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Biomedical subjects

M Nakazato

Publications and source records attributed to M Nakazato.

At least 73 records · Page 4Linked to original sources
PubMed1999

Narcolepsy in orexin knockout mice: molecular genetics of sleep regulation.

Neurons containing the neuropeptide orexin (hypocretin) are located exclusively in the lateral hypothalamus and send axons to numerous regions throughout the central nervous system, including the major nuclei implicated in sleep regulation. Here, we report that, by behavioral and electroencephalographic criteria, orexin knockout mice exhibit a phenotype strikingly similar to human narcolepsy patients, as well as canarc-1 mutant dogs, the only known monogenic model of narcolepsy. Moreover, modafinil, an anti-narcoleptic drug with ill-defined mechanisms of action, activates orexin-containing neurons. We propose that orexin regulates sleep/wakefulness states, and that orexin knockout mice are a model of human narcolepsy, a disorder characterized primarily by rapid eye movement (REM) sleep dysregulation.

Age of Onset↗
PubMed1999

Widespread distribution of orexin in rat brain and its regulation upon fasting.

Orexins A and B, novel hypothalamic peptides encoded by a single mRNA transcript, stimulate food intake. Two antisera specific for the individual rat orexins were prepared and sensitive RIAs developed. Orexin-A and -B are abundant in the rat hypothalamus, medulla-pons, and midbrain-thalamus, and moderately abundant in the cerebral cortex. No orexins were found in the adipose tissues or visceral organs studied. The major endogenous molecule of orexin-A is a 33-amino-acid peptide, and that of orexin-B a 28-amino-acid peptide. After a 48 h fast, the orexin-A and -B contents of the lateral hypothalamus exhibited a trend to increase, but the contents of other brain tissues significantly decreased as compared with the fed control rats. No circadian variations in the orexin contents were found in the brain. The extensive and abundant distribution of orexins in the brain and changes in their contents upon fasting suggest that they serve as neuromodulators and/or neurotransmitters that regulate feeding behavior through interaction with diverse neural networks.

Amino Acid Sequence↗
PubMed1999

Effect of lateral cerebroventricular injection of the appetite-stimulating neuropeptide, orexin and neuropeptide Y, on the various behavioral activities of rats.

The effect of lateral cerebroventricular injection of the appetite-stimulating neuropeptide, orexin and neuropeptide Y (NPY), on the behavior of rats was investigated. An immediate increase in face washing activity was observed after injection of orexin A or orexin B, but not NPY. Orexin A had a more potent effect on face washing behavior than orexin B. Grooming and burrowing activities also increased significantly after injection of orexin A, whereas, orexin B significantly increased burrowing and searching behavior. Feeding behavior and food consumption increased dramatically within 10 min of injection of NPY. Although the significant increase in feeding behavior was also observed after injection of orexin A, total food intake did not change significantly. These results suggest that orexin may be involved in the regulation of several other behavioral activities in rats, besides feeding.

Animals↗
PubMed1999

Down regulation of the prepro-orexin gene expression in genetically obese mice.

The gene expression of prepro-orexin, the precursor of orexin-A and orexin-B which are hypothalamic pepetides that are associated with feeding behavior, were examined in control (C57B1/6J) and obese (ob/ob and db/db) mice using in situ hybridization histochemistry. Orexins are identical with hypocretins that have been identified by directional tag PCR subtractive hybridization method. In situ hybridization histochemistry revealed that the expression of the prepro-orexin gene was significantly decreased in ob/ob and db/db mice compared with control mice. The gene expression of neuropeptide Y (NPY), a potent feeding stimulant, is known to be increased in ob/ob and db/db mice. The expression of the NPY gene in the arcuate nucleus was increased remarkably in ob/ob and db/db mice compared to that of control mice. An immunohistochemical study showed that orexin-A and orexin-B immunoreactive neurons exhibited in the lateral and posterior hypothalamic areas and the perifornical nucleus were distributed similarly in control, ob/ob and db/db mice. These results suggest that the regulatory mechanism of orexins/hypocretins may be different from that of NPY in genetically obese mice.

Animals↗
PubMed1999

Orexins, orexigenic hypothalamic peptides, interact with autonomic, neuroendocrine and neuroregulatory systems.

