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Biomedical subjects

H Hall

Publications and source records attributed to H Hall.

At least 19 recordsLinked to original sources

NK cells in autoimmune disease.

The role of NK cells in autoimmunity has not been extensively studied. Speaking for a disease-promoting role for NK cells in autoimmune diseases are recent results suggesting that IFN-gamma production by NK cells may help adaptive immune responses diverge in the direction of a Th1 response. NK cells may also be involved in direct killing of tissue cells, which could lead to acceleration of autoimmunity. However, NK cells have also been shown to protect from some autoimmune diseases. A possible reason for this discrepancy may lie in the capacity of NK cells also to produce Th2 cytokines, which could downregulate the Th1 responses that are common in autoimmune disorders. Thus there is at present no coherent view on the role of NK cells in autoimmunity, and more work is needed to clarify why NK cells in some cases aggravate disease and in some cases protect from disease.

Animals↗

Kinetics of the reaction of U(VI) with benzene-1,2-diphosphonic acid.

The kinetics of the reaction between dioxouranium(VI) and benzene-1,2-diphosphonic acid (BzDPA) has been investigated by stopped-flow spectrophotometry. The rate of reaction of uranyl ions with Arsenazo III (2,7-bis(2,2'-arsonophenylazo)-1,8-dihydroxynaphthalene-3,6-disulfonic acid) in 50:50 methanol-water solutions was also determined. Both formation and dissociation rate constants for the 1:1 complex between uranyl-BzDPA in acidic solutions were resolved. To gain insight into the effect of solvation on the progress of the reaction, the system was studied in triply distilled water, in 50:50 methanol-water, 80:20 methanol-water and in 50:50 tert-butanol-water as a function of temperature at pH 1.0. The rates of complex formation and dissociation reactions decrease as methanol substitutes for water in the medium and further decrease as tert-butanol replaces methanol as co-solvent. Activation parameters are most consistent with an associative process governing the progress of both complex formation and dissociation reactions. Introduction of alcoholic co-solvents results in notably more negative activation entropies for both complex formation and dissociation reactions, while the activation enthalpies are only slightly reduced in the mixed methanol-water medium. These results are compared with the kinetic features of other U(VI) systems.

Journal Article↗

The serotonin-2A receptor gene locus does not contain common polymorphism affecting mRNA levels in adult brain.

The serotonin-2A (HTR2A) receptor is a molecule of particular interest in biological psychiatry, as it is an important target for psychotropic drugs, and altered HTR2A expression has been found in several neuropsychiatric conditions, including depression and schizophrenia. Genetic association has been reported between a synonymous 102T/C polymorphism in the gene encoding HTR2A and a number of clinical phenotypes, including schizophrenia, clozapine response, psychotic symptoms in Alzheimer's disease and certain features of depression. Given that there are no known effects of the 102T/C polymorphism on the structure of the receptor, attention has switched to the possibility that the observations of both altered expression and genetic association point to functional sequence variants that alter expression of the HTR2A gene. Moreover, data have been presented recently suggesting that mRNAs containing the 102T- and C-alleles are differentially expressed. This suggests a direct effect of the variant itself on mRNA levels, or the influence of a distinct regulatory variant, such as the -1438A/G promoter polymorphism, with which it is in perfect linkage disequilibrium. The present study tested this hypothesis by employing a highly accurate quantitative allele- specific primer extension assay to measure the relative expression of brain mRNAs carrying each allele in 23 individuals heterozygous for the 102T/C polymorphism. Comparison between allele ratios derived from genomic DNA and mRNA from several cortical regions revealed that the 102C- and T-alleles are expressed identically. Furthermore, the absence of any interindividual variability in relative mRNA allele ratio suggests that the HTR2A locus is unlikely to contain common polymorphisms or epigenetic modification that alter HTR2A mRNA levels in adult brain, and essentially exclude such phenomena as a potential explanation for the altered expression and genetic associations that have been reported to date.

Adult↗

A laboratory model to evaluate cutout resistance of implants for pertrochanteric fracture fixation.

