Proliferative responses of CD4+ T-cell population to ovalbumin in patients with atopic dermatitis who are sensitive to hen eggs. Proliferative responses of peripheral blood mononuclear cells and T cells with monocytes to ovalbumin were significantly higher than those of B cells with monocytes to ovalbumin in patients with atopic dermatitis (AD) who were sensitive to hen eggs. The CD4+ T-cell/CD8+ T-cell ratios (the values obtained by dividing the maximum stimulation index of CD4+ T cells with monocytes to ovalbumin by the maximum stimulation index of CD8+ T cells with monocytes to ovalbumin) were significantly higher in AD patients sensitive to hen eggs than in non-atopic healthy controls. The proliferative responses of CD4+ T cells with monocytes to ovalbumin were more intensive than those of CD8+ T cells with monocytes in patients with AD sensitive to hen eggs compared with non-atopic healthy controls. These results suggest that the cells responding to ovalbumin are predominantly CD4+ T cells. However, there was no relationship between the stimulation index of proliferative responses of peripheral blood mononuclear cells to ovalbumin and RAST scores for hen eggs. Thus it is possible that the majority of the CD4+ T cells which respond to ovalbumin are not CD4+ helper T cells for IgE production.


Urinary glycylprolyl dipeptidyl aminopeptidase (GP-DAP) in insulin-dependent diabetic patients. Urinary glycylprolyl dipeptidyl aminopeptidase (GP-DAP) concentrations were determined in 36 insulin-dependent diabetic children aged 4-18 years with a duration of diabetes ranging from 1 month to 14 years. Abnormal urinary GP-DAP concentrations were found in 19 of the 36 patients. Twelve of 27 patients without microalbuminuria also had increased urinary concentrations of GP-DAP. There was a significant correlation between urinary GP-DAP and plasma fructosamine (r = 0.52, p < 0.001). Our data suggest that urinary GP-DAP may be used as a marker for diabetic nephropathy. However, there is also a possibility that increased urinary GP-DAP concentrations are functionally related to poor metabolic control. Longitudinal studies are needed to establish the clinical usefulness of urinary GP-DAP.


[The timely diagnosis of postoperative clostridial infection]. Authors report a case of clostridial myonecrosis postcholecystectomy. The causes of the occurrence of this severe, frequently lethal complication are reviewed, as well as the difficulties of an early diagnosis, the only factor able to save the patient. The curative treatment of the detected infection requires the initiation of a series of local and general measures and a permanent cooperation between the surgeon, the specialist in resuscitation and the infectionist.


[Retrospective studies on the relation between fibrocystic disease and cancer of the breast with therapeutic conclusions]. On the basis of 597 cases of benign and malignant tumoral, respectively dysplastic diseases of the breast, operated at the IInd Surgical Clinic of Tirgu-Mureş during the period 1977-1983, the authors consider the fibrocystic disease (FCD) of the II and III degree as a facultative precancerous lesion. The extremely high frequency of FCD associated to the mammary cancer in young women, as well as of intense epithelial proliferations and intermediate forms, of transition from typical epithelial hyperplasias to the atypical forms observed in some of these cases, is pointed out. The 4 cases (1.94%) of mammary neoplasms which occurred late after a mammary sectorectomy performed for FCD illustrate that the risk of mammary neoplasm in FCD is 7.4 times higher than the mammary cancer rate in women without FCD. In the extensive forms of FCD with marked epithelial proliferations, especially if additional risk factors are also present, the authors recommend a uni- or bilateral subcutaneous mastectomy, a uni- or bilateral simple mastectomy respectively. They deem necessary the setting up, in all departments of general surgery, of an adequate organizational framework for the dispensary care, the hormonal therapy and the control of women with FCD.


[Late reinterventions after the surgical treatment of gastric and duodenal ulcer]. A number of 87 reinterventions performed during a 5-year-period for late complications of the gastric and duodenal ulcer surgery are analysed. In most of them (64 cases), the cause of the reintervention was a postoperative ulcer. A long afferent loop (6 cases), the dumping syndrome (4 cases), the stenosis of the anastomosis opening (6 cases) and the primitive neoplasm of the gastric stump (7 cases) represented other causes of reintervention. The immediate postoperative results were very good and good in 69 cases. The risks related to the specific character of this surgery materialized themselves in 14 postoperative complications (anastomotic fistulas, haemorrhages from the anastomosis, stress ulcers etc.), which required iterative operations; the postoperative death rate attained 3.4%. The analysis of these postgastrectomy syndromes is an opportunity to discuss about the failure factors in the surgery of the gastric and duodenal ulcer, the possibilities of exploration and the principles which should guide the reparative therapy.


[Sclerosing cholangitis in the evolution of a surgical hepatic hydatid cyst]. The retrospective analysis of 3 clinical observations points out the etiopathogenetic, clinical and therapeutical aspects of the diffuse stenotic cholangitis, which can occur after the surgical treatment of the hepatic hydatid cyst. Although rare (2.9% of hydatid cysts, 13% of those which communicate with the bile ducts), the diffuse stenotic posthydatid cholangitis represents a severe postoperative complication in cases of median cysts, exerting a compression upon the convergence of hepatic ducts and communicating with the biliary tract. Its presence should be clinically suspected if a mechanical icterus with septic angiocholitis, sometimes associated with an external biliary fistula (from the residual cavity), occurs in the postoperative course of these patients, especially if the primary operation has excluded the remanance of an obstacle at the level of the main bile duct. The lesional substrate is comparable with that of the primitive sclerosing cholangitis, from which it differs through its clear relation with the primary treatment of the hepatic hydatid cyst, through the rapid course of stenotic lesions which, although diffuse, may become more marked in certain segments, as well as through the constant suprastenotic dilatation of the bile ducts. In the pathogenesis are involved the caustic action of some scolicide solutions (2 per cent formaldehyde solution, hypertonic salt solution) on the wall of the bile duct and the cystobiliary communication which predisposes to the peroperative occurrence o-a migration syndrome and of angiocholitis. It requires an early surgical reintervention in order to solve the cholestasis and angiocholitis: according to the morphological situation, we have the choice between disobstruction and trans-stenotic calibration drainage, on the one hand, and biliodigestive derivations in the hilum, which are more efficient, on the other. The prognosis is burdened with the vital risk of septic angiocholitis and with the early occurrence of a secondary biliary cirrhosis or of stenotic recurrences. Prophylaxis consists in the performance of a primary surgical treatment, adequate in median and communicating hydatid cysts, avoiding the "blind" intracystic administration of scolicide solutions, which exert a caustic action on the bile ducts.


[A choledochoduodenal fistula of ulcerous origin]. The case of a 75-year-old patient is reported in whom one of the rare complications of the duodenal ulcer, the choledochoduodenal fistula, has occurred as a result of the prolonged course of this disease. The case was solved by "indirect treatment" of the pathologic complex, with very good results confirmed over time.


[A scale of anesthetic-surgical risk].

[A scale of anesthetic-surgical risk]. An anaesthetic and surgical risk scale, which has also a predictive power regarding the lethality and the probability of occurrence of postoperative complications, is shown. The scale scores the constitutional taints, the extent of the operation, the age, the eventual emergency, the special anaesthetic risk. A second variant scores from the constitutional viewpoint each of the 7 organ systems and adds a lowering of the risk in case of an operation performed upon a system which is directly risk generating. The lethality figure was established by applying the scale to 1,945 patients operated and by recording the lethality distribution for each risk degree, after which the function which approximates most accurately the data score thus obtained is evaluated by the method of the least squares. In this way, a direct correlation of the calculated risk with the actual morbidity and complication rate, as well as with the lethality in the group of patients with complications is obtained. As the scale is achieved by the analysis of patients belonging to the field of general surgery and orthopaedics, it is valid only for this specialty.


Analysis of the agonist activity of fenoldopam (SKF 82526) at the vascular 5-HT2 receptor. 1. The 5-HT2 receptor agonist activity of fenoldopam (SKF 82526) was characterized in the rabbit isolated aorta preparation. 2. Fenoldopam was an agonist at the vascular 5-HT2 receptor with lower affinity and efficacy than the naturally occurring agonist 5-hydroxytryptamine (5-HT). Fenoldopam had an affinity (pKA) of 5.84 +/- 0.04 and efficacy (tau) of 0.57 +/- 0.04, whereas 5-HT had a pKA of 6.65 +/- 0.12 and tau of 2.66 +/- 0.41. 3. The constrictor effects of fenoldopam and 5-HT were competitively antagonized by the 5-HT2 antagonist, ketanserin, with pKB values of 8.81 +/- 0.11 and 8.83 +/- 0.10 respectively. 4. Prior incubation with fenoldopam produced a concentration-related rightward shift of a subsequent 5-HT concentration-response curve. This inhibition was specific for 5-HT since constrictor responses to angiotensin II were unaffected. 5. This study indicates that the D1 receptor agonist, fenoldopam, acts as an agonist at the vascular 5-HT2 receptor, but with an affinity and efficacy less than that of the naturally occurring agonist, 5-HT.


Regulatory role of brain angiotensins in the control of physiological and behavioral responses. Considerable evidence now indicates that a separate and distinct renin-angiotensin system (RAS) is present within the brain. The necessary precursors and enzymes required for the formation and degradation of the biologically active forms of angiotensins have been identified in brain tissues as have angiotensin binding sites. Although this brain RAS appears to be regulated independently from the peripheral RAS, circulating angiotensins do exert a portion of their actions via stimulation of brain angiotensin receptors located in circumventricular organs. These circumventricular organs are located in the proximity of brain ventricles, are richly vascularized and possess a reduced blood-brain barrier thus permitting accessibility by peptides. In this way the brain RAS interacts with other neurotransmitter and neuromodulator systems and contributes to the regulation of blood pressure, body fluid homeostasis, cyclicity of reproductive hormones and sexual behavior, and perhaps plays a role in other functions such as memory acquisition and recall, sensory acuity including pain perception and exploratory behavior. An overactive brain RAS has been identified as one of the factors contributing to the pathogenesis and maintenance of hypertension in the spontaneously hypertensive rat (SHR) model of human essential hypertension. Oral treatment with angiotensin-converting enzyme inhibitors, which interfere with the formation of angiotensin II, prevents the development of hypertension in young SHR by acting, at least in part, upon the brain RAS. Delivery of converting enzyme inhibitors or specific angiotensin receptor antagonists into the brain significantly reduces blood pressure in adult SHR. Thus, if the SHR is an appropriate model of human essential hypertension (there is controversy concerning its usefulness), the potential contribution of the brain RAS to this dysfunction must be considered during the development of future antihypertensive compounds.


Brain non-adenylated mRNAs.

