Biomedical subjects

K Yoshimura

Publications and source records attributed to K Yoshimura.

At least 181 records · Page 10Linked to original sources

PubMed1998

Isolation and characterization of novel Chlamydomonas mutants that display phototaxis but not photophobic response.

The unicellular green alga Chlamydomonas displays two distinct kinds of behavioral response to light: phototaxis, in which cells swim toward or away from the light source under constant illumination; and photophobic responses (also called stop responses or photoshock responses), in which cells transiently convert their flagellar waveform and swim backward upon sudden increase in light intensity. It has been suggested that the two responses partly share a common signal transduction pathway, but exactly how the different responses are produced has not been established. In this study, to help understand the molecular and cellular mechanisms that bring about the photophobic response, we isolated novel mutants (ppr1, ppr2, ppr3, and ppr4) that do not show the photophobic response. Importantly, these mutants retain the ability to display phototaxis, with almost the same sensitivities as in the wild type cell. Demembranated and reactivated flagellar axonemes of the ppr mutants were found to convert the bending patterns depending on the Ca2+ concentration, indicating that the axonemal mechanism for waveform conversion required for the photophobic response was unaffected by the mutations. In addition, measurements of electric currents in cell suspensions showed that these mutants generate normal photoreceptor currents (PRC) upon photostimulation, suggesting that they retain the normal activity of photoreception and the ionic channels that produce PRCs. However, the all-or-none flagellar current (FC), a Ca2+ current generated by PRC-induced depolarization of flagellar membrane, was absent or seriously impaired in the mutants. These findings clearly indicate that the all-or-none FC is necessary for the photophobic response but not for phototaxis. The isolation of the four genetically independent ppr mutants suggests that the generation of the FC is based on multiple components that are not used in the mechanism for phototaxis, and implies that the Chlamydomonas flagellar membrane possesses a voltage-dependent Ca2+-channel specifically used for generation of photophobic responses.

Animals↗

PubMed1998

Myosin heavy chain expression in skeletal muscle autografts under neural or aneural conditions.

BACKGROUND: Our purpose was to investigate (1) the heterogeneity of satellite cells derived from adult fast-twitch and slow-twitch skeletal muscles, (2) the influence of innervation on muscle regeneration, and (3) the differences between developmental myoblasts and satellite cells with regard to myosin heavy chain (MHC) expression. MATERIALS AND METHODS: Autografts under neural (nerve-intact graft; brief denervation interval) or aneural (aneural graft; prolonged denervation interval) conditions of the fast-twitch extensor digitorum longus (EDL) muscle or the slow-twitch soleus muscle were performed in adult rat hindlimbs. MHC expression during skeletal muscle regeneration was determined sequentially using immunocytochemistry. RESULTS: After grafting, most muscle fibers in the EDL and soleus underwent ischemic degeneration and regeneration; at the periphery of each muscle, a few adult fibers survived. All regenerating fibers initially expressed embryonic/fetal (developmental) MHC alone, and subsequently both developmental and fast MHC. During the first week, no expression of slow MHC was observed in regenerating fibers in either the EDL or the soleus. In nerve-intact grafts, regenerating fibers expressed slow MHC as early as the second week; under aneural conditions, no regenerating fibers expressed slow MHC even 4 weeks after grafting. On the other hand, some persisting fibers in aneural grafts could maintain expression of slow MHC 4 weeks after grafting; other fibers underwent MHC transformation induced by denervation. No significant difference in MHC expression during regeneration was observed for slow compared with fast muscles, under either neural or aneural condition. CONCLUSIONS: These data suggest that regenerating adult skeletal muscle fibers, derived only from satellite cells, cannot express slow MHC without motor innervation, and that persisting muscle fibers, derived from both myoblasts in fetal development and satellite cells, may be intrinsically distinct from regenerating fibers. Satellite cells derived from slow and from fast muscles may be a single, homogenous population and may be the same population as fetal (secondary) myoblasts with regard to MHC expression.

Animals↗

PubMed1998

Immunohistochemical analysis of clinically transplanted muscles.