OBJECTIVES: To establish a laboratory model of implant cutout, which can evaluate the effect of implant design on cutout resistance in a clinically realistic "worst case" scenario. SETTING: Orthopaedic biomechanics laboratory. DESIGN: Implant cutout was simulated in an unstable pertrochanteric fracture model, which accounted for dynamic loading, osteoporotic bone, and a defined implant offset. For model characterization, lag screw cutout was simulated in human cadaveric specimens and in polyurethane foam surrogates. Subsequently, foam surrogates were used to determine differences in cutout resistance between 2 common lag screws (dynamic hip screw, Gamma) and 2 novel blade-type implant designs (dynamic helical hip system, trochanteric fixation nail). MAIN OUTCOME MEASURES: Implant migration was continuously recorded with a spatial motion tracking system as a function of the applied loading cycles. In addition, the total number of loading cycles to cutout failure was determined for specific load amplitudes. RESULTS: Implant migration in polyurethane surrogates closely correlated with that in cadaveric specimens, but yielded higher reproducibility and consistent cutout failure. The cutout model was able to delineate significant differences in cutout resistance between specific implant designs. At any of 4 load amplitudes (0.8 kN, 1.0 kN, 1.2 kN, 1.4 kN) dynamic hip screw lag screws failed earliest. The gamma nail lag screw could sustain significantly more loading cycles than the dynamic hip screw. Of all implants, trochanteric fixation nail implants demonstrated the highest cutout resistance. CONCLUSIONS: Implant design can significantly affect the fixation strength and cutout resistance of implants for pertrochanteric fracture fixation. The novel cutout model can predict differences in cutout resistance between distinct implant designs.

Aged↗

Pathogenicity and antigenicity of a new influenza A (H5N1) virus isolated from duck meat.

Avian influenza A viruses are the ancestral origin of all human influenza viruses. The outbreak of highly pathogenic (HP) avian H5N1 in Hong Kong in 1997 highlighted the potential of these viruses to infect and cause severe disease in humans. Since 1999, HP H5N1 viruses were isolated several times from domestic poultry in Asia. In 2001, a HP H5N1 virus, A/Duck/Anyang/AVL-1/2001 (Dk/Anyang), was isolated from imported frozen duck meat in Korea. Because of this novel source of HP H5N1 virus isolation, concerns were raised about the potential for human exposure and infection; we therefore compared the Dk/Anyang virus with HP H5N1 viruses isolated from humans in 1997 in terms of antigenicity and pathogenicity for mammals. At high doses, Dk/Anyang virus caused up to 50% mortality in BALB/c mice, was isolated from the brains and lymphoid organs of mice, and caused lymphopenia. Overall Dk/Anyang virus was substantially less pathogenic for mice than the H5N1 virus isolated from a fatal human case in 1997. Likewise, Dk/Anyang virus was apathogenic for ferrets. Dk/Anyang virus was antigenically distinguishable by hemagglutination-inhibition (HI) assay from human H5N1 viruses isolated in 1997 and avian H5N1 viruses isolated in 2001 in Hong Kong. Nevertheless, prior infection with Dk/Anyang virus protected mice from death after secondary infection with HP human H5N1 viruses. These results indicate that compared with HP human H5N1 viruses, Dk/Anyang virus is substantially less pathogenic for mammalian species. Nevertheless, the novel source of isolation of this avian H5N1 virus must be considered when evaluating the potential risk to public health.

Animals↗

Pathogenesis of and immunity to a new influenza A (H5N1) virus isolated from duck meat.

The outbreak of avian influenza H5N1 in Hong Kong in 1997 raised concerns about the potential for the H5 subtype to cause a human pandemic. In 2001 a new H5N1 virus, A/Duck Meat/Anyang/AVL-1/2001 (A/Dkmt), was isolated from imported duck meat in Korea. The pathogenesis of this virus was investigated in mice. A/Dkmt virus had low infectivity but was lethal for mice at high doses, and at lethal doses, the virus replicated in the brains of infected mice. A/Dkmt virus cross-reacted poorly with ferret antisera raised against human H5N1 viruses, but prior infection with A/Dkmt virus protected mice from death after secondary infection with human H5N1 virus.

Animals↗

Synthesis, biological and autoradiographic evaluation of a novel Tc-99m radioligand derived from WAY 100635 with high affinity for the 5-HT(1A) receptor and the alpha1-adrenergic receptor.