Brain non-adenylated mRNAs. Most eukaryotic messenger RNA (mRNA) species contain a 3'-poly(A) tract. The histone mRNAs are a notable exception although a subclass of histone-encoding mRNAs is polyadenylated. A class of mRNAs lacking a poly(A) tail would be expected to be less stable than poly(A)+ mRNAs and might, like the histones, have a half-life that varied in response to changes in the intracellular milieu. Brain mRNA exhibits an unusually high degree of sequence complexity; studies published ten years ago suggested that a large component of this complexity might be present in a poly(A)- mRNA population that was expressed postnatally. The question of the existence of a complex class of poly(A)- brain mRNAs is particularly tantalizing in light of the heterogeneity of brain cells and the possibility that the stability of these poly(A)- mRNAs might vary with changes in synaptic function, changing hormonal stimulation or with other modulations of neuronal function. The mRNA complexity analyses, although intriguing, did not prove the existence of the complex class of poly(A)- brain mRNAs. The observed mRNA complexity could have resulted from a variety of artifacts, discussed in more detail below. Several attempts have been made to clone members of this class of mRNA. This search for specific poly(A)- brain mRNAs has met with only limited success. Changes in mRNA polyadenylation state do occur in brain in response to specific physiologic stimuli; however, both the role of polyadenylation and de-adenylation in specific neuronal activities and the existence and significance of poly(A)- mRNAs in brain remain unclear.


Differential sensitivities of the prostacyclin and nitric oxide biosynthetic pathways to cytosolic calcium in bovine aortic endothelial cells. 1. Bovine aortic endothelial cells were cultured in vitro, and shown to release both prostacyclin (PGI2; Kact = 24.1 nM) and endothelium-derived relaxing factor (EDRF, NO; Kact = 0.7 nM) in a concentration-dependent manner when exposed to bradykinin. 2. The bradykinin-dependent release of PGI2 (but not EDRF) was inhibited by 1 microM isoprenaline or 5 microM forskolin, and the inhibitory effect of isoprenaline could be reversed by the beta 2-adrenoceptor antagonist, ICI 118551. In contrast, isoprenaline had no capacity to inhibit PGI2 release stimulated by exogenous arachidonic acid. 3. Exposure of cells to bradykinin increased the cytosolic concentration of Ca2+ ions ([Ca2+]i; Kact = 4.8 nM), and the effect was inhibited by both 1 microM isoprenaline and 5 microM forskolin. 4. In similar experiments, exposure of cells to ionomycin also increased [Ca2+]i and the values of [Ca2+]i were calibrated in terms of the ionomycin concentration. In subsequent experiments involving exposure of endothelial cells to selected concentrations of ionomycin, it was possible to show that the biosynthesis of NO was triggered at ionomycin concentrations about one tenth of the required for PGI2 biosynthesis and that these corresponded to a [Ca2+]i threshold of 350 nM for PGI2 release while that for EDRF release was less than 200 nM. 5. These differences in Ca2+ ion sensitivity explain the selective inhibition of bradykinin-stimulated PGI2 biosynthesis (to the exclusion of NO biosynthesis) by isoprenaline or forskolin, both of which attenuate bradykinin-dependent increases in [Ca2+]i.


Blockade by antiarrhythmic drugs of glibenclamide-sensitive K+ channels in Xenopus oocytes. 1. The outward K+ current induced by KRN2391 (K+ channel opener) in Xenopus oocytes is blocked by glibenclamide. We have investigated the effects of various classes (I-IV) of antiarrhythmic drugs on this KRN2391-induced response. 2. All class I antiarrhythmic drugs (Na+ channel blockers) tested concentration-dependently suppressed KRN2391-induced responses with the rank order of potency (IC50 in microM), disopyramide (17.8) > aprindine (29.5) > propafenone (63.1) > ajmaline (145) > quinidine (151). Flecainide, SUN1165, lignocaine, mexiletine and procainamide were much less potent (IC50, 450- > 1000 microM) than quinidine. 3. The class II antiarrhythmic drugs (beta-blockers), timolol, (-)- and (+/-)- propranolol, and (+)- propranolol (a non-beta-blocker) inhibited KRN2391-induced K+ currents in a concentration-dependent manner with values for IC50 (microM) of 79, 131, 151 and 129, respectively, whilst butoxamine, oxprenolol, alprenolol, pindolol, nadolol, metoprolol and acebutolol were either weak (IC50, 300 microM-600 microM) or virtually inactive (IC50, > 1000 microM). 4. The class III antiarrhythmic drugs, amiodarone and (+)-sotalol scarcely affected KRN2391 responses. 5. All class IV drugs (Ca2+ antagonists) tested suppressed KRN2391-induced responses in a concentration-dependent manner with an IC50 of 6.3 microM for bepridil, 38 microM for prenylamine, 85 microM for verapamil and 135 microM for diltiazem. 6. In conclusion, antiarrhythmic drugs of classes I, II and IV potently blocked glibenclamide-sensitive K+ channels in Xenopus oocytes.


The local intracoronary administration of methylene blue prevents the pronounced antiarrhythmic effect of ischaemic preconditioning. Short periods of coronary artery occlusion (2 x 5 min) markedly reduce the severity of arrhythmias and the changes in ST-segment elevation and in the degree of inhomogeneity of conduction during a subsequent 25 min occlusion of the left anterior descending coronary artery in anaesthetized dogs. These changes were completely reversed if methylene blue (5 mg min-1) was infused into a side branch of the coronary artery throughout both the preconditioning and prolonged occlusions. These results suggest that the pronounced antiarrhythmic effects of preconditioning result from activation of guanylyl cyclase and result in increased levels of guanosine 3':5'-cyclic monophosphate.


Subclassification of release-regulating alpha 2-autoreceptors in human brain cortex. 1. Release-regulating alpha 2-autoreceptors in human brain were characterized pharmacologically in cortical slices from patients undergoing neurosurgery to remove subcortical tumours; the slices were prelabelled with [3H]-noradrenaline ([3H]-NA) and stimulated electrically (3 Hz, 2 ms, 24 mA) under superfusion conditions. 2. The stimulus-evoked tritium overflow was almost totally Ca(2+)-dependent and tetrodotoxin-sensitive. 3. Clonidine and oxymetazoline 0.01 to 1 microM inhibited in a concentration-dependent manner the evoked overflow of tritium. The two drugs were equipotent (EC50 = 0.03 microM) and their maximal effect was approx. 45%. Phenylephrine and methoxamine, up to 1 microM, did not affect tritium overflow. 4. Yohimbine (0.01-0.1 microM) shifted the concentration-response curve of clonidine to the right. The calculated pA2 value was 8.29. 5. Prazosin and 2-[2-[4-(o-methoxyphenyl)piperazine-1-yl]ethyl]-4,4- dimethyl-1,3(2H,4H)-isoquinolinedione (AR-C 239), tested at 0.3 microM, did not modify the concentration-response curve of clonidine. 6. The effect of clonidine was antagonized by (+)-mianserin (pA2 = 7.74), but not by up to 0.3 microM of the (-)-enantiomer. The concentration-response curve of clonidine was shifted to the right by the novel alpha 2-adrenoceptor antagonist, 5-chloro-4-(1-butyl-1,2,5,6-tetrahydropyridin-3-yl)-thiazole-2-ami ne (Z)-2-butenedioate (1:1) salt (ORG 20350) (pA2 = 7.55). 7. Yohimbine, (+)-mianserin and ORG 20350, but not prazosin and (-)-mianserin, increased the electrically-evoked tritium overflow, suggesting that autoreceptors may be tonically activated by endogenous NA. 8. Desipramine (1 microM) increased evoked tritium overflow from human cortex slices. The effect of clonidine (0.01- 1 g1M) on the evoked overflow of tritium was reduced in presence of 1 muM desipramine.9. It is proposed that autoregulation of NA release can occur in human cerebral cortex. The process involves activation of alpha 2-adrenoceptors which may be either the alpha2X or the alpha2D subtype.


Noradrenaline modulates smooth muscle activity of the isolated intravesical ureter of the pig through different types of adrenoceptors. 1. We have studied the effects of alpha- and beta-adrenoceptor agonists and antagonists on both phasic peristaltic activity and basal tone of the isolated intravesical ureter of the pig by means of isometric techniques in vitro. 2. Spontaneous phasic activity was exhibited by 21% of pig intravesical ureter preparations manifested as rhythmic contractions with average frequency and amplitude of 2.54 +/- 0.18 min-1 and 1.48 +/- 0.16 g (n = 31), respectively. 3. Adrenaline, noradrenaline and phenylephrine induced concentration-dependent increases in both phasic activity and basal tone of ureteral preparations, all three agonists being more potent in modifying ureteral phasic activity than baseline tone. B-HT 920, B-HT 933 and clonidine had no significant effect. 4. Phentolamine (10(-9)-10(-7) M) and prazosin (3 x 10(-11)-3 x 10(-8) M) significantly inhibited increases in both frequency of phasic activity and baseline tone induced by a submaximal dose of noradrenaline. Rauwolscine (10(-9)-10(-7) M) affected only the tone evoked by noradrenaline and higher concentrations of this antagonist were needed to block phasic activity. 5. Pretreatment of ureteral strips with the beta-adrenoceptor antagonist, propranolol (10(-6) M), significantly increased the maximum contraction evoked by noradrenaline. After incubation with phentolamine (10(-6) M), noradrenaline (10(-7)-10(-6) M) decreased phasic activity induced by prostaglandin F2 alpha (10(-5) M). Isoprenaline and salbutamol also abolished PGF2 alpha-induced phasic activity. Pafenolol (10(-6) M) and butoxamine (10(-6) M) blocked the inhibitory effect of noradrenaline, isoprenaline, and salbutamol on PGF2 alpha-induced phasic activity. 6. These results suggest that noradrenaline may modulate both phasic peristaltic activity and basal tone of pig intravesical ureter through both alpha- and beta-adrenoceptors.


Observations on calcium oxalate stone formers. Biochemical studies were performed on 22 adult patients with idiopathic recurrent calcium oxalate renal stone disease and 23 healthy controls. It was found that urinary glutamic-oxaloacetic acid transaminase (UGOT) and urinary glutamic-pyruvic acid transaminase (UGPT) activity was low and lactate dehydrogenase (LDH) and gamma-glutamyl transferase (gamma-GT) activity was high in the urine of calcium oxalate stone formers. No significant changes were observed in the activity of glutamic-oxaloacetic acid transaminase, glutamic-pyruvic acid transaminase and gamma-GT in their blood but a significant reduction was found in both LDH and alkaline phosphatase activity. It was concluded that the activity of UGOT and UGPT is reduced in patients with kidney stones.


Niacinamide blocks 3-acetylpyridine toxicity of cerebellar granule cells in vitro. 3-Acetylpyridine (3AP) is a potent neurotoxin when administered to laboratory animals. However, its neurotoxic effects have not been investigated extensively in vitro. Cultured cerebellar granule cells are killed by concentrations of 3AP of 0.1-1 mM (ED50 = 220 microM) but not by its 2-acetyl and 4-acetyl analogues. The toxicity of 3AP is enhanced by preexposure to subtoxic concentrations of N-methyl-D-aspartate (NMDA) and is unaffected by the NMDA receptor antagonists MK-801 or APV, as well as by deprenyl, mazindol, or tetrahydrofolic acid. However, 3AP toxicity is completely blocked by preincubating cerebellar granule cells with low concentrations of niacinamide. These data lead us to suggest that 3AP toxicity is due to the substitution of 3AP for niacinamide in the formation of niacinamide adenine dinucleotides (NAD(P)).


