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Biomedical subjects

S Nonaka

Publications and source records attributed to S Nonaka.

At least 55 records · Page 3Linked to original sources
PubMed1998

Rapid identification of Leishmania species from formalin-fixed biopsy samples by polymorphism-specific polymerase chain reaction.

The precise identification and classification of Leishmania species is important for public health surveillance since different species cause different clinical features of the disease. A highly specific polymerase chain reaction (PCR) panel was developed to enable the identification of the five major Leishmania species that cause New World cutaneous leishmaniases. The primers used for this panel were designed to distinguish the polymorphism in sequences of commonly amplified DNA bands of the parasites produced by arbitrarily primed PCR. These polymorphism-specific PCR diagnoses were performed with formalin-fixed biopsy specimens of the leishmanial lesions from four patients in Ecuador and one hamster skin lesion, and these lesions were determined to be caused by Leishmania (Viannia) panamensis, L. (Leishmania) mexicana, and L. (L.) amazonensis. The PCR panel may offer an important and practical approach to the standardized identification of Leishmania species in field examinations.

Animals↗
PubMed1998

Golgi vesiculation and lysosome dispersion in cells lacking cytoplasmic dynein.

Cytoplasmic dynein, a minus end-directed, microtubule-based motor protein, is thought to drive the movement of membranous organelles and chromosomes. It is a massive complex that consists of multiple polypeptides. Among these polypeptides, the cytoplasmic dynein heavy chain (cDHC) constitutes the major part of this complex. To elucidate the function of cytoplasmic dynein, we have produced mice lacking cDHC by gene targeting. cDHC-/- embryos were indistinguishable from cDHC+/-or cDHC+/+ littermates at the blastocyst stage. However, no cDHC-/- embryos were found at 8.5 d postcoitum. When cDHC-/- blastocysts were cultured in vitro, they showed interesting phenotypes. First, the Golgi complex became highly vesiculated and distributed throughout the cytoplasm. Second, endosomes and lysosomes were not concentrated near the nucleus but were distributed evenly throughout the cytoplasm. Interestingly, the Golgi "fragments" and lysosomes were still found to be attached to microtubules. These results show that cDHC is essential for the formation and positioning of the Golgi complex. Moreover, cDHC is required for cell proliferation and proper distribution of endosomes and lysosomes. However, molecules other than cDHC might mediate attachment of the Golgi complex and endosomes/lysosomes to microtubules.

Animals↗
PubMed1998

Chronic lithium treatment robustly protects neurons in the central nervous system against excitotoxicity by inhibiting N-methyl-D-aspartate receptor-mediated calcium influx.

Lithium is the most commonly used drug for the treatment of manic depressive illness. The precise mechanisms underlying its clinical efficacy remain unknown. We found that long-term exposure to lithium chloride dramatically protects cultured rat cerebellar, cerebral cortical, and hippocampal neurons against glutamate-induced excitotoxicity, which involves apoptosis mediated by N-methyl-D-aspartate (NMDA) receptors. This neuroprotection is long-lasting, occurs at therapeutically relevant concentrations of lithium with an EC50 of approximately 1.3 mM, and requires treatment for 6-7 days for complete protection to occur. In contrast, a 24-h treatment with lithium is ineffective. The protection in cerebellar neurons is specific for glutamate-induced excitotoxicity and can be attributed to inhibition of NMDA receptor-mediated calcium influx measured by 45Ca2+ uptake studies and fura-2 fluorescence microphotometry. The long-term effects of lithium are not caused by down-regulation of NMDA receptor subunit proteins and are unlikely related to its known ability to block inositol monophosphatase activity. Our results suggest that modulation of glutamate receptor hyperactivity represents at least part of the molecular mechanisms by which lithium alters brain function and exerts its clinical efficacy in the treatment for manic depressive illness. These actions of lithium also suggest that abnormality of glutamatergic neurotransmission as a pathogenic mechanism underlying bipolar illness warrants future investigation.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1998

Serum interleukin-2 receptor for the early diagnosis of rheumatoid arthritis.

