Mahidol University Annual Research Abstracts
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Thinkpad Mahidol University Annual Research Abstracts, Vol.28, 2001
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Laptop Computers 1Neuro-Behavioural Biology Center, Institute of Science and Technology for Research and Development, and 2Faculty of Public Health, Mahidol University, and 3Prasart Neurological Hospital and Institute, Ministry of Public Health, Thailand.
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Laptop Computer Key words: Sudden Unexplained Nocturnal Death Syndrome (SUNDS), Lai Tai, Monoamine oxidase enzymes A and B, Immunocytochemical staining, Human brainstem.
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Desktop Computer Previous investigations on the putative patho-physiological mechanism of Sudden Unexplained Nocturnal Death Syndrome (SUNDS or Lai Tai in local northeastern Thai dialect) in our center have consistently indicated that the cause of death in SUNDS may involve a genetic predisposing factor related to the deletion or mutation of the gene for the monoamine oxidase A (MAO-A) which is located on the human X-chromosome. The objective of the present investigation is to study the distribution and localization of MAO enzymes in the brainstem of the victims of SUNDS as compared to the normal adult Thai males. Four autopsy verified SUNDS brains from Thai northeastern males ages ranged between 22 and 47 years old who died with symptoms, and the autopsy and pathological findings consistent with the definition of SUNDS were obtained from the autopsy room of Ramathibodi Hospital. In addition, other four normal brains were obtained from adult Thai males who died from severe injuries or other diseases which did not appear to affect the brain. Serial cryostat cross sections of the brainstem were incubated with monoclonal antibodies specifically against MAO-A, MAO-B, and MAO-AB, and the presence of the MAO enzymes was demonstrated by the Horseradish Peroxidase (HRP) reaction products in neurons which were also counter stained with Nisslss stain to allow identification and localization of various specific neuronal groups. Immuno-positive HRP labeled neurons were consistently found in brainstem sections of all four SUNDS brains after incubation with specific monoclonal antibodies against MAO-A, MAO-B and MAO-AB. The distribution and localization of these labeled brainstem neurons from SUNDS victims were remarkably similar to those found in the normal adult Thai males. On the contrary, the density of neurons in the substantia nigra (SN), and the total count of neurons in the locus ceruleus (LC) , the sub-ceruleus nucleus (SC), the mid-line dorsal raphe (DR) nucleus, and the medullary (M) neurons from the brainstem of SUNDS victims were statistically significant ( p<0.01) lower than those in the corresponding areas of the normal adult Thai males. The finding clearly indicates that both MAO-A and MOA-B enzymes are expressed in all SUNDS brains obtained for the present study. It is quite possible that the monoclonal antibody against MAO-A used in the present study may not be specific toward the site of the mutation on the enzyme molecule. However, the significant reduction of the density or the total population of MAO labeled neurons in many areas of the brainstem which regulate vital functions such as sleep-arousal, cardiovascular and pulmonary functions, in SUNDS as compared to the normal brains suggest that the lower amount of MAO in SUNDS brain may be an important disposing factor during certain critical states of sleep which are related to high levels of biogenic monoamines in the brainstem beyond the catalytic capability of the well-known alternating pathway utilizing Catecho-O-Methy-Transferase (COMT) enzyme. Further definitive supporting evidence for this hypothesis in the future may come from direct assays of MAO enzymatic activities and levels of specific metabolites of monoamines in the brain of SUNDS as compared to normal brain.
Notebooks (Supported in Parts by Research Grants from Mahidol University and the Ministry of Public Health)
Lenovo Current Claim: There is a significant reduction of the density or the total population of MAO labeled neurons in many areas of the brainstem which regulate vital functions such as sleep-arousal, cardiovascular and pulmonary functions, in SUNDS as compared to the normal brains.
Hard Drive (Presented at the 3rd International Congress of the Asian Sleep Research Society on the New Millennium of Sleep Research in Asia, December 3-7, 2000, Bangkok, Thailand, and published in the Program and Abstracts page A172.)
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Laptop Parts DELTA- AND MU-OPIOID RECEPTORS IN BOVINE PINEALOCYTES (NO. 937)
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Hard Drives Saiphon Sawlom1, Piyarat Govitrapong1, Banthit Chetsawang1, Naiphinich Kotchabhakdi1 and M.Ebadi2
Electronics 1Neuro-Behavioral Biology Center, Institute of Science and Technology for Research and Development, Mahidol University, Salaya Campus, Nakornpathom 73170, Thailand; 2Department of Pharmacology, Physiology and Therapeutics, University of North Dakota, School of Medicine and Health Science, Grand Forks, ND 58202-9037, USA.
Canon Key words: melatonin, pineal gland, opioid receptor, N-acetyltransferase
Desktop Pc Several lines of evidence have reported that there are opioidergic nerve fibers innervating the pineal gland via opioid receptors (1). Our previous studies (2) have identified opioid receptors by using a nonselective radio-ligand, [3H]-Diprenorphine that revealed the existence of opioid receptors on bovine pinealocyte membranes. To further characterize and localize opioid receptor subtypes using radio-ligand-binding technique with specific receptor subtypes, [3H]-DPDPE was used as a selective ligand for delta opioid receptor subtypes and [3H]-DAMGO for mu opioid receptor subtypes. We have isolated the bovine pinealocytes by mechanical manipulation of cell extraction. The tissue-dependent specific binding in bovine pinealocytes increased linearly as a function of tissue concentrations,[3H]-DPDPE binding from 0.05 to 0.30 mg and [3H]-DAMGO binding from 0.10 to 0.35 mg. By Scatchard analysis of specific opioid binding on bovine pinealocytes, the results provide receptor density (Bmax) and association equilibrium constant (Kd) values as follows: [3H]-DPDPE binding was obtained Kd and Bmax as 1.286+0.61 nM and 553.095+24.24 fmol/mg protein and [3H]-DAMGO binding as 1.19+0.44 nM and 6.34+1.27 fmol/mg protein. Thus, the present results indicate that a majority of opioid receptor sites are delta opioid receptors, with a minority being mu opioid receptor sites and reveal an interaction among opioids and pineal gland.
Desktop Computers Acknowledgement: This work was supported by National Institute of Neurological Disorder and Stroke, USA, 1R01 NS-40160-1 Grant.
Think Pad (Presented at the 3rd International Congress of the Asian Sleep Research Society on the New Millennium of Sleep Research in Asia, December 3-7, 2000, Bangkok, Thailand, and published in the Program and Abstracts page A173.)
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