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K Sugasawa

Publications and source records attributed to K Sugasawa.

At least 37 records · Page 2Linked to original sources
PubMed1997

Analyses of neuro-otological complications after radiosurgery for acoustic neurinomas.

PURPOSE: To find out the optimum treatment parameters and the proper indications for treatment of acoustic neurinomas, univariate and multivariate actuarial analyses of neuro-otological complications after stereotactic radiosurgery for acoustic neurinomas were performed. METHODS AND MATERIALS: The subjects were 46 patients with acoustic neurinomas who underwent unilateral radiosurgery between June 1990 and June 1994 and were followed up at the University of Tokyo. Age ranged from 13 to 77 years (median, 54 years). Tumor diameter ranged from 0 to 25 mm (mean, 12 mm) at the cerebellopontine angle and from 2 to 15 mm (mean, 8.3 mm) in the internal auditory meatus. Maximum tumor doses ranged from 20 to 40 Gy (mean, 31.4 Gy), and peripheral doses from 12 to 25 Gy (mean, 16.8 Gy). One to eight isocenters were used (mean, 3.2). Median follow-up was 39 months. Eight events concerning neuro-otological complications were chosen, and the potential risk factors for them were analyzed by the actuarial analyses (univariate and multivariate). The events examined include hearing loss, vestibular function loss, facial palsy, and trigeminal nerve dysfunction. In order to point out potential risk factors for neuro-otological complications, univariate analyses were performed using both the Wilcoxon test and the log rank test, and multivariate analyses were performed with the Cox proportional hazards model. Variables nominated as potential risk factors were 1) demographic variables such as patient age and sex, 2) tumor dimensions, 3) treatment variables such as tumor doses and number of isocenters, and 4) pretreatment hearing levels. A variable with significant p-values (p < 0.05) in two or more of the three actuarial analyses (two univariate and one multivariate) was considered a possible risk factor. RESULTS: The possible variables that increase the risk for each event analyzed were: neurofibromatosis type II (NF2) and the number of isocenters for total hearing loss; experience of prior operation, the tumor diameter in the internal auditory meatus, and NF2 for hearing threshold elevation; peripheral tumor dose for vestibular function loss; patient age or midporus transverse tumor diameter (the two variables were correlated), and the number of isocenters for facial palsy; and the number of isocenters for trigeminal neuropathy. CONCLUSION: NF2 and the tumor diameter were the common risk factors for hearing loss in previous studies and ours. For the 5th/7th nerve dysfunction, the tumor diameter was the common risk factor. The risk of using more isocenters remains controversial. The difference in risk factors for hearing impairment and vestibular function loss suggests different mechanisms for the two. Further studies with larger populations and longer follow-up periods are required in order to draw conclusions on the risk factors in radiosurgery.

Adolescent↗
PubMed1997

Identification and characterization of XPC-binding domain of hHR23B.

hHR23B was originally isolated as a component of a protein complex that specifically complements nucleotide excision repair (NER) defects of xeroderma pigmentosum group C cell extracts in vitro and was identified as one of two human homologs of the Saccharomyces cerevisiae NER gene product Rad23. Recombinant hHR23B has previously been shown to significantly stimulate the NER activity of recombinant human XPC protein (rhXPC). In this study we identify and functionally characterize the XPC-binding domain of hHR23B protein. We prepared various internal as well as terminal deletion products of hHR23B protein in a His-tagged form and examined their binding with rhXPC by using nickel-chelating Sepharose. We demonstrate that a domain covering 56 amino acids of hHR23B is required for binding to rhXPC as well as for stimulation of in vitro NER reactions. Interestingly, a small polypeptide corresponding to the XPC-binding domain is sufficient to exert stimulation of XPC NER activity. Comparison with known crystal structures and analysis with secondary structure programs provided strong indications that the binding domain has a predominantly amphipathic alpha-helical character, consistent with evidence that the affinity with XPC is based on hydrophobic interactions. Our work shows that binding to XPC alone is required and sufficient for the role of hHR23B in in vitro NER but does not rule out the possibility that the protein has additional functions in vivo.

Amino Acid Sequence↗
PubMed1997

Two human homologs of Rad23 are functionally interchangeable in complex formation and stimulation of XPC repair activity.

