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Biomedical subjects

H Kobori

Publications and source records attributed to H Kobori.

At least 37 records · Page 2Linked to original sources

Fission yeast protein kinase C gene homologues are required for protoplast regeneration: a functional link between cell wall formation and cell shape control.

Two novel protein kinase C (n PKC) gene homologues, pck1+ and pck2+ were isolated from the fission yeast Schizosaccharomyces pombe (Toda et al. (1993) EMBO J. 12, 1987). We examined the functional differences of pck1+ and pck2+ in cell wall formation and actin organization of S. pombe. Regenerating protoplasts of a wild-type strain, single gene disruptants of pck1+ (delta pck1) and pck2+ (delta pck2) were used as a simple model to examine the functional links between PKC, cell wall formation and actin organization. Protoplasts of the wild-type strain and those of delta pck1 reverted to intact cells in osmotically stabilized liquid medium. A close spatial association between new cell wall formation and actin was observed in these two strains. In delta pck2, protoplasts did not revert to intact cells: (1) scarcely any new cell wall material was formed; (2) actin was not reorganized; and (3) nuclear division and an increase in the amount of cytoplasm were observed in the regenerating protoplasts. These findings demonstrate that the pck2+ gene has a function essential for protoplast regeneration but the pck1+ gene does not. Involvement of n PKCs in cell wall formation and actin organization was also clarified. The effect of staurosporine (a potent inhibitor of protein kinases) on regenerating protoplasts of the three strains confirmed the assumption that the pck2 protein is an in vivo target of staurosporine in the fission yeast.

Actins↗

Simultaneous quantitative determination of 3H, 14C, 32P, 35S, 51Cr, and 125I on the surface of a paper disc using combustion.

A paper disc that retained 3H, 14C, 32P, 35S, 51Cr, and 125I on its surface was combusted in a flow-type combustion flask filled with oxygen. The combustion products containing five radionuclides, except 14CO2, were collected by washing the inner wall of the flask with 0.5-M hydrogen peroxide containing excess NaHSO3. The 14CO2 that remained in the gas phase of the flask was introduced into another flask for absorption into an organic base solution. By determining the radioactivity of both the solutions containing the respective radionuclides by means of NaI(Tl) and liquid scintillation counters, the six radionuclides in the solutions were quantified. The detection limit of surface contamination with this method was compared with the limit specified by Japanese law for the surface contamination of controlled areas.

Fires↗

[An experimental study on development of gallstone disease after curative gastrectomy for cancer--postoperative alteration of the composition of canine gallbladder bile after subtotal gastrectomy with truncal vagotomy].

In connection with gallstone (GS) formation after gastrectomy for cancer, we examined alteration of the composition of gallbladder bile after subtotal gastrectomy with truncal vagotomy. Of 11 mongrel dogs with cholecystostomy, 3 underwent B-I and 4 B-II gastrectomy. Four dogs without gastrectomy were served as controls. Bile was collected for 12 months. Bile acids were quantified by GLC. The bile was cultured for bacteriology. TBA did not significantly differ among the three groups of the dogs. In the two gastrectomized groups, CDA (a secondary bile acid) kept a higher level and CA (a primary bile acid) maintained a lower concentration compared with those in the controls. A remarkable increase of non-conjugated bile acids was recognized in both gastrectomized groups. Lithogenicity was low for all of the 3 groups. All but 1 control dog incurred bile infection that persisted long. Black pigment stones containing calcium bilirubinate developed in 1 of the 3 B-I and 2 of the 4 B-II dogs but none in the controls. The bile infection seemed to be involved in the development of GS. As the alteration of bile composition and GS occurred solely in the gastrectomized dogs, subtotal gastrectomy with truncal vagotomy might precipitate the GS formation.

Animals↗

[Altered alpha atrial natriuretic peptide (ANP) concentrations in plasma, atria and hypothalamus in experimentally induced hyper- and hypothyroid rats].

