Immunoreactivity of an antibody to a beta/A4 sequence with Alzheimer paired helical filaments and tau protein.

beta/A4, a peptide that forms the extracellular amyloid fibrils of Alzheimer senile plaques, has also been proposed to be a component of Alzheimer paired helical filaments (PHFs). We compared BR88, an antiserum to amino acids 1-12 of beta/A4, with BR126, an antiserum to the sequence SEKLDFKDRVQS in tau protein, since tau protein is the only confirmed component of PHFs. In enzyme-linked immunosorbent assays (ELISAs), both antibodies reacted with pronase-treated PHFs better after PHFs were treated with guanidine. tau protein shares no sequence homology with beta/A4. Nevertheless, BR88 cross-reacted with human recombinant tau isoforms by ELISA and Western blot analysis with potencies comparable to those for anti-tau antibodies. BR88 reacted with a beta/A4 peptide as well on a molar basis as with tau protein and showed some reactivity to the tubulin-binding region of tau protein. In conclusion, the beta/A4 antiserum BR88 cross-reacts with tau protein, possibly explaining its reactivity with PHFs.


Beta-blockers, calcium channel blockers and the sulfhydryl-ACE inhibitors demonstrate protection against free-radical-mediated injury of cardiovascular cells and membranes.

During reperfusion of previously ischemic cardiac tissue, oxygen-centered free radicals are generated and may result in peroxidative injury of cardiovascular cells and membranes. Since the occurrence of reperfusion injury in patients is unpredictable, particularly in those patients with chronic ischemic coronary artery disease, silent ischemia and those predisposed to significant coronary spasm, it would be advantageous to provide continuing therapy with antioxidant agents.


Establishment of cytotoxic CD4+ T cell clones from cancer patients treated by local immunotherapy.

We have previously reported that the antitumor effect of OK-432, a Streptococcal preparation, is markedly augmented when injected intratumorally together with fibrinogen (Cancer, 69: 636-642, 1992). In order to elucidate the mechanism of the antitumor effects, we established T cell clones from regional lymph nodes of colorectal cancer patients who received this local immunotherapy. By culture of lymph node lymphocytes, in the presence of IL-2 and OK-432, 4 clones of T cells were established from 4 patients treated by local immunotherapy. These clones had a helper T cell phenotype (CD3+, CD4+, CD8-, CD56-, WT31+) and were successfully maintained for several months. The cells strongly expressed CD25 when stimulated with OK-432 and exhibited a high level of cytotoxic activity in part explained by the increased expression of ICAM-1 and LFA-1, and the release of TNF beta. These results suggest that the CD4+ T cells play a role in the antitumor mechanism of local immunotherapy.


[The effect of adrenoblockers on the neurons of the lateral hypothalamus under the action of pentagastrin].

To test the hypothesis of interaction pentagastrin (PG) noradrenaline (NA) in central neurochemical mechanisms of food motivation, we studied the activity of single neurons in lateral hypothalamus (LH) after s.c. PG injection. Following PG injection microiontophoresis (MIF) of propranolol prazosin was made. PG-induced changes were similar to neuronal activity in rabbits LH after 24-hour food deprivation in 59%. Propranolol-induced changes were following firing pattern in the process of food uptake in 68%. Prazosin MIF induced firing pattern of neuronal activity of saturated rabbits in 60%.


[Stress-inducing behavioral changes and the functions of the neurohormonal systems in monkeys of different social ranks].

As a result of intensive stressful stimuli hamadryas baboons developed, depending on their hierarchic status, depression-like states varying in severity. Their development correlated with a drastic release of biogenic amines, activation of pituitary-adrenal system and inhibition of hormonal function of the gonads. It is shown that dominant and low-rank monkeys, in spite of the differences in their initial psychoemotional states and in the blood levels of mediators and hormones, demonstrated marked disintegration in their individual and social behavior. There was a larger increase in the blood concentrations of dopamine, serotonin and cortisol and more significant inhibition of testosterone production under the influence of two-hour immobilization as compared to the subdominant animals.


[An analysis of the ionic regulation of the specific binding of 3H-diazepam depending on the phenotype of the emotional stress reaction].

It was shown that low NaCl concentrations (less than 50 mM) had more pronounced stimulatory effect on [3H]-diazepam ([3H]-DZ) binding in brain membranes of Balb/c (C) mice than in C57B1/6 (B6) mice. These interstrain differences disappeared after emotional stress in "open field" (OF) test. Low doses of diazepam (0.75 mg/kg) and hydazepam (1 mg/kg) induced anxiolytic effect in C mice and restored their normal [3H]-DZ binding level in the presence of NaCl. On the opposite, effects of the same doses of the benzodiazepines were not revealed either on the behavior in OF test or on stimulating properties of NaCl in B6 mice. Both benzodiazepines (10 mg/kg) induced similar behavior (sedative) and receptor (decrease of NaCl stimulating ability) in B6 and C mice. We made a conclusion that the ability of NaCl to increase [3H]-DZ binding is a physiological index which reflects hereditary differences in emotional-stress reactions and behavioral effects of benzodiazepine tranquillizers.


[The potential-dependent action of the opioid peptide dermorphin].

Dermorphin action was studied on cross-section strip of frog stomach muscle by a mechanographic recorder. The results show that dermorphin (10(-5)-10(-8) M) blocks acetylcholine effects, spontaneous activity and muscle contractions induced by direct electrical stimulation. All the above effects are hardly reversible. Dermorphin fails to block spontaneous and evoked activities if it is injected into the incubated medium during K(+)-depolarization (KCI--100 mM) of the muscle. Thus, dermorphin has voltage-dependent action. The discussion deals with dermorphin action on voltage-dependent Ca(2+)-channels of muscle cell membrane.


Takayasu's aortitis with renovascular hypertension.

We report a case of Takayasu's disease with severe renovascular hypertension in a girl from Eritrea. In the "burn-out" phase after the erythrocyte sedimentation rate had normalized, reconstructive vascular surgery was performed as further progression of the disease seemed unlikely. However, probably due to her growth, the graft rotated and a second operation was successfully performed.


Management of dental injuries.

Dental-related injuries, which range from chipped teeth to fractured jaws, are among the most common injuries seen in schools. This article describes the various injuries and provides guidelines, for the nurse's actions at the scene, to ensure the greatest potential for successful treatment. Guidelines are also given for informing other school and associated health personnel about dental emergencies.


PC-10 antibody to proliferating cell nuclear antigen (PCNA) is not related to prognosis in human breast carcinoma.

The PC-10 monoclonal antibody to PCNA was employed to analyze proliferative grade in conventionally-formalin fixed, paraffin-embedded tumour samples of 162 patients with primary breast carcinoma. To perform the immunocytochemical method, sections were not heated, were de-waxed using alcohol, and then immersed in a phosphate-buffered saline solution and in methanol with 0.5% hydrogen peroxide to block endogenous peroxidase activity. Immunostaining was performed by a streptavidin-biotin peroxidase substrate. A semiquantitative scoring system was used to evaluate the fraction of nuclei that were PCNA-positive. The score ranged from 0% to 75% with a median value of 25%, mean of 27.8 +/- 1.5. PCNA staining was significantly associated with oestrogen receptor-negativity (p = 0.011) and correlated, but not at a statistically significant level, with tumour size (p = 0.08). No significant association was observed between PCNA and node status, grading, DNA ploidy, progesterone receptor or menopausal status. Prognostic indices such as number of positive lymph nodes and DNA ploidy were significantly associated with relapse-free survival (RFS) and overall survival (OS). No significant correlation between PCNA nuclear immunostaining and RFS or OS was observed after a median follow-up of 4 years. Our results indicate that analysis of PCNA alone does not seem to be a useful marker in identifying patients at different prognosis in human breast cancer.


Multidrug resistance (MDR) gene expression in acute non lymphoblastic leukemia: sequential analysis.

Sequential evaluation of P-glycoprotein expression was performed in 29 patients with acute nonlymphoblastic leukemia using immunocytochemistry with the C219 antibody. At diagnosis, 32% of the patients exhibited more than 5% of the P-gp(+) leukemic cells. Under chemotherapy, 62% of the patients eventually expressed a subset of P-gp positive leukemic cells. After conventional doses of cytosine-arabinoside (Ara-C) and daunorubicin or mitoxantrone, positive P-gp cells were noted in 65% of the cases. This percentage was significantly higher (p = 0.002) than the proportion of positive cases (15%) observed after regimens containing either intermediate doses of Ara-C or cyclosporine A, a P-gp modulator.


Angiotensin II actions in paraventricular nucleus: functional evidence for neurotransmitter role in efferents originating in subfornical organ.

Angiotensin II (ANG) has been suggested to be the neurotransmitter utilised by subfornical organ (SFO) efferents projecting to the paraventricular nucleus (PVN). The PVN has been shown to be involved in mediating the cardiovascular response elicited by electrical stimulation of SFO. The possible role of ANG as a neurotransmitter in these pathways has been examined in the present study. The cardiovascular effects of ANG microinjection into the PVN were examined in urethane anaesthetized, male Sprague-Dawley rats. Microinjection of 20 ng or 50 ng ANG into PVN resulted in mean increases in blood pressure of 12.8 +/- 0.6 mmHg (P < 0.0005), and 16.2 +/- 1.4 mmHg (P < 0.0001) respectively, without effect on heart rate. These responses were significantly attenuated following systemic administration of losartan, an ANG type 1 receptor (AT1) antagonist (Control, +12.8 +/- 0.6 mmHg; post-losartan, +5.6 +/- 1.7 mmHg), but were unaffected by the AT2 receptor antagonist, PD123319 (Control, +10.8 +/- 1.6 mmHg; post-PD123319, +11.6 +/- 2.4 mmHg). Initial and later components of the biphasic pressor response elicited by electrical stimulation of SFO (200 microA, 10 Hz, 1 ms pulse width, 10 s) were also significantly attenuated by losartan, but unaffected by PD123319. The short latency increase in mean arterial pressure was 16.6 +/- 2.3 mmHg in comparison to a post-losartan increase of 9.3 +/- 1.6 mmHg (P < 0.001). Similarly, the secondary response consisted of a control increase of 9.6 +/- 1.3 mmHg and a post-losartan increase of 3.4 +/- 0.9 mmHg (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)


The effects of selective glutamate receptor antagonists on synchronized firing bursts in layer III of rat visual cortex.