XPC-hHR23B protein complex is specifically involved in nucleotide excision repair (NER) of DNA lesions on transcriptionally inactive sequences as well as the nontranscribed strand of active genes. Here we demonstrate that not only highly purified recombinant hHR23B (rhHR23B) but also a second human homolog of the Saccharomyces cerevisiae Rad23 repair protein, hHR23A, stimulates the in vitro repair activity of recombinant human XPC (rhXPC), revealing functional redundancy between these human Rad23 homologs. Coprecipitation experiments with His-tagged rhHR23 as well as sedimentation velocity analysis showed that both rhHR23 proteins in vitro reconstitute a physical complex with rhXPC. Both complexes were more active than free rhXPC, indicating that complex assembly is required for the stimulation. rhHR23B was shown to stimulate an early stage of NER at or prior to incision. Furthermore, both rhHR23 proteins function in a defined NER system reconstituted with purified proteins, indicating direct involvement of hHR23 proteins in the DNA repair reaction via interaction with XPC.

DNA Repair↗
PubMed1997

Postnatal development of the masseter muscles in the Japanese field vole Microtus montebelli, with special attention to differentiation of the fast-twitch oxidative fiber.

Postnatal development and differentiation of the masseter muscles consisting only of fast-twitch oxidative (FO) fibers in the adult Japanese field vole Microtus montebelli were studied using histochemical and electron microscopic techniques. The masseter muscles were composed of myotubes and muscle fibers at day 0 (birth day). Most muscle cells showed the strong reaction for myosin ATPase after both alkaline and acid preincubations. For NADH-dehydrogenase (NADH-DH), small granular diformazan deposits were recognized in the sarcoplasm. Afterwards, the masseter muscles consisted of myofibers and satellite cells at day 5. For myosin ATPase, weakly-reactive fibers after acid preincubation (fast-twitch fibers) increased in number. For NADH-DH, granular diformazan deposits in all the myofibers increased in size. Since all the myofibers had numerous sarcoplasmic reticula, and they reacted strongly after alkaline preincubation and weakly after acid preincubation for myosin ATPase at day 10 when the young start to take solid food, it seems that the masseter muscles become contractive fast. At day 15 (before weaning), all the myofibers showed the adult-like strong reaction for NADH-DH and had numerous well-developed mitochondria, thus they acquired the ability of the fast and sustained contraction. It is accordingly considered that the masseter muscles of the vole mature in a short time after birth because of adaptation for herbivorous food habit.

Adaptation, Physiological↗
PubMed1996

XPC and human homologs of RAD23: intracellular localization and relationship to other nucleotide excision repair complexes.

The xeroderma pigmentosum syndrome complementation group C (XP-C) is due to a defect in the global genome repair subpathway of nucleotide excision repair (NER). The XPC protein is complexed with HHR23B, one of the two human homologs of the yeast NER protein, RAD23 (Masutani at al. (1994) EMBO J. 8, 1831-1843). Using heparin chromatography, gel filtration and native gel electrophoresis we demonstrate that the majority of HHR23B is in a free, non-complexed form, and that a minor fraction is tightly associated with XPC. In contrast, we cannot detect any bound HHR23A. Thus the HHR23 proteins may have an additional function independent of XPC. The fractionation behaviour suggests that the non-bound forms of the HHR23 proteins are not necessary for the core of the NER reaction. Although both HHR23 proteins share a high level of overall homology, they migrate very differently on native gels, pointing to a difference in conformation. Gel filtration suggests the XPC-HHR23B heterodimer resides in a high MW complex. However, immunodepletion studies starting from repair-competent Manley extracts fall to reveal a stable association of a significant fraction of the HHR23 proteins or the XPC-HHR23B complex with the basal transcription/repair factor TFIIH, or with the ERCC1 repair complex. Consistent with a function in repair or DNA/chromatin metabolism, immunofluorescence studies show all XPC, HHR23B and (the free) HHR23A to reside in the nucleus.

Amino Acid Sequence↗
PubMed1996

Neuro-otological findings after radiosurgery for acoustic neurinomas.