In order to assess a possible involvement of thyroid hormone in atrial natriuretic peptide (ANP), experimentally induced hyper- and hypothyroid rats were employed, and the immunoreactive rat ANP (IR-ANP) concentrations in plasma, atria and brain regions including the hypothalamus were measured by a specific radioimmunoassay. Plasma IR-ANP concentration in hypothyroid rats was 14.5 +/- 2.9 (mean +/- SD) fmol/ml, significantly lower than that in control rats (p less than 0.05 vs control of 24.9 +/- 9.7 fmol/ml). Plasma IR-ANP concentration in hyperthyroid rats was 66.4 +/- 9.7 fmol/min, significantly higher than that in the controls (p less than 0.01). Atrial IR-ANP concentration in hyperthyroid rats was significantly lower than that in the controls (79.9 +/- 11.1 nmol/g vs 133.5 +/- 21.2 nmol/g (control), p less than 0.05), though no significant change was observed in atrial IR-ANP concentration in hypothyroid rats. While hypothalamic ANP concentration in hypothyroid rats was significantly lower than that in the controls (17.5 +/- 3.5 pmol/g vs 31.9 +/- 1.9 pmol/g (control), p less than 0.05), there was no significant change of that in the hyperthyroid rats. On reverse phase high performance liquid chromatography, the major peak in plasma and hypothalamus extract was thought to be identical to synthetic alpha-rat ANP (1-28). These results may suggest that in the hyperthyroid state an excessive amount of ANP is released from atria into the blood, and that in the central nervous system thyroid hormone involve ANP metabolism being different from the atrium.

Animals↗

[Immunoreactive 7B2 concentrations in plasma and cerebrospinal fluid in pathophysiological conditions and the responses to oral glucose load, intravenous LH-RH, TRH and arginine infusion].

The peptide, 7B2, originally isolated from pituitary, has been shown to be present in endocrine tumors of high concentrations in pancreatic islet tumors. Plasma from most of these patients showed very high 7B2 immunoreactivity (IR-7B2) though there is a lack of knowledge concerning physiological and pathological changes in plasma IR-7B2 levels in other conditions. To assess whether or not there is any alteration in circulating IR-7B2 levels due to age, sex or any specific condition, plasma levels of IR-7B2 were measured in the fasting state in 106 healthy subjects aged 3 months to 91 years, 101 diabetics, 28 patients with hyperthyroidism. 7 patients with primary hypothyroidism, 13 patients with liver cirrhosis, 43 patients with chronic renal failure, 35 patients with cerebral vascular accident, and 26 pregnant subjects. Twenty-four cord bloods were also included. The responses of circulating IR-7B2 to oral glucose, intravenous arginine infusion, volus thyrotropin (TRH) or volus luteinizing hormone-releasing hormone (LH-RH) injection were also evaluated. Particularly high IR-7B2 levels were found to exist in cord blood. Postnatally the concentrations decreased gradually with age to adult values (15.6 +/- 2.9pmol/liter (mean +/- SE) in 20's-60's), though plasma IR-7B2 levels again increased significantly in over 70's (37.1 +/- 3.2pmol/liter; P less than 0.01). There was no significant difference in plasma 7B2 levels in either sex. Among the pathological conditions studied, significantly high IR-7B2 levels were observed in patients with chronic renal failure (175.1 +/- 35.9pmol/liter). Some of the pregnant patients in their third trimester also showed high plasma IR-7B2 levels. A small but significant rise in plasma IR-7B2 was observed after a glucose load in control subjects and diabetics. Intravenous LH-RH exerted a rise in plasma 7B2 concentrations though arginine and TRH showed no significant effect on plasma IR-7B2 concentrations. Compared with the plasma concentrations, ten to fifty-fold high levels of IR-7B2 were observed in cerebrospinal fluid (CSF) from patients with cerebrovascular accidents or multiple sclerosis. These results suggested that the kidney plays a major role in 7B2 degradation and that LH-RH simulates IR-7B2 release from the pituitary gland. Whether reduced clearance or increased production was responsible for the IR-7B2 elevation in subjects under 10 years or over 70 years requires investigation. Furthermore, high levels of IR-7B2 in CSF might indicate its specific role for the central nervous system.

Administration, Oral↗

Actin is associated with the formation of the cell wall in reverting protoplasts of the fission yeast Schizosaccharomyces pombe.