Cortically evoked excitatory synaptic transmission in the cat red nucleus is antagonised by D-AP5 but not by D-AP7. Extracellular recordings were made from magnocellular red nucleus neurons (mRN) in alpha-chloralose (50 mg/kg, iv.) anaesthetised cats. Iontophoretically applied N-methyl-D-aspartate (NMDA) excited the neuronal firing which was antagonised by 4 selective NMDA receptor antagonists: 2-amino-5-phosphonopentanoate (AP5), 2-amino-7-phosphonoheptanoate (AP7), RS-4-(phosphonomethyl) piperazine-2-carboxylic acid (PMPC) and R-4-(3-phosphonopropyl) piperazine-2-carboxylic acid (CPP), whereas AMPA responses were uneffected. Monosynaptic excitatory responses were produced by stimulation of the sensorimotor cortex. These responses were reduced and often abolished by AP5 and PMPC but not by AP7 or CPP. It is postulated that two NMDA receptor subtypes exist on mRN neurones.


Characterization of dynorphin-containing neurons on dissociated dentate gyrus cell cultures. In the dentate gyrus, the synthesis of the opioid peptide, dynorphin, is modulated by a variety of stimuli. In order to elucidate the cellular and molecular mechanisms regulating the synthesis of dynorphin in the hippocampus, we have established a routine primary cell culture of dentate granule neurons and identified granule-like neurons by a characteristic marker, dynorphin, in these cultures. Cultures were prepared from 7-day-old rat pups and maintained in medium with 2% fetal bovine serum. These cultures contained approximately 20% neurons and survived for over 4 weeks. After 2 weeks in culture, neurons expressing dynorphin-A and its messenger RNA were detected using immunocytochemistry and in situ hybridization, respectively. In dentate cultures, enkephalin-, cholecystokinin-, neuropeptide Y- and substance P-positive cells were observed in addition to dynorphin-positive cells with immunocytochemistry. The results suggest that dentate gyrus cell cultures provide a valid in vitro model for studying molecular mechanisms regulating prodynorphin gene expression.


The N-methyl-D-aspartate antagonist MK-801 protects against serotonin depletions induced by methamphetamine, 3,4-methylenedioxymethamphetamine and p-chloroamphetamine. The non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 blocks the ability of D-methamphetamine (MA) to deplete striatal dopamine (DA). We now report that MK-801 attenuates decreases in serotonin (5-HT) concentration induced by MA and two other amphetamine analogues, 3,4-methylenedioxymethamphetamine (MDMA) and p-chloroamphetamine (PCA). Rats were injected with saline (1.0 ml/kg) or MK-801 (0.5, 1.0 or 2.5 mg/kg) followed by either saline (1.0 mg/kg), MA (4, 2 or 1 injection(s); 10.0, 20.0 or 40.0 mg/kg), MDMA (20.0 or 40.0 mg/kg) or PCA (5.0 or 10.0 mg/kg). In some experiments, two injections of MK-801 or saline were used. Seventy-two hours after the last injection rats were sacrificed and concentrations of 5-HT, 5-hydroxyindoleacetic acid (5-HIAA) and DA were determined in hippocampus and striatum. MA caused a depletion of 5-HT to 33% of control in hippocampus and to 50% of control in striatum after the 4 x 10.0 mg/kg dose regimen. When MK-801 (2.5 mg/kg) was co-administered with MA, concentrations of 5-HT did not differ from control levels in either brain region. MDMA depleted 5-HT to approximately 58% of control in hippocampus and 66% of control in striatum at the 40 mg/kg dose.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of Trypanosoma vivax infection on semen characteristics of Yankasa rams. Twelve Yankasa rams aged between 2 1/2 and 3 years with good semen characteristics were used in this 15-week study. Six rams were infected with Trypanosoma vivax, while six served as controls. The infected rams developed chronic trypanosomosis accompanied by fluctuating pyrexia, lethargy, anaemia, scrotal oedema and cachexia. There was a drastic and progressive deterioration in semen quality in all infected rams manifested by a decrease in volume or cessation of semen production, oligozoospermia, a sharp decrease in progressively motile sperm, elevated numbers of dead (eosinophilic) sperm and 100% morphological abnormalities of sperm in most animals. The rams were all deemed unfit for breeding by 3 weeks post-infection. Uninfected rams were healthy and had good semen characteristics throughout the investigation. The results show that rams infected with T. vivax may become infertile within a short interval due to rapid deterioration of semen characteristics and this trypanosome species may be an important causative agent of infertility in endemic areas.


APV prevents the elimination of transient dendritic spines on a population of retinal ganglion cells. Blockade of the N-methyl-D-aspartate (NMDA) receptors on retinal ganglion cells (RGCs) during development prevents the elimination of the exuberant spine-like processes in a population of Type I RGCs in hamsters. During the development of RGCs, exuberant dendritic spines have been observed which disappear during maturation. Blocking the NMDA receptors on developing RGCs with the antagonist, DL-2-amino-5-phosphonovaleric acid (APV) and the subsequent retention of some of the normally transient dendritic spines suggest that the morphological development of post-synaptic neurons may be affected by this treatment. Our result further suggests that the elimination of exuberant spines during normal development requires interactions between receptors on the spines and neurotransmitters released by the pre-synaptic inputs.


Long duration ventral root potentials in the neonatal rat spinal cord in vitro; the effects of ionotropic and metabotropic excitatory amino acid receptor antagonists. Long duration, primary afferent evoked ventral root potentials (VRP's) have been recorded in vitro from hemisected spinal cords prepared from 8-12-day-old rat pups. Single shock stimulation of a dorsal root at stimulus strengths sufficient to recruit C/group IV afferent fibres evoked a long duration (11.9 +/- 1.2 s) ipsilateral VRP in all preparations. This long duration VRP consisted of two components, (i) a slow wave, time to peak 137.0 +/- 5.1 ms, the amplitude of which was reduced to 8.7% of mean control value in the presence of the N-methyl-D-aspartate (NMDA) antagonist D-AP5 (40 microM), (ii) a prolonged wave with a time to peak of 2.0 +/- 0.2 s which was partially resistant to D-AP5 (40 microM). Both the slow and the prolonged waves were unaffected following superfusion with the metabotropic excitatory amino acid (EAA) receptor antagonist L-AP3 (100-200 microM). Low frequency (1-10 Hz) repetitive stimulation (20 s duration) of high threshold dorsal root afferents evoked a temporal summation of synaptic activity which generated a progressively depolarizing VRP. This cumulative VRP was graded with frequency of stimulation (0.89 +/- 0.13 to 1.25 +/- 0.19 mV). The cumulative VRP was followed by a post-stimulus depolarization which outlasted the period of repetitive stimulation by tens of seconds (47.6 +/- 8.4 to 91.2 +/- 19.9 s). In the presence of AP5 the amplitude of the cumulative VRP was depressed to 54.5 +/- 11.5% of control values when low frequency (1.0 Hz) stimulation was used. The proportion of the cumulative VRP resistant to D-AP5 increased as the frequency of stimulation was increased to 10 Hz. The decay time of the post-stimulus depolarization was unaffected by AP5. Neither the amplitude nor the post-stimulus depolarization of the cumulative VRP was affected by 200 microM L-AP3. It is suggested that both an AP5 sensitive and AP5 insensitive potential contribute to the long duration VRP evoked in the neonatal rat spinal cord following single shock high threshold afferent stimulation. Moreover, the AP5 insensitive prolonged depolarization is manifest following sustained low frequency stimuli and higher frequency inputs.


Tyrosine augments dopamine release in stimulated rat retina. Endogenous dopamine (DA) release was measured in perfused rat retinae. Perfusion with elevated potassium (40 mM K) resulted in a 5-6-fold increase in DA release over baseline or 11.6 +/- 0.9% of final tissue DA content. When the selective DA D2 receptor agonist quinpirole was added to the perfusion medium (at 1 and 10 microM), K-stimulated DA release was significant decreased compared to controls (to 7.0 +/- 1.6 and 6.14 +/- 1.4%, respectively). Addition of the D2 antagonist (+/-)-sulpiride (10 microM) significantly increased DA release to 19.1 +/- 1.3%. DA could be released with successive pulses of K; an initial 10 min pulse resulted in a 4-5-fold increase in endogenous DA release over basal levels or 11.4% of the final retinal tissue DA content and a 3-fold increase (a 9.3% fractional release) upon a second K stimulation given 50 min later. The ratio of stimulated DA release during the two K pulses was 0.82 +/- 0.04. When L-tyrosine (100 microM) was included in the medium throughout the perfusion, K2/K1 was increased to 1.14 +/- 0.13. Both tissue DA level and release were decreased by the tyrosine hydroxylase inhibitor, alpha-methyl-p-tyrosine (AMPT). At 10 microM AMPT K-stimulated DA release was reduced by 50% during the first pulse and completely abolished during the second K pulse. At 100 microM both basal and K-stimulated release were significantly reduced. Exposure of dark-adapted retinae to light in L-tyrosine-supplemented perfusion medium resulted in an increased release of DA compared to retinae perfused with tyrosine-free medium.(ABSTRACT TRUNCATED AT 250 WORDS)


Contribution of the central interaction between calcium and sodium to hemodynamic regulation in spontaneously hypertensive rats. The contribution of the central interaction between calcium and sodium to hemodynamic regulation was assessed in spontaneously hypertensive rats (SHRs) and Wistar-Kyoto (WKY) rats. The effect of a high calcium solution (Ca2+, 130 mg/dl, 10 microliters) infused into the cerebral ventricle (i.c.v.) on hemodynamic responses induced by a high sodium solution (Na+, 1,000 mEq/1, 10 microliters) i.c.v. and the mechanism by which high Ca2+ affects the hemodynamic responses induced by high Na+ i.c.v. were studied. High Na+ i.c.v. induced a pressor response with tachycardia in the SHRs, but induced a pressor response with reflex bradycardia in the WKYs. Prior treatment with high Ca2+ i.c.v. attenuated the pressor response induced by high Na+ i.c.v. (+55.6 +/- 4.4 to +33.1 +/- 3.2 mmHg, P < 0.01) and restored reflex bradycardia (+86.4 +/- 7.7 to -26.7 +/- 7.6 bpm, P < 0.01) in SHRs. Whereas prior treatment with high Ca2+ i.c.v. attenuated the pressor response (+35.7 +/- 2.0 to +22.2 +/- 4.0 mmHg, P < 0.05), it did not alter the degree of reflex bradycardia (-81.7 +/- 7.1 to -69.2 +/- 120 bpm, n.s.) in WKYs. Ganglionic blockade attenuated the pressor response (+56.9 +/- 3.5 to +42.9 +/- 2.3 mmHg, P < 0.05) and restored reflex bradycardia (+82.1 +/- 10.3 to -65.9 +/- 11.0 bpm, P < 0.01) in SHRs, whereas, inhibition of arginine vasopressin attenuated the pressor response without modification of the tachycardic response.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of glucose and fat antimetabolites on norepinephrine turnover in rat hypothalamus and brainstem. Rats were injected acutely with antimetabolites of either glucose (2 deoxy-D-glucose, 2DG), fat (methylpalmoxirate, MP or mercaptoacetate, MAC), or the combination of these agents, in dosages known to stimulate food intake. Norepinephrine (NE) turnover in hypothalamus and brainstem was determined after these treatments by the method of synthesis inhibition. Glucoprivation (2DG) increased NE turnover in hypothalamus, confirming previous studies. Fat antimetabolites alone had no effect on NE turnover, nor did a peripherally-acting fructose antimetabolite. Combination of MP and 2DG, but not MAC and 2DG, produced a greater NE turnover than 2DG alone. These data suggest that peripheral signals of metabolic emergency do not activate brain NE systems, except when these systems are already activated by an ongoing cerebral metabolic emergency. The role of hypothalamic NE in metabolic integration of feeding is discussed, and possible hemispheric differences.