This paper reports the synthesis, biological evaluation, in vitro and ex vivo autoradiography of the first Tc-99m ligand with subnanomolar affinity for the 5-HT(1A) receptor and a remarkably high affinity for the alpha1-adrenergic receptor. The neutral "3+1" mixed-ligand complex combines 4-(6-mercaptohexyl)-1-(2-methoxyphenyl)piperazine as monodentate and 3-(N-methyl)azapentane-1,5-dithiol as tridentate unit with oxotechnetium(V). The analogous rhenium complex was synthesized for complete structural characterization and used in receptor binding assays. In competition experiments both complexes display subnanomolar affinity for the 5-HT(1A) receptor (IC(50)0.24 nM for Re, 0.13 nM for Tc) but also very high affinities for the alpha1-adrenergic receptor (IC(50) 0.05 nM for Re, 0.03 nM for Tc). Biodistribution studies show a brain uptake in rat of 0.22% ID five minutes post injection. In vitro autoradiographic studies in rat brain and postmortem human brain indicate accumulation of the Tc-99m complex in brain areas which are rich in 5-HT(1A) receptors or in alpha1-adrenergic receptors. This in vitro enrichment can be blocked respectively by the 5-HT(1A) receptor agonist 8-OH-DPAT or by prazosin hydrochloride, an alpha1-adrenergic receptor antagonist. Ex vivo autoradiographic studies in rats show a slight accumulation of the Tc-99m complex in 5-HT(1A) receptor-rich areas of the brain, which could not be blocked, as well as in regions rich in alpha1-adrenergic receptors, which could be blocked by prazosin hydrochloride.

Animals↗

Synthesis and biological evaluation of technetium(III) mixed-ligand complexes with high affinity for the cerebral 5-HT(1A) receptor and the alpha1-adrenergic receptor.

Tc(III) and Re(III) complexes [M(NS(3))(CNR)] (M = Re, 99mTc, NS(3) = 2,2',2"-nitrilotris(ethanethiol), CNR = functionalized isocyanide bearing a derivative of WAY 100635) have been synthesized and characterized. Re was used as Tc surrogate for chemical characterization and in vitro receptor-binding studies. For two representatives subnanomolar affinities for the 5-HT(1A) as well as for the alpha1-adrenergic receptor were reached. Biodistribution studies in rats of the 99mTc complexes showed brain uptakes between 0.3 and 0.5% ID/organ (5 min p.i.). In vitro autoradiography of one 99mTc representative in sections of post mortem human brain indicate its accumulation in 5-HT(1A) receptor-rich brain regions. However, addition of the specific 5-HT(1A) receptor agonist 8-OH-DPAT as well as the alpha1-adrenoceptor antagonist prazosin could not substantially block this tracer accumulation. A preliminary SPET study in a monkey showed negligible brain uptake.

Animals↗

Autoradiographic evaluation of [11C]vinpocetine binding in the human postmortem brain.

The main objective ofthe study was to evaluate with autoradiographic technique whether or not [11C]vinpocetine, a compound widely used in the prevention and treatment of cerebrovascular diseases (Cavinton, Gedeon Richter Ltd., Budapest), binds to specific sites in the human brain in post mortem human brain sections. Binding was assessed under four conditions: the incubation was performed using Tris-HCl buffer with or without the addition of salts (0.1% (weight/vol) ascorbic acid, 120 mM NaCl, 5 mM KCl, 2 mM CaCl2 and 1 mM MgCl2), with or without the addition of excess (10 microM) unlabelled vinpocetine. Measurements on digitized autoradiograms indicated that [11C]vinpocetine labelled all grey matter areas in the human brain to a similar extent and no significantly heterogeneous binding could be demonstrated among cortical or subcortical regions. The addition of excess unlabelled vinpocetine lowered the binding slightly in all regions. Although these results indicate that [11C]vinpocetine does not bind to human brain transmitter receptors or transporters with a high affinity (Ki < 10 nM), it cannot be ruled out that the compound binds to receptors and/or transporters with lower affinity.

Autoradiography↗

Autoradiographic mapping of 5-HT(1B) and 5-HT(1D) receptors in the post mortem human brain using [(3)H]GR 125743.