To clarify the involvement of actin in the formation of the yeast cell wall, reverting protoplasts of Schizosaccharomyces pombe were used as a simple model system. Actin of reverting protoplasts was labeled with rhodamine-conjugated phalloidin and observed by conventional fluorescence microscopy and laser scanning confocal microscopy. A close spatial as well as temporal relationship between actin and cell wall formation was observed in protoplast reversion. That is, the site of actin 'dots' in the reverting protoplasts coincided with the site of new wall formation and the timing of rearrangement of actin coincided with the initiation of cell wall formation and with the timing of cell wall expansion. Treatment of reverting protoplasts with cytochalasin D (CD) further clarified the close relationship between actin and cell wall organization. The effect of CD was dose dependent. A high dose of CD caused the absence of actin as well as the complete inhibition of cell wall formation. A low dose of CD caused weakly stained unlocalized actin, which induced grossly aberrant cell wall deposition as well as substantial changes in the morphology of the reverting protoplasts. These results demonstrated that actin is associated with initiation of cell wall formation, the proper deposition of cell wall materials, and maintaining the normal morphology of reverting protoplasts. Scanning electron microscopy revealed the presence of a fibrillar net structure on the surface of non-treated control reverting protoplasts. However, the absence of a fibrillar network on the surface of reverting protoplasts was observed in the presence of a high concentration of CD. Lack of localization of microfibrils as well as poor development of the fibrillar network were also observed in the presence of a low concentration of CD. Recovery experiments confirmed the close relationship between actin and cell wall formation.

Actins↗

Immunoreactive 7B2 concentrations in rats with various endocrine conditions.

Changes in 7B2 immunoreactivity in the pituitary as well as in the other brain regions and gut after various endocrine situations were investigated. Gonadectomy and neonatal monosodium glutamate (MSG) treatment resulted in an appreciable increase in the pituitary 7B2 concentration, though 7B2 content in the MSG treated pituitary was not significantly different when calculation was performed on a per pituitary gland basis. The 7B2 concentration in the cerebellum, midbrain and cortex in thyroxine treated rats showed a significant increase, which might indicate possible thyroid hormone involvement in 7B2 metabolism in the brain. The pituitary 7B2 concentration during the estrous cycle did not change significantly. These results suggest that pituitary 7B2 may correlate to the pituitary gonadotropins and that brain 7B2 content may be modulated by thyroid hormones.

Adrenalectomy↗

Cell wall formation in regenerating protoplasts of Schizosaccharomyces pombe: study by high resolution, low voltage scanning electron microscopy.

The ultrastructure of regenerating cell wall in Schizosaccharomyces pombe protoplasts was studied with a high resolution, low voltage scanning electron microscope (LVSEM). In contrast to the transmission electron microscopy, the LVSEM images give three-dimensional information on the cell wall regeneration in yeast protoplasts. We found that, after only a few minutes of incubation, the protoplasts began to show protuberances in a unipolar manner, and a fibrilar network was formed asymmetrically which covered the whole surface of the protoplasts after 5 hr. The network consisted of microfibrils about 8 to 10 nm wide, forming flat and wavy bundles of various widths and lengths, up to about 200 nm wide and 1 micron long, mainly made of yeast glucan. Free ends of microfibrils were seldom found. Interfibrillar spaces were progressively filled with granular particles and finally the complete cell wall was formed after 12 hr. The fibrillar network was destroyed by the digestion with beta (1----3)-glucanase. When protoplasts were regenerating in the presence of aculeacin A, the fibrillar networks were not formed, resulting in incomplete cell wall formation. These observations suggest that beta-glucan is the main component of the microfibrils and that it plays an important role in the formation of the cell wall in S. pombe.

Antifungal Agents↗

Plasma and urine concentrations of atrial natriuretic peptide in patients with diabetes mellitus.

To examine whether plasma and urine concentrations of human atrial natriuretic peptide (hANP) are altered in patients with diabetes mellitus (DM), plasma and urine hANP concentrations were evaluated in 86 patients with diabetes mellitus using an extraction procedure. The mean recovery rate of extraction was 71.8 +/- 0.6% (mean +/- SEM). The major immunoreactive component of hANP in extracted plasma and urine appeared to be identical to synthetic alpha hANP as judged by reverse-phase high-performance liquid chromatography (HPLC). The patients were divided into three groups according to their renal complications. The patients in group 1 had no apparent abnormality in serum creatinine, serum or urine beta 2-microglobulin (beta 2-MG), or urine N-acetyl-beta-D-glucosaminidase (NAG); those in group 2 showed either beta 2-MG or NAG abnormality but no creatinine abnormality. The patients in group 3 were though to have an established diabetic nephropathy and showed a serum creatinine increase. Plasma ANP concentrations in groups 1, 2, and 3 were 10.7 +/- 2.1, 19.9 +/- 5.6, and 39.2 +/- 9.9 fmol/ml, respectively. These values in groups 2 and 3 were significantly higher than the control values (p less than 0.05 or p less than 0.01 versus 6.2 +/- 0.7 fmol/ml). Urine ANP concentrations in group 1 were also within normal range, though those in groups 2 and 3 markedly increased in comparison with normal values.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Pituitary, hypothalamic and pancreatic 7B2 concentrations in rats with streptozotocin-induced diabetes and spontaneously diabetic mice.