NMDA receptor mediates dopamine release in the striatum of unanesthetized rats as measured by brain microdialysis. We have studied the characteristics associated with the activation of the N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor on the release of dopamine (DA) in the striatum of awake rats as measured by brain microdialysis technique. NMDA dose-dependently stimulated the striatal DA release in Mg(2+)-free Ringer's solution. The stimulation was significant at 90 microM and the maximum observed effect was at the highest concentration tested (800 microM). The selective NMDA receptor antagonist, 2-amino-5-phosphonovalerate (AP5; 300 microM), blocked the stimulatory effect of NMDA. The NMDA-induced release of DA was reduced by 1.2 mM Mg2+ and totally blocked by 2.5 mM of the cation. Glycine (200 microM) potentiated the response evoked by 300 microM NMDA while 7-chloro-kynurenate (100 microM), an antagonist of the glycine site, reduced markedly this response. Neither atropine (100 microM) nor tetrodotoxin (TTX) (5 microM) prevented the stimulatory effect of NMDA. These results suggest that glutamate released from corticostriatal terminals presynaptically stimulates the release of DA via an NMDA receptor.


NMDA receptors assessed by autoradiography with [3H]L-689,560 are present but not enriched on corticofugal-projecting pyramidal neurones. Experimental lesions followed by binding of [3H]4-trans-2-carboxy-5,7-dichloro-4-phenylamino-carbonylamino-1,2 ,3,4- tetrahydroquinoline ([3H]L-689,560, a novel ligand that binds to the glycine modulatory site), [3H]glycine and [3H]glutamate (N-methyl-D-aspartate (NMDA) sensitive) to cryostat sections and quantitative autoradiography were used to investigate the cellular localization of the NMDA receptor complex in the neocortex of the rat. The lesions were produced by intrastriatal injections of either volkensin (2 and 6 ng) or ricin (10 ng): both are suicide transport agents but only the former is retrogradely transported in the CNS. The binding of [3H]L-689,560 was significantly reduced in rats receiving 2 or 6 ng volkensin in deep cortical layers of Fr1/Fr2 ipsilateral to the striatal lesion. Similar reductions were also seen in [3H]glycine and [3H]glutamate binding, but only in rats receiving 6 ng volkensin. Quantitative histological analysis had previously revealed a loss of large infragranular pyramidal neurones with sparing of both interneurones and supragranular pyramidal neurones. There were no significant reductions in binding of any ligand in the superficial layers. In cortical areas Par1/Par2, [3H]L-689,560 was also significantly reduced in deep layers but only in rats receiving 6 ng volkensin. Binding was also reduced in the superficial layers by contrast to Fr1/Fr2. [3H]Glycine and [3H]glutamate binding were unaffected in this area. Binding of [3H]L-689,560 was unaffected in any area following intrastriatal ricin injection. The present study indicates that the NMDA receptor complex is present on pyramidal cells forming the corticofugal pathways. This is discussed in terms of the 5-HT1A receptor which is enriched on these cells.


Inhibition of spinal opioid analgesia by supraspinal administration of selective opioid antagonists. The effect of intracerebroventricular administration of a selective mu- (CTOP) or delta- (ICI 174,864) opioid receptor antagonist on the antinociceptive effects produced by intrathecal administration of selective mu- (DAMGO), delta- (DPDPE) and kappa- (U50-488H) opioid receptor agonists was evaluated using the Randall-Selitto paw-withdrawal test, in the rat. While the intracerebroventricular administration of CTOP or ICI 174,864, alone, had no effect on nociceptive thresholds, intracerebroventricular administration of CTOP and ICI 174,864 produced marked antagonism of the antinociceptive effects of intrathecal DAMGO. The antinociceptive effects of intrathecal administration of DPDPE or U50,488H were not antagonized by intracerebroventricular administration of CTOP or ICI 174,864. These data suggest that, in the rat, along with the established descending antinociceptive pathways, there is an ascending antinociceptive control mechanism projecting from the spinal cord to the brainstem. The ascending antinociceptive control involves mu- and delta-opioid agonism at supraspinal sites and appears to be mediated selectively by mu-, but not by delta- or kappa-opioid agonism at the spinal level.


Effect of the neurotoxin AF64A on intrinsic and extrinsic neuronal systems of rat neostriatum measured by in vivo microdialysis. In the present in vivo microdialysis study the aziridinium ion of ethylcholine mustard, AF64A and the excitotoxin ibotenic acid were compared for their effects on extracellular striatal acetylcholine, choline, gamma-aminobutyric acid (GABA), dopamine and its metabolites, glutamate and aspartate, measured in the same perfusate sample, under basal and high KCL conditions. Ten days following unilateral striatal injections of AF64A (2 x 0.08 to 2 x 8 mM) there was a dose-dependent decrease in the extracellular striatal levels of acetylcholine and GABA, the two major intrinsic striatal neurotransmitter systems. No significant effects were observed on any of the monitored neurotransmitter systems following the lowest (2 x 0.08 mM) dose of AF64A, while at the intermediate (2 x 0.8 mM) dose, AF64A produced a unilateral > 50% and > 70% decrease in basal extracellular striatal acetylcholine and GABA levels respectively. The effects of K(+)-depolarization on extracellular acetylcholine and GABA levels were diminished by approximately 50%. At the highest dose (2 x 8 mM), extracellular striatal acetylcholine levels were non-detectable under basal conditions, while the GABA levels were decreased by > 50%, when compared with the contralateral intact side. However, at this dose, GABA levels were bilaterally decreased compared to levels observed in control animals. Basal extracellular striatal dopamine and glutamate levels, representing the two major extrinsic neurotransmitter systems innervating the neostriatum were only affected by the highest dose of AF64A. The excitotoxin ibotenic acid (2 x 28.4 mM) produced a strong unilateral decrease in extracellular striatal acetylcholine (> 80%) and GABA (> 90%) levels, without significantly affecting basal dopamine and glutamate levels.(ABSTRACT TRUNCATED AT 250 WORDS)


Dependence of carotid chemosensory responses on metabolic substrates. The dependence of the carotid chemosensory response to hypoxia on metabolic substrate and the hypothesis that lactic acidosis is essential for O2 chemoreception were tested. Effects of 3 types of substrate (glucose, glutamate and a mixture of amino acids) on the response to hypoxia (perfusate flow interruption) were measured (n = 33 carotid bodies). The response to nicotine (n = 25) was used to determine whether these effects were exclusive to the hypoxic response. The cat carotid body was perfused and superfused in vitro with modified Tyrode solution (pO2 > 400 Torr, pCO2 < 1 Torr, pH = 7.4) at 36 degrees C containing a given substrate for at least 15 min prior to flow interruption or nicotine injection. Without substrate, responses to flow interruption (n = 4) and nicotine (n = 2) were irreversibly depressed. With glucose, responses to flow interruption (n = 13) and nicotine (n = 8) increased in a concentration-dependent fashion. Glutamate (42 mM) alone (n = 11) or a mixture of amino acids (4.2 mM) plus 5.5 mM glucose (n = 12) substituted for 11 mM glucose (n = 10). Thus, glutamate (42 mM), or a mixture of amino acids (4.2 mM) or a high concentration of glucose (11 mM) can support chemosensory responses to flow interruption and nicotine. Since glutamate undergoes oxidative deamination to alpha-ketoglutarate without lactic acid production, O2 chemoreception does not depend on lactic acidosis.


[Interactions between GABAergic and cholinergic mechanisms involved in monocular optokinetic nystagmus in frogs]. The systemic administration of atropine, a muscarinic cholinergic antagonist, was found to suppress the Nasal-Temporal (N-T) component of the frog monocular optokinetic nystagmus (OKN), which had appeared following a prior injection of bicuculline and which does not exist in the normal animal. On the contrary, the administration of a nicotinic cholinergic antagonist (D-TC, alpha-BGT, Hexamethonium) following that of bicuculline has prolonged the duration of the induced N-T component. Thus, ACh was shown to attenuate or to reinforce the GABAergic inhibition of the N-T component through muscarinic receptors or nicotinic receptors respectively. These data point to the existence of strong interactions between these two neurotransmission systems involved in frog monocular OKN.


The presence of immunoreactive vertebrate bioactive peptide substances in hemocytes of the freshwater snail Viviparus ater (Gastropoda, Prosobranchia). 1. Using an immunocytochemical procedure a wide range of immunoreactive vertebrate bioactive peptides (BAPs) has been found in hemocytes of Viviparus ater: bombesin, calcitonin, CCK-8, CCK-39, GH, glucagon, insulin, oxytocin, neurotensin, secretin, serotonin, somatostatin, substance P, vasopressin, and VIP. 2. No immunostaining was observed for antigastrin and antithyroglobulin antibodies. 3. The presence of BAP-like molecules in hemocytes suggests a correlation between hemocyte and APUD cells and is evidence of a relationship between the neuroendocrine and the immune systems.


Nontraditional combination pharmacotherapy of hypertension. Traditional drug combinations that have additive hypotensive effect include double therapy with a diuretic and any other antihypertensive agent and triple therapy with a diuretic, direct vasodilator, and either a beta blocker or reserpine. Due to the availability of new classes of antihypertensive agents, other combinations of drugs are now increasingly used. The effectiveness of combination therapy for hypertension has been investigated; the results of these studies are reviewed.


Urticaria and angioedema.

Urticaria and angioedema. Urticaria and angioedema are commonly seen in the outpatient setting. Their pathogenesis involves complex cellular and humoral factors. Diagnosis depends on historical information such as duration of symptoms, exacerbating factors, and atopy. While many etiologic factors have been implicated, in most chronic cases no specific etiology is found. This article reviews physical and hereditary syndromes and discusses therapeutic regimens based on the duration and severity of symptoms.


Diagnostic value of a P-fimbriation test in determining duration of therapy in children with urinary tract infections. The patients were 117 children (aged 4 months to 14 years) with uncomplicated urinary tract infections caused by co-trimoxazole-sensitive Escherichia coli. The patients were randomly assigned to receive treatment with co-trimoxazole for 3 days (n = 58) or 7 days (n = 59). Urine was analyzed for bacteria before and immediately after treatment and again at 1 and 2 months. After 3 days' treatment, infection persisted in 14 of 31 patients with P-fimbriated strains of E coli and in 1 of 27 patients with non-P-fimbriated strains. After 7 days' treatment, infection persisted in 2 of 40 patients with fimbriated strains and in none of the 19 patients with nonfimbriated strains. One or 2 months after treatment, 3 days' treatment was rated successful in 26 of 27 patients with nonfimbriated strains and in none of the patients with fimbriated strains. Seven days' treatment was rated successful in all patients with nonfimbriated strains and in 32 of 40 patients with fimbriated strains. The results indicate that the length of treatment of urinary tract infections in children should be adjusted according to the presence of bacterial P-fimbriae in addition to the patients' clinical condition.