In the rat visual cortex in vitro, single-shock stimulations applied to the border between layer VI and the white matter evoke synchronized burst-firing by units in layer III. We have examined the effects of glutamate receptor antagonists on this activity, with antagonists applied via the bath to allow correlation of effects with concentrations. All synaptically driven components (recorded extracellularly as field potential 'S2' spikes, dipoles 'W1' and 'W2', and coinciding single-unit spikes) were inhibited by greater than 90% in 1.0 mM kynurenic acid and in 3 or 10 microM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, which selectively blocks AMPA/kainate receptors). S2 spike amplitudes were reduced by half in 0.7 microM CNQX. 2-Amino-5-phosphonovalerate (APV), a specific blocker of NMDA receptors, did not prevent S2 spike burst or horizontal spread of bursting within layer III. However, APV reduced the duration of synchronized bursts and the slower potentials which followed. In Mg(2+)-free medium, new components appeared which were APV-sensitive: (1) low amplitude spikes, distributed spatially like S2 spike, but recurring more slowly, and (2) slow potentials, distributed spatially like W1 and W2 potentials, but lasting for hundreds of milliseconds. The amplitudes of these spikes were reduced by half in 3 microM D-APV. Our data imply that: (1) glutamate receptors play a major role in mediating local, excitatory neurotransmission in the supragranular layers of neocortex, with NMDA and AMPA/kainate subtypes each subserving somewhat different functions; (2) AMPA/kainate receptors mediate rapid excitatory transmission between layer III neurons, responsible for driving the first 15 ms of synchronized bursts; (3) currents gated by NMDA receptors determine the duration of coherent firing bursts, and drive asynchronous neuronal firing following bursts; and (4) under conditions which circumvent block by extracellular Mg2+, activation of NMDA receptors greatly enhances and prolongs the response to single-shock stimulations. In vivo, activation of layer III neurons is likely to depend significantly upon currents gated by NMDA receptors whenever repetitively firing excitatory inputs summed over several tens of milliseconds provide enough depolarization to lift block by extracellular Mg2+.


Interaction between medullary and cervical regulation of renal sympathetic activity.

We have reported that electrical or glutamate stimulation of the dorsolateral surface of the cervical spinal cord elicits a 40-60% decrease in renal sympathetic activity (RSA) in anesthetized rats. Because evoked sympatho-inhibition was observed, however, only after transection of the cervical spinal cord at C1, we suggested that unidentified supraspinal neurons affect the regulation of RSA by cervical neurons. In the present experiments, we tested the hypothesis that the modulatory supraspinal neurons are located in the ventrolateral medulla by observing the effects of rostroventral, lateral, medullary (RVLM) injections of the GABAergic agonist, muscimol, on baseline RSA and on our ability to inhibit that activity by cervical stimulation. GABAergic inhibition in the RVLM of chlorolose anesthetized rats elicited changes in RSA that were similar to those observed after transection of the spinal cord, including a 41% decrease in mean arterial pressure and a 44% increase in RSA. Moreover, after muscimol inhibition of RVLM neurons, electrical or glutamate stimulation of the dorsolateral cervical spinal cord elicited a decrease in RSA in otherwise intact rats. These results suggest that neurons in the RVLM interact with neurons in the cervical spinal cord in the regulation of RSA.


Role of cervical neurons in propriospinal inhibition of thoracic dorsal horn neurons.

We previously reported that electrical or glutamate stimulation of the cervical spinal cord elicits a 40-60% decrease in renal sympathetic nerve activity (RSA) in the anesthetized rats. This sympatho-inhibition was possible, however, only after transection of the spinal cord at C1 or GABAergic inhibition of neurons in the rostral ventrolateral medulla. We postulated that cervical neurons inhibit RSA by inhibiting the activity of spinal interneurons that are antecedent to sympathetic preganglionic neurons (SPNs), and that these interneurons may be, in turn, excited by afferent signals. In this study, we tested the hypothesis that cervical neurons can inhibit visceroceptive thoracic spinal neurons. We recorded the spontaneous and evoked activity of 45 dorsal horn neurons responsive to splanchnic stimulation before, during, and after chemical or electrical stimulation of the cervical spinal cord in chloralose-anesthetized spinal rats. Cervical spinal stimulation that inhibited RSA also inhibited the spontaneous and/or evoked activity of 44 dorsal horn neurons. In addition to inhibiting splanchnic-evoked neuronal responses, cervical stimulation also inhibited responses, in the same neurons, evoked by noxious heat or light brushing of receptive dermatomes. We concluded that cervical neurons participate in propriospinal inhibition of afferent transmission and that this inhibitory system may be involved in controlling the access of afferent information to SPNs.


Disruption of synaptosomal calcium channel function by Lambert-Eaton myasthenic immunoglobulin is serum-dependent.

An autoantibody to nerve terminal Ca2+ channels has been suggested to mediate the pathogenesis of the neuromuscular disorder Lambert-Eaton Myasthenic Syndrome (LEMS). We demonstrated previously that in the presence of control human serum, immunoglobulins isolated from a patient with LEMS reduced flux of Ca2+ into isolated nerve terminals during depolarization. The objective of the present study was to determine the role of serum in reducing uptake of 45Ca2+ into rat brain synaptosomes by LEMS IgG. Depolarization-dependent uptake of 45Ca2+ through voltage-gated Ca2+ channels was determined using synaptosomes incubated with control (disease-free) and LEMS IgG with or without control human serum. In the absence of human serum, LEMS IgG did not reduce uptake of 45Ca2+ into synaptosomes. However, in the presence of control human serum (10% of total incubation volume), 45Ca2+ uptake was reduced significantly by LEMS IgG (2 and 4 mg/ml), but not by IgG from disease-free patients or by 10% (v/v) control human serum alone. This concentration of serum was found to be optimal; higher concentrations produced significant reductions in Ca2+ uptake, whereas at lower concentrations the serum/IgG combination was ineffective. The depressant effect of high concentrations of serum alone on 45Ca2+ uptake was mimicked by equal concentrations of bovine serum albumin suggesting that deficits in 45Ca2+ uptake produced by high concentrations of serum were due to increased protein binding of the radiolabel. Heat-inactivating the serum abolished its ability to interact with the LEMS immunoglobulins to depress 45Ca2+ uptake. This suggested a role for complement in this effect.(ABSTRACT TRUNCATED AT 250 WORDS)


Specificity of Lambert-Eaton myasthenic syndrome immunoglobulin for nerve terminal calcium channels.

Lambert-Eaton Myasthenic Syndrome (LEMS) is a presynaptic, neuromuscular disorder characterized by impaired nerve-evoked release of ACh. Repetitive nerve stimulation, which increases the probability of quantal release, improves the transmission defect. An autoantibody to Ca2+ channels of presynaptic motor nerve terminals is thought to mediate the pathogenesis of this disease. The goal of the present study was to examine the specificity of LEMS autoantibodies for nerve terminal Ca2+ channels as compared to other voltage-sensitive ion channels in nerve terminals, and to determine if non-specific membrane damage contributed to the pathogenesis of LEMS. The ion channel specificity of LEMS autoantibody was assessed by comparing the ability of acute application of IgG isolated from the plasma of a patient with LEMS to reduce depolarization-dependent uptake of 45Ca2+ and 22Na+ into or efflux of 86Rb+ from rat forebrain synaptosomes. The clinical diagnosis of LEMS was confirmed electrophysiologically by treatment of mice for 30 days with plasma (1.5 ml/day) taken from this patient. Characteristic reduction of quantal content elicited at 1 Hz and facilitation at 20 Hz was observed in mice treated with LEMS plasma compared to those treated with control plasma. One s, K(+)-stimulated 45Ca2+ uptake was inhibited 36.5 +/- 14.5% and 44.5 +/- 9.8% by acute application of 2 and 4 mg/ml LEMS IgG, respectively; IgG from patients with small cell carcinoma of the lung (SCC) had no effect on 45Ca2+ entry. The same concentrations of LEMS IgG affected neither voltage-dependent uptake of 22Na+ into veratridine-depolarized synaptosomes nor 86Rb+ efflux from K(+)-depolarized synaptosomes.(ABSTRACT TRUNCATED AT 250 WORDS)


[Pharmacologic profile of dithiadenoxide, a potential non-sedating H1-antihistaminic].

Dithiadenoxide, a product of metabolic S-oxidation of the H1-antihistaminic agent dithiaden (bisulepin, Dithiaden), is also an effective histaminic H1-antagonist in pharmacological experiments in in vivo conditions. In contrast to dithiaden, dithiadenoxide exerts a demonstrable inhibitory action as much as the subtoxic or sublethal dosage in experiments in mice and rats. Its acute toxicity in mice and guinea-pigs substantially decreases; acute toxicological findings in rats are not unambiguous. Other experimental findings demonstrate higher selectivity of pharmacological effects of dithiadenoxide.


Receptors for L-glutamate and GABA in the nervous system of an insect (Periplaneta americana).

The nervous system of the cockroach Periplaneta americana is well suited to studies of invertebrate amino acid receptors. Using a combination of radioligand binding and electrophysiological techniques, several distinct receptors have now been identified. These include an L-glutamate-gated chloride channel which has no known counterpart in the vertebrate nervous system, and a putative kainate/quisqualate receptor with pharmacological properties different from those of the existing categories of vertebrate excitatory amino acid receptors. GABA receptors have also been characterized in the cockroach nervous system. Bicuculline, benzodiazepines and steroids have revealed important differences between certain insect GABA-gated chloride channels and vertebrate GABA receptors. Identifiable neurones may facilitate the allocation of specific functions to amino acid receptor subtypes. In view of the existence of subtypes of amino acid receptors in insects, it is of interest to examine how this is reflected at the molecular level in terms of receptor subunit composition and amino acid sequence. Preliminary molecular cloning studies on insect GABA receptors are described.


An initial screen of a series of neuroactive peptides for activity on identified central neurones of Helix aspersa.

1. Intracellular recordings were made from identified neurones in the suboesophageal ganglia of Helix aspersa. Seven neuropeptides were tested for activity and their actions compared with acetylcholine and FMRFamide. 2. Three peptides isolated from nematodes, AF-1, AF-2 and PAN-1 had mainly inhibitory effects with thresholds of around 1 nM. This inhibition was due to an increase in potassium conductance. 3. The molluscan neuropeptides LSSFVRIamide, CARP and ACEP-1 were all active on certain neurones; the first two showed only inhibitory effects while ACEP-1 was mainly excitatory. The thresholds in each case were 0.1-10 microM. When norleucine replaced methionine in CARP, the potency was reduced by at least 100 times. 4. The echinoderm peptide, SALMF-1, only excited neurones but with a very low threshold, around 1.0 fM. 5. There was no obvious correlation between the action of these peptides and either acetylcholine or FMRFamide.


Inhibition of programmed cell death by cyclosporin.

1. Apoptosis is generally believed to occur as a consequence of activation of an internal, genetically controlled, program in the dying cells. 2. We have proposed an alternative hypothesis that, in hormone-induced apoptosis, the active participation of dying cells in the death process may be limited to expressing surface proteins that permit identification of cells destined to die by cytotoxic effector cells. 3. We investigated the effects of the immunosuppressant, cyclosporin, and the lysosomotropic agents, chloroquine and N-ethyl maleimide on thyroid hormone-induced regression of the bullfrog tadpole tail. 4. All three substances blocked regression. These results suggest that patency of the intracellular protein trafficking machinery is essential for programmed cell death in our system.


The effects of m-octopamine on salivary flow rates and protein secretion by rat submandibular glands.

