of histamine were shifted to the right by approximately 2 logarithmic units in the presence of 30 mumol/l cimetidine, but were not influenced in the presence of 0.3 mumol/l dimetindene. The H2-selective agonist impromidine (0.001-0.3 mumol/l) had similar actions as histamine on spontaneously active Purkinje fibers, while the H1-selective agonist 2-(2-pyridyl-)ethylamine was ineffective. It is concluded that the pronounced stimulatory action of histamine on spontaneous activity in sheep cardiac Purkinje fibers is exclusively mediated by H2 receptors.


Production of monoclonal antibodies to rotavirus suitable for detection of rotavirus in stool samples by one-step EIA.

Two stable hybridoma cell clones producing high amounts of monoclonal antibodies (MAb) to rotavirus have been selected. The MAbs were shown to recognize different epitopes of the major inner capsid rotavirus protein VP 6. The two types of MAb in question cross-reacted with human and animal group A rotaviruses. Due to their high affinity and good binding capacity to microtiter plates a simultaneous EIA was developed for detection of rotavirus in stool samples. The sensitivity and specificity of MAb one-step EIA was compared with an approved polyclonal sandwich EIA by testing 1309 stool samples.


Isolation and purification of HSV-1 specific transfer factor produced by HSV-1 immunized goat leukocyte dialysate.

A herpes simplex virus type 1 (HSV-1)-specific transfer factor (TF), was separated and purified from the leukocyte dialysate of goats immunized with HSV-1 using affinity chromatography on antigen-sorbent and reversed phase high performance liquid chromatography (RP-HPLC). The antigen-specific activities of the starting dialysate and the isolated TF component (s) were examined by 51Cr-labelled leukocyte adherence inhibition (51Cr LAI) assay. The analytical hydrophobic interaction HPLC (HI-HPLC) and isoelectric focusing (IEF) techniques were employed to evaluate the purity and the isoelectric point (PI) of isolated TF component(s). The experiments provided a two-step procedure for purifying the TF material from the starting dialysate. It seems that the purified active TF component (PTFC) was specific for HSV-1. The specific PTFC activity was increased 10,000-fold as compared with the activity of the dialysate. The active moiety appeared as a single band in the IEF gel as demonstrated by silver staining; it was hydrophilic and its PI was pH 4.48.


Chemical characterization of the purified component of specific transfer factor in the leukocyte dialysates from HSV-1 immunized goats.

The chemical characterization of the purified component responsible for HSV-1 specific transfer factor activity (PTFC) by high resolution analytical methods was performed. PTFC had a molecular weight of 6,000 dalton by the size-exclusion HPLC analysis; it showed a marked UV-absorbance spot at 254 nm and a fluorescent spot at 366 nm on the thin-layer plate by thin-layer chromatography which spots coincided at the same place of the plate. The amino acid composition and sequencing analyses showed that PTFC consisted of at least twelve different amino acids, but the amino acid sequence could not be determined. The combined results indicate that PTFC is a compound with a molecular weight of 6,000 dalton, composed of peptide and nucleotide-like material. The peptide is rich in aspartic acid and its N-terminal end may be blocked.


Comparison of two defective hepatitis A virus strains adapted to cell cultures.

The replication of two defective hepatitis A virus strains in cell culture was examined. The w.t. HAS-15 strain growing in FRhK-4 cells produced infectious icosahedral virions 27 nm in size as well as round shaped particles with lipids attached to their surface. The morphogenesis of HAV was membrane-dependent and the detected particles were in various degree of maturation. The MBB 11/5 strain growing in PLC/PRF/5 cells produced mainly noninfectious empty procapsids without RNA genome. The translation of viral proteins was uninhibited in both strains. The reason for restricted replication competence of both strains seemed to be different. In HAS-15, highly efficient encapsidation of the progeny RNA positive-strand lowered the formation of replicative intermediate forms. In MBB 11/5, nearly exclusive empty procapsid production gave evidence for the failure of the VPg primer protein attachment to viral RNA. Changes in the efficacy of viral genome replication were a result of the adaptation of HAV to propagation in vitro.


Binding rate of rabies virus to alpha interferon primed mouse neuroblastoma cells.

Mouse neuroblastoma cells (MNA) were primed with 20 I.U. of alpha interferon (Mu IFN-alpha) prior to exposure to the Challenge Virus Standard strain of rabies virus (CVS). Saturation of CVS receptor sites occurred when 0.71 microgram of 3H-CVS protein bound to 20,000 MNA cells. After 180 min incubation time, the amount of viral protein attributed to specific binding was estimated to be 0.45 microgram. Mu IFN-alpha treatment of MAN cells did not increase the number of specific cell receptor sites to CVS but it did significantly increase the rate that CVS was able to bind to cell receptors. The amount of time required to reach saturation of MNA cell receptors to CVS decreased from 120 min in untreated cells to 30 min in Mu IFN-alpha primed MNA cells. Although treatment with Mu IFN-alpha increased the binding rate of rabies virus to MNA cells, the virus was unable to complete a productive infection 48 hrs after the Mu IFN-alpha was removed.


Detection of tick-borne encephalitis virus in ixodid ticks collected in natural foci by time-resolved fluoroimmunoassay.

Time-resolved fluoroimmunoassay (TR-FIA) was used for the first time for evaluation of infestation of ixodid ticks with tick-borne encephalitis virus. Comparison of TR-FIA results with those obtained in enzyme immunoassay and by virus isolation confirmed the high efficacy of the method in question. Positive results of TR-FIA coincided with the data of virus isolation in 83.6% cases, the level of false-negative results did not exceed 1.2%, the overall time consumption amounted to about 1.2 hr.


Immunoenhancement and suppression induced by adenovirus in chicken.

Chickens injected intravenously (i.v.) with human adenovirus type 6 (Ad6) reveal a 2-17-fold increase in the number of plaque-forming cells producing antibody (Ab) against sheep red blood cells (SRBC) 2-6 days after virus infection. Further, polyclonal B-cell activation has been demonstrated by the quantitation of immunoglobulin-producing cells (IgPC) and cells producing immunoglobulin (Ig) of IgM isotype (IgPC mu) in the spleen of chicken inoculated with Ad6. Ad6 infection in chicken results in immunosuppression against SRBC when this unrelated antigen is given after virus infection. It seems that coincidence occurs between the B-cell mitogenic activation and the immunosuppression caused by Ad6, as the most pronounced change in both activities appears on the fourth day following virus infection. These findings suggest that the B-cell mitogenicity of the virus contributes to the impairment of the humoral immune response to SRBC.


Two-way cross-protection between West Nile and Japanese encephalitis viruses in bonnet macaques.

Cross-protection between Japanese encephalitis (JE) and West Nile (WN) viruses was tested in bonnet macaques (Macaca radiata) immunized either with JE virus (JEV) or WN virus (WNV). JEV immunized monkeys were challenged by intranasal (i.n.) route with WNV and vice versa. Four control unimmunized monkeys were similarly infected either with WNV or JEV. Two of three control monkeys infected with WNV, developed paralysis followed by death. Virus was recovered from the central nervous system (CNS) of the both dead control monkeys and the histopathological examination of CNS revealed changes suggestive of viral encephalitis. The control monkey infected with JEV developed encephalitis and the virus was recovered from the blood and CNS. All the 3 JEV-immunized monkeys withstood WNV challenge, whereas only 2 of the 5 WNV immunized monkeys withstood the challenge with JEV. Out of 3 WNV-immunized monkeys surviving challenge with JEV, 2 revealed symptoms suggestive of mild encephalitis followed by complete recovery. The third monkey died on the 60th day post-infection (p.i.) without any symptoms and virus was recovered only from the olfactory lobe. These studies indicate that the immunization with JEV protects the bonnet macaques against WNV, whereas the WNV immunization only reduces the severity of the disease due to JEV.


Effect of acyclovir on selected immune functions.

The toxicity of acyclovir (ACV) produced by Lachema, Brno was compared with that of Zovirax, Wellcome. The in vitro suppressive effect of both substances was found equal and concentration dependent. The primary humoral antibody response was more sensitive to ACV than the cellular (blast transformation) response. However, spleen cells of drug-treated mice (either with the domestic compounds or Wellcome origin) differed neither in blast transformation test nor in the secretory antibody response. None of the drugs when given in adequate therapeutic dose did significantly influence the cell mediated response or antibody formation by spleen cells. Summing up, the acute immunotoxicity of both compounds was low; in this respect acyclovir Lachema did not differ from Zovirax Wellcome.


Serological diagnosis of parainfluenza virus infections: verification of the sensitivity and specificity of the haemagglutination-inhibition (HI), complement-fixation (CF), immunofluorescence (IFA) tests and enzyme immunoassay (ELISA).

Using four serological tests paired sera were examined of 117 patients with acute respiratory diseases, in whom parainfluenza viruses (PIV) infection was demonstrated by virus isolation, and of 41 patients with typical clinical mumps symptoms. Comparative analysis showed the high sensitivity of IFA and ELISA. A significant rise of antibodies in convalescent sera with homologous antigen of PIV was found in nearly 100 percent of cases. Only the sera of youngest children with high titres of persisting maternal antibodies remained without seroconversion. Cross heterologous antibody responses could be found by means of ELISA in 45% and by IFA in 10%, of patients who in the past experienced infection with one or more PIV or mumps virus--apart from homologous antibody reaction. HI and CF test proved to be less sensitive for detection of postinfections antibodies, especially in primoinfections with PIV types 1 and 2.