The value of measuring soluble interleukin-2 receptor (sIL-2R) in the sera of patients with joint pain as a predicting parameter for the future development of rheumatoid arthritis (RA) was examined. sIL-2R was measured by the ELISA method. Sixty-four patients with joint pain (suspected RA: sus-RA) but no bone or joint destruction were enrolled over 2 years and 47 were selected for the study. Eleven patients whose diagnosis was sus-RA after a year of observation were successively followed-up for 5 years. Two-thirds of the patients whose sIL-2R levels were higher than those of normal healthy adults (< 82 pmol/l; mean +2SD) developed RA within a year. On the other hand, one-quarter of the patients with normal levels of sIL-2R also developed RA within a year. The presence of two or three of the following three items in patients with joint pain without any bone and joint destruction was thus indicated to be useful for the early diagnosis of RA: elevated CRP level (> or = 1.0 mg/dl), positive rheumatoid factor (RF) (> or = 30 IU/ml) and an elevated sIL-2R level (> or = 100 pmol/l). Sensitivity and specificity were 72.7% and 96.0%, respectively. The probability of development of RA is expressed as P = 1/[1 + exp(2.673 - 0.01784 x sIL-2R - 0.4398 x CRP - 0.004835 x RF)], with R2 = 0.3083 and p<0.0005. On the other hand, the sIL-2R levels did not correlate with any future bone or joint changes within a year of observation. The above criteria may therefore hopefully justify the early treatment of patients with joint pain using drugs that can modify the patients' immune function. However, the validity of these criteria still need to be examined more thoroughly in the future.

Adult↗
PubMed1998

Facial nerve parasympathetic preganglionic afferents to the accessory otic ganglia by way of the chorda tympani nerve in the cat.

The distribution of accessory otic ganglia and connections between the ganglia and the chorda tympani nerve were investigated in the cat in order to determine the parasympathetic preganglionic facial nerve afferents to the otic ganglia using whole mount acetylthiocholinesterase (WATChE) histochemistry. The otic ganglia consist of a single main prominent ganglion and many small accessory ganglia lying on a plexus around the origins of the branches of the mandibular nerve and near the junction of the chorda tympani nerve and lingual nerve. In cell analysis of Nissl-stained preparations, the neurons composing the accessory otic ganglia were morphologically similar to the main otic ganglion neurons. Connecting branches from the chorda tympani nerve to the peripherally located accessory otic ganglia were found and they were not stained by WATChE histochemistry. WATChE-positive connecting branches from the ganglia to the inferior alveolar, lingual, and mylohyoid nerves were also found in the same preparations. The WATChE histochemistry on various autonomic nervous tissues revealed that autonomic postganglionic nerve fibers are selectively stained darkly and that preganglionic fibers remain unstained. Therefore, it is considered that the WATChE-negative connections from the chorda tympani nerve consist chiefly of autonomic preganglionic fibers, whereas the WATChE-positive connections to the branches of the mandibular nerve are mainly postganglionic fibers. This suggests that some of the facial nerve parasympathetic preganglionic fibers in the chorda tympani nerve are mediated in the accessory otic ganglia and then join the branches of the mandibular nerve to supply the target mandibular tissues.

Acetylcholinesterase↗
PubMed1998

The augmentation of intrinsic laryngeal muscle activity by air-jet stimulation of the nasal cavity in decerebrate cats.

The purpose of this study was to examine the functional roles of nasal afferents in modulating the activity of the intrinsic laryngeal muscles. The electromyographic activities of the intrinsic laryngeal muscles and major respiratory muscles were recorded in cats during nasal air-jet stimulation. The activities of brainstem respiratory neurons were also recorded to determine which neurons transmit nasal afferent signals to the intrinsic laryngeal motoneurons. These axonal projections were identified by antidromic activation evoked by stimulation to the spinal cord at C4 level and the laryngeal nerve. The length of the respiratory cycle was prolonged and the diaphragmatic activity was decreased during air-jet stimulation of the nasal cavity. In contrast, the activities of both the intrinsic laryngeal adductor and abductor muscles were increased. Examination of the laryngeal reflexes revealed increase in the activities of intrinsic laryngeal motoneurons during both respiratory phases. Most of the respiratory neurons recorded decreased their peak firing rate during air-jet stimulation, reflecting decreased diaphragmatic activity; however, the peak firing rate of the bulbospinal expiratory neurons in the portion of the ventral respiratory group caudal to the obex did not decrease during stimulation. These findings demonstrate the nasal air-jet stimulation decreases the activities of major inspiratory muscles in order to avoid inspiration of foreign bodies into the nasal cavity and augments the activities of intrinsic laryngeal muscles, enabling prompt elicitation of the laryngeal airway reflex. Our findings also suggest that the nasal afferents suppress the major inspiratory activities by way of brainstem inspiratory neurons, but that the activities of intrinsic laryngeal muscles are controlled through undetermined pathway(s) other than the pathway through respiratory neurons.