The distribution of 5-HT(1B) and 5-HT(1D) receptors in the human post mortem brain was examined using whole hemisphere autoradiography and the radioligand [(3)H]GR 125743. [(3)H]GR 125743 binding was highest in the substantia nigra and the globus pallidus. Lower levels were detected in the striatum, with the highest densities in the ventromedial parts. In the amygdala, the hippocampus, the septal region and the hypothalamus, lower [(3)H]GR 125743 binding was observed, reflecting low densities of 5-HT(1B/1D) receptors. In the cerebral cortex, binding was similar in most regions, although restricted parts of the medial occipital cortex were markedly more densely labeled. Binding densities were very low in the cerebellar cortex and in the thalamus. Two methods were used to distinguish between the two receptor subtypes, the first using ketanserin to block 5-HT(1D) receptors and the second using SB 224289 to inhibit 5-HT(1B) receptor binding. The autoradiograms indicated that in the human brain, the 5-HT(1B) receptor is much more abundant than the 5-HT(1D) receptor, which seemed to occur only in low amounts mainly in the ventral pallidum. Although [(3)H]GR 125743 is a suitable radioligand to examine the distribution of 5-HT(1B) receptors in the human brain in vitro, the selectivities of ketanserin and SB 224289 are not sufficiently high to give definite evidence for the occurrence of the 5-HT(1D) receptor in the human brain.

Basal Ganglia↗

Molecular properties of fibrin-based matrices for promotion of angiogenesis in vitro.

The molecular properties of fibrin-based matrices, such as fibrillar structure and covalent modifications with adhesion domains, influence the angiogenic behavior of human umbilical vein endothelial cells (HUVECs) in vitro. The fibrillar structure of fibrin-based matrices was influenced by pH but not by covalent incorporation of exogenous adhesion domains. Native fibrin-based matrices polymerized at pH 10 formed organized and longitudinally oriented fibrin fibrils, which provided a good angiogenic substrate for endothelial cells. Furthermore, upon covalent incorporation of the model ligand L1Ig6, which binds to the integrin most prominently expressed on the surface of angiogenic endothelial cells, alpha(v)beta3, these matrices became angiogenesis-promoting when polymerized at physiological pH. The amount of incorporation of L1Ig6 into the matrices depended on the fibrinogen concentration on all three fibrin chains. Soluble forms of L1Ig6 diffused rapidly out of the matrix. Most important, L1Ig6-modified matrices were very specific in inducing the angiogenic phenotype of HUVECs, whereas control cells did not differentiate on these matrices. Our results indicate that artificial extracellular matrices can influence cell behavior in two ways. One way is based on the three-dimensional fibril structure of the matrix molecules themselves, and the other is due to providing specific binding sites for direct cell-matrix interactions that lead to the activation of second-messenger cascades and thus promoting angiogenic differentiation.

Binding Sites↗

Measurement of the striatal dopamine transporter density and heterogeneity in type 1 alcoholics using human whole hemisphere autoradiography.

Dopaminergic mechanisms are involved in the positive reinforcing and addicting effects of alcohol. Positron emission tomography (PET) and single photon emission tomography (SPET) studies have indicated alterations in striatal dopamine transporters (DAT) and in presynaptic dopamine (DA) function in alcoholics, although also contradictory results have been reported. Normal variations in blood flow, metabolism, and receptor densities are apparently important to brain function. Such variations are known to decrease during pathophysiological processes, such as epilepsy, whereas normal receptor distributions are broadly heterogenous. We evaluated the densities and heterogeneities of striatal DAT in 8 adult-onset, Cloninger type I alcoholics and 10 controls using [125I]N-(3-iodoprop-2E-enyl)-2beta-carbomethoxy-3beta- (4'-methylphenyl)nortropane ([125I]PE2I) as a ligand for human postmortem whole hemisphere autoradiography, which provided high resolution images of the brain when compared with in vivo PET and SPET. The mean density and heterogeneity of DAT were markedly lower in the alcoholics. A significant linear correlation existed between DAT density and heterogeneity, as well as between DAT densities in the nucleus accumbens and in the dorsal striatum (caudate and putamen) in alcoholics, but not consistently in controls. The observed low DAT density and heterogeneity in the dorsal striatum suggest that type 1 alcoholics may have a dysfunctional DA system. These data indicate that human whole hemisphere autoradiography with the analysis of binding heterogeneity may be a relevant tool to measure pathological processes in the brain.

Alcoholism↗

Long-term follow-up of functional outcomes and radiographic changes at adjacent levels following lumbar spine fusion for degenerative disc disease.