We determined the immunohistochemical distributions of orexin-A and orexin-B, hypothalamic peptides that function in the regulation of feeding behavior and energy homeostasis. Orexin-A and -B neurons were restricted to the lateral and posterior hypothalamus, whereas both orexin-A and -B nerve fibers projected widely into the olfactory bulb, cerebral cortex, thalamus, hypothalamus, and brainstem. Dense populations of orexin-containing fibers were present in the paraventricular thalamic nucleus, central gray, raphe nuclei, and locus coeruleus. Moderate numbers of these fibers were found in the olfactory bulb, insular, infralimbic and prelimbic cortex, amygdala, ventral, and dorsolateral parts of the suprachiasmatic nucleus, paraventricular nucleus except the lateral magnocellular division, arcuate nucleus, supramammillary nucleus, nucleus of the solitary tract, and dorsal motor nucleus of the vagus. Small numbers of orexin fibers were present in the perirhinal, motor and sensory cortex, hippocampus, and supraoptic nucleus, and a very small number in the lateral magnocellular division of the paraventricular nucleus. Intracerebroventricular injections of orexins induced c-fos expression in the paraventricular thalamic nucleus, locus coeruleus, arcuate nucleus, central gray, raphe nuclei, nucleus of the solitary tract, dorsal motor nucleus of the vagus, suprachiasmatic nucleus, supraoptic nucleus, and paraventricular nucleus except the lateral magnocellular division. The unique neuronal distribution of orexins and their functional activation of neural circuits suggest specific complex roles of the peptides in autonomic and neuroendocrine control.

Animals↗
PubMed1999

Familial Amyloidotic Polyneuropathy: domino liver transplantation.

BACKGROUND/AIMS: The primary cause of Familial Amyloidotic Polyneuropathy is a variant transthyretin gene on chromosome 18. Progressive polyneuropathy followed by fatal cardiac and renal failure commonly manifest during middle age. Within 10 years after onset of clinical symptoms, affected individuals usually die due to malnutrition or heart failure. Currently, liver transplantation is the only available therapeutic option. METHODS: We performed liver transplantation in two patients with Familial Amyloidotic Polyneuropathy carrying the transthyretin-30 mutant. Two patients aged more than 50 years received the two explanted amyloidotic livers. This procedure is called Domino liver transplantation. We report the outcome in the studied subjects and analyze the metabolic consequences of this procedure. RESULTS: We determined the serum half-life of transthyretin-30 as 2.25 days using daily monitoring of transthyretin-30 levels. An affected amyloidotic patient had an increased serum concentration of lipoprotein(a) of 78 mg/dl before transplantation. The tumor patient, who received the organ from this affected patient, developed an almost identical serum concentration of lipoprotein(a) after liver transplantation, confirming the liver as the primary site of synthesis of this lipoprotein. CONCLUSION: Once Domino liver transplantation has been performed, the impact of the liver-dependent metabolism of specific proteins of interest can be studied.

Adult↗
PubMed1999

Thyroglobulin regulates follicular function and heterogeneity by suppressing thyroid-specific gene expression.

Thyroglobulin (TG) is the primary synthetic product of the thyroid and the macromolecular precursor of thyroid hormones. TG synthesis, iodination, storage in follicles, and lysosomal degradation can each modulate thyroid hormone formation and secretion into the circulation. Thyrotropin (TSH), via its receptor (the TSHR), increases thyroid hormone levels by upregulating expression of the sodium iodide symporter (NIS), thyroid peroxidase (TPO), and TG genes. TSH does this by modulating the expression and activity of the thyroid-specific transcription factors, thyroid transcription factor (TTF)-1, TTF-2, and Pax-8, which coordinately regulate NIS, TPO, TG, and the TSHR. Major histocompatibility complex (MHC) class I gene expression, which is also regulated by TTF-1 and Pax-8 in the thyroid, is simultaneously decreased; this maintains self tolerance in the face of TSH-increased gene products necessary for thyroid hormone formation. We now show that follicular TG, 27S > 19S > 12S, counter-regulates TSH-increased thyroid-specific gene transcription by suppressing the expression of the TTF-1, TTF-2, and Pax-8 genes. This decreases expression of the TG, TPO, NIS and TSHR genes, but increases class I expression. TG action involves an apical membrane TG-binding protein; however, it acts transcriptionally, targeting, for example, a sequence within 1.15 kb of the start of TTF-1 transcription. TG does not affect ubiquitous transcription factors regulating TG, TPO, NIS and/or TSHR gene expression. TG activity is not duplicated by thyroid hormones or iodide. We hypothesize that TG-initiated, transcriptional regulation of thyroid-restricted genes is a normal, feedback, compensatory mechanism which regulates follicular function, regulates thyroid hormone secretion, and contributes to follicular heterogeneity.