Pretreatment of the host cell with 1-(S)-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine (HPMPC) is sufficient for its antiviral effect.

1-(S)-(3-Hydroxy-2-phosphonylmethoxypropyl)cytosine (HPMPC) inhibits efficiently human cytomegalovirus (HCMV) replication. The drug exhibited marked antiviral effect in vitro not only at single application after virus adsorption, but also if given 48 hr post-infection. The inhibition of HCMV replication by HPMPC occurred after mere pretreatment of the host cells with the drug followed by its removal from the medium before virus infection. No free or host cell bound infectious virus was detected in the drug pretreated (1.5 micrograms/ml) virus infected cells. This effect was drug concentration dependent.


Detection of influenza B virus in throat swabs using the polymerase chain reaction.

An assay protocol based on exploiting the polymerase chain reaction (PCR) for the direct detection of influenza B virus in throat swabs is described. By the use of PCR with nested primers, it was possible to detect the virus in throat swabs. Dilution experiments showed that as little as 1 plaque forming unit of virus was sufficient for detecting the HA gene by the PCR. All throat swab samples from which influenza B virus had been isolated by conventional methods were also positive by the PCR method.


Detection of viral DNA polymerase activity in salmon tumour tissue induced by herpes virus, Oncorhynchus masou virus.

DNA polymerase activities were surveyed in tumour tissue and normal tissue of cherry salmon (Oncorhynchus masou). High activity of DNA polymerase alpha was detected in the tumour tissue but not in the normal tissue. This indicates that the tumour cells replicate prosperously. Viral DNA polymerase activity was detected only in the tumour tissue, indicating that Oncorhynchus masou virus (OMV) DNA should replicate there. DNA polymerase beta activity was of same level in both tissues. This is the first evidence that herpesvirus DNA polymerase was detected in tumour tissue in association with herpesvirus.


Experimental infection of domestic cats by cowpox virus.

Infection of young domestic cats by cowpox virus isolated from sick rodents (family Muridae) revealed their high susceptibility to the virus; a severe disease with 100% lethality developed after oral inoculation as well as upon skin scarification. The disease in dermally infected animals was accompanied by eruptions on the site of inoculation. High concentration of the virus was detected in lungs of animals infected by either of inoculation routes. The data testify the possible participation of domestic cats as intermediate hosts in the circulation of cowpox virus.


Urinary incontinence in the elderly: pharmacologic therapies.

Treatment of acute urinary incontinence should be directed toward the underlying cause, such as infection, medication side effect, atrophic vaginitis, anxiety, depression and restricted mobility. Pharmacologic treatment depends on identification of one of the four subtypes of chronic urinary incontinence: stress, urge, overflow or mixed. Stress incontinence responds to alpha-adrenergic agents, which increase sphincter tone. Urge incontinence is the most common type of incontinence in the elderly; it can be treated with anticholinergic agents, smooth muscle relaxants, estrogen replacement therapy in women and, possibly, calcium antagonists. Overflow incontinence is caused by neurologic deficits, such as diabetes, or outflow obstruction, such as from prostatic enlargement, urethral stricture and tumors. Anticholinergic agents and alpha-adrenergic agents should be considered only after existing outflow obstruction is surgically corrected or intermittent catheterization is unsuccessful.


Update on drug therapy for HIV and related infections in adults.

Antiretroviral therapy with zidovudine is indicated in patients with CD4 cell counts below 500 per mm3 (500 x 10(6) per L). Patients intolerant of zidovudine and those with advanced human immunodeficiency virus infection may benefit from newer antiretroviral agents, such as didanosine (ddl) or zalcitabine (ddC). Prophylactic therapy for Pneumocystis carinii pneumonia is indicated in patients with CD4 cell counts below 200 per mm3 (200 x 10(6) per L), in patients with CD4 cell counts less than 20 percent of the total lymphocytes and in patients with a prior history of P. carinii infection. In addition, prophylaxis is often initiated if thrush is present, even when CD4 cell counts are above 200. Trimethoprim-sulfamethoxazole is the drug of choice for Pneumocystis prophylaxis; aerosolized pentamidine is reserved for patients unable to tolerate trimethoprim-sulfamethoxazole. Oral candidiasis is treated with nystatin suspension, clotrimazole troches, ketoconazole or fluconazole, with fluconazole used for resistant or more invasive infection. Finally, acyclovir is used to treat herpes zoster or herpes simplex virus infection.


Further evidence for dominant inheritance at the chromosome 15q11-13 locus in familial Angelman syndrome.

Eleven patients with Angelman syndrome (AS) and their parents from 5 families have been studied with high resolution chromosome analysis and molecular probes from region 15q11-13 in an attempt to elucidate the mode of inheritance in familial AS. No deletions were detected. All families were informative with a combination of different short arm cytogenetic markers. All sets of sibs inherited the same maternal chromosome 15, but in 3 families sibs inherited different paternal 15s. Analysis of 6 polymorphic DNA markers supported the conclusion that AS sibs inherit the same maternal 15, but often different paternal 15s. These data make autosomal recessive inheritance at a 15q11-13 locus very unlikely and support the hypothesis that familial AS is due to maternal transmission of a mutation within 15q11-13.


Deletions and microdeletions of 22q11.2 in velo-cardio-facial syndrome.

Velo-cardio-facial syndrome (VCFS), an autosomal dominant disorder, is characterized by cleft palate, cardiac defects, learning disabilities and a typical facial appearance. Less frequently, VCFS patients have manifestations of the DiGeorge complex (DGC) including hypocalcemia, hypoplastic or absent lymphoid tissue and T-cell deficiency suggesting that these 2 conditions share a common pathogenesis. Here, we report the results of cytogenetic and molecular studies of 15 VCFS patients. High-resolution banding techniques detected an interstitial deletion of 22q11.21-q11.23 in 3 patients. The remaining 12 patients had apparently normal chromosomes. Molecular analysis with probes from the DiGeorge Chromosome Region (DGCR) within 22q11 detected DNA deletions in 14 of 15 patients. In 2 families, deletions were detected in the affected parent as well as the propositus suggesting that the autosomal dominant transmission of VCFS is due to segregation of a deletion. Deletions of the same loci previously shown to be deleted in patients with DGC explains the overlapping phenotype of VCFS and the DGC and supports the hypothesis that the cause of these two disorders is the same.


Onset of action and duration of effect of formoterol and salmeterol compared to salbutamol in isolated guinea pig trachea with or without epithelium.

Formoterol and salmeterol are 2 newly developed beta 2-adrenoceptor agonists for inhalation with prolonged duration of effect compared with currently available beta 2-agonists. The clinical duration of effect has been suggested to be correlated to a decreased washability in vitro, i.e. decreased effect due to continuous washing of the organ bath. In this study we compared the washability and the onset of the relaxatory effect of formoterol (0.01 microM), salmeterol (0.05 microM) and salbutamol (0.1 microM) in isolated guinea pig trachea contracted with carbachol (0.1 microM). We also evaluated a possible influence of the epithelium on onset of action and/or duration of effect. A significant relaxatory effect of formoterol and salmeterol remained after washing, whereas no effect of salbutamol remained. We also found a rapid onset of action for both salbutamol and formoterol, but a significantly slower onset for salmeterol. Both the washability and the onset of action were found to be independent of the presence of the epithelium.


Amplified assay of alkaline phosphatase using flavin-adenine dinucleotide phosphate as substrate.

A simple to use, robust, quantitative, and extremely sensitive colorimetric assay for alkaline phosphatase (EC 3.1.3.1), designed to be used as a detection system in diagnostic assays employing antibodies or gene probes, is described. This technology is based on the novel principle of prosthetogenesis, according to which a purpose-designed substrate (a prosthetogen) for a primary analyte-linked enzyme label is hydrolyzed to produce a prosthetic group for a detector enzyme system. The prosthetogen employed here is a derivative of FAD which is phosphorylated at the 3'-position of the ribose ring (FADP), the label enzyme is alkaline phosphatase, and the detector is a D-amino-acid oxidase/horseradish peroxidase-coupled system. Essentially each turnover of every molecule of alkaline phosphatase produces a molecule of D-amino-acid oxidase for detection. Thus enormous amplification of the initial signal is achieved in short time periods because of the relatively high turnover number of alkaline phosphatase for FADP. The system can be formatted as a stable, preformed, freeze-dried preparation containing all analytical components, which is reconstituted simply by addition of buffer solution. This methodology can quantitate less than 0.1 amol of alkaline phosphatase in 30 min at 25 degrees C using microtiter plates.


Immunocytochemical and in situ hybridization evidence for the coexistence of GABA and tyrosine hydroxylase in the rat locus ceruleus.