BACKGROUND: Although a number of studies have examined the morphology and function of experimentally transplanted muscles, immunohistochemical evaluation of clinically transplanted muscles has not been reported. The purpose of this study was to examine clinically transplanted muscles at long periods after transplantation with biochemical markers specific for satellite cell activation and muscle regeneration. MATERIALS AND METHODS: Nine biopsies of muscles transplanted to the paralyzed face were examined. In five cases, the gracilis muscles were transplanted about 1 year after cross face nerve grafting. The other four cases underwent one-stage latissimus dorsi (LD) muscle transplantation. Twelve to 162 months after transplantation, muscle biopsies were harvested in nine cases. In eight cases, secondary corrections of facial expression including debulking of the grafted muscle were required, while another muscle was transplanted in one case because of the failed first operation. As control, six specimens of normal LDs were examined as well. Monoclonal antibodies were employed to visualize myosin heavy chain (MHC) isoforms (slow, fast, and embryonic) and MyoD protein. RESULTS: Although one specimen exhibited only small, atrophic fibers indicating failed reinnervation, the remaining eight specimens showed regularly distributed fibers and type grouping indicating successful reinnervation. There was no statistically significant difference in fiber area and lesser diameter between normal LDs and transplanted LDs. However, even in these successfully reinnervated muscles, intermediate and small fibers expressing embryonic MHC and small cells expressing MyoD were observed, suggesting that satellite cells were activated for repair of the adjacent fibers. CONCLUSIONS: Muscle adaptation (presumably to denervation), which is a regenerative change accompanied by activation of satellite cells, was still seen even long periods after transplantation. It is concluded that, in microneurovascular human skeletal muscle transfers, there is a wide variation in the time required for reinnervation of individual muscle fibers, and it may be that human muscle fibers cannot be properly reinnervated after denervation has continued for a certain period such as 12 months.

Adult↗

PubMed1998

Obstruction of the right ventricle outflow tract during right cardiac prolapse.

We observed cardiac prolapse following right pneumonectomy in which the patient experienced a 90 degree rotation of the heart and compression of right ventricular outflow tract from behind by the aorta, which led to an increase in right atrial pressure and a decrease of pulmonary artery pressure and cardiac output. Compression of the right ventricular outflow tract by the base of the aorta was observed as an additional shock inducing factor in right cardiac prolapse.

Carcinoma, Squamous Cell↗

PubMed1998

High urinary excretion level of citrate and magnesium in children: potential etiology for the reduced incidence of pediatric urolithiasis.

It is well known that the incidence of calcium oxalate (CaOX) urolithiasis is much lower in children than in adults [2, 21]. One purpose of this study was to compare the inhibitory activity on CaOX crystal growth and nucleation of urine from children (ufC) with that of urine from adults (ufA). Another was to measure low molecular weight urinary substances related to CaOX lithiasis, including citrate and magnesium, which have been identified as stone inhibitors. The excretion volume per body weight of uric acid, phosphorus, magnesium and citrate was all significantly higher in 24-h ufC than in 24-h ufA, but that of calcium and oxalate was not. The growth inhibitory activities against CaOX crystals of ufC and ufA were measured in a whole urine system. The diameter of the crystals produced in this system was smaller for ufC (3.68 microm) than for ufA (4.66 microm). We also examined the metastable limit for CaOX with fresh spot urine, which was 3.15 mmol/l in ufC and 0.41 mmol/l in ufA. These results indicate that ufC has a more powerful inhibitory effect on CaOX crystal growth and nucleation than ufA. We also found that the excretion rate of citrate and magnesium in ufC was much higher than in ufA. We suggest that these two stone inhibitors are very likely to elevate the inhibitory activity of ufC against CaOX crystal growth and nucleation. The lower incidence of CaOX lithiasis in children might thus be partly attributed to citrate and magnesium.

Adult↗

PubMed1998

Arterial reconstruction: justified for patients with intermittent claudication?