Suicide risk associated with drug and alcohol addiction. The association of alcohol and drugs with suicidal thinking and behavior is both causal and conducive. The subjective state of hopelessness is key to the disposition to actual suicide. Alcohol and drugs are influential in providing a feeling of hopelessness by their toxic effects, by disruption of interpersonal relationships and social supports, and, possibly, by manipulating neurotransmitters responsible for mood and judgment. Because alcoholism and drug addiction are leading risk factors for suicide and suicidal behavior, any alcoholic or drug addict should be assessed for suicide, especially if actively using alcohol or drugs.


Gonocytes of male rats resume migratory activity postnatally. In the testis of the neonatal rat, maturation of germ cells, or gonocytes, lays the foundations for spermatogenesis which will begin later in postnatal development. One of the most critical and yet least understood of the events that occur during the immediate neonatal period is relocation of gonocytes from the more central part of the seminiferous cord, where they are surrounded by Sertoli cells, to its periphery, where they contact the basement membrane. For the current study, we examined this change in gonocyte position by identifying some of the cellular mechanism involved, with the aim of determining whether movement of gonocytes to the basement membrane in vivo and development of cellular processes by these cells in vitro represents a resumption of migratory activity similar to that displayed by their fetal ancestors and by other motile cells. First, we used either thiamine pyrophosphatase cytochemistry or the fluorescent probe nitrobenzoxadiazole ceramide to visualize the Golgi complex in gonocytes and found that (1) this organelle matures and apparently enlarges in vivo with a time course paralleling movement of gonocytes to the basement membrane and undergoes similar changes in vitro that correlate with gonocyte process formation, and (2) the Golgi complex is located in perinuclear cytoplasm facing the apparent direction of gonocyte movement in vivo and in cytoplasm near the cellular process in the great majority of elongated gonocytes in coculture. Next we used two drugs, brefeldin A and monensin, which have in common their ability to disrupt the Golgi complex, and found that both drugs prevent process formation by gonocytes in a manner that is completely reversible. We also tested the involvement of the cytoskeleton in gonocyte elongation by utilizing nocodazole to disrupt and taxol to stabilize microtubules, as verified by alpha-tubulin immunofluorescence. Inclusion of the drug abolished (taxol) or substantially diminished (nocodazole) the ability of gonocytes to elongate in a reversible manner. We also found that the Golgi complex was intact in the presence of taxol and that microtubules were intact in the presence of both Golgi complex-specific drugs. Thus, our findings indicate that (1) both the Golgi complex and microtubules are involved in development of processes by gonocytes and (2) neither structure is sufficient by itself to allow these cells to elongate. Taken together, our data provide new evidence suggesting that the cellular mechanism utilized by postnatal gonocytes in relocating to the basement membrane are those mediating active migration.(ABSTRACT TRUNCATED AT 400 WORDS)


An amphicrine pancreatic cell line: AR42J cells combine exocrine and neuroendocrine properties. The permanent cell culture line AR42J, derived from a rat pancreatic acinar carcinoma, is widely used for functional studies of exocrine pancreatic acinar cells. We now present evidence that these cells are amphicrine in that they contain zymogen granules as well as small (40-80 nm) neuroendocrine (NE) vesicles and typical neurotransmitters. Using the small NE vesicle-specific markers synaptophysin and "protein S.V.2", including synaptophysin cDNA probes, we have found that AR42J cells synthesize these proteins and contain vesicles harboring these proteins with biophysical properties similar to those of small NE vesicles. NE properties of these cells are further indicated by the presence of considerable amounts of stored amino acids (gamma-aminobutyric acid (GABA), glycine, glutamate) and by the presence of the GABA-synthesizing enzyme, glutamic acid decarboxylase. Finally, intermediate filament (IF) protein typing showed only cytokeratins 8 and 18, indicating that AR42J cells possess an IF protein complement indistinguishable from that of acinar and islet cells. Our results document the unusual case of a permanent cell line with combined exocrine and neuroendocrine properties that may be indicative of a derivation from a cell with multipotential character.


Inhibition of respiratory burst activity in alveolar macrophages by bisbenzylisoquinoline alkaloids: characterization of drug-cell interaction. The objective of this study was to investigate the effects of various bisbenzylisoquinoline (BBIQ) alkaloids on respiratory burst activity of alveolar macrophages and to characterize the interaction of these drugs with alveolar phagocytes. BBIQ alkaloids were chosen for study because they exhibit a wide range of antifibrotic potencies in a rat model, with tetrandrine being very effective and tubocurarine being ineffective. These drugs inhibited zymosan-stimulated oxygen consumption with a potency sequence of tetrandrine (TT) approximately fangchinoline (FA) > berbamine (BE) approximately cepharanthine (CE) approximately cycleanine (CY) >> tubocurarine (TU). This inhibition of respiratory burst activity could not be attributed to a drug-induced decline in the ATP content of these pneumocytes. Drug binding to alveolar macrophages was directly dependent on temperature and drug concentration. The sequence for binding capacity was FA > TT approximately BE approximately CY > CE >> TU. Therefore, there was no simple relationship between binding capacity and inhibitory potency. Binding capacity was not related to lipophilicity of these alkaloids. In addition, tetrandrine failed to bind to metabolically dead cells or sonicated macrophage preparations. These data suggest that the interaction of BBIQ alkaloids with phagocytes is not simply nonspecific binding to membrane lipids. Alteration of the cytoskeletal system with vinblastine, taxol, or cytochalasin B decreased tetrandrine binding by approximately 33% when added separately and by 93% when added jointly. Pre-exposure of alveolar macrophages to stimulants increased the ability of BBIQ alkaloids to inhibit both oxygen consumption and superoxide release. These data suggest that the mechanism by which BBIQ alkaloids inhibit activation of phagocytes involves microtubules and bules and microfilaments. Pre-exposure of macrophages to stimulants would change the conformation of cytoskeletal components and may make these structures more susceptible to drug interaction.


Molecular detection of genetic defects in congenital adrenal hyperplasia due to 21-hydroxylase deficiency: a study of 27 families. Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase (21-OHase) deficiency is inherited as an autosomal recessive trait. Patients can present with the salt wasting, simple virilizing or a non-classical form of the disease. The gene for P450C21, the enzyme carrying 21-OHase activity, has been mapped to the major histocompatibility complex on chromosome 6p. Using molecular hybridisation techniques we have studied the genetic defect in 27 families with one or more affected offspring diagnosed and treated at the University Hospital of Essen. DNA samples were digested with restriction endonuclease TaqI, PvuII, BglII, and EcoRI and analysed by Southern blot hybridisation with the cDNA probe pC21/3c. Eleven of 40 haplotypes associated with the salt wasting form were found to have a large deletion of 30 kb affecting the 5' end of the active 21-OHase gene and the 3' end of the closely linked pseudogene. Results in another 11 cases are compatible with gene conversion; 18 cases were not informative. The 30 kb deletion was associated with a combination of the HLA antigens Bw47 and DR7 in 7 of 11 cases. In the haplotypes with gene conversion, no linkage disequilibrium to HLA antigens was found. No apparent gene alterations were detected in simple virilizing and non-classical haplotypes. The direct detection of the genetic defect in 55% of the salt wasting haplotypes may help to improve predictive testing in families with CAH.


Research in 'prostatitis syndromes': the use of alfuzosin (a new alpha 1-receptor-blocking agent) in patients mainly presenting with micturition complaints of an irritative nature and confirmed urodynamic abnormalities. A double-blind, placebo-controlled study in 20 patients with 'prostatitis syndromes' and confirmed urodynamic abnormalities using the alpha 1-receptor-blocking compound alfuzosin is reported. Flow rate recordings are probably the most reliable and useful objective variables in this type of investigation. There is a significant beneficial effect in the group using an active compound concerning maximal flow (p = 0.01), flow time (p = 0.03) and time to maximal flow (p = 0.01). However, compared with the group using a placebo, only the change in the maximal flow rate appeared to be significantly different. Subjective effects were more pronounced in the alfuzosin group, but the spurious nature of the subjective observations is stressed. Based on objective parameters, alfuzosin seems to be effective compared to placebo in the treatment of these patients with micturition complaints of an irritative nature and urodynamic abnormalities, while only minor side effects were noticed.


Comparative assessment of proliferating cell nuclear antigen immunostaining and of nucleolar organizer region staining in transitional cell carcinomas of the urinary bladder. Correlation with other conventional prognostic pathologic parameters. The expression of two cell proliferation indices, the proliferating cell nuclear antigen (PCNA), using the monoclonal antibody PC-10 in the immunoperoxidase method, and the nucleolar organizer regions (NORs), using the colloid silver nitrate staining technique, was assessed in formalin-fixed paraffin-embedded material of 50 transitional cell urinary bladder carcinomas. A relationship was found between the histologic grade and each of the two indices used. A significant difference was observed, particularly between carcinomas of grade II and III (p < 0.001). A relationship was also demonstrated between each of PC-10 and AgNOR scores and the clinical stage, but it was attributed mostly to the close correlation of the latter with the histologic grade of these tumors. The linear correlation coefficient between PC-10 and AgNOR scores was 0.757 (p < 0.001). Our results suggest that PC-10 and AgNOR scores may be important prognostic indices in urinary bladder cancer.


Uptake of radioligands by rat heart and lung in vivo: CGP 12177 does and CGP 26505 does not reflect binding to beta-adrenoceptors. The biodistribution of (-)-4-(3-t-butylamino-2-hydroxypropoxy)-[5,7-3H-benzimidazol-2-one (CGP12177, a non-selective beta-adrenoceptor antagonist) and 1-[2-(3-carbamoyl-4-hydroxy)-(5-3H-phenoxy)]-2-propanol methanesulfonate, (CGP26505, a beta 1-adrenoceptor antagonist) was studied in rats pretreated with various alpha- and beta-adrenoceptor blocking drugs (5 min before 3H injection, in dosages at which the drugs demonstrated the expected selectivity). Cardiac and pulmonary radioactivity were measured after 10 min, when specific binding was maximal. Uptake of [3H]CGP12177 was linked to binding to beta-adrenoceptors since it was not affected by prazosin or yohimbine, and was equally well inhibited by propranolol, unlabelled CGP12177 and isoprenaline. Moreover, atenolol and CGP20712A inhibited [3H]CGP12177 uptake in heart (predominantly beta 1-adrenoceptors) more potently than ICI 118,551, while in lungs (predominantly beta 2-adrenoceptors) ICI 118,551 was more potent than atenolol or CGP20712A. In contrast, [3H]CGP26505 uptake in the target organs was equally effectively inhibited by propranolol and ICI 118,551, and significantly lowered by alpha-adrenoceptor antagonists. We conclude that [11C]CGP12177, but not [11C]CGP2605 will be suitable for positron emission tomography imaging of beta-adrenoceptors in animals.