We have demonstrated the coexistence of GABA-like and tyrosine hydroxylase-like immunoreactivities (GABA-LI and TH-LI, respectively) in the same neurons of the rat locus ceruleus (LC). The profiles of these cells were labeled by alternately immunostaining adjacent sections for GABA-LI or TH-LI by the avidin-biotin-peroxidase complex method or the peroxidase-anti-peroxidase method after perfusion (either Zamboni's fixative or PPG), and observation at light and electron microscopic levels. For light microscopy, pairs of adjacent sections of more than 590 (Zamboni's) and 260 (PPG), and for electron microscopy, 40 ultrathin sections cut from adjacent semithin plastic sections (Zamboni's), were examined. GABA-LI was found in 80% (1,309/1,642 in total) of small and medium-sized neurons, uniformly scattered throughout the LC. Observations unequivocally show that the majority of GABA-ergic neurons are also noradrenergic. Several neurons are neither noradrenergic nor GABA-ergic, while other noradrenergic neurons do not show GABA-LI. It is shown that astrocytes, but not oligodendrocytes, contain GABA. In situ hybridization using a probe DNA fragment of the glutamic acid decarboxylase (GAD) cDNA, amplified by the polymerase chain reaction, detected GAD mRNA signals in many neurons throughout the LC, supporting the presence of a GAD/GABA system in the LC. Multiple "classical" transmitters, including GABA, serotonin, and noradrenaline, coexist in many LC neurons and may contribute to its widely diverging projections throughout the entire CNS.


Sustained improvement in asthma with long-term use of formoterol fumarate.

Inhaled formoterol fumarate, a long acting beta 2 agonist, produces bronchodilatation that is sustained for approximately 12 hours. To determine whether such bronchodilator effects are maintained and asthma control sustained during chronic administration, we monitored airflow indices and clinical control in 112 asthmatic patients who self administered formoterol twice daily for periods ranging from 9 to 12 months. Subjects were recruited immediately following completion of a 3-month double-blind comparison of formoterol, 12 micrograms, bid versus salbutamol, 200 micrograms, qid. Assessments were conducted at baseline, 3, 6, and 9 months, where baseline represents the final visit of the 3-month comparative study. Patients were asked to complete diary cards and twice daily PEFR for a 2-week period before each assessment. Throughout the follow-up study, there was no indication of worsening of asthma control or deteriorating lung function. For the patients who continued to receive formoterol, the previous improvement in asthma control and lung function was maintained at the level reached in the 3-month study. There was an improvement in flow rates and asthma symptoms in the group switched from salbutamol to formoterol by the first clinic visit. This improvement in asthma control and lung function was maintained over the subsequent 6 months. Formoterol was well tolerated during the study period. We conclude that prolonged use of formoterol fumarate twice daily results in sustained improvement in symptoms and flow rates in asthma with no evidence of tachyphylaxis.


Detection of beta-blocker use in people with asthma.

Although there have been numerous reports of adverse outcomes for people with asthma who are placed on beta-blockers, there has been no description of how often people with asthma receive prescriptions for beta-blockers. Despite the fact that pharmacy claims are available and can be used for clinical evaluation, there has been no description of a practical surveillance or warning system to recognize and reduce the rate of beta-blocker use in people with asthma. This study used administrative claims data to estimate the prevalence of patients with asthma who also had prescriptions for beta-blockers. Chart audit was used to supplement our understanding of the causes of the problem and its consequences. In the calendar year 1989, in a large midwestern group practice that contracts with a single health maintenance organization (HMO), 3,170 HMO patients presumed to have asthma were identified. Of those 3,170 patients, 44 or 1.4% also had filled prescriptions for beta-blockers. The occurrence of beta-blocker use varied by age group: from less than 1% in patients below 30 years of age, rising to 8.9% in patients aged 60 to 69. Two of the patients with asthma who had prescriptions for beta-blockers were hospitalized for asthma in the study period. In 61% of the cases, different physicians managed the asthma care from those who prescribed the beta-blockers. In the remaining 39%, one physician was responsible for both the asthma care and beta-blocker prescription. We conclude prescribing beta-blockers for individuals with asthma is not uncommon. Current systems of administrative claims data permit the development of warning systems to help avert adverse outcomes.


Oxatomide modifies methacholine- and exercise-induced bronchoconstriction in asthmatic children.

Oxatomide is a potent inhibitor of both the release and effects of allergic mediators and is similar to calcium antagonists in chemical structure. It prevents histamine release by inhibiting not only the increase in calcium intake, but also intracellular calcium release. We investigated its effect on methacholine-induced and exercise-induced bronchoconstriction in asthmatic children. Methacholine challenges were performed after oral administration of 0.88 mg/kg oxatomide or placebo in nine asthmatic children in a double-blind placebo-controlled study. Respiratory thresholds were improved in seven patients and log PC20 in the oxatomide group (6.65 +/- 1.34 micrograms/mL) was significantly higher than that in the placebo group (5.74 +/- 1.04 micrograms/mL) (P < .05). Exercise challenges were performed after oral administration of 1.5 mg/kg oxatomide or placebo in eight asthmatic children in a double-blind placebo-controlled study. Oxatomide produced acute bronchodilatation with 6.1% improvement on an average in FEV1. The mean maximal % fall obtained by oxatomide was 13.5%, while that by placebo was 22% (P < .05). These results indicate that oxatomide reduces nonspecific bronchial hyperresponsiveness.


Polymorphisms of a scrapie-associated fibril protein (PrP) gene and their association with susceptibility to experimentally induced scrapie in Cheviot sheep in the United States.

The duration of the incubation period for scrapie, a fatal transmissible neurodegenerative disorder of sheep and goats, is mainly determined by the Sip gene, which has 2 alleles (sA--susceptible and pA--resistant). A diagnostic test is not available to detect scrapie in live animals. We analyzed genomic DNA extracted from frozen sheep brains collected from Cheviot sheep of the United States that had been inoculated with the SSBP/1 scrapie inoculum. Digestion of the DNA with EcoRI or HindIII followed by the addition of a scrapie-associated fibril protein (PrP)-specific marker probe, yielded fragments of 6.8 (e1) and 4.0 (e3) kb, or 5.0 (h1) and 3.4 (h2) kb, respectively. Fragments e1 and h2 were associated with the histopathologic diagnosis of scrapie, and fragments e3 and h1 were associated with survival. A valine/alanine polymorphism within the PrP coding region that resulted in a BspHI site was further used to determine the genotype of these Cheviot sheep. Digestion of polymerase chain reaction fragments with BspHI resulted in an undigested fragment b- (0.840 kb), digested fragments b+ (0.460 and 0.380 kb), or both types of fragments. Survival time of b+/b+ homozygous sheep was significantly (P < 0.01) shorter (218 +/- 26.0 days) than survival time for b-/b- sheep (> 700 days after inoculation). Results indicated that b+ and b- are markers for the Sip sA and pA alleles, respectively. The intermediate duration of the incubation period for heterozygous sheep (b+/b-; 342.9 +/- 25.3 days) indicated that the Sip sA allele is expressed codominantly to the Sip pA allele.


Serum secretory leukoprotease inhibitor levels to diagnose pneumonia in the elderly.

In pneumonia in the elderly, one occasionally encounters difficulties in evaluation with respect to both clinical observation and treatment. Thus a simple serum indicator is indicated. We measured secretory leukoprotease inhibitor (SLPI) concentrations in sera to see whether this can provide a useful indicator for pneumonia, especially in the elderly. Serum samples from patients over 65 yr of age, with (n = 54) or without (n = 87) pneumonia, and from healthy, young (n = 16) and aged (n = 188) control subjects were assayed using ELISA for human SLPI. Comparisons were made between groups with clinical diagnoses of either definite or probable pneumonia and among cases with various other respiratory diseases, including bronchial asthma, chronic obstructive pulmonary disease, and lung cancer. The mean SLPI concentration in patients with pneumonia was significantly higher than in patients without pneumonia or in healthy controls. The data suggest that the measurement of SLPI can provide a useful indicator for pneumonia to be used in clinical evaluation.


[Treatment of recurrent ulcers after parietal cell vagotomy. Analysis of 18 cases].

One hundred and twenty seven patients had a follow-up of more than 4 years after parietal cell vagotomy for duodenal, pyloric or prepyloric ulcer. The recurrence rate is 15.8% (20 on 127). Eighteen patients who have recurred, had a follow-up of 2 years or more after treatment of the recurrent disease. Ten patients had a early recurrence (within 2 years); eight had a late one (more than 2 years). One patient had been operated as an emergency for perforation. Seventeen patients had first received a medical treatment. This treatment was sufficient for 10 out of 17. Seven patients were reoperated (3 partial gastrectomy), 4 partial gastrotomy associated with truncal vagotomy. Seven out of the 10 early recurrences was reoperated and only one of the 8 late recurrences was reoperated. The early recurrences seem to be more serious that the late ones. Medical treatment is always prescribed as first line therapy and a partial gastrectomy alone or with vagotomy is necessary in unsuccessful cases.


Results of surgery in primary hyperparathyroidism.