Animals↗
PubMed1999

Growth of moulds inoculated into commercial mineral water.

The growth of mould spores of Penicillium sp. and Cladosporium sp. inoculated in a commercial mineral water product was studied. The strains had been isolated as fungal foreign bodies in commercial mineral waters. In product A, which was not originally sterilized and was contaminated with psychrophilic bacteria, the inoculated mould spores of the strains did not grow; no increases in viable colony counts or beta-glucans concentration in the samples were observed during storage. In a sterilized product A, inoculated spores of the strains grew into visible foreign bodies. The viable colony counts and the beta-glucans concentration in the samples increased during storage. These results showed that in a sterilized mineral water product, mould spores could grow into visible foreign bodies.

Cladosporium↗
PubMed1999

A case of hereditary amyloidosis transthyretin variant Met 30 with amyloid cardiomyopathy, less polyneuropathy, and the presence of giant cells.

Transthyretin-Met 30 (TTR-Met 30) is a variant of transthyretin and is usually associated with familial amyloid polyneuropathy. It is rare that patients with TTR-Met 30 will primarily develop amyloid cardiomyopathy. This report presents a patient with late-onset TTR-Met 30 who primarily developed amyloid cardiomyopathy, with less amyloid polyneuropathy in the peripheral nervous system than is usually seen. An autopsy was performed, and histological examination revealed many foreign-body giant cells and macrophages in the area of amyloid deposition that was found in nearly all of the organs.

Aged↗
PubMed1999

Late-onset familial amyloid polyneuropathy type I (transthyretin Met30-associated familial amyloid polyneuropathy) unrelated to endemic focus in Japan. Clinicopathological and genetic features.

Clinicopathological and genetic features were assessed on 35 Japanese families affected by late-onset familial amyloid polyneuropathy type I (transthyretin Met30-associated familial amyloid polyneuropathy, FAP TTR Met30) whose siblings were unrelated to endemic Japanese foci. In these patients (50 years or older), the most common initial symptom was paraesthesias in the legs. Autonomic symptoms were generally mild and did not seriously affect daily activities. The male-to-female ratio was extremely high (10.7 : 1). A family history was evident in only 11 out of 35 families, and other patients were apparently sporadic. The rate of penetrance was very low. Symptomatic siblings of familial cases showed a late age of onset, male preponderance and clinical features similar to those of the probands. Asymptomatic carriers, predominantly female, were detected relatively late in life. The geographical distribution of these late-onset, FAP TTR Met30 cases was scattered throughout Japan. In three autopsy cases and 20 sural nerve biopsy specimens, neurons in sympathetic and sensory ganglia were relatively preserved. Amyloid deposition was seen in the peripheral nervous system, particularly in the sympathetic ganglia, dorsal root ganglia and proximal nerve trunks such as sciatic nerve. These abnormalities were milder than those seen in typical early-onset FAP TTR Met30, as observed in two Japanese endemic foci of this disease. While axonal degeneration was prominent in myelinated fibres, resulting in severe fibre loss, unmyelinated fibres were relatively preserved. Our cases of late-onset FAP TTR Met30 showed features distinct from those of typical early-onset FAP TTR Met30 that occurred in the two Japanese endemic foci. Factors responsible for clinicopathological differences between these two forms of FAP TTR Met30 need to be identified.

Action Potentials↗
PubMed1999

[A study of human beta-defensin-1 and human beta-defensin-2 in airway mucosal defense].