The objective of this study was to evaluate the effects of arterial reconstruction in patients with intermittent claudication. A total of 243 patients (305 limbs) underwent lower extremity vascular reconstruction at our institution from 1979 to 1995. They were assessed by physical examination, pulse volume recordings, segmental pressure, Duplex ultrasonography, and intravenous subtraction arteriography to evaluate the effects of arterial reconstruction. Surviving patients (220 limbs) were enrolled for evaluation of outcome during the follow-up period. There were 59 deaths during the follow-up period. The cumulative life-table 5-year patency rates were 90% +/- 3%, 73% +/- 6%, and 74% +/- 10% for aortoiliac, infrainguinal, and aortofemorodistal arterial reconstructions, respectively. Among 129 repairs in the aortoiliac region, Fontaine stages I, II, and III were found in 109 limbs (84.5%), 17 limbs (13.2%), and 2 limbs (1.5%), respectively. There was one (0.7%) minor amputation. There was Fontaine stage I in 50 limbs (76.9%), Fontaine stage II in 14 limbs (21.5%), and Fontaine stage III in 1 limb (1.5%) for repairs in the infrainguinal region. Among the 26 aortofemorodistal repairs, there was Fontaine stage I in 21 limbs (81%) and Fontaine stage II in 5 limbs (19%). There was a statistically significantly higher incidence of Fontaine stage I than Fontaine stage II or III in aortofemoral, infrainguinal, and aortofemorodistal arterial reconstructions (p < 0.0001). Arterial reconstruction for patients at Fontaine stage II offered benefits and improved quality of life at follow-up. Arterial reconstruction for patients at Fontaine stage II offers benefits and improved quality of life at follow-up. It was concluded that the significant improvement in quality of life after arterial reconstruction warrants continued use of the procedure in patients with intermittent claudication.

Adult↗

PubMed1998

The role of CD4+ and CD8+ T-cells in host morbidity and innate resistance to angiostrongylus cantonensis in the mouse.

Strain-dependent differences in host morbidity and mortality due to Angiostrongylus cantonensis infection have been established between C57BL/6 and BALB/c mice; C57BL/6 mice show rapid worm killing with low morbidity, whereas BALB/c mice indicate slow worm killing with high morbidity and mortality. To determine the possible roles of CD4+ and CD8+ T-cells in host morbidity and innate resistance to A. cantonensis infection we treated C57BL/6 and BALB/c mice with anti-CD4 or anti-CD8 monoclonal antibody and examined the changes in host morbidity and worm-killing activity. Our study indicates that anti-CD4 antibody treatment interferes with worm killing and improves the morbidity of A. cantonensis-infected BALB/c mice, whereas anti-CD8 antibody treatment fails to improve the morbidity. Tumor necrosis factor-alpha (TNF-alpha, or cachectin) production in infected mice was not correlated with host morbidity. Anti-IL-5 monoclonal antibody treatment also failed to affect the morbidity of infected BALB/c mice, although their worm-killing activity was restrained as shown in anti-CD4-treated mice. These findings clearly indicate that the morbidity of infected BALB/c mice is regulated by some unknown CD4+ T-cell-dependent mechanism but not by an IL-5-, eosinophil-, or TNF-alpha-dependent mechanism.

Angiostrongylus cantonensis↗

PubMed1998

Negative association between asthma and variants of CC16(CC10) on chromosome 11q13 in British and Japanese populations.

The gene encoding Clara cell-derived inflammatory molecule CC16 has been cited as a candidate gene for atopic asthma on chromosome 1lq13. A genetic association study was performed with variants of the CC16 gene on chromosome 1lq13 in relation to asthma in British (n=275) and Japanese (n=300) populations. No significant association was found between asthma and CC16 genotypes, irrespective of atopic status in these two populations. These data suggest that CC16 might not be the major locus for asthma on 11q13.

Asthma↗

PubMed1998

Purification and characterization of ascorbate peroxidase in Chlorella vulgaris.

Chlorella vulgaris contained only one isoform of ascorbate peroxidase (AsAP) as the hydrogen peroxide (H2O2)-scavenging system except for catalase at a specific activity of 3.3 +/- 0.2 units/mg protein. The activity of glutathione peroxidase was not detected in the extracts from cells grown in the absence and presence of sodium selenite. We detected the activity of monodehydroascorbate reductase involved in the regeneration of ascorbate, but we failed to detect the dehydroascorbate reductase activity. AsAP has been purified to electrophoretic homogeneity from Chlorella cells. The enzyme was a monomer with a molecular mass of 32 kDa using gel filtration and SDS-polyacrylamide gel electrophoresis. The enzyme showed higher specificity with ascorbate than with pyrogallol. The K(m) values of the enzyme for ascorbate and H2O2 were 111 +/- 8.9 and 20 +/- 2.5 microM, respectively. When the enzyme was diluted with the ascorbate-deleted medium, the half inactivation time was approximately 15 min. The absorption spectra of the purified enzyme and the inhibition by cyanide and azide showed that it is a hemoprotein. The enzyme was markedly inhibited by 0.2 mM p-chloromercuribenzoate. The enzyme cross-reacted by immunoblotting with the monoclonal antibody raised against Euglena cytosolic AsAP. The amino acid sequences in the N-terminal region of Chlorella AsAP showed no significant similarity to any other AsAPs from higher plants and algae.