The pituitary protein, 7B2 has been demonstrated in the pancreas and is known to be present in very high concentrations in pancreatic endocrine tumors. To investigate whether any changes in 7B2 concentrations might be present in the pancreases affected by different types of diabetes and whether the diabetic state itself might affect pituitary and hypothalamic 7B2 concentrations, streptozotocin (STZ)-treated rats and mice with natural-onset diabetes (obese hyperglycemic, or ob/ob, mice and non-obese diabetic, or NOD, mice) were employed. A significant reduction in pancreatic 7B2 concentrations was found in STZ-treated rats. The pancreatic 7B2 concentration was significantly high in ob/ob mice (p less than 0.05, versus the concentration in their lean littermates, and the decrease observed in older NOD mice, appeared to parallel their insulin reserve. Pituitary and hypothalamic 7B2 concentrations were similar in STZ-treated and control rats. A reduction in pituitary and hypothalamic 7B2 concentrations was observed in older NOD mice (both p less than 0.01 versus respective values in younger mice). In ob/ob mice, a significant reduction was also found in hypothalamic 7B2 concentration (p less than 0.01 versus that in control mice). Gel permeation chromatography showed that 7B2 immunoreactivity comprised two molecular components, and the relative proportion in the pancreas differed from that in other tissues. In the pancreas, a smaller molecular component was predominant (elution coefficient, 0.62).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Changes in the plasma and urine alpha human atrial natriuretic peptide (alpha hANP) concentration in patients with thyroid disorders.

In order to assess the possible involvement of thyroid hormone in alpha human atrial natriuretic peptide (alpha hANP), we investigated the plasma and urine ANP concentration in patients with primary hyperthyroidism and hypothyroidism. Plasma and urine were extracted through Sep-Pak C18 cartridges and the urine ANP concentration was corrected by urine creatinine (cre. mg/dl) and expressed as fmol/mg.cre.. The plasma ANP concentration in patients with untreated hyperthyroidism (32.3 +/- 7.0 fmol/ml; n = 22) was higher than in normal subjects (p less than 0.01 vs control; 6.2 +/- 0.7 fmol/ml). After restoration to euthyroidism, the plasma ANP concentration (patients with treated hyperthyroidism) fell to normal (8.9 +/- 1.9 fmol/ml). The plasma ANP concentration in patients with untreated hypothyroidism (14.1 +/- 3.0 fmol/ml; n = 7) was higher than normal, but in two of them there was mild renal dysfunction and an incomplete right blundle branch block in the electrocardiogram. It was possible that these factors contributed to the observed increase in plasma ANP. However, a significant positive correlation was found between plasma ANP and free thyroxine (n = 40, r = 0.449; p less than 0.01) and free triiodothyronine (n = 40, r = 0.546; p less than 0.01). The urine ANP concentration in patients with untreated hyperthyroidism was markedly higher than in normal subjects (p less than 0.01), but in untreated hypothyroidism not significantly different from normal.

Adolescent↗

Circulating concentrations of immunoreactive peptide 7B2 in certain pathophysiological conditions, and response to oral glucose load.

A peptide, 7B2, originally isolated from pituitary, is present in endocrine tumors, with high concentrations in pancreatic islet tumors. Plasma from most of these patients showed very high immunoreactivity to 7B2 (IR-7B2). To assess whether or not there is any alteration in circulating 7B2 concentrations due to age, sex, etc., we measured concentrations in plasma in 96 fasting healthy subjects, ages three months to 91 years; in patients with various other conditions, including pregnancy; and in cord blood. The response of circulating IR-7B2 to oral glucose was also evaluated. We found particularly high IR-7B2 concentrations in cord blood. Postnatally the concentrations were lower and decreased gradually with age to values for adults [15.6 (SE 2.9) pmol/L], increasing again significantly (P less than 0.01) in persons older than 70 years [37.1 (SE 32) pmol/L]. There was no significant sex-related difference in values for plasma. For the pathological conditions studied, we observed significantly supranormal values in patients with chronic renal failure [175.1 (SE 35.9) pmol/L]. Some of the pregnant patients in their third trimester also showed high values. A small but significant increase in plasma IR-7B2 was observed after a glucose load, both in control subjects and diabetics. Perhaps the kidney plays a major role in 7B2 degradation.