1. m-Octopamine given i.v. or i.p. was a potent sialogogue for rat salivary glands. 2. Salivation in response to i.v. m-octopamine was completely abolished by prazosin and phenoxybenzamine. 3. The alpha-type of proteins were secreted in response to all doses of i.v. and i.p. m-octopamine and these were converted into the beta-type with prazosin, but not with yohimbine. 4. m-Octopamine stimulated both alpha- and beta-adrenoceptors and was a much more selective alpha 1-agonist than was the p-isomer.


Cardiac activity of some peptide hormones in the frog, Rana tigrina.

1. The cardiac responses of isolated frog (Rana tigrina) atria to peptide hormones were studied. 2. Calcitonin gene-related peptide (CGRP), arginine vasotocin (AVT), bovine parathyroid hormone fragment (bPTH-(1-34)) and oxytocin (OXY) produced dose-related positive chronotropic and inotropic responses; atrial natriuretic peptide (ANP) was negative chronotropic and inotropic; cholecystokinin (CCK), vasoactive intestinal peptide (VIP) were without effects. 3. The dose-related responses under bPTH-(1-34) stimulation but not CGRP or AVT were attenuated in the presence of ANP (300 ng/ml, approximately 0.98 x 10(-7) M). As expected ANP decreased the basal AR and AT responses of the isolated atria and the inhibitory effects were dose-dependent. 4. As shown previously, propranolol blocked the atrial tension stimulated by bPTH (1-34) but did not alter the cardiac responses to CGRP and AVT. 5. In the presence of beta-adrenergic blocker (propranolol 10(-7) M) or ANP (10(-7) M), the AR and AT changes under ISO stimulation in the frog were also decreased. 6. These cardiac changes suggest the cardiac inhibitory effects of ANP are related to beta-adrenoceptor activity and ANP might be a beta antagonist.


Effects on rat renal osmolytes of extended treatment with an aldose reductase inhibitor.

1. The mammalian renal medulla uses sorbitol, myo-inositol, betaine and glycerophosphorylcholine as intracellular osmolytes. 2. Sorbitol synthesis was inhibited by feeding male Wistar rats the aldose reductase inhibitor sorbinil at 40 mg/kg/day for 71 d, and renal inner medullas were extracted for analysis. 3. Aldose reductase activities and sorbitol contents were greatly reduced in sorbinil-treated animals, while betaine contents increased significantly (with no other osmolytes changing). 4. The betaine increase compensated for the sorbitol decrease such that the total organic osmolytes maintained the same ratio to sodium contents as controls. 5. These results are identical to the pattern previously reported for sorbinil treatment of rats for 10 d, but not for 21 d.


Adenylate cyclase from Hirudo medicinalis segmental ganglia: modulation by physiological and non-physiological agents.

1. In Hirudo medicinalis segmental ganglia GTP is essential for the full expression of the stimulatory action of serotonin on the adenylate cyclase activity. The amine, in turn, increases the overall affinity of the enzymatic system for GTP. 2. GTP gamma S and Gpp(NH)p, non-hydrolysable analogues of GTP, dose-dependently enhance the basal enzyme activity, but impair the stimulatory effect of serotonin. 3. Fluoride ions biphasically modulate the leech adenylate cyclase both in the absence and in the presence of GTP. The ion effect is also influenced by non-physiological guanine nucleotides.


Responses of the rectum and oesophagus of the snail Helix aspersa to purine nucleotides and nucleosides.

1. Purine compounds were examined for pharmacological activity in the rectum and oesophagus of the garden snail Helix aspersa. 2. In the rectum, adenosine, AMP, ADP and ATP (above 10 microM) and acetylcholine (above 1 nM) consistently caused concentration-dependent contractions. The slope of the dose-response curve for ADP in the rectum was significantly steeper than for the other purine compounds. The contractile responses to the nucleotides and acetylcholine, but not adenosine, were selectively potentiated by physostigmine (1 microM). Atropine (1 microM) and tubocurarine (30 microM) failed to block the responses to the purines or acetylcholine. 3. In the oesophagus, adenosine, AMP, ADP and ATP (above 10 microM) and acetylcholine (above 1 nM) caused concentration-dependent contractions that were antagonised by atropine (1 microM). Tubocurarine (30 microM) failed to block the responses to the purine compounds or acetylcholine. Physostigmine (1 microM) potentiated the responses to ADP and acetylcholine but not ATP, AMP or adenosine. 4. In both the rectum and the oesophagus, the synthetic analogues of purine compounds including 2-chloroadenosine, alpha,beta-methylene ATP and 2-methylthio ATP were inactive up to a concentration of 100 microM. 5. Electrical field stimulation of the rectum and oesophagus produced consistent contractions which were unaffected by atropine (1 microM), tubocurarine (30 microM) or physostigmine (1 microM). These responses were not modulated by any of the purine compounds or their stable analogues. 6. The responses obtained appear novel even within known invertebrate purinergic systems, suggesting a differentiation of purinoceptor subtypes in this species.(ABSTRACT TRUNCATED AT 250 WORDS)


Adenosine-theophylline interactions on the lipolytic response to beta-adrenergic agonists in porcine adipose tissue.

1. Many beta-adrenergic agonists did not stimulate porcine adipose tissue slice lipolysis unless theophylline, an antagonist of the adenosine receptor was added to the incubation medium. 2. In contrast to previous results, theophylline itself was an effective lipolytic agonist in tissue from many pigs. 3. Lipolysis was partially inhibited by adenosine or phenylisopropyl adenosine. Lipolysis was marginally stimulated by adenosine deaminase or 8-phenyltheophylline. 4. The data suggest that the mechanism of theophylline stimulation of lipolysis was only partially through antagonism of the adenosine receptor.


Inhibition of slow TTX-insensitive inward current by the anticonvulsant carbamazepine in an identified neuron of Lymnaea stagnalis.

1. Pentylenetetrazol (PTZ), induces tonic depolarization and bursting activity in an identified neuron, B1, of Lymnaea stagnalis. This is due in part to activation of a slow, tetrodotoxin-insensitive inward sodium current. 2. Carbamazepine (CBZ) reversed the effect of PTZ on both membrane potential and inward current, after a delay of up to 5 min. CBZ alone had no effect on voltage or current responses. 3. These results suggest that CBZ blocks the slow sodium current, possibly via a decrease in intracellularly stored calcium ions.


Alterations in rat alveolar surfactant system induced by treatment with carbicron (O-[(2-butenoic acid)-N,N-dimethylamide-3-yl]-O,O-dimethylphosphate).

1. Intoxication of rats with carbicron (O-([2-butenoic acid)-N,N-dimethylamide-3-yl]-O,O-dimethylphosphate) induced a reduction of the total phospholipids and phosphatidylcholine in lung alveolar surfactant. 2. The lipid transfer protein activity was inhibited due to carbicron treatment. 3. No alterations were observed in phospholipase A2 activity in the alveolar surfactant of intoxicated animals. The structural order parameter (SDPH) of bilayer liposomes, prepared from surfactant phospholipids of carbicron-treated rats also remained unchanged.


Effect of temperature reduction on the reactivity of the mouse vas deferens to adrenergic drugs.

1. Dose-response curves for noradrenaline, phenylephrine and clonidine were determined isometrically on the mouse vas deferens at 26 degrees C, 15 degrees C and compared to the one obtained at 37 degrees C. 2. In the presence of noradrenaline, reducing temperature induced an increase of both maximal developed tension and sensitivity to the drug. Reduction by 50% of the extracellular calcium concentration abolished the maximal contraction potentiation. 3. When reducing temperature to 26 degrees C, the maximal contraction was increased and depressed in the presence of phenylephrine and clonidine respectively. 4. The results suggest (a) that cooling increases the reactivity of mouse vas deferens by activation of alpha 1 adrenoceptors and depresses it by activation of alpha 2 adrenoceptors (b) that calcium ions could play an important role in the potentiation of the maximal contraction.


Feeding behavior and brain acetylcholinesterase activity in bream (Abramis brama L.) as affected by DDVP, an organophosphorus insecticide.

1. Bream (Abramis brama) were exposed to sublethal concentration of organophosphorus insecticide DDVP and the amount of food consumed and brain acetylcholinesterase (AChE) examined in exposed fish. 2. Exposure to DDVP resulted in decreased amount of food consumed and inhibited brain AChE activity. 3. Intraperitoneal injection of the fish with cholinergic drugs, atropine and TMB-4 recovered the feeding efficiency in exposed fish. TMB-4 recovered brain AChE activity as well. 4. The results revealed that cholinergic system in fish brain constitutes biochemical mechanism controlling feeding behavior in fish.


The effect of mercury on chloride secretion in the shark (Squalus acanthias) rectal gland.

1. Mercuric chloride inhibited chloride secretion in a dose dependent way in isolated perfused rectal glands. The effect was readily apparent at a concentration of 10(-6) M and profound and irreversible at 10(-4) M. 2. The dithiol dithiothreitol (DTT) 10(-2) M completely prevented the effect of 10(-6) M mercuric chloride, reduced that at 10(-5) M and 10(-4) M, and made the inhibition at the latter concentration reversible. 3. Two organic mercurials, mersalyl and meralluride, that are effective diuretics in the mammalian kidney, and p-chloromercuribenzoyl sulfonic acid (PCMBS), that has no diuretic activity, had no effect on chloride secretion by the rectal gland. 4. The effect of mersalyl was not modified by lowering the pH of the solution perfusing the glands. 5. These results indicate that inorganic mercury and organic mercurials do not share the same mechanism of action. 6. The absence of an effect of organic mercurials on chloride transport in the rectal gland suggests that its effect on another chloride transporting epithelia, the thick ascending limb of the loop of Henle, is not mediated by inhibition of the chloride cotransporter or Na+, K(+)-ATPase, common to both epithelia.


Accumulation and excretion of aluminium and iron by the terrestrial snail Helix aspersa.

1. The snail Helix aspersa was fed one 24 hr meal containing Al, Fe or both together in barley flour pellets. Accumulation and distribution within the digestive gland, kidney, crop and remaining soft tissues were examined over the subsequent 30 days using atomic absorption spectroscopy (A.A.S.). 2. The digestive gland contained significantly (P < 0.05) elevated levels of Al and Fe for 8 and 12 days. The digestive gland is the major sink for both Al and Fe in Helix. 3. The kidney rapidly accumulated Al and Fe but the increase was short-lived. The kidney may therefore be involved in the elimination of metal not incorporated into the digestive gland. 4. Iron was absorbed by the crop but Al was not. This may indicate a route of uptake of Fe into the digestive gland not shared with Al. 5. No obvious pattern of accumulation of Al and Fe were seen in the remaining soft tissues or the blood of Helix. 6. Aluminium is present in the faeces for 12 days suggesting that Al is released relatively slowly. 7. Presence of both Al and Fe in the feed induced a change in the pattern of accumulation in the digestive gland but not in the kidney, crop and remaining soft tissues. 8. The distribution of Al is discussed in relation to the suggestion that Al follows the ferretin pathway during accumulation.


Some effects of aluminium on rat brain protein synthesis.