In a prospective study 92 consecutive patients with biochemically proved primary hyperparathyroidism underwent initial neck exploration at Oulu University Hospital. The incidence of multiglandular disease was 34%. 23 patients (25%) underwent a simultaneous thyroidectomy. The cure rate after initial exploration was 91.3%, ectopic parathyroid glands and multiglandular disease being the most common causes of failure. Simultaneous thyroidectomy increased somewhat but not significantly the complication risk. There was a slight tendency for serum calcium concentrations to increase during the mean follow-up of 2.3 +/- 1.5 years. Four patients with persistent hypercalcaemia underwent a successful reoperation during that time. Thus the overall cure rate was 95.6%, but 5.4% of the patients required permanent medication for hypocalcaemia. We conclude that the most common causes for failed initial exploration were ectopic parathyroid glands and multiglandular disease. The incidence of multiglandular disease was unusually high in this series. Because simultaneous thyroidectomy increased somewhat the complication risk of initial neck exploration, the indications for this additional procedure should be carefully considered. The results of parathyroid surgery were good and dependent on how many patients underwent reoperation during the follow-up. A consequence of tendency for serum calcium concentrations to increase during the follow-up could be that a definite cure cannot always be attained in cases of primary hyperparathyroidism.


Nigral damage and dopaminergic hypofunction in mesencephalon-immunized guinea pigs.

To support a potential role for immune mechanisms in the destruction of substantia nigra (SN) neurons, guinea pigs were immunized with bovine mesencephalon containing SN neurons. After immunization no clinical signs of basal ganglia dysfunction appeared. However, pathological examination revealed evidence of neuronal damage in the SN in 8 of 17 guinea pigs immunized with bovine mesencephalon. No nigral pathology was noted in animals immunized with spinal cord gray matter or Freund's adjuvant alone. Accompanying the SN damage in mesencephalon-immunized guinea pigs was a 25% decrease in tyrosine hydroxylase activity in the SN and a 27% decrease in dopamine content in the striatum. Deposits of IgG were detected by immunohistochemical techniques in sections of SN from mesencephalon-immunized guinea pigs and in sections of human SN after exposure to serum from mesencephalon-immunized guinea pigs. These data document the antigenicity of SN and suggest the possibility that immune mechanisms can contribute to basal ganglia pathology.


Paternal uniparental disomy of chromosome 15 in a child with Angelman syndrome.

Angelman and Prader-Willi syndromes are clinically distinct neurobehavioral disorders most commonly resulting from large deletions of chromosome 15q11-q13. The deletions arise differentially during maternal or paternal gametogenesis, respectively. A subgroup of patients with either syndrome have no apparent deletion, and because many such patients with Prader-Willi syndrome display inheritance of two copies of chromosome 15 from the mother only (uniparental disomy; UPD), we suggested that paternal UPD might be found in patients with Angelman syndrome. We report here clinical, cytogenetic, and molecular evidence on the 1 patient with paternal UPD for chromosome 15 who was found in our study population. This represents, to our knowledge, the first patient with paternal UPD to be studied with DNA probes from the chromosome 15q11-q13 critical region. In contrast to our findings for patients with Prader-Willi syndrome, in which maternal UPD was common, our data demonstrate that paternal UPD is infrequent in patients with Angelman syndrome.


Overview of principles and current uses of DNA probes in clinical and laboratory medicine.

Fundamental considerations of deoxyribonucleic acid (DNA) probe hybridization reaction including concept of stringency is reviewed. Restriction fragment length polymorphism (RFLP) and its detection using DNA probes are discussed together with the importance of RFLP in genetic linkage analysis. Methodological considerations, such as limitations in sample preparation and improvements effected in techniques, cloning DNA, and labelling of DNA probes, are addressed. Principles of hybridization technology and blotting techniques are reviewed. Amplification of DNA by the polymerase chain reaction (PCR), labelling with PCR, detection of PCR products and separation procedures prior to use of PCR are discussed. Other amplification methods and in-situ hybridization (ISH) procedures are reviewed. Selected current applications of DNA probes in the area of infectious diseases, genetic diseases, HLA typing, paternity and forensic testing are briefly discussed. Emphasis is placed on the use of DNA probes in the area of oncology. Background on activation of proto oncogene, loss of tumor suppressor genes, chromosomal translocation, and chromosome analysis are provided for understanding DNA probe assays for ph1 positive leukemias and applications of Interphase cytogenetic technique in oncology. Finally, analytical strategies for detection of tumor suppressor genes are addressed.


Electrocardiographic changes during acute treatment of hypertensive emergencies with sodium nitroprusside or fenoldopam.

Electrocardiograms are routinely obtained before and during the acute treatment of hypertensive emergencies, usually to rule out "ischemic changes." Despite a few anecdotal reports of electrocardiographic changes, little is known about the incidence and significance of such changes, or their relationship to the treatment used. We prospectively analyzed 12-lead electrocardiograms from 21 patients admitted for hypertensive emergencies (average blood pressure, 222 +/- 4/140 +/- 3 mm Hg). Patients were randomly assigned to treatment with sodium nitroprusside (n = 11) or the dopamine receptor agonist fenoldopam mesylate (n = 10). Electrocardiograms were obtained at baseline and within 30 minutes of reaching goal blood pressure (diastolic blood pressure, 100 to 110 mm Hg). There was no significant effect of either drug treatment on PR interval, QRS duration, QT interval, or R-wave amplitude, and no major ST-segment changes were noted. During treatment with either drug, the average T-wave amplitude decreased in all leads except aVR. New T-wave inversions in lead V4 occurred in two and four patients after fenoldopam and nitroprusside treatment, respectively. There were no clinically apparent episodes of myocardial ischemia in any patient. Even in the absence of obvious myocardial ischemia, a decrease in T-wave amplitude, including T-wave inversion, occurs commonly during acute blood pressure reduction in hypertensive emergencies, an observation that may be explained by the accompanying acute changes in cardiac chamber volumes.


Number and DNA content of hypertrophic spermatogonia in normal and cryptorchid human testes.

The number and the DNA content of hypertrophic spermatogonia were studied in normal and cryptorchid testes. The number of hypertrophic spermatogonia in cryptorchid testes is higher than in control testes at the same age. This finding suggests alterations in the spermatogonia of cryptorchid males. The results show a polyploid DNA content in more than 80% of hypertrophic spermatogonia in both normal and cryptorchid testes. There were hypertrophic spermatogonia with a DNA content between 2c and 4c (0.5%) and between 4c and 8c (9%). These spermatogonia might be cells in the S phase of the cell cycle.


Alpha- and beta-adrenoceptor cross-talk in the regulation of glycogenolysis in dog and guinea-pig liver.

The dog liver glycogenolytic response to isoprenaline (EC50 = 3 x 10(-9) M) was selectively blocked by 10(-5) M of practolol, but not by butoxamine. In contrast, the glycogenolytic response to isoprenaline (EC50 = 3 x 10(-7) M) was inhibited by 10(-6) M of butoxamine, but not by practolol, in the guinea-pig liver. This suggests that, in the dog, the isoprenaline response is dominated by beta 1-adrenoceptors, while in the guinea-pig beta 2-receptors control such response. Glucose release from dog and guinea-pig liver slices was also stimulated by amidephrine (EC50 = 10(-6) M in the dog and 4 x 10(-5) M in the guinea-pig). Both prazosin and yohimbine blocked this response. The effectiveness of clonidine as a glucose-mobilizing agent could only be established in the dog liver. Prazosin showed greater activity than yohimbine in antagonizing the response to both agonists. In the dog, low concentrations of alpha-adrenoceptor agonists (10(-9) M), that failed to modify the basal glucose release per se, selectively depressed the isoprenaline response. Prazosin, but not yohimbine, reversed this inhibitory effect. It is concluded that glucose release from the dog liver is regulated by two opposite mechanisms that seem to be associated to alpha 1-adrenoceptors (inhibitory) and to beta 1-adrenoceptors (stimulatory).


A specific RFLP type associated with the occurrence of sheep scrapie in Japan.

We have investigated restriction fragment length polymorphism (RFLP) on the PrP gene and the frequencies of RFLP patterns in 35 healthy Suffolk sheep randomly collected. According to the combinations of PrP encoding DNA fragments generated by restriction enzymes Eco RI and Hind III, the RFLP patterns were classified into six types and designated as types I to VI. The frequencies of these types were as follows: I, 8.6%; II, 11.4%; III, 17.6%; IV, 11.4%; V, 28.6%; and VI, 22.9%. In 10 sheep diagnosed as having natural scrapie, RFLP types, I, III, IV, and V were determined. To examine the correlation between the RFLP type and the occurrence of scrapie, the frequencies of RFLP types in sheep infected with natural scrapie were compared with those in healthy sheep. It was found that the frequency of type I in the sheep with natural scrapie was 70%, about eight times higher than that in randomly collected healthy sheep. In the 13 experimentally infected sheep that had been used for other purposes, however, no relationship between the RFLP type and onset of scrapie was found.


Coexistence of presynaptic beta 1- and beta 2-adrenoceptors in splenic strips from young rats and betaxolol-induced beta 1-stereoselective antagonism.