Antagonism of vecuronium by one of its metabolites in vitro. The neuromuscular-blocking agent vecuronium bromide undergoes hydrolysis to three pharmacologically active metabolites (3-desacetyl, 17-desacetyl and 3,17-desacetyl vecuronium) which might modify the neuromuscular-blocking action of their parent compound. In order to elucidate the possible role of the interaction between vecuronium and its metabolites in the complications reported after long-term use of vecuronium in intensive care unit (ICU) patients, the relative potency of vecuronium, 3-desacetyl and 3,17-desacetyl vecuronium was determined in the rat hemidiaphragm in vitro and the mode of interaction of the above-mentioned compounds investigated. Dose-response relationships were established for each substance alone and for combinations of vecuronium with its metabolites. The relative potency at the EDmax50 levels (% maximal effect) were in the order of 1:1.2:27 for vecuronium, the 3-desacetyl derivative and the 3,17-desacetyl derivative, respectively. The mode of interaction characterized by isobolographic and algebraic (functional) analysis showed vecuronium and 3-desacetyl vecuronium to interact in an additive fashion while the combined effect of the parent compound and its 3,17-desacetyl derivative was less than additive, indicating antagonism.


Stimulation by corticotropin-releasing factor of the release of immunoreactive dynorphin A from mouse spinal cords in vitro. Corticotropin-releasing factor (CRF) has been shown to release endogenous opioid peptides from several rat brain regions. Since we have demonstrated previously that the actions produced by intrathecally administered CRF in mice involve spinal kappa opioid receptors, experiments were conducted in this study to test the possibility that CRF may release dynorphin A, a putative endogenous kappa opioid agonist, from the mouse spinal cord. Using a superfusion system in vitro, mouse spinal cords were superfused with aerated (95% O2, 5% CO2) Krebs-Ringer buffer. Fractions of superfusion were collected and dynorphin A levels in each fraction were monitored by radioimmunoassay. The presence of CRF in the perfusion buffer stimulated significantly the release of immunoreactive dynorphin A. The releasing rate of immunoreactive dynorphin A returned to the basal level after withdrawing CRF from the superfusion buffer. The stimulatory effect of CRF on the release of immunoreactive dynorphin A was abolished by alpha-helical CRF-(9-41), a CRF receptor antagonist, indicating that the dynorphin-releasing effect of CRF was mediated by CRF receptors in the spinal cord. Also the dynorphin-releasing effect of CRF was a concentration-related phenomenon, with an estimated EC50 value of 5.3 nM. The results from this study support the hypothesis that intrathecally administered CRF may produce its effects by releasing endogenous dynorphin from the terminals of dynorphin-containing neurons in the spinal cord. This study also provides evidence to support the notion that there is a close communication between CRF- and opioid peptide-containing neuronal pathways in the central nervous system.


Attenuation of the inhibitory effect of dynorphin on dopamine release in the rat nucleus accumbens by repeated treatment with methamphetamine. Dynorphin (1-100 nM) dose dependently inhibited both spontaneous and electrically evoked endogenous dopamine (DA) release from slices of the nucleus accumbens of untreated rats. When this inhibitory effect was compared, it was significantly reduced in rats pretreated (for 9 days) with methamphetamine (6 mg/kg per day i.p.) relative to rats treated with saline. These findings indicate that dynorphin inhibits DA release from the nucleus accumbens, and that treatment with methamphetamine reduces the modulatory action of dynorphin on DA release. It is possible that repeated administration of methamphetamine leads to attenuation of the inhibition of DA release from the nucleus accumbens via presynaptic dynorphin-sensitive receptors.


Mixed agonist-antagonist properties of umespirone at neostriatal dopamine receptors in relation to its behavioral effects in the rat. In normal rats treated with the inhibitor of cerebral decarboxylase, NSD-1015 (100 mg kg-1 i.p.), umespirone (1.9-30.0 mumol kg-1 s.c.) produced an increase in neostriatal DOPA (dihydroxyphenylalanine) accumulation, whereas decreased DOPA accumulation was obtained in reserpine-pretreated (5 mg kg-1 s.c., -18 h) animals. The latter effect was statistically significant only in the ventral, limbic, portion of the neostriatum. Neostriatal 5-hydroxytryptophan (5-HTP) accumulation was decreased in the reserpine-treated animals but not in normal controls. DOPA accumulation in the neocortex was not affected by umespirone treatment in either preparation, whereas 5-HTP accumulation was decreased in the reserpine-treated animals. Spontaneous locomotor activity was suppressed by umespirone at doses that did not affect treadmill locomotion (7.9-31.2 mumol kg-1 s.c., -30 min), and there were no signs of catalepsy at doses ranging from 31.2-249.6 mumol kg-1 s.c. up to 2 h after injection. Thus, umespirone behaves as a mixed dopamine receptor agonist/antagonist and also displays 5-HT receptor agonist properties. This biochemical profile was associated with sedation, as observed in the open-field, at doses which did not affect treadmill locomotion or induced catalepsy.


Multidimensional behavioral analyses show dynorphin A-(1-13) modulation of methamphetamine-induced behaviors in mice. The effects of intracerebroventricular (i.c.v.) injection of dynorphin A-(1-13) on methamphetamine-induced behavioral alterations in mice were determined by using multidimensional behavioral analyses. Methamphetamine (0.3, 1.0 and 3.0 mg/kg s.c.) produced a marked increase in linear locomotion, circling, rearing and/or grooming behaviors. The behavioral effects of methamphetamine (1.0 mg/kg s.c.) were almost completely antagonized by pretreatment with the dopamine D2 receptor antagonist, S(-)-sulpiride (3.0 and/or 10.0 mg/kg i.p.), but not with the dopamine D1 receptor antagonist, SCH 23390 (0.01 or 0.03 mg/kg i.p.). Although dynorphin A-(1-13) (3.0 or 12.5 micrograms i.c.v.) alone did not produce any significant effects on behavior, the methamphetamine (1.0 mg/kg s.c.)-induced increase in circling ipsilateral to the injection side was markedly enhanced by dynorphin A-(1-13) (12.5 micrograms i.c.v.). In contrast, the peptide (12.5 micrograms i.c.v.) inhibited the methamphetamine (1.0 mg/kg s.c.)-induced increase in rearing, whilst the increase in grooming remained unchanged. The effects of dynorphin A-(1-13) (12.5 micrograms i.c.v.) were fully reversed by the opioid antagonist, Mr 2266 (5.6 mg/kg s.c.). These results suggest that the unilateral administration (i.c.v.) of dynorphin A-(1-13) inhibits the activity of dopamine-elicited neurotransmission, resulting in an increase in ipsilateral circling and in a decrease in rearing.


Transmitter-like 3,4-dihydroxyphenylalanine is tonically released by nicotine in striata of conscious rats. Microdialysis and high performance liquid chromatography with an electrochemical detector were applied to compare the characteristics of nicotine-evoked release of endogenous 3,4-dihydroxyphenylalanine (DOPA) from striata of conscious rats and those of the release of dopamine (DA). Dialysates were collected every 20 min 3-8 h after the start of perfusion. Nicotine was perfused for 20 min through a probe. (+/-)-Nicotine (100-300 microM) constantly and repeatedly released DOPA and DA over a similar time course in a dose-dependent manner. The ratio of the DOPA and DA release evoked was approximately 1:3. The (+/-)-nicotine (200 microM)-induced DOPA release was mecamylamine (500 microM)-sensitive, tetrodotoxin (100 nM)-sensitive and Ca2+ (removal plus 12.5 mM Mg2+ addition)-dependent. The (+)-isomer produced no DOPA release. These characteristics of DOPA release were almost the same as those of DA release. Furthermore, mecamylamine alone inhibited the basal release of DOPA but not of DA. Nicotine released stereoselectively endogenous DOPA via nicotinic acetylcholine receptors from striata of freely moving rats in a manner similar to transmitter DA. These acetylcholine receptors function tonically for the release of DOPA. These findings are further support for our proposal that DOPA is an endogenous neuroactive substance.


Effects of ibutilide fumarate, a novel antiarrhythmic agent, and its enantiomers on isolated rabbit myocardium. Ibutilide fumarate is currently in Phase II clinical trials for the treatment of life-threatening cardiac arrhythmias. The cardiovascular effects of ibutilide and its d- and l-stereoisomers, U82208E and U82209E were tested in an isolated rabbit myocardium system. In a series of repeated measures experiments, threshold, effective refractory period, force of contraction, conduction time and rate were measured at various pacing frequencies in isolated papillary muscles, ventricular muscle strips and right atria exposed to 10(-7), 10(-6) and 10(-5) M drug. Although there were occasional instances where one form had a greater or lesser effect on a given parameter, overall there was little pharmacological difference between the racemic mixture and its constituent forms. At the highest dose, effective refractory periods at 1 and 3 Hz increased by 18-32 ms, conduction times measured at 3 Hz increased by 27-30% and atrial rate decreased by 19-32%, while threshold and force of contraction were generally unaffected. In this study there were no clear cut pharmacologic differences between the three forms of this class III antiarrhythmic agent. Parallel studies to determine pA2 values of ibutilide and sotalol demonstrated that ibutilide possesses weak beta-adrenoceptor blocking properties.


Studies on the location of catecholamine receptors in canine sympathetic ganglia. Receptors mediating catecholamine-induced inhibition were studied in cardiac ganglia of pentobarbital-anesthetized dogs. Using selective agonists and antagonists the presence of three receptor subtypes was verified: alpha 1- and alpha 2-adrenoceptors and dopamine D2 receptors. Activation of alpha 1-adrenoceptors or dopamine D2 receptors reduced the response to preganglionic nerve stimulation but not to direct stimulation of the nicotinic acetylcholine receptors of the principal ganglion cells: response to both types of stimulation were reduced by activation of ganglionic alpha 2-adrenoceptors. These results suggested that two inhibitory systems were present in canine sympathetic ganglia and mediated the effects of exogenous catecholamines. One system involved alpha 1-adrenoceptors and dopamine D2 receptors located proximal to the synapse of the pre- and postganglionic neurons and the other involved alpha 2-adrenoceptors located distal to the intraganglionic synapse.


The adrenoceptor agonist, SDZ NVI 085, discriminates between alpha 1A- and alpha 1B-adrenoceptor subtypes in vas deferens, kidney and aorta of the rat. The potency of the alpha 1-adrenoceptor agonist (-)-(4aR, 10aR)-3,4,4a,5,10,10a-hexahydro-6-methoxy-4-methyl-9-methylthio-2H -naphth [2,3-b]-1,4-oxazine (SDZ NVI 085) was investigated both in isolated vas deferens and perfused kidney of the rat, two tissues with alpha 1A-adrenoceptor subtype characteristics, and in the rat thoracic aorta, in which the contribution of different alpha 1-adrenoceptor subtypes mediating contraction is controversial. In vas deferens and kidney, SDZ NVI 085 evoked smooth muscle contraction and vascular constriction and was of similar potency to L-phenylephrine. Contractions of vas deferens in response to (-)-noradrenaline and SDZ NVI 085 were resistant to chloroethylclonidine treatment (3 x 10(-5) M), sensitive to (+/-)-isradipine (10(-8) M) and competitively antagonized by 5-methyl-urapidil (pA2 = 9.04 and 8.82, respectively). The potencies of a number of alpha 1A-/alpha 1B-adrenoceptor-discriminating antagonists to reverse renal vasoconstriction due to either (-)-noradrenaline or SDZ NVI 085, and their affinities in vas deferens correlated significantly with their pKi values at alpha 1A binding sites in rat cortex. In rat aorta, SDZ NVI 085 up to 5 x 10(-4) M failed to evoke contraction. The affinities of subtype-selective antagonists determined in aorta correlated significantly with the pKi values at alpha 1B binding sites but differed from pKi values at alpha 1A sites in rat cortex. Thus, the contractile alpha 1-adrenoceptor in rat aorta can be best characterized as B subtype. SDZ NVI 085 might be a selective alpha 1A-adrenoceptor agonist and thus be used as a new tool either to detect (rat vas deferens and kidney) or exclude (rat aorta) a contribution of alpha 1A-adrenoceptors functionally involved in smooth muscle contraction.