1. Infant rats and rabbits received intraperitonal aluminium (Al) chloride (5, 10 or 20 mg Al/kg body weight) every third day from one to four weeks of age. 2. When the polysomal fraction was tested in a protein synthesizing system, a significant increase in the incorporation of [14C] leucine, [14C] phenylalanine, or [35S] methionine into proteins in vitro was observed at the higher doses in rats but not rabbits. 3. The incorporation of [35S]methionine into brain ferritin was measured using polysomal mRNA or mRNA "stored" in the ribonucleoprotein (RNP) particle fraction. 4. The results suggest that Al exposure causes the mobilization of ferritin mRNA from the latter fraction to the polysomal fraction for increased ferritin synthesis.


Potentiation by m-cresol on transepithelial sodium transport across frog skin induced by insulin.

1. In this study we found that insulin mixed with m-cresol, normally used as pharmaceutical preparation, shows an earlier and larger stimulating effect on transepithelial sodium transport than insulin alone. 2. The action displayed by the m-cresol seems to be specific for insulin because m-cresol mixed with ADH, a hormone known to stimulate sodium transport, failed to show the potentiation seen for insulin. 3. It is proposed that m-cresol could facilitate the interaction with its receptor by modifying the insulin molecule. 4. This finding could be of biological and pharmacological importance.


Characteristics of outward current induced by application of dopamine on a snail neuron.

1. A closer characterization of the potassium channel opened by the application of dopamine (DA) on an identified Helix pomatia neuron was attempted. The effect of K+ channel blockers (TEA and 4-AP) on the DA-induced current was examined. The results indicate that the channel opened by DA does not share the pharmacological properties of other snail neuron K-channels. 2. The I-V relation for IDA was successfully fitted by the Constant Field equation except below the reversal potential where the current was smaller than expected. The assumption that DA binding is voltage-sensitive is supported by the increment of the Hill coefficient with hyperpolarization (from nH approximately equal to 1 to nH approximately equal to 2). 3. The presence of the phosphodiesterase inhibitor IBMX does not affect the DA induced outward current. However, the assumption that the snail neurons' DA receptor belongs to the D2 class is in contrast to the antagonistic effects of ergot alkaloids which, in mammalian neurons, are competitive antagonists of D1 receptors. 4. The examination of the voltage-sensitivity of the blocking action of the ergot alkaloid (Bromoergocryptinine) revealed that it does not compete with DA for the same binding site as in mammalian D1 receptors.


[The effect of antipsychotic drugs on dopamine-receptors on caudate calf].

Using radioligand receptor binding assay (RRA), dopamine receptor in the calf caudate were identified with 3H-Spiperone as radioactive ligand. The total number of maximal binding sites (Bmax), the equilibrium dissociation affinity (Kd), the concentrations of drug required to inhibited 50% specific binding (Ic50) and competitive inhibition constant (Ki) of antipsychotics in calf caudate were determined by sturative binding experiment and inhibitive binding experiment. The result showed the mean Bmax was 0.659 pmol/mg protein membrane and Kd was 0.31 nmol/L. Scatchard analysis denotes that it was hyperbole and there were two binding positions in calf caudte. Antipsychotics can inhibit 3H-Spiperone binding specifically the order of affinity constants (1/Ki) was Haloperdol and sulpiride, Chlorpromazine and Clozapin, Tranquis and Taractan. This abilities of drugs to compete with 3H-Spi binding will be used as basis of RRA for measuring blood levels of antipsychotics.


DNA polymerase delta and epsilon holoenzymes from calf thymus.

Replication of singly-DNA primed M13 DNA by DNA polymerase (pol) delta completely relies on the simultaneous addition of proliferating cell nuclear antigen (PCNA), replication factor C (RF-C) and replication protein A (RP-A) (or E. coli single-strand DNA binding protein, SSB). Pol epsilon core alone is able to synthesize the products on singly-primed ssDNA. However, DNA synthesis by pol epsilon was stimulated up to 10-fold upon addition of the three auxiliary proteins PCNA, RF-C and SSB. This stimulation of pol epsilon by PCNA/RF-C/SSB appears to be the superposition of two events: pol epsilon holoenzyme (pol epsilon, PCNA, RF-C) synthesized longer products than its pol epsilon core counterpart, but elongated less primers. Furthermore, we analyzed the cooperative action of pol alpha/primase with pol delta or pol epsilon holoenzymes on unprimed M13 DNA. While pol delta displayed higher dNMP incorporation than pol epsilon, when a single primer was preannealed to DNA, pol epsilon was more efficient in the utilization of the primers synthesized by pol alpha/primase. Under these conditions both longer products and a higher amount of dNMP incorporation was found for pol epsilon holoenzyme, than for pol delta. Our data support the hypothesis of pol delta as the leading and pol epsilon as the second lagging strand replication enzyme.


[Pathophysiology and gene abnormalities of endocrine tumors].

Various functioning and non-functioning tumors arise from endocrine glands in both the sporadic and familial forms and pathophysiology of the tumors is variable due to differences in the sort of tumor-bearing endocrine organs and in the amount of hormones released. In this paper, gene abnormalities in growth hormone (GH)-secreting pituitary adenoma, ectopic GHRH-producing tumor, multiple endocrine neoplasia (MEN) and ectopic parathyroid hormone (PTH)-producing tumor are documented in relation to etiology and pathophysiology. GH-secreting pituitary adenoma is heterogeneous in clinical features, pathological findings and GH responses to various secretagogues. A point mutation of codon 201 of Gs alpha gene was observed in 2 out of 45 GH-secreting pituitary adenomas (4.4%), but no point mutation of Gi2 alpha gene was found. Pituitary tumors may occur at any stage of differentiation from the totipotent cells to mature anterior pituitary cells, and the mutations of Gs alpha and H-ras genes as well as loss of heterozygosity (LOH) found on chromosome 11 in some adenomas must be involved in their tumorigeneses. Since 1959, 34 patients with ectopic GHRH-producing tumor associated with acromegaly have been reported. In our case of MEN type 1, the paradoxical rise of plasma GH after TRH or glucose administration disappeared after resection of the tumor. The tumor cells showed neither rearrangement nor amplification of GHRH gene and 20 oncogenes including ras, myc, and erb. Only LOHs of HRAS1 and D11S151 were detected in this tumor, but no point mutation was found in HRAS1 gene. Therefore, a kind of tumor suppressor gene may be involved in the tumorigenesis of the tumor in addition to inactivation of MEN-1 locus. In MEN-1 patients, we reported LOH on chromosomes 1, 9, 11 and 16, while we reported point mutation as being present only in Gs alpha gene on chromosome 20. This point mutation was found specifically in GH-secreting pituitary adenoma but not in hyperplastic parathyroid and pancreas adenoma. These data suggest that in MEN-1 patients tumorigenesis occurs and advances from hyperplasia and adenoma to cancer during multistep changes of genes such as inactivation of MEN-1 gene and other tumor suppressor genes and activation of oncogenes. Ectopic PTH-producing tumor was first reported by us in 1989, and this was followed by 2 papers. These patients showed a disturbance of consciousness and high levels of serum calcium and plasma PTH.(ABSTRACT TRUNCATED AT 400 WORDS)


Fishes of District Sundargarh, Orissa, with special reference to their potential in mosquito control.

An extensive fish fauna survey was carried out in Sundargarh, a malaria-endemic district in Orissa, during 1988 to 1990 to identify and evaluate the indigenous larvivorous fishes for mosquito control. In all, 57 species belonging to 19 families under 6 orders were found in the local water bodies. On laboratory evaluation against anopheline and culicine larvae, six potential larvivorous fishes, viz. Aplocheilus panchax, Oryzias melastigma, Oreochromis mossambicus, Gambusia affinis, Danio (B.) rerio and Esomus danricus were selected. Feasibility of mass multiplication of these fishes in village ponds for operational use is being studied.


Search for enteric microbial pathogens in patients with ulcerative colitis.

Microbial pathogens were sought in faeces of patients with active ulcerative colitis and again after 3 months treatment. 64 patients were examined during their first episode of ulcerative colitis and 30 with relapse of chronic disease. At presentation, bacterial pathogens were not found; 1 patient had cryptosporidiosis. In 10 patients treatment appeared to result in some loss of colonisation resistance as evidenced by colonisation with beta-haemolytic streptococci, Staphylococcus aureus, candida and Clostridium difficile. Unidentified cytotoxic activity was present in the faeces of 4 patients at presentation and 2 patients during or after treatment. We conclude that enteric infection is an uncommon finding in patients with active ulcerative colitis.


Effect of gastrin receptor antagonists on gastric acid secretion and gastrin and somatostatin release in the rat stomach.

The effects of two recently developed gastrin receptor antagonists, PD 136450 and L-365,260, on pentagastrin-stimulated acid secretion were investigated in rats. PD 136450 at a dose of 6 mg/kg s.c. (9.6 mumol) completely abolished acid secretion induced by pentagastrin. The inhibition of 18 mg/kg PD 136450 s.c. lasted for at least 8 h and was still effective after 14 days of treatment (18 mg/kg s.c. every 8 h). Acute application of L-365,260 at a dose of 3.8 mg/kg, which is equimolar (9.6 mumol) to 6 mg/kg PD 136450 reduced acid responses slightly. However, when L-365,260 was administered intravenously at a dose of 3 mg/kg, this antagonist completely abolished the pentagastrin-stimulated acid secretion. Furthermore, the effect of PD 136450 on endogenous gastric somatostatin and gastrin releases was tested in the isolated, vascularly perfused rat stomach. PD 136450 perfused at a concentration of 1 microM slightly increased somatostatin secretion after stimulation with a high dose of isoproterenol (10(-7) M). There was no effect of PD 136450 on basal or acetylcholine-stimulated gastrin secretion.


Effects of progesterone on some brain neurotransmitters in intact rats.

Effects of progesterone on four neurotransmitters (viz, noradrenaline, 5-HT, dopamine and histamine) of brain were seen in rats with intact ovaries. It was found that progesterone lowers the noradrenaline concentration in medulla, pons, midbrain, hypothalamus, thalami and pituitary, uniformly, when the rats were killed within 4 hours of progesterone injection. At longer intervals (48 hrs) effects of progesterone were seen when progesterone in heavy dose was administered to rats pretreated with estrogen. It is likely that one of the modes of action of the oral contraceptives may be the reduction of noradrenaline content in selected areas of brain, by progesterone. It is also suggested, therapeutic usage of progesterone carries the risk of development of depression in the user.


Development and trends in the drug treatment of essential hypertension.