The pharmacological properties of facilitatory presynaptic beta-adrenoceptors were characterized using three kinds of beta-agonists and antagonists in superfused strips from young Sprague-Dawley rats, loaded with [3H]noradrenaline. Isoproterenol (10(-9) M to 10(-7) M) concentration-dependently facilitated [3H] release evoked by transmural field stimulation at 5 Hz. Salbutamol (10(-8) M and 10(-7) M) and prenalterol (10(-8) M to 10(-6) M) also facilitated the stimulation-evoked [3H] release. Pretreatment with ICI 118,551 (10(-6) M) completely antagonized the concentration-facilitation curve for isoproterenol (10(-9) M to 10(-7) M). Atenolol (10(-6) M) antagonized only the isoproterenol (10(-9) M)-induced facilitation. dl-Betaxolol (10(-6) M) antagonized the isoproterenol (10(-9) M and 10(-8) M)-induced facilitation and the antagonistic action for isoproterenol (10(-8) M) was more marked than that of atenolol, whereas the same concentration of d-betaxolol produced no antagonism. Thus, presynaptic beta 1- and beta 2-adrenoceptors coexist on noradrenergic neurons innervating splenic strips of young rats. Betaxolol is a useful tool to characterize presynaptic beta 1-adrenoceptors.


The Canadian Forces Airsickness Rehabilitation Program, 1981-1991.

Airsickness is a significant obstacle in the training of some student pilots. When conventional therapy fails, desensitization therapy may be indicated. Using experience gained by the RAF and USAF, the Canadian Forces (CF) began such a program in 1981. This paper reports program results from 1981-1991. Following subject identification, treatment consists of three phases: biofeedback relaxation therapy, ground-based desensitization training and in-flight desensitization therapy with a pilot-flight surgeon. Employing a definition of cure used by the RAF, success was compared with that of the RAF and USAF programs. A total of 22 student pilots have undergone rehabilitation, 17 of whom have been successfully treated for a success rate of 77%. This is comparable to success rates of other programs. It is reaffirmed that desensitization is a valid clinical tool in treatment of airsickness.


6-[N,S-dimethyl-N'-cyanothioureidomethyl]-6,11-dihydro-5H- dibenz[b,e]azepine hydrochloride (Fran 12): a histamine and 5-hydroxytryptamine antagonist with pressor properties.

We have synthesized and examined some of the pharmacological properties of 6-[N,S-dimethyl-N'-cyanoisothioureidomethyl]-6,11-dihydro-5H- dibenz(b,e)azepine hydrochloride (Fran 12), a derivative of 6-methylaminomethyl-6,11-dihydro-5H- dibenz[b,e,]azepine. In the guinea-pig isolated ileum, Fran 12 (10(-7)-10(-5) M) caused parallel rightward shifts of the concentration-response curves to histamine. A Schild plot gave a pA2 of 7.48, with a slope not significantly different from -1.0. In the rat stomach fundus strip and in endothelium-denuded aortic rings, Fran 12 inhibited contractile responses to 5-hydroxytryptamine in a non-competitive manner. In both chloralose-anaesthetized and pithed rats, it inhibited pressor responses to 5-hydroxytryptamine. It had no effect on depressor responses to 5-hydroxytryptamine in anaesthetized rats. In pithed rats, Fran 12 (0.25-2 mg/kg, i.v.) produced dose-dependent increases in blood pressure. These were not inhibited by i.v. phentolamine, prazosin, yohimbine, propranolol, methysergide, pentolinium or atropine but were inhibited by verapamil. These results indicate that Fran 12 is a histamine and 5-hydroxytryptamine antagonist which also exerts pressor effects via a peripheral action. The pressor action does not appear to be mediated via effects on alpha 1- or alpha 2-adrenoceptors, muscarinic or nicotinic cholinoceptors or 5-hydroxytryptamine receptors, although calcium channel activation may play a role.


The spontaneously hypertensive rat (SHR) as an animal model of childhood hyperactivity (ADHD): changed reactivity to reinforcers and to psychomotor stimulants.

Childhood hyperactivity (attention-deficit hyperactivity disorder, ADHD) is a behavior disorder affecting 2-6% of grade-school children. The main symptoms are attention problems and hyperkinesis. The disorder is commonly treated with psychomotor stimulants, usually methylphenidate hydrochloride (ritalin) or d-amphetamine. Neither the cause of the disorder nor the basis of the effectiveness of the drug treatment is well understood. Differences in reinforcement processes have been implicated as part of the underlying problem. The main purpose of the present research was to investigate reinforcement processes and motor characteristics with and without stimulant medication in SHR, as an animal model of ADHD, and WKY controls, its normoactive progenitor strain. SHR behavior turned out to be more sensitive to immediate reinforcement and proportionately less sensitive to delayed reinforcement when compared to the behavior of WKY, as demonstrated by systematic changes in rates of responding throughout fixed-interval schedules of reinforcement of bar-presses by water. The psychomotor stimulants weakened the control by immediate reinforcers and strengthened the control by delayed reinforcers, with the effect of the drugs being more pronounced in WKY than in SHR. The results are consistent with clinical observations that ADHD children are less willing than others to accept "delayed gratification" and that methylphenidate increases the control of delayed reward over their behavior.


Effects of medetomidine, an alpha-2 adrenoceptor agonist, and atipamezole, an alpha-2 antagonist, on spatial memory performance in adult and aged rats.

The effects of a novel, highly selective alpha-2 agonist, medetomidine, and its antagonist, atipamezole, were studied on the working memory of rats performing a spatial delayed alternation task. Testing was performed in two stages, at the age of 8.3 months (mean) and again when the rats were 17.6 months (mean). A low dose (3 micrograms/kg) and a high dose (30 micrograms/kg) of medetomidine improved the performance of the old rats in the memory task but had no effect on the young rats. The dose-response curve of medetomidine resembles that of guanfacine, another alpha-2 agonist. At the low dose of medetomidine (3 micrograms/kg) the animals showed no signs of sedation. Since medetomidine even at a low dose has a beneficial effect on the memory performance of old rats, it could be a good candidate for the treatment of age-associated memory dysfunction.


Protective effects of verapamil on cis-platinum and carboplatinum nephrotoxicity in dehydrated and normohydrated rats.

The aim of the study was to compare the nephrotoxic effects of cis-platinum (CDDP) and carboplatinum (CBDCA) and the protective potential of verapamil in rats. CDDP (3 mg/kg) and CBDCA (60 mg/kg) were administered intraperitoneally in a single dose on day 1. Verapamil (0.1 mg/kg) was administered 30 min before i.p. injection. Rats were assayed daily for urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG), alkaline phosphatase (AP) and gamma-glutamyl-transferase (GMT), on day 3 they were assayed for creatinine clearance. CBDCA appears to be less nephrotoxic than CDDP. Verapamil shows protective effects against the nephrotoxicity of platinum derivatives in both groups of rats.


A chemiluminescence fiber-optic biosensor system for the determination of glutamine in mammalian cell cultures.

A chemiluminescence fiber-optic biosensor system has been developed for determining glutamine in hybridoma cell cultures producing monoclonal antibodies against viral surface antigens. Glutaminase and glutamate oxidase (GLO) were immobilized onto aminopropyl glass beads via glutaraldehyde activation separately and packed in a column. Two separate columns containing immobilized GLO and catalase were placed upstream to eliminate endogenous glutamate. In the presence of ferricyanide, luminol reacted with hydrogen peroxide released from the enzymatic reactions to produce a chemiluminescence (CL) light signal which was detected and quantitated with a fiber-optic system. In combination with flow injection analysis it was possible to process samples virtually identically, thus avoiding difficulties in reproducing the CL signal. There was an excellent linear relationship between the CL response and standard glutamine concentration in the range 10(-6) to 10(-3) M. A complete analysis could be performed in 2 min including sampling and washing. Each immobilized enzyme column was stable for at least 300 repeated analyses without any loss of activity. When the biosensor system was used for the determination of glutamine in spent mammalian cell cultures, the values obtained compared well with those of high-performance liquid chromatography, thus validating the applicability of the CL fiber-optic system.


Further data on the development of SRIF-like immunoreactive nerve cell populations in the chick embryo brain stem. I. Medulla and pons.

Further immunocytochemical analysis of the neuroblasts with SRIF-like immunoreactivity (ir) was carried out on the chick embryo medulla and pons. 5 or 100 microns rombencephalon sections were obtained from 60 White Leghorn chick embryos at stages (E = Embryonic days) ranging from E4 1/2 to E18 and incubated with rabbit polyclonal antibodies against synthetic cyclic Somatostatin-14, according to PAP-DAB technique. In the medulla and pons the ir appeared as from E12. From E12 to E13 1/2-E14 the ir distribution gradually changed. From E14 to E18 numbers and spatial arrangement of the positive neuroblast groups did not show substantial changes; in these respects the ir distributional pattern proved to be markedly different from the one observed by the Authors in adult animals. Moreover, from E13 to E15 the positive neuroblast density appeared to be higher than that of positive neurons in adults. These results are consistent with a possible SRIF local regulative role.


Further data on the development of SRIF-like immunoreactive nerve cell populations in the chick embryo brain stem. II. Midbrain tegmentum.

Further immunocytochemical analysis of the neuroblasts with SRIF-like immunoreactivity (ir) was carried out on the chick embryo midbrain tegmentum. 5 or 100 microns mesencephalon sections were obtained from 60 White Leghorn chick embryos at stages (E = Embryonic days) ranging from E4 1/2 to E18 and incubated with rabbit polyclonal antibodies against synthetic cyclic Somatostatin-14, according to PAP-DAB technique. In the midbrain tegmentum the ir appeared as from E12. From E12 to E13 1/2-E14 the ir distribution gradually changed. From E14 to E18 numbers and spatial arrangement of the positive neuroblast groups did not show substantial changes; in these respects the ir distributional pattern proved to be similar to the one observed by the Authors in adult animals. From E17 to E18 a decrease in the positive neuroblast density appeared to occur, particularly in a ventrally placed group. These results are consistent with a possible local regulative role of the SRIF.