Amino Acid Sequence↗

PubMed1998

A case of poliomyelitis-like syndrome.

The authors report the case of a boy aged 4 years who had sudden abdominal pain and inability to walk on the day before admission to hospital and who developed abdominal distention and difficulty urinating. On admission, the abdominal skin reflexes, knee jerks, cremaster and anal reflexes were absent and power in the lower extremities was reduced. Spinal fluid examination showed a cell count of 383/mm3, with 95% neutrophils and 5% lymphocytes; spinal fluid protein of 44 mg/dl; and glucose 75 mg/dl. Serological studies did not reveal any significant antibodies for polio virus type 1, 2, 3, various ECHO viruses, Coxsackie types A4, A7, A9, B1 or B5. However, the titer of Coxsackie virus antibody type A10 was 128 in the acute phase and only 32 in the recovery phase 4 weeks later. Magnetic resonance scans were performed on the second day; the findings were normal in the brain, but interesting lesions were revealed in the thoracic cord with both T1-weighted images and T2-weighted images. Neurological symptoms improved asymmetrically.

Child, Preschool↗

PubMed1998

Focal pachypolymicrogyria in three siblings.

The malformation of focal pachypolymicrogyria might be the manifestation of an X-linked recessive disorder according to the results of this study. Three siblings revealed focal pachypolymicrogyria on magnetic resonance imaging (MRI) and had a strong family history of epilepsy and mental retardation. All three siblings had the same mother; the father of Patient 1 was not related to the mother, but the father of Patients 2 and 3 was related to her. The MRI of the father of Patients 2 and 3 demonstrated focal pachypolymicrogyria. The mother's MRI was normal. In this family, epilepsy or mental retardation was found mainly in the males (Patient 3 was an exception), and they were all born to female members of this family, not male. Patient 3 was probably a homozygote with an X-linked recessive inheritance, and therefore, she demonstrated the most severe clinical findings.

Adolescent↗

PubMed1998

Analysis of T cell populations and IL-3 mRNA expression in mesenteric lymph node cells and intestinal intraepithelial lymphocytes in Strongyloides ratti-infected mice.

T cell populations and IL-3 mRNA expression were analysed in mesenteric lymph node cells and intestinal intraepithelial lymphocytes (IEL) in Strongyloides ratti-infected mice. On days 7 and 12 post-infection, 2.6 times as many mesenteric lymph node cells were present in S. ratti-infected mice compared with uninfected mice. Although the percentages of CD3+, CD4+ and CD8+ cells decreased during infection, the absolute numbers of these cell types increased on day 7 due to an overall increase in the mesenteric lymph node cell number. The CD4/CD8 ratio in IEL was increased on day 5, whereas no significant change in the CD4/CD8 ratio was observed in the mesenteric lymph node cells. Expression of IL-3 mRNA, which is an important cytokine for the induction of murine mucosal mastocytosis and S. ratti-expulsion, was examined in mesenteric lymph nodes and IEL of uninfected and infected mice. IL-3 mRNA was detected in mesenteric lymph nodes of S. ratti-infected mice but not detected in the lymph nodes of uninfected mice. IL-3 mRNA was detected in IEL from both infected and uninfected mice with an 20-fold increase in expression in IEL of infected mice. Overall, IL-3 mRNA levels were higher in IEL than in mesenteric lymph nodes following S. ratti-infection. Expression of IL-4, IL-10, stem cell factor (SCF or c-kit ligand) and IFN-gamma mRNA was also examined in these two tissues. IL-10 mRNA was not detected in any tissue examined and IFN-gamma mRNA levels were unaltered as a result of an S. ratti-infection. Elevated expression of mRNA for SCF (5-fold) and IL-4 (20-fold) was observed in the mesenteric lymph nodes of infected mice. In contrast, SCF mRNA levels were similar in IEL of uninfected and infected animals and only a modest increase in IL-4 mRNA was observed in IEL of infected mice.