A brief survey is given of the development of the drug therapy of essential hypertension over five decades, followed by a discussion on newer antihypertensive drugs and principles. Virtually all levels and elements of the sympathetic nervous system can be modulated by drugs in such a manner that elevated blood pressure is lowered, for instance with the use of central alpha 2-adrenoceptor agonists (clonidine, alpha-methyldopa), ganglioplegic drugs, peripheral adrenergic neurone blockers, and alpha- and beta-adrenoceptor antagonists. The beta-blockers have been maintained as a very major group of antihypertensive therapeutics. Among alpha-adrenoceptor antagonists, only the selective alpha 1-blockers are useful as antihypertensive drugs, although some interest has developed in newer alpha 2-adrenoceptor antagonists which are selective for postsynaptic alpha 2-adrenoceptors. Diuretics, in spite of some criticism, are still a cornerstone in antihypertensive drug treatment. Their position has been reinforced by the results of the recent Systolic Hypertension in the Elderly Program, Swedish Trial in Old Patients with Hypertension and Medical Research Council (Elderly) trials. Calcium antagonists have been extended by several new dihydropyridines which appear to be more vasoselective than the classical compounds. Various new ACE inhibitors have been introduced, but so far they have not clearly offered major advantages over the established compounds. Renin inhibitors and angiotensin II-receptor antagonists are being studied for their potential as antihypertensive drugs and they are also important because they may modulate the renin-angiotensin-aldosterone system. Inhibitors of neutral endopeptidase will cause an accumulation of endogenous atrial natriuretic peptide, and hence lower blood pressure by counteracting several effects of the renin-angiotensin-system. Potassium channel openers are vasodilators and potential antihypertensive drugs with additional anti-ischaemic activity.


The 'serotonin/norepinephrine link' beyond the beta adrenoceptor.

C6 rat glioma cells were utilized as a model system to probe the 'serotonin/norepinephrine link' at the level of preproenkephalin (PPE) gene expression. The beta adrenoceptor mediated increase in PPE mRNA was attenuated by the selective beta 1 adrenoceptor antagonist metoprolol which blocked the isoproterenol induced cyclic AMP generation by 97%. The subtype nonspecific antagonist propranolol blocked both the isoproterenol induced increase in cyclic AMP and the increase in the PPE mRNA steady-state levels. Serotonin (5-HT) had no effect on the density of beta adrenoceptors or their down-regulation by isoproterenol and did not alter the PPE gene expression in the absence of the beta signal. However, 5-HT significantly deamplified the beta signal mediated enhancement of the PPE mRNA thus indicating that the aminergic link occurs beyond the beta adrenoceptor.


Muscle-derived differentiation factor increases expression of the tyrosine hydroxylase gene and enzyme activity in cultured dopamine neurons from the rat midbrain.

Our earlier work demonstrated that certain populations of brain neurons which do not synthesize catecholamine (CA) neurotransmitters in vivo, will, when grown in culture with muscle-derived differentiation factor (MDF), unexpectedly express the gene for the CA biosynthetic enzyme tyrosine hydroxylase (TH). In this paper, we sought to determine whether MDF could also regulate TH expression in those neurons which normally synthesize CA neurotransmitters. Incubation of cultured dopamine neurons from the ventral midbrain with MDF elevated the levels of TH mRNA and TH enzyme activity 5- to 40-fold higher than that measured in control cultures. Sympathetic neurons were unaffected by a similar MDF treatment. Unlike the 2-day critical period for MDF-responsivity in non-CA neurons. CA neurons remained susceptible to MDF's influence over an extended developmental interval (E14-18), suggesting that MDF may be important for TH gene regulation in brain CA neurons even differentiation is complete. Because of these unique properties, MDF may provide a unique opportunity to explore ways in which the TH gene might be directly manipulated in these cell populations in order to correct the CA imbalances that occur in certain neurological diseases and disorders.


Hypothalamic neuropeptide expression after food restriction in Zucker rats: evidence of persistent neuropeptide Y gene activation.

The Obese Zucker rat is a model of genetic obesity characterized by hyperphagia, hyperinsulinemia and other endocrine abnormalities. In order to elucidate pathogenetic mechanisms contributing to disturbed feeding behavior in these animals, the effect of food restriction on three hypothalamic neuropeptides involved in the control of food intake was studied. Eighteen male obese and 18 lean Zucker rats were randomly divided into two groups: half of the animals were food-restricted for 2 weeks, while the other half served as controls and were fed ad libitum. The levels of preproneuropeptide Y (preproNPY), preprocorticotropin releasing factor (preproCRF) and preprosomatostatin (preproSOM) mRNAs were determined using in situ hybridization technique. In addition, plasma insulin and corticosterone concentrations were analyzed. Food restriction significantly increased the expression of preproNPY mRNA in the arcuate nucleus in both Zucker phenotypes, while the expressions of preproCRF mRNA in the paraventricular nucleus (PVN) and preproSOM mRNA in the periventricular nucleus (PeV) were not altered. The expression of preproNPY mRNA was significantly greater in control obese animals compared to control lean animals. Food restriction lowered plasma insulin levels, but did not change plasma corticosterone levels. It is concluded that food restriction specifically activates NPY gene transcription in the arcuate nucleus the response being similar in both Zucker phenotypes. The results suggest that orexigenic NPY plays a role in the adaptation to altered feeding status.


Analysis of the human tyrosine hydroxylase promoter-chloramphenicol acetyltransferase chimeric gene expression in transgenic mice.

To investigate cis-elements responsible for catecholaminergic (CAnergic) neuron-specific expression of the tyrosine hydroxylase (TH) gene, we produced lines of transgenic mice carrying 5.0-kb, 2.5-kb and 0.2-kb fragments from the 5'-flanking region of the human TH gene fused to a reporter gene, chloramphenicol acetyltransferase (CAT), and designated them as TC 50, TC 25, and TC 02, respectively, and reporter gene expression in transgenic mice was analyzed by CAT assay by immunocytochemistry with anti-CAT antibody. High-level CAT expression was observed in the brain and adrenal gland using the 5.0-kb promoter of the TC 50 mice, but ectopic expression was consistently observed in several somatic tissues, e.g. thymus, colon, and testis. In brain, expression was achieved in CAnergic neurons with the largest construct (5.0 kb), but not with 2.5 kb or 0.2 kb of 5' flanking sequence. However, TC 50 mice also expressed CAT immunoreactivity in non-CAnergic neurons. In the TC 25 line CAT immunoreactivity was detected only in some non-CAnergic neurons. In the TC 02 line no CAT immunoreactivity was detected in any of the tissues examined. These results indicate that the 5.0-kb DNA fragment of the TH gene upstream region contains activity to express CAT in CAnergic neurons and surprisingly, lacks some regulatory elements attenuating ectopic expression, and that the 2.5-kb and 0.2-kb fragment are not sufficient for the proper expression. We discuss the presence of the tissue-specific regulatory elements in the structure portion of the TH gene and/or 3'-flanking region.


A world united against AIDS. Report on the Nursing Satellite Conference and the Eighth International Conference on AIDS, Amsterdam, The Netherlands, 19-24 July, 1992.

The dramatic increase of new HIV/AIDS cases in women and their babies continues to require midwives to be vigilant in their efforts to identify clients at risk for HIV. The role of STDs in HIV transmission and disease progression is evident. Research is lacking on the effect of AIDS drugs, pregnancy and birth on the HIV-infected woman. Prevention, through safer sex, is a critical education strategy to consider. Data suggesting intrapartum transmission of HIV is particularly interesting and warrants further study. Development of the international framework for future clinical vaccine trials is underway. Initial studies on the ability of HIV vaccines to limit perinatal transmission may provide some encouraging future results.


Molecular genetics and the ontogeny of pigment patterns in mammals.

The conclusion that animal development is guided by a hierarchical system of gene expression and interaction has gained considerable support from recent molecular genetic studies on fruit flies (Drosophila melanogaster) and mice (Mus musculus). They demonstrate that the patterns of organization revealed by terminal differentiation of cells is anticipated by a myriad of transient prepatterns that channel the developing embryo toward its genetically-programmed target. The numerous white spotting mutants in mice exhibit some of the most dramatic and variable patterns of cutaneous melanin pigmentation. Until recently, the mechanisms of action of white spotting genes and their relationship to the developmental genetic hierarchy remained unknown. It now appears that certain white spotting genes may encode growth factors essential for melanoblast development. Others may be related to homeobox genes that play a number of developmental roles, the primary one being the determination of regional organization along the anterior-posterior axis of the early embryo. The patterns of homeobox gene expression are consistent with several of the developmental models for white spotting in mice and other mammals. It is evident that white spotting genes are not solely concerned with the terminal differentiation of melanoblasts into melanocytes. They are heterogeneous with regard to action and level of expression within the developmental hierarchy.


Neuroleptic treatment is an unlikely cause of elevated dopamine in thalamus of schizophrenic subjects.

Previous studies have indicated a marked increase in the dopamine/norepinephrine ratio in thalami of schizophrenic patients compared with those of control subjects. Since these results all came from patients who were receiving neuroleptic drugs, the possibility exists that the increased dopamine concentrations are an effect of medication. To address this question, similar analyses were done on thalami from Huntington's Disease patients who had received neuroleptic treatment. The results showed no differences between the thalami of Huntington's Disease patients and controls, strongly suggesting that chronic treatment with neuroleptic drugs does not result in an increase of endogenous dopamine in the thalami of human subjects.


Comparison of brain tumor growth kinetics by proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine (BrdU) labeling.

The bromodeoxyuridine (BrdU) labeling study provides valuable cell kinetic information for individual tumors that could suggest the prognosis of each patient who had a tumor. Recently, a monoclonal antibody against the proliferating cell nuclear antigen (PCNA or cyclin), a nuclear protein expressed in proliferating cells, was developed which could be used on formalin fixed, paraffin embedded tissue. The purpose of this study was to compare the cell kinetic data obtained by the BrdU labeling study and the PCNA method in the same patient. The relationship between labeling indices of BrdU incorporated into S-phase and PCNA expressed by cycling cells was investigated in 31 patients with brain tumors. Both of the labeling indices showed good correlation with histological grade of the tumor. The values of the PCNA labeling index (LI) were parallel but higher than those of the BrdU LI, and the relation PCNA LI = 2.2 x BrdU LI + 0.8 (r2 = 0.86) was obtained. The results of this study show that PCNA could replace the BrdU method for identifying the proliferating cells, and the major advantages of PCNA method is that it could be done without any pretreatment and avoid injection of the teratogenic agent for diagnostic purpose.


[Changes in the hypophyseal-thyroid system under narcotic action in the parturients of a group at high risk of perinatal pathology].

Eighty-seven parturients were examined to disclose the function of the thyroid during narcotic exposure in labor and in cesarean section in cases of high risk of giving birth to infants with perinatal abnormalities. Neurotropic agents were found to inhibit the fetal hypophyseo-thyroid system in cases with a history of intrauterine hypoxia, thus saving this system from exhaustion. Employment of various types of general anesthesia in cesarean section in this patient population provides an adequate maternal neurovegetative protection from surgical aggression.


The carbohydrate epitope 3-fucosyl-N-acetyllactosamine is developmentally regulated in the human cerebellum.