Human beta-defensin-1 (hBD-1) and human beta-defensin-2 (hBD-2) are new members of the defensin family. In this study, we investigated their gene expressions in the respiratory epithelial surface of the human lung and their bactericidal activities against Escherichia coli. Both hBD-1 and hBD-2 gene transcripts were detected in human lung tissue and cellular component of bronchoalveolar lavage fluid by reverse transcription-polymerase chain reaction. Synthetic hBD-1 and hBD-2 had dose-dependent bactericidal activities against E. coli. The concentration for the 50% colony reduction of hBD-2 was 0.46 nmol/ml, that for HNP-1 2.15 nmol/ml, and hBD-1 99.3 nmol/ml under conditions nearly the same as in human bronchial airway surface liquid. We prepared an antiserum against hBD-2 and established a highly sensitive radioimmunoassay and quantified plasma hBD-2 concentrations in normal subjects and patients with pneumonia. The plasma concentration of hBD-2 in normal individuals was 8.3 +/- 0.9 fmol/ml (mean +/- SE). The mean hBD-2 plasma concentration for the 12 patients with bacterial pneumonia in the acute stage was 34.2 +/- 3.4 fmol/ml, significantly higher than normal individuals, and returned to the normal range after recovery. The presence of hBD-1 and hBD-2 in the airway tract and their bactericidal activity suggest that they function in innate airway mucosal defense.

Adult↗
PubMed1999

Sympathetic and cardiovascular actions of orexins in conscious rats.

The novel hypothalamic peptides orexin-A and orexin-B are known to induce feeding behavior when administered intracerebroventricularly, but little is known about other physiological functions. The renal sympathetic nerves play important roles in the homeostasis of body fluids and the circulatory system. We examined the effects of intracerebroventricularly administered orexins on mean arterial pressure (MAP), heart rate (HR), renal sympathetic nerve activity (RSNA), and plasma catecholamine in conscious rats. Orexin-A (0.3, 3. 0 nmol) provoked an increase in MAP (94.3 +/- 0.7 to 101.9 +/- 0.7 mmHg and 93.1 +/- 1.1 to 108.3 +/- 0.8 mmHg, respectively) and RSNA (28.0 +/- 7.0 and 57.9 +/- 12.3%, respectively). Similarly, orexin-B (0.3, 3.0 nmol) increased MAP (93.9 +/- 0.9 to 97.9 +/- 0.9 mmHg and 94.5 +/- 1.1 to 105.3 +/- 1.7 mmHg, respectively). Orexin-A and -B at 3.0 nmol also increased HR. In other conscious rats, a high dose of orexin-A and -B increased plasma norepinephrine. Plasma epinephrine only increased with a high dose of orexin-A. These results indicate that central orexins regulate sympathetic nerve activity and affect cardiovascular functions.

Animals↗
PubMed1999

Plasma and urine levels of uroguanylin, a new natriuretic peptide, in nephrotic syndrome.

Uroguanylin, a new natriuretic peptide originally isolated from urine, stimulates the membrane guanylate cyclase C receptor. No information, however, is available on the plasma and urine levels of uroguanylin in nephrotic syndrome (NS), the state associated with sodium and water retention. Using a sensitive radioimmunoassay, we measured the plasma and urine concentrations of immunoreactive (ir-)uroguanylin in NS patients and compared them with those of patients with non-nephrotic glomerulonephritis. Plasma ir-uroguanylin, blood pressure and the cardiothoracic ratio were higher, and urine excretion of ir-uroguanylin was lower in the NS patients. Plasma ir-uroguanylin in the NS patients significantly decreased during remission as compared with findings on admission. There was a significant inverse correlation between the concentration of plasma ir-uroguanylin and that of serum total protein or albumin. Moreover, fluid retention in the NS patients was correlated with the changes in plasma ir-uroguanylin between admission and remission, indicative that the plasma concentration increases with the severity of the nephrotic state. Taking into account its potent natriuretic effect, these findings suggest that uroguanylin may function in the pathophysiological mechanism in NS.

Adult↗
PubMed1999

Enterochromaffin-like cells, a cellular source of uroguanylin in rat stomach.