There are very few studies with more than 20 years' follow-up of lumbar spine fusions for disc degeneration. Currently, there is a lot of interest in the subject of degenerative changes above the level of fusion; this study is concerned with such changes in the very long term (30 years). Twenty-eight patients showing sound fusion on radiographs following posterior midline spinal fusion performed by a single surgeon between 1968 and 1970 were compared with an age- and gender-matched group of 28 patients who had undergone surgery for degenerative disc disease without fusion during the same period, by the same surgeon and using similar criteria for evaluation (Short Form 36 and Oswestry Disability Index; functional testing using self-paced walk and timed up-and-go; flexion and extension lateral radiographs of the lumbar spine). In this study, the incidence of radiographic changes at levels above the level of previous involvement was twice as high in the fusion group as in the non-fusion group. However, there was no statistically significant difference between the two groups in the outcomes measured using validated scales and functional testing. The study emphasises the importance of complete evaluation of these patients using validated outcome measurement instruments against the background of radiographic changes and subjective assessment of back pain. It also shows that radiographic changes do not necessarily mean functional impairment in all patients following lumbar spine fusion for degenerative disc disease.

Case-Control Studies↗

An overview on functional receptor autoradiography using [35S]GTPgammaS.

[35S]GTPgammaS binding autoradiography is a novel method to study the distribution and function of neurotransmitter receptors in tissue sections. This technique unifies the advantages of receptor-autoradiography and [35S]GTPgammaS binding, providing anatomical and functional information at the same time. Due to these two main features, it can also be called 'functional autoradiography'. [35S]GTPgammaS binding has long been used to study the first step of the intracellular signaling pathway, but until the mid 1990s it has only been performed on cell membrane extracts. Functional autoradiography evolved from this biochemical assay and ligand autoradiography, and is based on the increase in guanine nucleotide exchange at G-proteins upon agonist stimulation. With the technique, activation of G-protein-coupled receptors upon agonist binding can be detected, and, at the same time, the location of activated receptors can also be visualized. Thus only those presumably active G-protein-coupled receptors are visualized that can be involved in signal transduction. In the past 5 years the technique has become more and more frequently used in neuroscience, and it has been adapted to several receptors in different species, including also the human brain. [35S]GTPgammaS binding autoradiography can be used to describe the distribution of G-protein-coupled receptors. Some inferences on their coupling efficiency can also be drawn. Besides the localization of ligand binding sites, it provides information on the action of the ligand on the receptor: agonists, antagonists, and inverse agonists can clearly be distinguished. Moreover, [35S]GTPgammaS binding autoradiography can successfully be combined with other in vitro assays, like receptor autoradiography, in situ hybridization histochemistry, or even with biochemical and electrophysiological experiments. This review presents an overview on the history and the development of this technique. Its main advantages and limitations are summarized, together with a few basic technical questions. A number of experiments performed with [35S]GTPgammaS binding autoradiography so far, and some possible applications for the future, are also reviewed.

Animals↗

New halogenated [11C]WAY analogues, [11C]6FPWAY and [11C]6BPWAY--radiosynthesis and assessment as radioligands for the study of brain 5-HT1A receptors in living monkey.