The carbohydrate epitope 3-fucosyl-N-acetyl-lactosamine (CD15) is involved, as a constituent of glycoconjugates, in cell-cell interactions and cell sorting during rodent CNS morphogenesis. The present study was designed to test whether CD15 is also involved in the development of the human CNS. Human cerebellar hemispheres and vermes from the 24th week of gestation (wg) to the 26th postnatal month (pnm) and from adults were investigated for CD15 immunoreactivity, using the monoclonal antibody MMA. Our findings establish that the carbohydrate moiety is developmentally regulated in neuronal and glial cells during their differentiation. First, the parallel fibers of granule cells are CD15+ during the epoch of synaptogenesis with Purkinje cell dendrites. Second, a subpopulation of neurons from the dentate nucleus is transiently CD15+ from the 32nd wg until the 15th pnm. Third, at the onset of myelination (around the 35th wg), CD15 immunoreactivity is discernible in the cytoplasm of young oligodendrocytes. Immunoreactivity on protoplasmic astrocytes of the inner granular layer and on fibrous astrocytes of the white matter progressively increases during fetal development. In addition, the CD15 epitope is persistently present on Bergmann glial processes and ependymal cells. Within the three subdivisions of the cerebellum, i.e., hemispheres, vermis, and flocculonodular lobe, the CD15 expression follows a different timing of morphogenesis. For example, diminution of immunoreactivity in the parallel fibers occurs first in the phylogenetically older flocculonodular lobe and vermis, and later in the phylogenetically younger hemispheres. This study shows that in the human cerebellum the distribution of CD15 undergoes marked developmental changes. This epitope may also act in cell-to-cell recognition, and perhaps could play a role in controlling CNS development.


Acute ethanol administration and the metabolism of glutamine by skeletal muscle of the rat: implications for ethanol-induced reductions in protein synthesis.

The effect of acute ethanol administration (75 mmol/kg) on the metabolism of glutamine by skeletal muscle of the rat was studied in order to investigate the hypothesis that the concentration of this amino acid in muscle controls the rate of protein synthesis. Ethanol administration was without effect on the concentration of glutamine in EDL (extensor digitorum longus) and plantaris muscles (Type II fibre-rich muscles), but increased the concentration in soleus muscle (Type I fibre-rich muscle). The rate of release of glutamine was increased from EDL muscle, but unchanged from soleus muscle of treated animals. Ethanol administration was without effect on the maximal activity of glutamine synthetase in both soleus and EDL muscles. It is concluded that changes in the concentration of glutamine in muscle cannot explain the ethanol-induced changes in the rate of protein synthesis. Nevertheless, the increase in the concentration of glutamine in soleus muscle following ethanol administration is of interest and may be mediated via modulation of the glutamine transporter and/or the activity of glutamine synthetase in vivo.


Studies of nonsedative antihistamines. II. Assessment of its antihistaminic potency.

An analysis is made of the effectiveness of terfenadine at dosages of 60 and 120 mg, of cetirizine at a dose of 10 mg, and of loratadine and ebastine at a dose of 10 mg in inhibiting the papule induced by 20 mcg of intradermal histamine. Although all produced significant inhibition, cetirizine and terfenadine 120 mg showed a significantly greater inhibition coefficient than loratadine.


Changes in markers of alcohol intake in man below 'safe' drinking levels.

In a group of 343 working men, only 34 of whom regularly drank more than 60 ml of ethanol per day, logistic regression was used to determine the combination of biomarkers which best discriminated between those who regularly drank less than 30 ml or 30 ml or more of ethanol daily. The index consisted of apolipoprotein A-II, uric acid, gamma-glutamyl transpeptidase and mean corpuscular volume. Even with the relatively low level of alcohol intake reported in these subjects (assessed using a 7-day retrospective alcohol diary), the groups with higher or lower intake of alcohol were separated with a sensitivity of 68%, a specificity of 74% and a positive predictive value of 71%. Systolic blood pressure, adjusted for age and body mass index, was also linearly related to alcohol intake, and when included in the biomarker index improved the proportion correctly classified from 71% to 75%. Using the biomarkers only, 94% of subjects regularly drinking at least 80 ml of ethanol equivalent per day and 100% of the non-drinkers were correctly classified.


HER-2/neu oncogene expression, DNA ploidy and proliferation index in breast cancer.

HER-2/neu gene expression, DNA ploidy and proliferation index were studied in 250 cases of breast cancer. Expression of HER-2/neu was determined by using an antibody to the HER-2/neu receptor. Ki-67 antibody was used to determine the proliferation index of the breast cancers, and the Feulgen method was used to assess DNA amounts in the tumor cells. Histochemical staining was quantitated by image analysis. Of the cancers studied, 72 were positive for overexpression of HER-2/neu protein; of these, 62 (86%) possessed near-tetraploid DNA content, and 47 (65%) had more than one G0G1 stem line (polyploid) of DNA distribution. Cells from the cases negative for HER/2-neu overexpression contained DNA amounts that ranged from diploid to varying degrees of aneuploid. A significant difference in the amounts of cellular proliferation in HER-2/neu overexpressing cancers was found between those that expressed the HER-2/neu receptor on their membranes and those that exhibited mainly cytoplasmic receptors.


Measurement of the immunoperoxidase staining of macrophages within liver granulomata of mice infected with Mycobacterium tuberculosis.

A quantitative image analysis technique developed for the measurement of the extent of macrophage activation and epithelioid cell differentiation was performed on mice infected experimentally with Mycobacterium tuberculosis. The granulomatous inflammatory response within the liver reached a peak at day 23 and declined by day 33. Animals of strain B10.BR (H-2k) showed an increased granuloma fraction as compared to Balb/k (H-2k) mice, thus confirming the influence of non-H2 genes in the control of granuloma formation in mice. Using a monoclonal antibody against CD11b/CD18 (Mac1;CR3), we observed two subpopulations of macrophages within the granulomata. The small, darkly staining cells at the periphery of granulomata appear to be newly recruited macrophages. Larger, paler staining cells toward the center of granulomata represent activated and mature epithelioid macrophages. Using a semiautomated image analyzer (Quantimet 970), we measured the relative numbers of these macrophage subpopulations. There were more activated macrophages (epithelioid cells) associated with the increased granuloma fraction in the B10.BR mice than in the Balb/k. However, similar numbers of newly recruited peripheral macrophages were found in both Balb/k and B10.BR strains. This technique has shown qualitative as well as quantitative differences in the granulomatous inflammatory response in this murine model of tuberculosis in strains of mice with quite different antibody repertoires to mycobacterial antigens.


Minimal growth factor requirements of human fetal kidney in serum- and glucose-free culture.

The addition of insulin plus transferrin to Leibovitz's L-15 medium was previously shown to restore important cellular functions in a serum-free system developed in our laboratory for human fetal kidney explants. The objective of the present study was to compare the effectiveness of this insulin plus transferrin combination with one used in other in vitro systems whereby serum is substituted by a mixture of five hormones (insulin, transferrin, hydrocortisone, triiodothyronine and prostaglandin E1). In fetal kidney it was found that the combination of insulin plus transferrin was as effective as the five-hormone mixture on DNA synthesis after 5 days of culture and was even more effective in younger fetuses (10-13 weeks) compared with older fetuses (16-19 weeks). However, protein synthesis was more sensitive to the five-hormone combination. Selective deletion of individual hormones showed that insulin is the essential factor for the growth of cultured kidney explants. Differentiation of brush border membranes in nephrons, as evaluated by alkaline phosphatase and gamma-glutamyl-transferase activities, was not significantly modified by either of the two combinations. The present results indicate that insulin plus transferrin represents the optimizing condition for our culture model. The response to supplements varies according to fetal age and possibly to tissue proliferation states and/or cell type.


The baculovirus Autographa californica nuclear polyhedrosis virus genome includes a papain-like sequence.

The published DNA sequence that includes the gene for the envelope glycoprotein gp67 of the baculovirus Autographa californica nuclear polyhedrosis virus also contains a segment that shows 32% amino acid identity to papain, a cysteine endopeptidase from the papaya plant (Carica papaya). The viral papain-like sequence is apparently affected by a frame-shift mutation, but otherwise appears capable of encoding a functional enzyme. Catalytically essential amino acids appear to be conserved, as do disulphide bridges. The overall structure of the putative protein is similar to that of papain, as judged by hydropathy profile. Secondary structure prediction using a consensus of seven methods indicates that the putative viral enzyme retains the alpha/beta domain structure of papain, including the long helix beginning with cysteine-25. Infecting SF9 cells with the virus did not lead to a detected increase in cysteine endopeptidase activity, and a cysteine endopeptidase inactivator appeared to have no effect on infectivity. Irrespective of whether the sequence encodes a functionally active cysteine endopeptidase, this is the first example of a papain-related sequence in a viral genome.


Activated oncogenes and putative tumor suppressor genes involved in human breast cancers.

Cytogeneticists first proposed that the karyotypic abnormalities identified on chromosomes 1, 3, 6, 11, 13, 16, 17, and 18 supported a genetic basis for breast cancer. Such abnormal banding patterns, however, may represent either loss-of-function or gain-of-function molecular events. RFLP analyses have since confirmed that 20-60% of primary and spontaneous human breast tumors exhibit allelic losses on these same chromosomes, although the exact genes involved at these chromosomal sites remain largely unknown. Knowledge gained about the Rb-1 and p53 tumor suppressor genes at 13q14 and 17p13 in breast and other human tumors supports the paradigm that for any chromosomal locus, allelic loss associated with a mutation in the remaining tumor allele signifies an involved tumor suppressor gene. Given this paradigm, there are nearly a dozen putative breast tumor suppressor genes under active investigation, with most investigators now focusing on various chromosome 17 loci. Among the known proto-oncogenes found activated in breast cancer, amplification of c-erbB-2 at 17q21 is the most widely studied and clinically significant gain-of-function event uncovered to date, occurring in about 20% of all primary breast tumors. The involvement of this overexpressed membrane receptor has engendered interest in related tyrosine kinase receptors, such as EGFR, IR, and IGF-I-R, as well as their respective ligands, which may be overexpressed in a greater fraction of tumors, contributing to the autocrine and paracrine regulation of breast cancer growth and metastasis. New attention is being given to the potentially oncogenic function of structurally altered nuclear transactivating steroid hormone receptors, such as ER, whose overexpression has long been used to determine endocrine therapy and prognosis for individual breast cancer patients. While c-myc was one of the first known proto-oncogenes to be found amplified and overexpressed in human breast cancers, the actual incidence and clinical significance of its activation remain disputed and in need of further study. Lastly, we can expect greater clarification about the importance of various 11q13 genes found coamplified in nearly 20% of primary breast cancers, and pursuit into the intriguing possibility that a cyclin-encoding gene represents the overexpressed locus of real interest in this amplicon. Virtually all of these important genetic abnormalities identified thus far are associated with but not restricted to human breast cancers. The absence of identifiable molecular defects relating to the tissue specificity of this malignancy must be considered a substantial gap in our basic understanding of breast carcinogenesis.(ABSTRACT TRUNCATED AT 400 WORDS)


Involvement of G proteins, cytoplasmic calcium, phospholipases, phospholipid-derived second messengers, and protein kinases in signal transduction from mitogenic cell surface receptors.