Animals↗

PubMed1998

Expression of Tamm-Horsfall protein in stone-forming rat models.

OBJECTIVES: To examine the expression of Tamm-Horsfall protein (THP) and calcium oxalate deposition in three rat models to clarify whether THP plays an active role in crystal formation or whether crystals induce the secretion of this protein. MATERIALS AND METHODS: A stone-forming rat model (model 1) with marked tubular dilatation in an entire kidney was produced by rendering Wistar rats (aged 8 weeks) hyperoxaluric and hypercalciuric, through compulsorily feeding with 0.12 mL of 5% ethylene glycol (in two doses daily) and 0.5 microgram of vitamin D3 every other day. Two other rat models were also produced. Model 2 comprised stone-forming rats with minimal tubular dilatation, achieved by giving rats the same dose of ethylene glycol once daily, and model 3 comprised stone-free rats with marked tubular dilatation achieved by unilateral ureteric ligation. The rats' kidneys were resected after 4 weeks and all resected kidneys immunohistochemically stained with an antibody to THP. Simultaneously, the location of calcium oxalate (CaOx) crystals was established with von Kossa staining. The relation between crystals and the secretion of THP was also assessed in vitro. Cultured renal epithelial cells (NRK-52E) were stained with an antibody to THP after they had been cultured for 72 h in a medium containing CaOx crystals. RESULTS: In model-1 kidneys with both tubular dilatation and many crystals, there was local and intense expression of THP in many renal tubules. CaOx crystals and the intense expression of THP tended to occur in the same renal tubules. In model 2 kidneys with little tubular dilatation, only a few renal tubules expressed THP strongly and the location of the crystals rarely coincided with that of THP expression. In model 3 kidneys with marked tubular dilatation but no crystals, THP was expressed strongly in many renal tubules. The expression of THP in cultured NRK-52E cells was not stimulated by CaOx crystals. CONCLUSIONS: The results from the in vivo models suggest that THP did not initiate crystal formation and the strong expression of THP was induced not by crystals but by renal tubular damage caused by tubular dilatation. From the close association of THP and crystals in model 1 kidneys, this protein might play a secondary role as an adhesive, promoting stone formation.

Adjuvants, Immunologic↗

PubMed1998

Use of the disposable stapler to insure proper fixation of a transferred muscle in treatment of facial paralysis.

The authors introduce the use of a disposable stapler to create a stable muscle end in transferring a free neurovascular muscle segment for the treatment of facial paralysis. This procedure allows the firm fixation of the fragile soft end of a muscle transferred to the nasolabial region of a paralyzed face, and avoids adhesion and tethering of the muscle end to the skin, which may result in inaccurate positioning of a newly-created nasolabial fold and an asymmetrical smile. Transection and hemostasis at both ends of the harvested muscle segment also can be achieved simultaneously. Although the disposable stapler incurs additional cost, it can greatly improve the result of a free neurovascular muscle transfer for the treatment of facial paralysis, as well as simplify harvesting of the muscle segment. No complications and problems have been encountered in stapling the muscle.

Disposable Equipment↗

PubMed1998

Comparison of clinical outcomes of laparoscopic and conventional open adrenalectomy.

From September 1992 to November 1996, 28 patients underwent laparoscopic adrenalectomy at Osaka University Medical Hospital. They were compared with 25 instances of conventional open surgery performed between May 1990 and April 1996 at the same institution. Laparoscopic adrenalectomy was performed via either a transperitoneal or a retroperitoneal approach. The mean operative time of 375 minutes for laparoscopic adrenalectomy was significantly longer than that of 133 minutes for open surgery. The average hospital stay for laparoscopic surgery was significantly shorter than that of conventional open adrenalectomy. The convalescent period was also significantly shorter in the patients who had laparoscopic adrenalectomy. There was no statistical difference in blood loss during the operation or the number of doses of analgesics administered after operation in the two groups. We conclude that laparoscopic adrenalectomy is one of the options to be selected in surgically managing adrenal tumors. Laparoscopic adrenalectomy could become a standard operative procedure as instruments and techniques of laparoscopy improve significantly.