Animals↗
PubMed1998

Association of human papillomavirus type 16 with malignant melanoma.

We report a case of malignant melanoma associated with human papillomavirus (HPV) in a 37-year-old woman. The patient has had numerous brown papular and nodular tumors, 5 to 30 mm in diameter, on her left leg for > 15 years, some of them coalescing rapidly in the last 12 months to a multilobulated black nodule diagnosed as malignant melanoma by histology and immunohistochemistry. HPV type 16 DNA was detected in the melanoma specimen by reverse transcriptase polymerase chain reaction (rt-PCR) and in situ hybridization (ISH) of the tumor tissues. This is the first report of melanoma associated with HPV 16.

Adult↗
PubMed1998

A preliminary study aimed at the detection of Leishmania parasites in subjects with cutaneous leishmaniasis using polymerase chain reaction.

As a basic study for future diagnosis of cutaneous leishmaniasis, we tried to detect Leishmania parasites representing different species in the subgenera Leishmania and Viannia from subject patients with cutaneous leishmaniasis by using the polymerase chain reaction (PCR) with the subgenus Viannia specific primer. Four out of the 14 specimens revealed an amplified DNA of 70 bp specific for the subgenus Viannia (L. braziliensis complex). No bands were detected in the rest of the specimens belonging to the subgenus Leishmania and unclassified groups. The base sequences of the amplified DNA corresponded with those of the L. (V). braziliensis kinetoplast minicircle. We concluded that PCR using the present primer specific for the subgenus Viannia would be useful in detecting Leishmania parasites in lesions of cutaneous leishmaniasis caused by the L. braziliensis complex.

Animals↗
PubMed1998

Comparative studies of the detection rates of Leishmania parasites from formalin, ethanol-fixed, frozen human skin specimens by polymerase chain reaction and Southern blotting.

In this study, detection rates of Leishmania parasites from human skin were compared among three different types of specimens, formalin-fixed, ethanol-fixed, and frozen, by polymerase chain reaction (PCR) and Southern blotting. For this purpose, we used biopsy specimens collected from 19 leishmaniasis patients and performed PCR and Southern hybridization with the probe specific for Leishmania (Viannia) braziliensis complex. Among these 19, 16 specimens were from cutaneous leishmaniasis (CL), one, diffuse cutaneous leishmaniasis (DCL) and 2, mucocutaneous leishmaniasis (MCL) and were formalin-fixed and paraffin-embedded. The causative agents for one case of CL and one case of DCL were already identified as L. (Leishmania) complex. Six specimens of CL were preserved in 100% ethanol. Two specimens of MCL were frozen tissues. PCR using the formalin-fixed and paraffin-embedded specimens revealed positive bands at 70 bp in 9 (47.4%) out of 19 specimens of CL, MCL and DCL. Southern blotting detected the signals in 12 (63.2%) out of the 19. PCR using the 100% ethanol-fixed specimens revealed positive bands in 4 (66.7%) out of 6, and Southern blotting also detected the signals in 4 (66.7%) out of the 6. PCR and Southern blotting using 2 frozen specimens of MCL were always positive (100%). Although we failed to detect significant differences by Chi-square test between the results from the formalin-fixed, paraffin-embedded specimens and those from 100% ethanol-fixed ones, we concluded that ethanol-fixed specimens, convenient for transportation and storage, would be more useful for diagnosis of leishmaniasis by PCR in a developing country.

Adolescent↗
PubMed1998

Phacomatosis pigmentovascularis type IIb associated with Sturge-Weber syndrome and pyogenic granuloma.

A case of phacomatosis pigmentovascularis (PPV) in a 6-year-old girl with Sturge-Weber syndrome, pyogenic granuloma, and other complications is described. It is relatively rare that a complete form of Sturge-Weber syndrome was associated with PPV. A review of the literature on PPV, focusing on total number of reported cases and etiological speculations, is presented. To our knowledge, a total of 118 cases of PPV, including the present one, have been reported to date. Regardless of many speculations, the true etiology remains unknown. The average "density" of mast cells (MCs) per mm2 appearing in the central region of the pyogenic granuloma was calculated to be 86.3/mm2 and that in the adjacent nevus flammeus was 37.9/mm2. The "density" of mast cells in pyogenic granuloma separately calculated from ten other cases was 105.5 +/- 28.6/mm2 (mean +/- SD), compared with that in normal skin, 6.85 +/- 4.9/mm2 (n = 20). There was a significant difference between the two, indicating that MCs are closely associated with angiogenesis in pyogenic granuloma.