Uroguanylin is an endogenous peptide ligand for guanylyl cyclase-C, an apical membrane receptor predominantly located in the gastrointestinal epithelium. It regulates intestinal and renal fluid and electrolyte transport through the second messenger, cyclic GMP. Uroguanylin messenger RNA and the peptide are present in rat stomach, but the cellular source has not been identified. We separated gastric mucosal cells by size into seven fractions (F1-F7) and enriched endocrine cells into F1-F3 using counterflow elutriation. Uroguanylin messenger RNA and peptide were found in F1-F3 by Northern blot analysis and an RIA specific for rat uroguanylin. Uroguanylin-producing cells were identified as endocrine cells by immunocytochemical methods using antisera for uroguanylin, prouroguanylin, and chromogranin A, as well as by in situ hybridization cytochemistry. Double-staining showed that uroguanylin and histamine are colocalized in enterochromaffin-like (ECL) cells that release histamine, leading to the stimulation of gastric acid secretion from parietal cells. Uroguanylin is synthesized in ECL cells. These findings should contribute to elucidating the physiological functions of ECL cells and the cyclic GMP-mediated gastric ion transport mechanism.

Animals↗
PubMed1999

Usefulness of MALDI/TOF mass spectrometry of immunoprecipitated serum variant transthyretin in the diagnosis of familial amyloid polyneuropathy.

A matrix-assisted laser desorption ionization/time-of-flight (MALDI/TOF) mass spectrometry (MS) system was used to detect variant transthyretin (TTR) in immunoprecipitated serum TTR molecules obtained from 6 patients with familial amyloid polyneuropathy (FAP) who were already proven not to have ATTR Val30Met. This simple and quick method showed six different patterns of mass spectra of TTR-related immunoprecipitates from these patients, and in each patient the clearly identified characteristic doublet-shaped ion peaks consisted of normal and variant TTR apart from each other peak with a mass difference between them. DNA sequencing confirmed that the patterns of variant TTR corresponded respectively to ATTR Val30Leu, ATTR Phe33Val, ATTR Asp38Ala, ATTR Ser50Arg, ATTR Ala97Gly and ATTR Ala97Ser. ATTR Asp38Ala and ATTR Ala97Ser are previously unknown variants of TTR leading to the development of FAP. ATTR Phe33Val was found in a Chinese FAP patient and ATTR Ala97Ser in a Taiwanese. Serum analysis using immunoprecipitation and MALDI/TOF MS system can provide useful information when investigating FAP patients with diverse types of variant TTR.

Adult↗
PubMed1999

Bioreactor operated production of lipase: castor oil hydrolysis using partially-purified lipase.

A highly stable lipase from Pseudomonas aeruginosa KKA-5 was produced by batch cultivation technique employing shake flask and 5 L-bioreactor. The bioreactor was run at different airflow rates. Low airflow rates (1 and 3 L/min), did not lead to effective growth and lipase production. Growth increased by about one order and lipase production increased by about 6 times, at an airflow rate of 5 L/min. Lipase production occurred during decelerated cell growth. A highly stable lipase was produced which retained its activity in the running bioreactor, even after a period of one month. This stable lipase was partially-purified using ammonium sulphate precipitation technique. Castor oil was hydrolyzed using 300U crude and partially-purified lipase, each. Approximately 21-fold, partially-purified lipase could hydrolyze 81% castor oil within a period of 96 hr, where as only 63% hydrolysis was obtained, in 216 hour, when crude lipase was used.

Bioreactors↗
PubMed1998

Identification of human beta-defensin-2 in respiratory tract and plasma and its increase in bacterial pneumonia.

Human beta-defensin-2 (hBD-2), a novel antimicrobial peptide, was originally isolated from human skin. We found that synthetic hBD-2 has high bactericidal activity against Escherichia coli under conditions nearly the same as in human bronchial airway surface liquid. We prepared an antiserum against hBD-2 and established a highly sensitive radioimmunoassay (RIA). Reverse-phase high-performance liquid chromatography coupled with the RIA showed that hBD-2 in patients with human lung, bronchoalveolar lavage flid, and plasma. The plasma concentration of hBD-2 in patients with bacterial pneumonia was 32.1 +/- 3.7 fmol/ml (mean +/- SE), 3.9-fold that of normal individuals. By reverse transcription-polymerase chain reaction, the hBD-2 gene transcript was detected in the respiratory epithelial surface of human lung. Human beta-defensin-2 seems to function in airway mucosal defense. Our findings provide a clue to elucidate its pathophysiological significance in respiratory infection.

Adult↗