Some putative mitogenic signal transduction mechanisms involving G proteins, calcium, phospholipases, and protein kinases have been discussed. Several elements in this signal transduction scheme are not yet well understood and require further experimental investigation. With regard to the heptahelix receptors, exactly how do they activate PLA2? Is PLA2 activation linked to mitogenic pathways? Is this via stimulation of protein kinase C or perhaps another mechanism? How do heptahelix receptors activate tyrosine phosphorylation, and is it important in their ability to stimulate cell growth? With regard to the various phospholipases that are thought to be regulated by receptor-mediated stimuli, only PI-PLC beta and PI-PLC gamma are well characterized. PLA2, PC-PLD, and PC-PLC require further study in regard to determination of molecular structure and elucidation of mechanisms of phospholipase activation (e.g., what are the molecular mechanisms whereby tyrosine kinases and Ras affect PC-PLC?). The protein kinase C dependent and protein kinase C independent mechanisms that enable mitogenic stimuli to activate the Erk/MAP kinase are enigmatic at this time. How Raf-1 activates SRE-containing gene promoters (such as the fos promoter) is also not known. However, given the current rapid rate of progress in this field, it is likely that a much more complete understanding of the mitogenic signal transduction process will soon be obtained.


DNA binding by the Myc oncoproteins.

The c-Myc protein is a potential activator of transcription, with the ability to bind in a heterodimer form with Max to DNA sequences containing the core hexanucleotide sequence CAC(G/A)TG. These properties are shared with L-Myc, a homologous oncoprotein expressed in small cell lung carcinoma cells; with N-Myc, expressed in neuroblastoma cells; and with avian v-Myc, the c-Myc homolog expressed by a chicken retrovirus. The c-Myc, and probably v-Myc, proteins also have nonspecific DNA binding function, which may improve the kinetics of specific DNA binding. Curiously, this domain appears not to be conserved in L-Myc or N-Myc [22]. The data that have accumulated to date are consistent with a model in which a c-Myc/Max heterodimer positively regulates the transcription of growth-related genes, with Max homodimer functioning as a negative regulator of the same genes (Fig. 4) [55]. Max is expressed constitutively at low levels, whereas c-Myc is expressed at low levels in quiescent cells, but high levels of c-Myc are induced by mitogenic stimulation [56]. Thus, in proliferating cells c-Myc/Max heterodimers might bind to the regulatory elements of growth-related genes, where the c-Myc TAD might stimulate transcription. Conversely, in quiescent cells with little c-Myc present, Max homodimers might predominate. They might bind to exactly the same regulatory elements, but due to the apparent absence of a TAD in Max [36], transcription might be repressed. Validation of this model will require the demonstration of clear regulation of a physiological promoter of a growth-related gene by c-Myc/Max. Although it is widely believed that Myc proteins function as transcriptional activators, this hypothesis has only been conclusively supported recently [57, 58]. A theory that c-Myc plays a role in DNA replication is not as well substantiated at this point. It is even possible that Myc might be involved in both transcription and replication. Although the function of these fascinating proteins has been enigmatic for a decade, the rate of progress in our understanding of Myc function is accelerating. Such progress will undoubtedly lead to a deeper appreciation of this protein, which lies at the crossroads of cellular proliferation and oncogenesis.


Oncogenes in human lung cancer.

The rapid pace of research in the genetics of human cancer will predictably render any review of the topic out of date by the time of its publication. Prospects for the near future will likely include the identification of a chromosome 3p gene(s) linked with the development of familial renal cancer and, perhaps, also lung cancer. In addition, the availability from the Human Genome Project of an increasing number of well-characterized markers will accelerate the search for additional human recessive oncogenes. Many questions still remain about the etiology of lung cancer and how to apply this information for patient care. For example, identification of the cell of origin for small cell and non-small cell lung cancers will facilitate our understanding of the development of these tumors and improve the possibilities for future preventive strategies. In addition, we now realize that these cancers arise from the sequential accumulation of multiple genetic mutations (Table 3; Fig. 1). Therefore, a central question is which of these targets are essential for the process of carcinogenesis, and whether there is a critical temporal order for this process with a defined premalignant phase in a discrete field of bronchial tissue. In addition, are there genetically inherited susceptibilities to the development of lung cancer (either directly or via variabilities in carcinogen metabolism) that could be accurately identified in the general population? Finally, is there a rate-limiting mutation and will the genetic correction of this defect suffice to restore growth regulation, or will the replacement of multiple gene products be required for tumor suppression? We are already witnessing the beginnings of the use of molecular diagnostic markers as a research tool for assigning prognostic information. The expression of neuroendocrine markers in non-small cell lung cancer has recently been applied as an indicator of the potential response to combination chemotherapy [15]. Similar methods are being applied to the expression of tumor suppressor genes or the presence of somatic mutations in dominant oncogenes such as the ras gene. However, the clinical benefit of this prognostic information with currently available treatment programs is still uncertain.(ABSTRACT TRUNCATED AT 400 WORDS)


[Clinical and anatomopathologic study of 800 cryptorchid testis].

662 children (800 cryptorchid testes) has been studied in order to check if the age and location were important factors in the degree of the lesion found in the biopsy. We have related in this survey the tubular fertility index (IFT) and tubular diameter (DT) to the age and location of the testes and we have not proved statistically their relationship, for which reason we do not advise to plan the age of surgery in terms of possible anatomopathologic damage.


Regional expression of the Wnt-3 gene in the developing mouse forebrain in relationship to diencephalic neuromeres.

During early vertebrate development, a series of neuromeres divides the central nervous system from the forebrain to the spinal cord. Here we examine in more detail the expression of Wnt-3, a member of the Wnt gene family of secreted proteins, in the developing diencephalon, in comparison to the expression of the homeobox gene Dlx-1. In 9.5-day mouse embryos, Wnt-3 is expressed in a restricted area of the diencephalon before any morphological signs of subdivisions appear. Around embryonic day 11.5, Wnt-3 expression becomes restricted to one of the neuromeres of the diencephalon, the dorsal thalamus. Dlx-1 is expressed in a non-overlapping area immediately anterior to and abutting the Wnt-3 expressing domain, corresponding to the ventral thalamus. In addition, Wnt-3 is expressed in the midbrain-hindbrain region. In the adult mouse, Wnt-3 and Dlx-1 are expressed in subsets of neural cells derived from the original areas of expression in the diencephalon. Taken together, our results suggest that Wnt-3 and Dlx-1 provide positional information for the regional specification of neuromeres in the forebrain. The continued expression of these genes in the adult mouse brain suggests a distinct role in the mature CNS.


Pancreaticogastrostomy after pancreatoduodenectomy.

The aim of this study was to evaluate the place of pancreaticogastrostomy (PG) in reducing pancreatic fistula after pancreatoduodenectomy. From January 1988 to June 1991, 32 consecutive patients (mean age, 57 years) were operated on, 25 for malignant disease (78%). The pancreatic remnant was normal in 17 patients (53%) and sclerotic in the others. There was one operative death (3.1%) unrelated to PG. Post-operative complications occurred in five patients (16%). Only two complications were related to PG: 1 patient had anastomotic intra-gastric bleeding and was reoperated on, 1 patient with a normal pancreatic remnant developed a pancreatic fistula (3.1%) treated conservatively. Reported series of PG, as well as our results, demonstrates that PG is associated with a dramatic decrease of both pancreatic fistula and mortality rates. The risk of anastomotic haemorrhage can be reduced by preventative ligation of submucosal gastric vessels. In conclusion, PG appears as a simple and reliable method of management of the pancreatic remnant after pancreatoduodenectomy.


[Detection of the HIV p24 antigen on lymphocyte membranes using flow cytometry].

The evaluation of the presence of p24 antigen on the membrane of peripheral blood mononuclear cells from 31 HIV infected individuals is presented. The study was performed by indirect immunofluorescence and flow cytometry and the data were analyzed by the Kolmogorov-Smirnov test. Values obtained [D/s(n)] result from the comparison of the fluorescence histograms of each sample with a control one. Cases showing p24 Ag on peripheral blood mononuclear cells also presented percentages of CD3, HLA-DR positive cells significantly higher than p24 negative ones. In addition, D/s(n) values were superior in symptomatic patients than in asymptomatic ones, which indicate the existence of a correlation between flow cytometry results, viral replication and clinical course. Nevertheless in this study, as well and in previous ones, a high degree of cross reactivity between the anti-p24 monoclonal antibody employed and normal lymphocytes has been observed. This reactivity is localized preferentially in the CD4 positive subset.


Effects of 2-deoxy-D-glucose on the glucose metabolism in Saccharomyces cerevisiae studied by multinuclear-NMR spectroscopy and biochemical methods.

The effects of various concentrations of deoxyglucose (DG) on the aerobic metabolism of glucose in glucose-grown repressed Saccharomyces cerevisiae cells were studied at 30 degrees C in a standard pyrophosphate medium containing 4.5 10(7) cells/ml. 31P-nuclear magnetic resonance (NMR) spectroscopy was used to monitor DG phosphorylation and the formation of polyphosphates. The production of soluble metabolites of glucose was evaluated by 13C- and 1H-NMR and biochemical techniques. The cells were aerobically incubated with 25 mM of glucose and various concentrations of DG (0, 5 and 10 mM) in order to determine the DG concentration leading to optimum of 2-deoxy-D-glucose 6-phosphate (DG6P) formation without over-inhibiting the synthesis of other metabolites. The production of DG6P increased by about 25% when the external DG concentration was doubled (from 5 to 10 mM). The formation of polyphosphates (polyP), on the other hand, was found to be mainly conditioned by the DG concentration. The amount of polyP decreased by a factor of four upon addition of 5 mM DG and became undetectable in the presence of 10 mM DG. The glucose consumption and the production of soluble metabolites of [1-13C]glucose were then evaluated as a function of time in both the absence and presence of 5 mM DG. The effect of DG is to decrease the glucose consumption and the formation of polyphosphates, ethanol, glycerol, trehalose, glutamate, aspartate and succinate while stimulating the formation of arginine and citrate. Upon co-addition of 25 mM glucose and 5 mM DG, the ratio between the initial rates of glucose consumption (0.16 mM/min) and DG6P production (0.027 mM/min) is about (5.9 +/- 1.2), not very different from the ratio of the initial concentration of glucose and DG (= 5.0). Therefore, hexokinase can phosphorylate deoxyglucose as well as glucose. However, after 100 min of incubation, the glucose concentration in the external medium decreased by about 64% while only 10% of DG was phosphorylated. DG6P was formed and quickly reached the limiting value about 30 min after co-addition of glucose and DG. Nevertheless, when the maximum quantity of DG6P was obtained, the DG consumption became negligible. By contrast, the glucose consumption and the production of ethanol and glycerol, although substantially reduced by about 42%, varied linearly with time up to 80 min of incubation. Thus even in the presence of an excess of DG, glycolysis is only slowed but not gradually or completely inhibited by DG.(ABSTRACT TRUNCATED AT 400 WORDS)


Effects of beta-blocking drugs in alcohol withdrawal: a double-blind comparative study with propranolol and diazepam.