Biopsy, Needle↗
PubMed1998

[Anti-proliferative effects of heating on the human prostatic carcinoma cells in culture].

Prostatic cancers are well-known to be sensitive to heat stress. However, the mechanism by which the cancer cells are killed by high temperature remains poorly understood. The present study was undertaken to determine the anti-proliferative effects of heat stress on the prostatic cancer cells in culture. Heat shock at 43 degrees C inhibited the cell growth of three different prostatic cell lines. Flow cytometrical analysis using BrdU and PI showed a decrease in the proportion of cells in an S phase, accompanied by cell accumulation in G1 and G2, in both JCA-1 and PC-3 but not in LNcap. Both JCA-1 and PC-3 presented a strong expression of hsp70 at 37 degrees C. The heat shock caused apparent enhancement of the expression of hsp70 through the cell cycle. A treatment at 43 degrees C for 8 hours resulted in not only an apparent increment of positive hsp70 cells, but cells with subdiploid DNA content in LNcap. Flow cytometrical analysis by FITC-labeled Annexin V showed increment of apoptotic cells at 43 degrees C for 8 hours in LNcap cells. The results suggest that apoptosis is an important pathway of heat-induced killing of these cells. In conclusion, the cell growth of prostatic cancers may be affected by the temperature through relationship of the cell cycle and hsp70.

Apoptosis↗
PubMed1998

Lithium protects rat cerebellar granule cells against apoptosis induced by anticonvulsants, phenytoin and carbamazepine.

We have studied the neuroprotective actions of lithium against various insults in cultured cerebellar granule cells of rats. The anticonvulsants, phenytoin and carbamazepine, have been shown to induce apoptosis of cerebellar granule cells at high concentrations. Here we found that co-presence of LiCl (1-10 mM) dose-dependently protected against phenytoin (20 microM)- and carbamazepine (100 microM)-induced neuronal apoptosis as assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide metabolism, morphological inspection, chromatin condensation and DNA fragmentation. These neuroprotective effects were not prevented by inclusion of myoinositol nor mimicked by a potent inositol monophosphatase inhibitor, suggestive of a mechanism independent of inositol monophosphatase blockade. Lithium also significantly protected against apoptosis of cerebellar granule cells induced by aging of the cultures. Additionally, lithium suppressed death of cerebellar granule cells exposed to a low concentration of extracellular potassium. In contrast, it had no protective effect on cell death induced by Ca++ ionophores, a Na+ channel opener, a protein kinase inhibitor, a nitric oxide donor or H2O2. Thus, lithium has robust neuroprotective effects against apoptotic cell death induced by multiple insults with limited selectivity. These actions provide a new avenue to study the molecular and cellular mechanisms of this drug.

5'-Nucleotidase↗
PubMed1997

Lombard reflex during PAG-induced vocalization in decerebrate cats.

The Lombard reflex occurs when a speaker increases his vocal effort while speaking in the presence of ambient noise. The purpose of this study was to clarify whether the Lombard reflex can be evoked during controlled vocalization in an animal model. In decerebrate cats, repetitive electrical stimulation was applied to the midbrain periaqueductal gray (PAG) to evoke vocalization. Pure tone auditory stimulation was delivered through a loudspeaker. The activities of the laryngeal adductor muscle, diaphragm and external oblique abdominal muscle and the voice intensity were measured during PAG stimulation, in the presence and absence of the auditory stimulation. To clarify the effects of the auditory laryngeal reflex on the activity of laryngeal adductor motoneurons, the amplitude of the laryngeal reflex evoked by single shock stimulation of the superior laryngeal nerve was also measured during respiration, in the presence and absence of auditory stimulation. The sound made by the cats due to PAG-induced vocalization was augmented by exposure to auditory stimulation, and the activities of the laryngeal adductor muscle and external oblique abdominal muscle were also augmented. During respiration, auditory stimulation also increased the amplitude of the laryngeal reflex evoked in the laryngeal adductor muscle. These results demonstrate that the essential neuronal mechanisms for evoking the Lombard reflex exist within the brainstem.