Adolescent↗

Heat-labile alkaline phosphatase from Antarctic bacteria: Rapid 5' end-labeling of nucleic acids.

A heat-labile alkaline phosphatase has been purified to near homogeneity from HK47, a bacterial strain isolated from Antarctic seawater. The active form of the enzyme has a molecular weight of 68,000 and is uniquely monomeric. The optimal temperature for the enzymatic activity is 25 degrees C. Complete and irreversible thermal inactivation of the enzyme occurs in 10 min at 55 degrees C. By using this heat-labile enzyme for dephosphorylation followed by a 10-min heat treatment, rapid end-labeling of nucleic acids by T4 polynucleotide kinase has been achieved.

Journal Article↗

Bacterial plasmids in antarctic natural microbial assemblages.

Samples of psychrophilic and psychrotrophic bacteria were collected from sea ice, seawater, sediments, and benthic or ice-associated animals in McMurdo Sound, Antarctica. A total of 155 strains were isolated and tested for the presence of plasmids by DNA agarose gel electrophoresis. Thirty-one percent of the isolates carried at least one kind of plasmid. Bacterial isolates taken from sediments showed the highest plasmid incidence (42%), and isolates from seawater showed the lowest plasmid incidence (20%). Plasmids were significantly more frequent in the strains which had been first isolated from low-nutrient media (46%) than in the strains which had been isolated from high-nutrient media (25%). Multiple forms of plasmids were observed in two-thirds of the plasmid-carrying strains. A majority of the plasmids detected were estimated to have a mass of 10 megadaltons or less. Among 48 plasmid-carrying strains, 7 showed antibiotic resistance. It is concluded that bacterial plasmids are ubiquitous in natural microbial assemblages of the pristine marine ecosystem of Antarctica.

Journal Article↗

[Effects of gamma-oryzanol on the hypothalamo-pituitary axis in the rat].

The effects of gamma oryzanol (gamma-OZ), a ferulic acid of triterpene alcohol, on the synthesis and release of the growth hormone (GH) and prolactin (PRL) in vitro and turnover rates of hypothalamic catecholamines were investigated. A single subcutaneous injection of 20 mg/kg of gamma-OZ suppressed GH synthesis and PRL release 1 hour after the injection. gamma-OZ increased medial basal hypothalamic (MBH) dopamine (DA) content, and the DA content was decreased by a treatment of alpha-methyl-p-tyrosine (alpha MpT), a tyrosine hydroxylase inhibitor, indicating increased synthesis and release of DA in MBH by gamma-OZ. gamma-OZ did not alter norepinephrine (NE) content in MBH, while the NE content was significantly decreased, indicating unchanged synthesis and increased release of NE in MBH by gamma-OZ. These results may explain the previous data concerning the changes in serum levels of GH and PRL by gamma-OZ and also suggest that gamma-OZ can affect the synthesis and/or release of at least two hypothalamic neurotransmitters, DA and NE, resulting in the alterations of anterior pituitary hormone synthesis and/or release.

Animals↗

Estimation of inter-electrode distance for body surface mapping: application of Fourier analysis to potential distribution due to the epicardial breakthrough.

For the purpose of determining an appropriate distance of lead points for constructing body surface maps, Fourier analysis was performed on potential distribution reflecting the epicardial breakthrough, and inter-electrode distance necessary for sampling of harmonics contributing to the body surface potential distribution was determined by application of sampling theorem. Potential distribution was simultaneously recorded with an interval of 4 mm for 250 msec along the vertical (head to foot) and horizontal (right to left) line crossing a second-minimum appearing on the chest surface of a healthy adult, and the data obtained from 9.6 cm along the respective lines underwent Fourier transform. Relative contribution of the second harmonics to the original wave forms increased with time lapse after QRS initiation, in accordance with reductions of the first harmonics component, and attained the maximum (32.5% of total power spectrum without d-c component) at the instant of occurrence of the second-minimum. A similar tendency was observed among higher harmonics, but their contribution was comparatively low (within 10%). Inter-electrode distance determined by sampling theorem was 4.8 cm for the first harmonics and 2.4 cm for the second harmonics. In conclusion it is proposed that placement of electrodes with an inter-electrode distance of 2.4 cm is necessary for acquiring clinically important data on the epicardial breakthrough.

Electrocardiography↗