Alcohol withdrawal is associated with a decrease in gamma-aminobutyric acid neurotransmission. This explains the efficacy of benzodiazepines. However, an increase in adrenergic activity may also play a part in alcohol withdrawal symptoms, suggesting a potential efficacy of beta-blocking drugs. A double-blind comparative study of propranolol and diazepam was carried out in 28 patients suffering from moderate uncomplicated alcohol withdrawal. Patients were treated for 15 days with either 75 mg of propranolol or 30 mg of diazepam. The results show that both drugs at the dosages used are equipotent in reducing physical withdrawal symptoms and anxiety symptoms. This suggests that most likely the central as well as the peripheral effects determine the clinical usefulness of propranolol in the management of alcohol withdrawal. However, propranolol is ineffective in preventing major motor seizures, suggesting that different neurobiological mechanisms underlie the alcohol withdrawal symptoms.


[The immunotropism of sunflower oil and starch in peroral administration to mice].

It has been shown that per os introduction of sunflower oil and starch to adult thymectomized mice resulted in reconstitution of the number of Thy-I+ spleen cells similar of the glutamic acid effect. Glutamic acid restored the immune response to sheep red blood cells (SRBC), however, sunflower oil and starch were not effective. The influence of these compounds on the immune response to SRBC in normal and sham-thymectomized mice was different: glutamic acid stimulated it, starch had no influence on the immune response, sunflower oil suppressed it.


Intracellular glutathione and its metabolizing enzyme activities in a metastatic variant melanoma cell line.

Levels of glutathione peroxidase (GSH-PX), glutathione-S-transferase (GST), gamma-glutamyl transpeptidase (gamma GTP) and glutathione reductase (GR) activities in the B16-F10 metastatic melanoma cell line are higher than those in non-metastatic B-16 murine melanoma cells. An inverse relationship was observed between the level of reduced glutathione (GSH) and metastatic capacity. Interferon (IFN), an antitumour and antimetastic agent, reduced the experimental metastatic capacity of B16-F10 cells while increasing the intracellular GSH content. This was associated with a fall in activity of GSH-metabolizing enzymes. These results suggest a correlation of intracellular GSH and its metabolizing enzymes with malignant transformation.


Biological and enzymatic features of human melanoma clones with different invasive potential.

Cell clones derived from a human melanoma metastasis selected for different integrin profiles were examined in vitro for invasive potential and biological and biochemical features potentially related to this process. Clones which expressed high levels of integrins showed high invasive potential, extracellular matrix degradation, and adhesion to gelatin-coated substrates. A correlation was also found between invasiveness and intracellular and extracellular plasminogen activator activity. Heparanase and collagenase type IV activities were apparently unrelated to invasiveness. gamma-Glutamyl transferase (GGT) activity was high in highly invasive clones, whereas melanin content was high in slightly invasive clones. Heterogeneity was also observed in cellular parameters such as cell dimensions, growth features and DNA index. The intrinsic biological and biochemical heterogeneity of a cell population derived from a single metastasis may be responsible for the different behaviour of clones, regardless of their invasive potential. Since slightly invasive cells are more differentiated than highly invasive cells, malignancy and differentiation are inversely correlated in such human melanoma clones.


DNA fingerprinting in the Chinese with an oligonucleotide probe (GTG)5.

DNA fingerprinting is a very powerful tool that enables specific identification of individuals. In order to study fingerprinting patterns in the Chinese, the oligonucleotide probe designed by Epplen et al was used. Peripheral blood leukocyte DNA was digested with restriction enzyme Hinf I and probed by (GTG)5. Normal females and males were tested; all revealed very different DNA fingerprints. Most of the discernible bands in the gel were polymorphic; there was no specific association between any polymorphic band and the sex of the individual tested. In situ gel-hybridization was used with the oligonucleotide probe, and after electrophoresis, all procedures could be completed within one working day. Three patients who had received bone marrow transplants from their respective siblings had the same fingerprints as their donors. The inheritance of all bands from either the father or mother proved parents' authenticity. DNA fingerprinting with a oligonucleotide probe is obviously a useful technique. Clinical application may include engraftment monitoring, zygosity determination and paternity testing.


Serum interleukin-2 and soluble interleukin-2 receptor in renal transplant recipients.

Interleukin-2 (IL-2) and soluble interleukin-2 receptor (sIL-2R), released during T-lymphocyte activation, were measured in serial samples of serum from 32 patients with renal allografts and other uremic patients. Patients undergoing chronic hemodialysis had elevated sIL-2R levels (1,801.93 +/- 753.23 U/mL) which dropped after stable renal transplantation (822 +/- 438 u/mL). However, these values were higher than those of a normal control group (397.3 +/- 84.5 u/mL, p < 0.01). Marked elevation of sIL-2R (1,503.78 +/- 640 u/mL) was noted in patients with acute rejection episodes compared to those in a stable allograft condition (p < 0.02) and those with cyclosporine nephrotoxicity (793.2 +/- 245.2 u/mL, p < 0.01), but returned to a stable level after successful anti-rejection treatment (745.91 +/- 345.8 u/mL, p < 0.01). Acute tubular necrosis and infection also showed a comparable rise in the sIL-2R level. The increase in sIL-2R with rejection was found one to four days earlier than the clinical diagnosis of acute rejection. There was a marked rise in the serum IL-2 level of uremic and post-transplant patients when compared to normal subjects (34.76 +/- 32.16 u/mL and 9.3 +/- 12.7 u/mL vs 4.38 +/- 3.38 u/mL, p < 0.001), but no significant differences were found between the IL-2 level of patients with acute rejection and cyclosporine nephrotoxicity or acute tubular necrosis (3.74 +/- 4.51 u/mL, 1.57 +/- 1.25 u/mL and 6.73 +/- 6.3 u/mL, p > 0.05). The diagnostic value of sIL-2R assay was more meaningful than that of IL-2.(ABSTRACT TRUNCATED AT 250 WORDS)


Microencapsulated pancreatic islets: a pathologic study.

Dog pancreatic islets isolated by an enzymatic digestion method were encapsulated in an alginate-poly L-lysine-alginate membrane. These microencapsulated pancreatic islets were cultured in vitro to study their ability of insulin secretion. Portions of these in vitro-cultured microencapsulated pancreatic islets were taken out for a viability dye exclusion study as well as for pathologic studies to correlate them with insulin secretion ability. We found that there was a strong correlation between them. Good insulin-secreting microcapsules showed well-preserved cell membranes and beta-cell granules. An in vitro culture for one to two days in RPMI-1640 made the islets more stable, the cellular surface became smoother and the beta-granules were in better shape. The microencapsulated pancreatic islets were also injected into the peritoneum of streptozotocin-induced diabetic CDF1 mice. Blood glucose levels dropped and stayed low for up to 60 days. But, when non-encapsulated dog pancreatic islets were used, the blood glucose levels remained low for only about 14 days. A small portion of the injected microcapsules were washed out at specific times for pathologic study. Up to 28 days after injection, only a few of the injected microcapsules showed pericapsular cellular infiltrate. However, after 56 days, most of the microcapsules showed dense pericapsular cellular infiltrate. Immunohistochemical analysis of these infiltrates showed that the majority of cells were fibroblasts and macrophages. Most of the cells located in the inner portion of the infiltrate were fibroblasts, while the macrophages were located mainly on the outer portion. Both scanning and transmission electron microscopy showed that the surface of the microcapsule outer wall was much smoother than the inner wall. The size of the microcapsules was approximately 0.6-0.8 mm and the thickness of the wall measured around 10 nm. The smaller the microcapsule is, the less chance there is of rupture with release of the xenographic islets. Once the wall of the transplanted microcapsules was ruptured, the inner surface showed more increased inflammatory cell and fibroblast infiltration than the outer surface.


Reproducibility of the measurement of insulin sensitivity by the modified insulin suppression test.

A significant increase in insulin resistance has been implicated in many human diseases and the absence of a simple, accurate, reproducible measurement of in vivo insulin sensitivity has become a major concern. In order to evaluate the reproducibility of insulin sensitivity measured by the modified insulin suppression test, 12 healthy young Chinese men were subjected to the same test two weeks apart. After three days on a standard diet and an overnight fast, somatostatin (350 micrograms/h), insulin (25 mU/m2/min) and glucose (240 mg/m2/min) were infused concomitantly for three hours. A steady state plasma glucose (SSPG) concentration achieved during the last 30 minutes of infusion represented the measurement of insulin sensitivity. Comparisons between the metabolic clearance rate of insulin (MCRi), plasma total triglyceride and lipoprotein cholesterol fractions on two different days were carried out. The results indicated that mean SSPG concentrations on Day 1 (5.73 +/- 0.43 mmol/L) correlated with mean SSPG concentrations on Day 14 (5.51 +/- 0.38 mmol/L; r = 0.82, p < 0.002). The relationship slope did not differ from 1 (0.74, p > 0.05), the intercept was close to the origin (1.24 mmol/L, p > 0.05) and the mean coefficient of intra-individual variation was 10.3%. There was no difference between the MCRi for Day 1 and Day 14 (529 +/- 26 vs 526 +/- 24 mL/m2/min, p = NS), with a mean coefficient of intra-individual variation of 6.9%.(ABSTRACT TRUNCATED AT 250 WORDS)


Congenital hypothyroidism in Taiwan: experience before mass screening.

The clinical data of 91 patients with congenital hypothyroidism, not detected by neonatal screening, were reviewed. Our results disclosed that congenital hypothyroidism in Taiwan is more common in girls than boys, with a female to male ratio of 1.8:1. The majority of our patients were from a first pregnancy. Only 4.8% of our patients had a history of prolonged gestation, and the median value of birth weight in these patients was 3,300 g. Both of these data are comparable to those of the normal population. Short stature, constipation, dry skin and periorbital edema were the most common symptoms and signs. An ectopic thyroid gland was the most common type of congenital hypothyroidism in our series. The conditions of most of our patients were detected after the age of three months; poor mental prognosis was expected. However, it is hoped that neonatal screening programs for congenital hypothyroidism in Taiwan can improve such situations.


Morbidity and mortality trends of pulmonary tuberculosis in Taiwan.

An epidemiologic study of pulmonary tuberculosis was carried out to examine the secular trend and geographic variations of the incidence, prevalence and mortality of pulmonary tuberculosis in Taiwan. The prevalence and incidence rates declined-steadily from 1910 to 1980, but have remained unchanged during the last decade. Mortality also showed an 8.0% annual decrease from 1974 to 1986, but has remained constant without any apparent changes in recent years. Mortality for pulmonary tuberculosis is much higher in the aboriginal areas, where patients have a poor adherence to treatment and where a high prevalence of alcohol abuse exists. The mortality for pulmonary tuberculosis in Taiwan is higher than that in most developed countries, such as the U.K. and the U.S.A., but is lower than that in mainland China and the Republic of Korea.