Acoustic Stimulation↗
PubMed1996

Modulation of vomiting by the medullary midline.

Studies were conducted to investigate the possible role of the brainstem midline region as a source of facilitatory input for the vomiting process. Experiments were conducted using the "fictive vomiting' model in decerebrate, paralysed cats. Dysfunction of the medullary midline region produced by localized injections of the neurotoxin kainic acid abolished or greatly attenuated fictive vomiting. In addition, some respiratory-related midline neurons were found to fire in synchrony with co-active phrenic and abdominal nerve discharge during fictive vomiting. These experiments demonstrate the importance of the medullary midline for the normal occurrence of the vomiting process. An explanation for the post-lesion elimination of vomiting is that the lesions remove an important source of facilitatory input to spinal respiratory motoneurons and/or to the brainstem circuitry that mediates vomiting.

Animals↗
PubMed1996

Activities of expiratory neurones of the Bötzinger complex during vocalization in decerebrate cats.

Repetitive electrical stimulation of the midbrain peri-aqueductal grey (PAG) terminates quiet breathing and initiates inspiration that precedes vocalization. To understand the neuronal mechanisms underlying this phenomenon, activities of expiratory neurones (n = 39) of the Bötzinger complex (BOT) were examined in decerebrate cats. Most augmenting expiratory (E-aug) neurones (20/22) of the BOT, including 15 bulbospinal neurones, decreased their activities (9/20) or ceased to discharge (11/20) after the onset of stimulation of the PAG. This suggests that suppression of E-aug neurones of the BOT, which project to phrenic motoneurones, results in disinhibition of these neurones, and, in turn, terminates expiration and initiates inspiration preceding vocalization.

Animals↗
PubMed1996

Quantitative analysis of ferrochelatase mRNA in blood cells of erythropoietic protoporphyria patients.

Ferrochelatase (FC; heme synthetase, EC 4.99.1.1.) catalyses the synthesis of heme from protoporphyrin IX, the final step in the heme synthetic pathway. The hereditary deficiency of this enzyme gives rise to erythropoietic protoporphyria (EPP). We developed a rapid, non-radioactive means of measuring human FC mRNA levels in the EPP patients. It is based on the reverse transcriptase-polymerase chain reaction (RT-PCR) performed on the RNA obtained from peripheral blood. The amplified DNA was detected by agarose gel electrophoresis with ethidium bromide staining and the fluorescent intensity was measured by scanning densitometry applied directly to Polaroid 665 negative film. The relative expression level of FC mRNA, compared with that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA, was estimated at several points in the exponential phase of PCR cycles or at a point in the exponential phase of PCR performed on serially diluted the cDNA samples. The estimate of the FC mRNA by this method correlated well with the level of the FC mRNA measured by Northern blotting in the EB virus-transformed lymphocytes of the same patients. The level of the FC mRNA appeared to vary among the patients in whom a decreased level of enzymatic activity was indicated.

Adolescent↗
PubMed1996

Functional role of ventral respiratory group expiratory neurons during vocalization.

The activities of expiratory neurons of the caudal ventral respiratory group (cVRG) (n = 31) were recorded extracellularly during vocalization in unanesthetized, decerebrate cats. Vocalization was induced by electrical stimulation (0.2 ms, 20-80 microA, 100 Hz, lasting for 2-5 s) of the periaqueductal gray (PAG). The firing rates of more than two-thirds of the neurons (24/31) increased during periods of vocalization induced by stimulation of the PAG, with an increase in abdominal muscle activities. The axons of most of these neurons (19/24) projected to the contralateral lumbar spinal cord, and one-third of the bulbospinal neurons (6/19) were orthodromically activated by stimulation of the PAG. Even after sectioning of the axons of bulbospinal cVRG neurons by midsagittal lesion between the obex and the C1 spinal cord, stimulation of the PAG still induced adduction of the vocal cords and vocalization. However, activities of abdominal muscles during vocalization were abolished. These results indicate that the efferent signals for vocalization from the PAG may be relayed at the cVRG neurons and converted into adequate activation of these neurons, contributing to the activation of the abdominal muscles for vocalization.

Abdominal Muscles↗