The NMDA receptor antagonist MK-801 prevents imipramine-induced supersensitivity to quinpirole. Chronic treatment with imipramine enhanced the locomotor stimulant response to quinpirole (0.3 mg/kg s.c.), a dopamine D2/D3 receptor agonist. Chronic, but not acute, blockade of the NMDA receptor with the non-competitive antagonist MK-801 (0.3 mg/kg i.p.) prevented the imipramine-induced potentiation of the quinpirole effect. The results suggest that NMDA receptors play a role in the development of supersensitivity to dopamine receptor agonists produced by chronic antidepressant treatment.


Effect of human thyroid stimulating autoantibodies on the radioiodine uptake of the mouse thyroid gland. Administration of sera from patients with active Graves' disease to nude mice bearing human thyroid xenografts provides the opportunity to compare directly the effects of TSAb on thyroid tissue of different species. The results presented in this paper, namely the fact that only one of 8 Graves' sera increased the RAIU of the nu/nu mouse thyroids modestly while all sera increased the RAIU in the transplanted human tissue considerable, clearly show a species specific effect of TSAb.


Recombinant TSH-receptor for determination of TSH-receptor-antibodies. We have expressed the complete coding sequence of the human TSH-R in eucaryotic cells. Limiting dilution enabled us to select two clones, JP09 and JP26, which have formed the basis of binding and bioassays respectively, for TSH-R antibodies. Results obtained in the binding assay correlated well with those obtained in the TRAK (Henning Berlin) assay, while the bioassay was at least as sensitive as measurements made with FRTL5 or human thyroid cells. These cell lines provide a reliable source of human TSH-R which will be useful in routine diagnosis.


Modulation of leukemic cell sensitivity to lymphokine-activated killer cytolysis: role of intercellular adhesion molecule-1. The role of CD11/CD18 leukocyte adhesion molecules and their ligands in mediating non-major histocompatibility complex (MHC) restricted lymphocyte cytotoxicity is controversial. In order to examine the role of target cell intercellular adhesion molecule-1 (ICAM-1; CD54), a ligand of lymphocyte function-associated antigen (LFA-1) (CD11a/CD18), we exposed the human leukemia cell line, HL-60, to a variety of agents implicated in modulating ICAM-1 expression and/or sensitivity to lymphocyte cytolysis. Exposure of HL-60 cells to retinoic acid (RA), interferon (IFN)-alpha, IFN-beta, and IFN-gamma induced protection from lymphokine-activated killer (LAK) cytolysis. Only RA and IFN-gamma induced ICAM-1 expression. Tumor necrosis factor and vitamin D3, which also induced ICAM-1 expression, increased HL-60 sensitivity to LAK lysis. Granulocyte-macrophage colony-stimulating factor also increased sensitivity to LAK lysis; ICAM-1 was not induced. The state of cellular differentiation and expression of class I and II MHC antigens also did not correlate with sensitivity to LAK cytolysis. Exposure of untreated HL-60 cells and HL-60 cells expressing ICAM-1 to monoclonal antibody (mAb) versus ICAM-1 did not modulate LAK sensitivity. Exposure of LAK cells to mAb versus LFA-1 partially inhibited cytolysis; mAb versus CD18 inhibited cytolysis more completely. HL-60 cells were resistant to natural killer lysis; exposure to the various experimental agents did not alter sensitivity. We conclude that leukemic cell sensitivity to LAK cytolysis can be modulated by a variety of agents. Although our results suggest a role for leukocyte CD11/CD18 adhesion molecules in LAK cytolysis, the poor correlation between ICAM-1 expression and sensitivity to LAK lysis suggest that interactions other than LFA-1/ICAM-1 conjugation may be more central to the processes involved.


Adhesion receptors are differentially expressed on developing thymocytes and epithelium in human thymus. The thymic microenvironment consists of a network of interrelated cells of epithelial, fibroblastic, endothelial, and hemopoietic origin. Within this environment, the development of specific T-lymphocyte subpopulations partially depends on the selective interaction of T-cell precursors with such cells. Human thymic epithelial cell strains, generated with a defective retroviral vector containing simian virus 40 (SV40) large T antigen and the neomycin resistance gene or by transfection with an SV40 plasmid defective in the origin of replication, provide useful tools for understanding the mechanisms contributing to the control of T-cell maturation. Because interepithelial, epithelial-macrophage, and lymphocyte-epithelial cell interactions are important for thymocyte differentiation, the distribution of integrin and nonintegrin adhesion receptors on these cells and on developing thymocytes in vivo and in vitro has been examined in detail. Our results indicate that the transformed human thymic epithelial cell strains express the common very late antigen (VLA)-beta 1 receptor and unique alpha chains VLA-2, VLA-3, and VLA-6. The cells are also positive for LFA-3 and ICAM-1 and weakly express beta 3, beta 4, and VNR alpha. They do not express the Leu-cellular adhesion molecules (CAM). This phenotypic profile on cultured thymic epithelium generally corresponds to the distribution of integrin and other receptor molecules on thymic epithelial cells in tissue sections. The majority of thymocytes also express the integrin VLA-beta 1 and -beta 2 chains as well as VLA-4, VLA-6, and LFA-1 alpha(L). Three-color flow cytometric analyses show differential levels of expression of these adhesion receptors on human thymocyte subsets. Taken together with the immunohistochemical localization of extracellular matrix molecules, these studies suggest that both the distribution of receptor-ligand pairs and the level of expression of adhesion molecules may influence T-cell development within the thymus.


Functional characterization of mouse granulocytes and macrophages produced in vitro from bone marrow progenitors stimulated with interleukin 3 (IL-3) or granulocyte-macrophage colony-stimulating factor (GM-CSF). Bone marrow from C3H/ouj mice was depleted to < 1% of CD11b+ granulocytes and macrophages using paramagnetic beads coated with sheep anti-rat antibodies. CD11b- cells, enriched three- to fourfold in colony-forming cells, were stimulated in liquid culture with interleukin 3 (IL-3) or granulocyte-macrophage colony-stimulating factor (GM-CSF). Cultures stimulated with IL-3 or GM-CSF increased cell numbers fourfold at 7 days, with the CD11b+ population increasing to 63% +/- 9% (n = 5) with IL-3 or 96% +/- 1% (n = 4) cells with GM-CSF. Functional responsiveness of the granulocytes and macrophages was assessed by flow cytometry in an oxidative burst assay using dichlorofluorescein (DCF) and a quantitative phagocytosis assay using opsonized fluorescent beads. Granulocytes and macrophages, identified by light scatter characteristics and allophycocyanine staining of CD11b, were assayed simultaneously with granulocytes from fresh mouse bone marrow and peripheral blood. GM-CSF-generated CD11b+ cells had higher oxidative responses than similar populations produced in response to IL-3. The oxidative burst of these in vitro generated CD11b+ populations was similar to the equivalent fresh bone marrow population. Oxidative burst responses of peripheral blood phagocytic cells could not be adequately measured in this system. Peripheral blood CD11b+ cells were the most phagocytic, followed by GM-CSF-stimulated CD11b+ cells; IL-3-stimulated and bone marrow CD11b+ cells were the least phagocytic. These data demonstrate that functional granulocytes can be produced in vitro using growth factors and that GM-CSF produces a more responsive cell than IL-3.


No genetic differences between affected and unaffected members of a German family with Leber's hereditary optic neuropathy (LHON) with respect to ten mtDNA point mutations associated with LHON. In order to investigate possible synergistic influences of different mtDNA mutations on penetrance and severity of Leber's hereditary optic neuropathy (LHON), a large German LHON pedigree is characterized with respect to 10 different mutations associated with LHON. All members of the family carry three different mtDNA mutations (at nucleotide 4,216, 11,778 and 13,708) in a homoplasmic form, regardless of whether or not they are clinically affected. Testing for another 7 mutations reveals negative results in all family members. Hence, the variable disease expression of the family members cannot be explained by varying combinations of LHON-associated mtDNA mutations.


POU-specific domain of Oct-2 factor confers 'octamer' motif DNA binding specificity on heterologous Antennapedia homeodomain. The bipartite DNA binding domain of the POU family of transcription factors contains a 'POU-specific' domain unique to this class of factors and a 'POU homeodomain' homologous to other homeodomains. We compared DNA binding of the Oct-2 factor POU domain and the Antennapedia (Antp) homeodomain with a chimeric Oct-2/Antp protein in which the distantly related Antp homeodomain was substituted for the Oct-2 POU homeodomain. The Oct-2/Antp chimeric protein bound both the octamer and the Antp sites efficiently, indicating that DNA binding specificity is contributed by both components of the POU domain.


Neuropeptide Y: a novel neuroendocrine peptide in the control of pituitary hormone secretion, and its relation to luteinizing hormone. Evidence that establishes neuropeptide Y (NPY) as an important neuromessenger in the regulation of anterior pituitary hormone secretion is reviewed. In particular, NPY plays a critical role in stimulating the episodic, basal pattern of luteinizing hormone (LH) release, as well as the preovulatory surge of LH release in several species. The stimulatory effect of NPY on LH secretion is dependent upon the presence of gonadal hormones and involves amplification of the response of other interacting stimulatory signals. NPY acts at the level of the median eminence to excite the release of LH-releasing hormone (LHRH) via a mechanism that leads to the mobilization of intracellular Ca2+. These stimulatory LHRH responses are mediated by Y1NPY receptors. Moreover, NPY activates postsynaptic messenger pathways that complement and reinforce those affected by norepinephrine, which is another major neuroregulator of LHRH secretion and which is released as a cotransmitter with NPY in the median eminence. Additionally, NPY is released into the hypophyseal portal blood for transportation to the anterior pituitary where it enhances the release of LH in response to LHRH. This facilitatory, modulating effect at the pituitary level involves an allosteric increase in LHRH binding to its receptor leading to augmented influx of Ca2+ from the extracellular space. There is evidence that gonadal steroids regulate NPY neurosecretion in a site-specific manner, and that alterations in NPY secretion may occur in part via a direct action of the steroids on NPY neurons in the brainstem and hypothalamus and in part through an indirect effect involving removal of the inhibitory influence of endogenous opioid peptides. These findings are integrated into an overall hypothesis for induction of the preovulatory LH surge on proestrus requiring an interplay between NPY and other neuronal networks. In aged male rats, due to the inability of hypothalamic NPY neurons to respond appropriately to the trophic effects of androgens, NPY neurosecretion is diminished. Further, a review of the literature reveals that NPY may modulate the secretion of other pituitary hormones through a similar combination of hypothalamic and pituitary actions.