Platelet-derived growth factor (PDGF) A-chain mRNA heterogeneity generated by the use of alternative promoters and alternative polyadenylation sites.

Expression of the platelet-derived growth factor (PDGF) A-chain gene is known to give rise to multiple transcripts of different sizes. Northern blot analysis, using a set of DNA probes corresponding to various parts of the human PDGF A-chain gene, indicated heterogeneity in both the 5' and 3' end of the transcripts. The 3' heterogeneity was shown to be the result of differential use of three polyadenylation signals. S1-nuclease protection- and chloramphenicol acetyltransferase (CAT)-assays, revealed the 5' heterogeneity to be the consequence of two alternative promoters. Whereas the upstream promoter contained typical TATA- and CAAT-boxes, the downstream promoter lacked a TATA-box but seemed to be composed of several GC-boxes.


Resistance of HIV-1 to AZT might also involve the cellular expression of multidrug resistance P-glycoprotein.

Resistance of tumor cells to the antigrowth activity of several cytotoxic compounds has been associated with the expression of the so-called multidrug resistance protein or P-glycoprotein. This article addresses the question whether the expression of such protein could also affect the sensitivity of HIV to AZT. Our data indicate that this possibility does exist. In fact, multidrug-resistant CEM VBL100 cells, which express high levels of P-glycoprotein, are less sensitive to both the antiproliferative activity and the antiviral action of AZT. Additionally, our data suggest that this phenomenon is specifically mediated by P-glycoprotein since trifluoroperazine, which is known to circumvent multidrug resistance due to the action on P-glycoprotein, increases the intracellular accumulation of AZT and affects the sensitivity of HIV to AZT. Although the biological and clinical significance of these observations has still to be established, this study suggests that cellular factors, other than virus itself, should be taken into account to address the phenomenon of drug resistance of HIV.


Complex correlations between the morphology, electrophysiology and peptide immunohistochemistry of guinea-pig enteric neurones.

Neuroanatomical, electrophysiological and immunohistochemical techniques were used to describe correlations between soma morphology and electrophysiological properties in two groups of guinea-pig enteric neurones posing particular challenges. Lucifer Yellow-staining of 542 myenteric plexus neurones of duodenum revealed a great diversity of neuronal morphology. The distribution was: Dogiel Type I 27%, Dogiel Type II 54%, Stach Type IV 9%; 10% were unclassified. Correlations were sought in 59 of these cells between morphology and electrophysiological properties but no particular association was recognised. Dynorphin A(1-8)-like immunoreactivity (Dyn A(1-8)-IR) was found in up to 90% of identified submucous neurones of guinea-pig ileum. Of 62 S-neurones, 41 showed 'weak' and 19 had 'intense' Dyn A (1-8)-IR. There was no evidence of Dyn A(1-8)-IR in 2 S-neurones, nor in 8/8 AH-neurones. As for 11/16 vasoactive intestinal peptide- (VIP-) IR neurones, there was a strong correlation between the presence of 'weak' Dyn A(1-8)-IR and the occurrence of inhibitory (IPSPs) and slow excitatory synaptic potentials (EPSPs) (13/16 cells tested), which were never observed in neurones with 'intense' Dyn A(1-8)-IR (16/16) or neuropeptide Y (NPY)-IR (8/8). Similarly, 7/7 neurones with 'weak' Dyn A(1-8)-IR, but not those (7/7) with 'intense' Dyn A(1-8)-IR, hyperpolarised or showed a conductance change to noradrenaline. It was concluded that dynorphin A(1-8)-like-IR was contained in two populations of submucous neurone that are anatomically, immunohistochemically, electrophysiologically and pharmacologically distinct and closely related to those containing VIP and NPY. Furthermore, as in the myenteric plexus throughout the small intestine, opioid peptides are not expressed in Dogiel Type II cells.


Comparison of the efficacy of esmolol and alfentanil to attenuate the hemodynamic responses to emergence and extubation.

To define the ability of esmolol and alfentanil to control the hemodynamic changes associated with extubation and emergence. Randomized, double-blind, placebo-controlled study. General surgery operating rooms at a university hospital. Forty-two ASA physical status I and II patients without history of cardiac or pulmonary disease undergoing surgery not involving the cranium or thorax. Patients were given either a bolus dose of normal saline followed by an infusion of normal saline, a bolus dose of alfentanil 5 micrograms/kg followed by an infusion of normal saline, or a bolus dose of esmolol 500 micrograms/kg followed by an infusion of esmolol 300 micrograms/kg/min. Emergency and extubation resulted in significant increases in heart rate (HR) and blood pressure (BP) in the placebo group. Alfentanil controlled the responses to emergence but prolonged the time to extubation (p < 0.05). Esmolol significantly controlled the responses to emergence and extubation (p < 0.05). Emergence and extubation after inhalation general anesthesia result in significant increases in BP and HR in healthy patients. An esmolol bolus dose and subsequent infusion significantly attenuated these responses. A small bolus dose of alfentanil minimized the responses to emergence but prolonged the time to extubation and was no longer protective at that point.


Assessment of the anticholinergic effect of the new antihistamine mizolastine in healthy subjects.

1. Twelve healthy subjects were enrolled in a double-blind placebo controlled cross-over study in order to assess the possible anticholinergic effects of four doses of a new antihistamine compound, mizolastine, compared with hyoscine butylbromide (HBB) used as a reference anticholinergic drug. 2. Although mizolastine, a potent and selective H1-receptor blocker has no affinity for muscarinic receptors and does not antagonize the effects of carbachol in rodents, a study was initiated to investigate its effects on various effectors possessing muscarinic receptors (eye, heart, sweat gland, salivary gland). 3. HBB (40 mg, s.c.) impaired accommodation, decreased salivary flow and inhibited cardiac sinus arrhythmia. Pupil diameter and maximum constriction speed, carbachol-induced skin sweating and Valsalva ratio were unaffected. 4. Mizolastine (5, 10, 20, 40 mg p.o.) did not affect any parameter at any time point, demonstrating a lack of anticholinergic effect.


Polyarteritis nodosa presenting with hypertensive encephalopathy.

Polyarteritis nodosa (PAN) is a rare cause of malignant hypertension. As far as we are aware, hypertensive encephalopathy has not been described in the literature as a presenting feature of PAN.


Chromosomal localisation of two putative 11p oncosuppressor genes involved in human ovarian tumours.

In this study, 44 primary or metastatic human ovarian tumours were tested for allelic deletions on the short arm of chromosome 11. Analysis of 12 polymorphic loci by Southern blotting evidenced loss of heterozygosity (LOH) in at least one locus in 41% of cases. Moreover, two hot spots of deletions were tentatively mapped on 11p13 and 11p15.5. Our results demonstrated that LOH at 11p is a common event in ovarian carcinomas and were indicative of the possible existence in 11p of two oncosuppressor genes involved in ovarian carcinogenesis. The similarity observed with 11p allelic losses in Wilms tumours, clustered in 11p13 and 11p15.5 too, suggests that deletion and possibly inactivation of the same growth regulatory genes (WT genes) could also contribute to development of the malignant phenotype in ovarian carcinomas. Finally, a statistically significant association (P = 0.005) between 11p deletions and hepatic involvement was suggested by the analysis of distribution of 11p LOH relative to different clinical and pathological parameters of the tumour patients.


Identification of a retinoic acid responsive enhancer 3' of the murine homeobox gene Hox-1.6.

The putative vertebrate morphogen retinoic acid (RA) has been shown to induce expression of many mammalian homeobox genes in cell lines, suggesting expression of this gene family in developing vertebrate embryos may be controlled in part by RA. Using the teratocarcinoma cell line F9 as a model system, we have studied the RA-response of the murine homeobox gene Hox-1.6. RA treatment of F9 cells causes the appearance of a DNAse I hypersensitive site 3' of Hox-1.6, approximately 5 kb downstream of the Hox-1.6 promoter, and this site has been shown to reflect the presence of an RA-responsive enhancer 3' of the gene. The RA-responsiveness of the enhancer is controlled by a retinoic acid responsive element (RARE) identical to the RARE of the retinoic acid receptor (RAR) beta gene; however, other sequences also influence the activity of the enhancer, suggesting the presence of binding sites for novel proteins which regulate Hox-1.6 expression. Experiments with Hox-1.6 minigenes in which lacZ expression is controlled by the Hox-1.6 promoter and enhancer demonstrate that it is the 3' enhancer which confers RA responsiveness on the endogenous promoter, as constructs which lack the enhancer, or the RARE alone, do not respond to RA. Our results support the idea that RA is an endogenous vertebrate morphogen; identification of the RA-responsive enhancer downstream of Hox-1.6 demonstrates that RA directly controls the transcription of at least one member of a gene family that determines tissue identity in the vertebrate embryo.


Studies on concentrations of NA and HVA and activity of DBH in the serum from schizophrenic patients, first-degree relatives and normal subjects.