[Carbonyl-(11)C]WAY-100635 ([(11)C]WAY) is an established radioligand for the study of brain serotonin(1A) (5-HT(1A)) receptors in living animals and humans with positron emission tomography (PET). There is a recognised need to develop halogenated ligands for 5-HT(1A) receptors, either for labelling with longer-lived fluorine-18 for more widespread application with PET or with iodine-123 for application with single photon emission tomography (SPET). Here we used autoradiography and PET to assess two new halogenated analogues of WAY, namely 6BPWAY and 6FPWAY [N-(2-(1-(4-(2-methoxyphenyl)-piperazinyl)ethyl))-N-(2-(6-bromo-/fluoro-pyridinyl))cyclohexanecarboxamide] as prospective radioligands, initially using carbon-11 as the radiolabel. Labelling of 6BPWAY and 6FPWAY with carbon-11 was accomplished by acylation of the corresponding secondary amine precursors with [carbonyl-(11)C]cyclohexanecarbonyl chloride. After incubation of human brain crysections with [(11)C]6BPWAY or [(11)C]6FPWAY, the highest accumulation of radioactivity was observed in cortical areas and the hippocampal formation. Both radioligands had high nonspecific binding. There was a rapid accumulation of radioactivity in the monkey brain after intravenous injection of [(11)C]6BPWAY and [(11)C]6FPWAY. High accumulation of radioactivity was observed in the frontal and temporal cortex and the raphe nuclei, areas known to contain a high density of 5-HT(1A) receptors. The ratios of radioactivity in receptor-rich temporal cortex to that in receptor-poor cerebellum at peak equilibrium were 1.9 (at 10 min) and 3.0 at (at 20 min) for [(11)C]6BPWAY and [(11)C]6FPWAY, respectively. In pretreatment experiments with high doses of unlabelled WAY, the level of radioactivity in the frontal and temporal cortex and the raphe nuclei was reduced to the same level as in the cerebellum. Radioactive metabolites of [(11)C]6FPWAY appeared at a rate similar to those for [(11)C]WAY, with 17% of the radioactivity in plasma represented by unchanged radioligand after 40 min. Radioactive metabolites of [(11)C]6BPWAY appeared much more slowly. At 40 min after injection 45% of the radioactivity in plasma still represented unchanged radioligand. The results indicate that 6-pyridinyl radiohalogented analogues of WAY are new leads to radioligands for PET or SPET.

Aminopyridines↗

Dopamine D(2)/D(3)-receptor and transporter densities in nucleus accumbens and amygdala of type 1 and 2 alcoholics.

Alcohol acts through mechanisms involving the brain neurotransmitter dopamine (DA) with the nucleus accumbens as the key zone for mediating these effects. We evaluated the densities of DA D(2)/D(3) receptors and transporters in the nucleus accumbens and amygdala of post-mortem human brains by using [(125)l]epidepride and [(125)I]PE2I as radioligands in whole hemispheric autoradiography of Cloninger type 1 and 2 alcoholics and healthy controls. When compared with controls, the mean binding of [(125)I]epidepride to DA D(2)/D(3) receptors was 20% lower in the nucleus accumbens and 41% lower in the amygdala, and [(125)I]PE2I binding to DA transporters in the nucleus accumbens was 39% lower in type 1 alcoholics. These data indicate that dopaminergic functions in these limbic areas may be impaired among type 1 alcoholics, due to the substantially lower number of receptor sites. Our results suggest that such a reduction may result in the chronic overuse of alcohol as an attempt to stimulate DA function.

Adult↗

The scientific study of unusual rapid wound healing: a case report.

This paper examines in a Western medical setting the claim made by a Middle Eastern school of Sufism that its members can attain unusually rapid wound healing from deliberately caused bodily damage. The demonstration involved a Sufi practitioner inserting an unsterilized metal skewer through one side of the cheek area of the face (lateral buccal) and out through the other side. The insertion was observed by Western scientists. The left facial puncture healed within 2 minutes; the right facial puncture was three-quarters healed after 8 hours. According to the practitioner, the piercing was not associated with subjective pain. Radiological, immunological, and electroencephalograph (EEG) recordings were taken before and after the insertion. A random event generator (REG) was also run during the demonstration. Radiological and axial CT images documented the presence of the metal skewer through the cheeks. EEG findings were inconclusive. Immunological recordings showed no alterations. There was a trend toward REG non-randomness and, therefore, negentropy or increased complexity and order, which may be associated with the re-establishment of tissue integrity. Such rapid wound healing has therapeutic implications for new treatments of serious medical and pain-related problems as well as implications for the study of "paranormal" healing phenomena.

Adult↗

Infection of a child in Hong Kong by an influenza A H3N2 virus closely related to viruses circulating in European pigs.

Influenza virus A/Hong Kong/1774/99, isolated from a young child with mild influenza, was shown to be similar in its antigenic and genetic characteristics to H3N2 viruses circulating in pigs in Europe during the 1990s and in particular to be closely related to viruses isolated from two children in the Netherlands in 1993. Similar viruses had previously not been identified outside Europe. Although there is little evidence as to how the child contracted the infection, it appears likely that pigs in southern China were the source of infection. Characteristics shared with the European swine viruses include resistance to the anti-influenza drugs amantadine and rimantadine. Thus not only does this incident once again highlight the potential of pigs as a source of novel human influenza viruses, but also indicates the potential for emergence of amantadine-resistant human viruses.

Amantadine↗