Unexplained infertility: a review of diagnosis, prognosis, treatment efficacy and management. The dilemma of unexplained infertility posed by Southam in 1960 remains today: despite advances in the diagnostic assessment of infertility, many couples still have no explanation for their infertility. Even the most sophisticated evaluation of semen, ovulation and genital tract competence cannot reveal all of the possible defects in the complex process leading to conception. Because it arises from these shortcomings in our knowledge of fertilization and from our inability to utilize all of the current knowledge, unexplained infertility is a challenge for both biological and clinical research. This paper attempts to summarize some clinical issues in the management of unexplained infertility: the prevalence of the disorder, problems in the definition and possible explanations for the existence of this diagnostic category. It reviews outcome-based clinical publications as a guide to decision-making on what diagnostic tests to use, provides a summary of the untreated prognosis and evaluates study results that may serve as a basis for treatment decisions in this puzzling diagnostic category of infertility.


Maternal weight, weight gain during pregnancy and pregnancy outcome. To evaluate the effects of abnormal maternal weight or weight gain on pregnancy outcome. Records for 191 mothers with abnormal prepregnancy weight (> or = 20%) above, or under, ideal body weight for height) or weight gain > or = 20 kg, or < or = 5 kg during pregnancy were reviewed. The control group consisted of 166 mothers with normal prepregnancy weight and normal weight gain during pregnancy. Data on mothers and their infants were analyzed by one-way analysis of variance. Obese women and mothers with excessive weight gain during pregnancy had an increased incidence of induced labor (P < or = 0.05) and tendency for emergency cesarean sections during the delivery. Obese women had more large-for-date babies than controls (P < or = 0.05). Weight gain < or = 5 kg during pregnancy was most common in slightly obese women and did not carry any special obstetric or neonatal risk. Underweight women had a significant risk for delivering a small-for-data baby. Obese women and women with excessive weight gain during pregnancy need special follow-up and counseling during pregnancy and delivery. Underweight women may need prepregnancy nutritional counseling to guarantee normal fetal growth. In developed countries suboptimal weight gain (< or = 5 kg) during pregnancy seems not to need any medical intervention.


Potential relationship between dengue fever and neural tube defects in a northern district of India. A sudden increase in number of births of newborns with neural tube defects (NTD) was observed from June, 1989 to September, 1989 in Medical College and Hospital, Rohtak and various other government and private hospitals of the district of Rohtak. Out of a total 4785 deliveries whose records were collected, there were 87 newborns with NTD with an incidence of 18.18/1000 births which was three times higher than the previous incidence of 6.8/1000 births in the preceding 4 years. There was an epidemic of dengue fever in this area from September, 1988 to December, 1988 affecting almost one member from each family. This coincided with the period of their first trimester. Of these, 18 patients suffered clinically from dengue fever, 21 patients had positive dengue fever history in their family members, 21 patients had positive history in their neighbors. The cluster of NTD appears to be due to dengue virus infection.


A trial of labor after cesarean section in patients with or without a prior vaginal delivery. To determine the outcome of labor in women with a previous cesarean section, with or without prior vaginal delivery. Records were reviewed for 1065 women with a previous cesarean section at 'Virgen Macarena' Hospital who were attended for a subsequent labor. Chi-squared tests demonstrated that women with previous vaginal delivery (n = 346) had a significantly higher rate of vaginal delivery after a trial of labor (95.24%) than those without previous vaginal delivery (n = 719) (82.95%). All the ruptures of uterine scar (n = 4) were found in women without previous vaginal delivery. It appears that a cesarean section in a multiparous woman is not a determinant fact in her reproductive history and the risk of rupture of uterine scar did not appear to be present.


Use of low molecular weight heparin for prophylaxis and treatment of thromboembolism in pregnancy. Low molecular weight heparin (LMWH) preserves the antithrombotic action but not the anticoagulant activity of heparin. LMWH is safe, does not cross the placenta and is administered as a single daily injection. We report our experience with 6 pregnant women given LMWH for treatment or prophylaxis of thromboembolism. The drug was successfully given to 5 women for periods of 6 weeks--6 months and no thromboembolic complications occurred during pregnancy or pueperium. There were no hemorrhagic complications and no excessive bleeding was observed during delivery. The sixth patient relapsed after 6 weeks of therapy. This patient also showed resistance to standard heparin administered intravenously at a very high dose. LMWH should be considered an alternative to standard heparin in pregnant women requiring antithrombotic prophylaxis and therapy.


Ovarian function after conservational ovarian surgery: a long-term follow-up study. To determine the long-term effects of conservational ovarian surgery on subsequent ovarian function. Medical, surgical and menstrual records of 87 reproductive age women who have undergone pelvic surgery for indications including endometriosis, infertility, and pelvic pain between June 1982 and June 1989 were retrospectively reviewed. Sixty-seven patients had ovarian surgery during their procedure; 26 ovarian cystectomies (OC), 19 partial ovarian resections (OR) and 22 ovariolyses (OL). Twenty patients had no ovarian surgery (NOS). The mean follow-up period was comparable in all groups (OL 32 +/- 15 months, OR 34 +/- 26 months, OC 41 +/- 30 months and NOS 37 +/- 21 months). The mean time to onset of the first postoperative menses was not significantly different among the four groups (OL 24 +/- 7 days, OR 24 +/- 13 days, OC 29 +/- 26 days and NOS 22 +/- 6 days). The cumulative conception rates were not significantly different (OL 23%, OR 37%, OC 19% and NOS 25%). Menstrual disturbances, defined as perceived deviations from preoperative patterns, did not appear to be related to the type or extent of ovarian surgery and occurred in comparable frequencies among patients with or without ovarian surgery (OL 23%, OR 37%, OC 23% and NOS 20%). In the majority of cases, menstrual disturbances occurred either in the early postoperative months and were self-limited, or much later in which case they were related to recurrent endometriosis. Premature ovarian failure has so far occurred in one patient suggesting an incidence comparable to that in the general population. It is concluded that in this group of patients, conservational ovarian surgery had no significant effects on ovulatory and menstrual function over a prolonged follow-up period.


Scanning electron microscopy of endometriotic lesions in the pelvic peritoneum and the histogenesis of endometriosis. To determine whether the epithelium of an endometriotic lesion has eutopic endometrial glandular epithelium morphology or not for the histogenesis of endometriosis. Scanning electron microscopy (SEM) of the endometrium was done for 25 cases. Histologically proven endometriotic lesions of 10 of these 25 cases were processed for SEM. Peroneum samples of another 15 cases without histological endometriosis were also studied. Three of 10 cases with histologically proven endometriotic lesions revealed endometrium-like morphology by SEM, whereas normal appearing peritoneum without histological endometriosis in 15 cases showed no endometrium-like structures by SEM. Although abovementioned results did not support retrograde menstruation and implantation, they suggested a possibility of derivation from the endometrium.


The feasibility of suppressing ovarian activity following the end of amenorrhoea by increasing the frequency of suckling. To determine the feasibility of suppressing ovarian activity by increasing the frequency of suckling episodes. Prospective study was carried out with 19 exclusively breastfeeding volunteers. Ten subjects (experimental group) increased the suckling episodes by minimum 50% per day from the beginning of the first postpartum menses. Nine controls continued breastfeeding as before. Estradiol, progesterone, LH, FSH and prolactin were measured in blood samples, drawn twice a week up to the second postpartum menses or for 60 days, by RIA. Student's t-test was employed. The higher suckling frequency prevented ovulation in 7 of 10 cases examined according to the plasma progesterone concentration (< 9.5 nmol/l). Significantly higher average prolactin value could also be found in the experimental group (1038 (527) munits/l vs. 518 (245) munits/l; P < 0.05). Results suggest that an earlier initiation of the increase in breastfeeding frequency may delay the resumption of ovulation. Frequent, full time lactation may reduce the risk of pregnancy.


Prenatal diagnosis of recurrent Meckel syndrome. We report a rare case of Meckel-Gruber syndrome in a woman who had three affected offsprings in the past with similar condition. Ante-natal ultrasonographic diagnosis and management are presented.


Fetal macrosomia. ACOG Technical Bulletin Number 159--September 1991. When macrosomia exists, shoulder dystocia is a primary obstetric concern. Current methods for estimating birth weight prior to delivery are imprecise, and macrosomia often cannot be predicted. Certain conditions or combinations of conditions should increase the index of suspicion for shoulder dystocia. When these conditions are identified, consideration should be given to the management of potential shoulder dystocia. Despite due care and appropriate clinical judgment, however, injury during the birth process cannot always be prevented.


Ectopic expression of the Drosophila homeotic gene proboscipedia under Antennapedia P1 control causes dominant thoracic defects. A deletion mutation in the Antennapedia Complex of Drosophila melanogaster, Df(3R)SCBXL2, induces both dominant and recessive loss-of-function phenotypes. The deletion is associated with diminished function of proboscipedia (pb), a homeotic gene required for mouthparts formation. Df(3R)SCBXL2 also has associated dominant thoracic defects related to diminished expression of the homeotic Antennapedia (Antp) gene copy on the homologous chromosome. This is shown to be a consequence of ectopic pb expression in the thorax. Newly juxtaposed Antp sequences provide the pb gene on the deletion bearing chromosome with a second promoter, Antp P1, in addition to its own. Ectopic pb protein expression occurs under Antp P1 control, by alternate splicing, and results in diminished accumulation of Antp protein in the imaginal disc cells where Antp P1 is normally expressed. The analysis of this mutant chromosome thus demonstrates that pb protein is capable of participating in the negative regulation of a more posteriorly expressed homeotic gene, as well as serving a homeotic "selector" function in the head.


Polymorphism and divergence in the Mst26A male accessory gland gene region in Drosophila. Drosophila males, like males of most other insects, transfer a group of specific proteins to the females during mating. These proteins are produced primarily in the accessory gland and are likely to influence the female's reproduction. The results of studies of DNA sequence polymorphism and divergence in two genes coding for male accessory gland proteins of Drosophila are reported here. The Mst26Aa and Mst26Ab transcription units are tandemly arranged in a approximately 1.6-kb segment in Drosophila sechellia, Drosophila mauritiana and Drosophila simulans as they were reported to be in Drosophila melanogaster. The DNA sequences of 10 alleles from D. melanogaster and one allele each from the three sibling species reveals a high degree of amino acid replacement variation. A substantial part of the variation is due to insertion/deletion differences. Possible functional significance of these amino acid sequence changes is discussed. Statistical analyses based on the neutral theory of molecular evolution show that the distribution of polymorphism over the 1.6-kb region is inconsistent with the pattern of divergence between the species. The amount of 4-cutter restriction map polymorphism in a larger sample of 75 alleles from the same D. melanogaster population is similar to that obtained from the DNA sequence of the 10 alleles (a pairwise average of 0.007 difference per site). The 6-cutter restriction map survey of a 18-kb region containing the Mst26A genes indicates that polymorphism in the region flanking these genes maybe higher. The failure of polymorphisms and divergence in the Mst26A region to conform to the expectations of a simple mutation-drift-equilibrium model indicates that selection in or near this region has played a role in the history of these genes.