The serum noradrenaline (NA), homovanillic acid (HVA) and dopamine beta-hydroxylase (DBH) have been examined in neuroleptic-free and -treated patients, healthy first-degree relatives of the patients and normal subjects. Analysis of variance (ANOVA) revealed significant differences in the concentrations of serum NA(F = 2.91, p < 0.05) and HVA (F = 3.58, p < 0.05), and in the activity of serum DBH (F = 2.77, p < 0.05) among the four groups. The serum NA was significantly higher in neuroleptic-free patients (475 +/- 220 pg/ml, n = 18), than in healthy first-degree relatives (343 +/- 189 pg/ml, n = 37, p < 0.05) or in normal subjects (354 +/- 111 pg/ml, n = 17, p < 0.05), and it also was significantly higher in neuroleptic-treated patients (442 +/- 223 pg/ml, n = 58) than in healthy first-degree relatives (p < 0.05) or in normal subjects (p < 0.05). There was a trend towards high serum HVA in neuroleptic-free patients (11.3 +/- 6.3 ng/ml, n = 17) compared with the other three groups. The serum DBH activity was high in neuroleptic-free patients (31.2 +/- 15.6 nmol/min/ml, n = 17), and significantly in comparison with those treated with neuroleptic drugs (21.6 +/- 10.9 nmol/min/ml, n = 56, p < 0.05). There was a significant negative correlation between HVA concentration and DBH activity in the serum from neuroleptic-free patients (r = -0.64, n = 16, p < 0.01), and there appeared to be three subgroups with alterations of serum DBH activity and HVA concentration.(ABSTRACT TRUNCATED AT 250 WORDS)


Smoking and schizophrenia.

Several studies have shown that patients with schizophrenia have an extremely high prevalence of smoking, almost 90%, compared to only 33% in the general population and 45-70% in patients with other psychiatric diagnoses. The reasons for the high prevalence of smoking among schizophrenics is unknown, but it is likely that smoking behavior in schizophrenia may be a complex process, related to numerous interrelationships between the psychopathological, biochemical, and neuropharmacological aspects of smoking and of schizophrenia.


Thermodynamic effects of active-site ligands on the reversible, partial unfolding of dodecameric glutamine synthetase from Escherichia coli: calorimetric studies.

Dodecameric glutamine synthetase (GS) from Escherichia coli undergoes reversible, thermally induced partial unfolding without subunit dissociation. A single endotherm for Mn.GS (+/- active-site ligands) in the presence of 1 mM free Mn2+ and 100 mM KCl at pH 7 is observed by differential scanning calorimetry (DSC). Previous deconvolutions of DSC data for Mn.GS showed only two two-state transitions (with similar tm values; 51.6 +/- 2 degrees C), and indicated that cooperative interactions link partial unfolding reactions of all subunits within the Mn.enzyme dodecamer [Ginsburg, A., & Zolkiewski, M. (1991) Biochemistry 30, 9421]. A net uptake of 8.0 equiv of H+ by Mn.GS occurs during partial unfolding, as determined in the present DSC experiments conducted with four buffers having different heats of protonation at 50 degrees C. These data gave a value of 176 +/- 12 kcal (mol of dodecamer)-1 for delta Hcal corrected for buffer protonation. L-Glutamine and L-Met-(SR)-sulfoximine stabilize the Mn.GS dodecamer through the free energies of ligand binding, and these were shown to be partially and totally released, respectively, from the 12 active sites at high temperature. Ligand effects on Tm values from DSC were similar to those from spectral measurements of Trp and Tyr exposures in two subunit domains. Effects of varying [ADP] on DSC profiles of Mn.GS were complex; Tm is increased by low [ADP] and decreased by > 100 microM free ADP. This is due to the exposure of an additional low-affinity ADP binding site per GS subunit at high temperature with log K1' = 4.3 and log K2' = 3.6 at 60 degrees C relative to log K' = 5.5 for ADP binding at 30 degrees C, as determined by isothermal calorimetric and fluorescence titrations. Moreover, delta Hcal at > 27% saturation with ADP (corrected for ADP binding/dissociation) is approximately 80-100 kcal/mol more than in the absence of ligands. Changes in domain interactions could result from ADP bridging subunit contacts in the dodecamer. Each of the active-site ligands investigated here produces different effects on DSC profiles without uncoupling the extremely cooperative, partial unfolding reactions in the Mn.GS dodecamer.


The sensitive determination of abanoquil in blood by high-performance liquid chromatography/atmospheric pressure ionization mass spectrometry.

A method is described for the determination of abanoquil in human blood. The method is based on high-performance liquid chromatography (HPLC)/atmospheric pressure positive ion chemical ionization mass spectrometry, using (2H3)abanoquil as internal standard. Multiple reaction monitoring is employed for selectivity and sensitivity, which enables quantification over the range 10-500 pg ml-1 with acceptable precision and accuracy. This assay methodology illustrates the versatility of atmospheric pressure ionization/tandem mass spectrometry, in conjunction with HPLC, for the separation and quantification of drugs in the subnanogram per millilitre range.


Cytokine expression and endothelial cell and lymphocyte activation in human cardiac allograft rejection: an immunohistochemical study of endomyocardial biopsy samples.

We used monoclonal antibodies and immunohistochemical staining of frozen tissue sections to study the expression of cytokines in human cardiac allograft rejection. The 113 endomyocardial biopsy samples were stained for interleukin (IL)-2, IL-6, and interferon-gamma. The findings were compared to expression of the endothelial cell adhesion molecule ICAM-1, and the lymphocyte receptor for the adhesion molecule VCAM-1, VLA-4. Four biopsy samples from patients with idiopathic cardiomyopathy served as controls. IL-2 was not expressed in lymphocytes of controls and only occasionally in mild or moderate cellular rejection, humoral rejection, and Quilty lesions. IL-2 expression was prominent in severe cellular rejection. Interferon-gamma expression increased in proportion to the severity of cellular rejection and was not expressed in other conditions. IL-6 staining, which was only observed in occasional cases, was mild. Cytokine and adhesion molecule expression tended to increase with the severity of cellular rejection. This study shows that cytokine expression can be documented in human allograft endomyocardial biopsy samples with immunohistochemical techniques. The findings support the concept of an important role for cytokines in human cardiac allograft rejection.


Laboratory and phase I studies of new cancer drugs.

New drug discovery continues to follow the time-honored paths of screening and novel target identification combined with analogue development and serendipity. The agents selected here reflect these various approaches. They include drugs already showing significant antitumor activity, eg, anthrapyrazoles, temozolomide, camptothecin analogues, and taxotere, and updated information is provided on their development. Other drugs just entering or currently in phase I trials include rhizoxin, which seems capable of overcoming multidrug resistance; the novel bioreductive agent EO9 [corrected]; and bryostatin, a highly potent protein kinase C agonist. Sequence specificity could well prove to be an important factor in the development of DNA-interactive agents, and information is provided on the progress with distamycin mustard and the new cyclopropylpyrroloindole analogues carzelesin and adozelesin.


Cytotoxicity of adriamycin, idarubicin, and vincristine in acute myeloid leukemia: chemosensitization by verapamil in relation to P-glycoprotein expression.

A 4-day colorimetric tetrazolium dye (MTT) assay was used to assess the cytotoxicity of adriamycin (ADM), vincristine (VCR), and idarubicin (IDA) in blasts isolated from 37 patients with newly diagnosed and pretreated acute myeloid leukemia (AML). The effect of verapamil (VRP) as a chemosensitizer was studied in relation to the expression of the membrane efflux pump P-glycoprotein (PGP) as determined by a semiquantitative flow-cytometric proceder. A slight positive correlation was found between the fraction of cells expressing PGP and the ID50 values for ADM and VCR, but not between cellular PGP content and sensitivity to IDA. The overall data showed no significant sensitization effect of VRP. However, in specimens with more than 10% cells expressing PGP, 2 microM VRP sensitized cells to ADM and VCR significantly. The median of sensitization ratios (SRs), i.e., the ratios of cytotoxic drug ID50 in the absence/presence of VRP, were 1.89 and 2.0, respectively. No sensitizing effect of VRP on the cytotoxicity of IDA was observed. Related to the clinical status, the median fraction of PGP-positive blasts was elevated fourfold in pretreated patients (n = 16) in comparison to patients with de novo AML (n = 19). No differences in ID50 values were observed between newly diagnosed and pretreated patients. However, SRs for ADM and VCR were higher in samples of pretreated patients compared with de novo AML. PGP-mediated cellular drug resistance may thus be circumvented in leukemic blasts by application of chemosensitizers or, potentially, alternative anthracyclines.


PCR-determined expression of the MDR1 gene in chronic lymphocytic leukemia.