Adolescent↗

PubMed1998

Molecular characterization and physiological role of a glyoxysome-bound ascorbate peroxidase from spinach.

cDNAs encoding two cytosolic and two chloroplastic ascorbate peroxidase (AsAP) isozymes from spinach have been cloned recently [Ishikawa et al. (1995) FEBS Lett. 367: 28, (1996) FEBS Lett. 384: 289]. We herein report the cloning of the fifth cDNA of an AsAP isozyme which localizes in spinach glyoxysomes (gAsAP). The open reading frame of the 858-base pair cDNA encoded 286 amino acid residues with a calculated molecular mass of 31,507 Da. By determination of the latency of AsAP activity in intact glyoxysomes, the enzyme, as well as monodehydroascorbate (MDAsA) reductase, was found to be located on the external side of the organelles. The cDNA was overexpressed in Escherichia coli (E. coli). The enzymatic properties of the partially purified recombinant gAsAP were consistent with those of the native enzyme from intact glyoxysomes. The recombinant enzyme utilized ascorbate (AsA) as its most effective natural electron donor; glutathione (GSH) and NAD(P)H could not substitute for AsA. The substrate-velocity curves with the recombinant enzyme showed Michaelis-Menten type kinetics with AsA and hydrogen peroxide (H2O2); the apparent Km values for AsA and H2O2 were 1.89 +/- 0.05 mM and 74 +/- 4.0 microM, respectively. When the recombinant enzyme was diluted with AsA-depleted medium, the activity was stable over 180 min. We discuss the H2O2-scavenging system maintained by AsAP and the regeneration system of AsA in spinach glyoxysome.

Amino Acid Sequence↗

PubMed1998

One-stage transfer of the latissimus dorsi muscle for reanimation of a paralyzed face: a new alternative.

The two-stage method combining neurovascular free-muscle transfer with cross-face nerve grafting is now a widely accepted procedure for dynamic smile reconstruction in cases with long established unilateral facial paralysis. Although the results are promising, the two operations, about 1 year apart, exert an economic burden on the patients and require a lengthy period before obtaining results. Sequelae such as hypoesthesia, paresthesia, and conspicuous scar on the donor leg for harvesting a sural nerve graft also cannot be disregarded. To overcome such drawbacks of the two-stage method, we report a refined technique utilizing one-stage microvascular free transfer of the latissimus dorsi muscle. Its thoracodorsal nerve is crossed through the upper lip and sutured to the contralateral intact facial nerve branches. Reinnervation of the transferred muscle is established at a mean of 7 months postoperatively, which is faster than that of the two-stage method. In our present series with 24 patients, 21 patients (more than 87 percent) believed that their results were excellent or satisfactory, which also compares well with the results of the two-stage method combining free-muscle transfer with cross-face nerve graft.

Adolescent↗

PubMed1998

Incidence of lymph node metastasis and its impact on long-term prognosis in clinically localized prostate cancer.

BACKGROUND: Pelvic lymph node dissection (PLND) is an important staging method for men with clinically localized prostate cancer. We report our experience with staging PLND and the impact of lymph node metastasis on long-term prognosis. METHODS: One hundred forty-eight consecutive patients who underwent staging PLND for clinically localized prostate cancer were retrospectively studied. Patients were evaluated for the presence and number of lymph node metastases, treatment (prostatectomy vs. radiotherapy), and endocrine therapy, and analyzed with respect to disease progression and survival. The mean follow-up period was 52.9 months (range, 2.3 to 165.8 months). RESULTS: Thirty-two patients (21.6%) had pelvic lymph node metastases, the incidence of which markedly decreased from 32.3% in 1982 to 1987 to 6.7% in 1994 to 1997. The intervals to disease progression and cancer death were significantly shorter in patients with positive lymph nodes (P < 0.001). In stage D1 disease, patients who underwent a radical prostatectomy tended to be free of progression longer than those receiving radiotherapy or conservative therapy (P = 0.0546). Other factors, such as early endocrine therapy, the extent of lymph node involvement and the Gleason score of the primary tumor did not predict disease progression or survival. CONCLUSION: These data suggest a decreasing trend in the incidence of lymph node metastasis in the PSA era. Although longer disease-free intervals were observed in radical prostatectomy-treated patients, the impact of an aggressive approach to stage D1 disease awaits further studies.

Adenocarcinoma↗