To determine the role the multiple drug-resistance (MDR 1) gene plays in chronic lymphocytic leukemia (CLL), we measured the expression of the MDR 1 gene in 30 patients with this disease. A rapid, highly sensitive, and nonradioactive technique based on the polymerase chain reaction (PCR) was used for that purpose. In this technique, called differential PCR, the target (MDR 1) and a reference gene (beta 2-microglobulin) are co-amplified by PCR from random hexamer-primed cDNA in the same reaction vessel. The level of target gene expression is reflected in the ratio between the intensities of the two resulting PCR product bands, as measured by high-performance liquid chromatography (HPLC). MDR 1 gene expression was detectable in 29/30 (97%) patients with CLL, with a median expression level of 0.36 U (human placenta = 1 U). There was no correlation between expression of the MDR 1 gene and clinical stage, time from diagnosis, absolute lymphocyte count, several lymphocyte surface markers, or prior treatment in the patients analyzed. Immunocytochemical studies of the same material using the monoclonal antibody C219 showed a very low or undetectable expression of the P-glycoprotein in the lymphocytes of all patients studied, whereas granulocytes were significantly more immunoreactive. We conclude that the level of expression of the MDR 1 gene in CLL is generally low, that the removal of granulocytes is important in studies of expression of MDR 1 mRNA in CLL, and that differential PCR provides a rapid and reliable method for quantifying the amount of a specific mRNA, even in very small samples of total RNA.


Effect of tissue perfusion and oxygenation on accumulation of collagen in healing wounds. Randomized study in patients after major abdominal operations.

To compare the accumulation of collagen in standardised wounds in patients who had abdominal operations and whose postoperative fluid replacement was decided either clinically or by measurements of subcutaneous oxygen tension (PscO2). Prospective randomised trial. University Hospital. Twenty-nine patients with cancer or inflammatory bowel disease who were undergoing abdominal operations. Silicone rubber catheter placed in the upper arm to measure PscO2. Two tubes of expanded polytetrafluoroethylene (ePTFE) implanted subcutaneously parallel to the silicone rubber catheter to measure the amount of collagen accumulated. Amount of postoperative fluid replacement required in each group, and measurements of hydroxyproline/cm of the ePTFE tubes. The group treated according to PscO2 measurements received more fluid on the day of operation than the group treated according to clinical criteria (p < 0.05). They had accumulated more collagen in their ePTFE tubes by day 7 (p < 0.05). Replacement of fluid according to measurements of PscO2 rather than by clinical criteria results in improved accumulation of collagen in healing wounds.


Ultrasonographic screening for abdominal aortic aneurysm.

To assess the prevalence of abdominal aortic aneurysm in a selected group of men over the age of 60, and define main risk factors. Population based screening study. Private Norwegian health maintenance organisation. 500 men over the age of 60 years. General examination by a general practitioner, together with measurements of blood glucose and serum cholesterol concentrations. Abdominal scan with a B-mode ultrasound imager. An increase in the diameter of the aorta of more than 150% over the diameter at the origin of the superior mesenteric artery, or maximum diameter of more than 29 mm. Correlation with history of smoking, serum cholesterol concentration, and general health. 29 patients (5.8%) had small, and 12 (2.4%) had large, abdominal aortic aneurysms. There was a significant association between aortic aneurysm and history of smoking (p < 0.01), poor health (defined as coexistent hypertension, cardiovascular disease, or diabetes mellitus) (p < 0.01), and increasing age (p = 0.025). There was no association with hypercholesterolaemia. Ultrasonic screening of groups at risk followed by elective operation may reduce mortality of abdominal aortic aneurysm.


Association between perioperative blood transfusion and postoperative infection in patients having elective operations for gastrointestinal cancer.

To assess the effect of blood transfusion on the development of infection after elective operations for gastrointestinal and pancreatic cancer. Prospective open study. 285 consecutive patients admitted with gastric, colorectal, and pancreatic cancer, 215 of whom were suitable for the study. Sex, age, total iron binding capacity, measurement of haemoglobin and serum albumin concentration, assessment of immune response by delayed hypersensitivity skin test, and weight loss were recorded, as were duration of operation, operative blood loss, and pathological TNM stage of the disease. Sixty patients (28%) developed infections postoperatively. Univariate analysis showed that the development of infection was significantly associated with the amount of blood transfused (p < 0.001), operative blood loss (p < 0.05), and length of operation (p < 0.01), but not weight loss (p = 0.07) or delayed hypersensitivity response (p = 0.08). Multiple logistic analysis showed that blood transfusion was a significant independent variable only when more than 1,000 ml were transfused (odds ratio 6.5, p < 0.05). Transfusion of more than 1,000 ml of blood is an independent risk factor in the development of infection postoperatively in patients undergoing operations for gastrointestinal cancer.


Value of computed tomography in preoperative evaluation of resectability and staging in oesophageal carcinoma.

To evaluate computed tomography (CT) as a method of staging patients with oesophageal carcinoma. Retrospective study. Ullevaal Hospital, Oslo, Norway. 85 patients who presented with oesophageal carcinoma between February 1982 and December 1989. Assessment by CT was made in all patients. Correlation with findings at operation, and histological examination of operative specimens. From the findings at operation in the 46 patients who were operated on (resection, n = 39; exploration only, n = 7) the sensitivity, specificity, and accuracy of CT staging of the growth were 80%, 91%, and 88%, and when histological confirmation of infiltration was taken into consideration the figures were 25%, 84%, and 54%, respectively. The sensitivity, specificity, and accuracy of CT detection of lymph node metastases were 22%, 95%, and 55%. Staging of oesophageal cancer with CT before operation is useful in judging resectability, but the smaller tumours should be assessed with caution. CT is unreliable in the diagnosis of the depth of infiltration and the presence of lymph node metastases compared with histological examination.


Serum interleukin-6 and C reactive protein responses in patients after laparoscopic or conventional cholecystectomy.

To investigate the acute phase inflammatory response to surgical trauma after laparoscopic and conventional cholecystectomy. Prospective open study. University Hospital in The Netherlands. 21 patients with symptomatic cholelithiasis admitted for elective cholecystectomy who had had no previous upper abdominal operations. 12 patients underwent conventional, and 9 patients laparoscopic, cholecystectomy. Circulating interleukin-6 (IL-6) and C reactive protein concentrations were measured 6, 12, 24 and 48 hours after operation. Changes in IL-6 and C reactive protein concentrations, and comparison of operative blood loss and length of stay in hospital. Those treated by laparoscopic cholecystectomy lost significantly less blood (median 60 compared to 100 ml) and spent significantly fewer days in hospital (median 2 compared with 7 days), (p < 0.01 in each case). The only changes in circulating IL-6 concentrations were seen in patients over the age of 60 years who underwent conventional cholecystectomy. There were significant differences in C reactive protein concentrations between the two operations at both 24 and 48 hours after the operation (p < 0.01 in each case). We conclude that laparoscopic cholecystectomy reduces the acute phase inflammatory response compared with the conventional operation; there seems to be no relevant correlation between plasma concentrations of IL-6 and C reactive protein; the presence of IL-6 does not affect the response of C reactive protein to trauma; and the response of IL-6 to trauma is age dependent.


Early carcinoma of the gallbladder.

To clarify the definition and clinicopathological characteristics of early cancer of the gallbladder. Open study. University hospital. 78 patients with pathological stage T1a and 11 with pT1b tumours. Of those with pT1a tumours, 51 were treated by simple, and 27 by radical, cholecystectomy. For those with pT1b tumours the figures were 5 and 11, respectively. There were two recurrences, both in patients with pT1a tumours, histological examination of which had shown tumour present at the surgical margins. There were no cases of neurovascular invasion but only one case of lymphatic invasion that was mild and restricted to the lamina propria in a patient with a pT1b tumour. The outcome in the rest was excellent. Most pT1 cancers spread locally. Complete resection results in cure regardless of the scope of the operation done. The main determinant of prognosis is the presence of tumour at the resection margins. All pT1 cancers can therefore be regarded as early cancers of the gallbladder.


Routine early endoscopic cholangiography, sphincterotomy and removal of common duct stones in acute gallstone pancreatitis.

To study the efficacy, safety and timing of endoscopic retrograde cholangiography (ERC) and sphincterotomy in patients with acute gallstone pancreatitis. Open study in Tampere University Hospital, Finland. 45 consecutive patients with acute gallstone pancreatitis who underwent ERC, with or without sphincterotomy. The results of early, compared with late, ERC with or without sphincterotomy. ERC was successful in all 45 patients. Ampullary impacted stone was found in eight. Common duct stones were found in 21 (47%) and sphincterotomy was successful in 19 of these (90%). Nine patients developed complications (20%), five of the nine in whom severe disease had been predicted (56%) and four of the 36 in whom mild disease had been predicted (11%, p < 0.01). Three patients required operations for necrotising pancreatitis, in two of whom sphincterotomy had failed. There was no difference in outcome between the 21 patients who had ERC with or without sphincterotomy within 72 hours (median 48 h) of the onset of symptoms and the 24 in whom it was delayed for a median of 144 hours. ERC and sphincterotomy may be done safely as a routine in patients with acute gallstone pancreatitis, and delay for a median of six days (range 3-14) from the onset of symptoms did not seem to affect the outcome in our patients.


Delorme's operation for rectal prolapse in elderly or unfit patients.

To evaluate the results after Delorme's operation for rectal prolapse in elderly and unfit patients. Retrospective analysis and telephone interview. Akademiska sjukhuset, Uppsala, Sweden. A consecutive series of 14 women (median age 82, range 32-92) operated on for rectal prolapse 1987-1992 and followed up after a median of 18 months. There were no serious postoperative complications. Continence was improved in 6/11 incontinent patients. The recurrence rate was 3/14 (21%). Delorme's operation is an attractive alternative in elderly or unfit patients, but is not recommended for younger and low risk patients because of the recurrence rate.