OBJECTIVE: To evaluate the neuro-otological complications in patients after radiosurgery for acoustic neurinomas. DESIGN: Inception cohort, retrospective study. SETTING: University hospital. PATIENTS: A consecutive sample of 46 patients with acoustic neurinomas who underwent unilateral gamma knife radiosurgery at the University of Tokyo, Japan, between June 1990 and June 1994 were followed up by otolaryngologists for more than 3 months. INTERVENTION: Gamma knife stereotactic radiosurgery. MAIN OUTCOME MEASURES: Neuro-otological examinations including pure tone audiometry, auditory brain stem response, and caloric test. RESULTS: Tumor growth occurred in 2 patients (4.3%). Seven (18%) of the 38 patients with preserved hearing of any extent became deaf within 1 year. In cases of gradual hearing loss, the average deterioration rate was approximately 8 dB per year. Abnormalities of auditory brain stem response preceded deafness in 2 patients. Caloric response, preserved before treatment in 13 patients, disappeared 4 to 13 months after treatment (median, 8 months) in 9 (69%) of them, whereas their hearing was preserved. Delayed facial palsy and persistent trigeminal neuropathy occurred in 10 (22%) and 7 (15%) of the 46 patients, respectively. Severe facial palsy tended to persist. CONCLUSIONS: The rates of neuro-otological complications of radiosurgery are almost comparable with those previously reported from other institutions. The deafness within 1 year after treatment might be attributed to a lesion in the cochlear nerve. Hearing loss did not parallel vestibular function loss. The persistent severe facial palsy contrasts with previously reported findings. Considering the serious facial nerve complications that occurred in some of our patients, further study to disclose the risk factors for neurological dysfunction would be needed for radiosurgery to become a true, safe alternative to microsurgery.

Adolescent↗
PubMed1996

Auditory findings after stereotactic radiosurgery in acoustic neurinoma.

Auditory function after gamma knife stereotactic radiosurgery (GK) was evaluated in 22 patients with newly diagnosed unilateral acoustic neurinoma who had had some measurable hearing preoperatively. No tumor growth was observed in any patients. The preservation rates for useful hearing (n = 13) and any measurable hearing (n = 22) were 62% and 86%, respectively, at 2 years after GK. Postoperative hearing dysfunction was attributed to progressive cochlear impairment in 13 patients and to progressive injury in the retrocochlear auditory pathway in 5 patients. Hearing tended to be well preserved when caloric response was unchanged postoperatively. Preservation of hearing was not associated with the initial tumor volume, decrease of tumor size, maximum tumor dose, or peripheral tumor dose.

Journal Article↗
PubMed1996

HHR23B, a human Rad23 homolog, stimulates XPC protein in nucleotide excision repair in vitro.

A protein complex which specifically complements defects of XP-C cell extracts in vitro was previously purified to near homogeneity from HeLa cells. The complex consists of two tightly associated proteins: the XPC gene product and HHR23B, one of two human homologs of the Saccharomyces cerevisiae repair gene product Rad23 (Masutani et al., EMBO J. 13:1831-1843, 1994). To elucidate the roles of these proteins in "genome-overall" repair, we expressed the XPC protein in a baculovirus system and purified it to near homogeneity. The recombinant human XPC (rhXPC) protein exhibited a high level of affinity for single-stranded DNA and corrected the repair defect in XP-C whole-cell extracts without extra addition of recombinant HHR23B (rHHR23B) protein. However, Western blot (immunoblot) experiments revealed that XP-C cell extracts contained excess endogenous HHR23B protein, which might be able to form a complex upon addition of the rhXPC protein. To investigate the role of HHR23B, we fractionated the XP-C cell extracts and constructed a reconstituted system in which neither endogenous XPC nor HHR23B proteins were present. In this assay system, rhXPC alone weakly corrected the repair defect, while significant enhancement of the correcting activity was observed upon coaddition of rHHR23B protein. Stimulation of XPC by HHR23B was found with simian virus 40 minichromosomes as well as with naked plasmid DNA and with UV- as well as N-acetoxy-2- acetylfluorene-induced DNA lesions, indicating a general role of HHR23B in XPC functioning in the genome-overall nucleotide excision repair subpathway.

Animals↗
PubMed1996

Pharmacologic modulation of delayed-type hypersensitivity in mice.

In order to determine the mast cell requirements in murine delayed-type hypersensitivity (DTH), the effects of the antihistamine chlorpheniramine, the inhibitor of mast cell degranulation repirinast (CAS 73080-51-0, MY-5116), the lipoxygenase and cyclooxygenase inhibitor BW-755C (3-amino-1-[m-(trifluoromethyl)phenyl]-2-pyrazoline), and the cyclooxygenase inhibitors ibuprofen, diclofenac and phenylbutazone on murine DTH reactions were examined. When chlorpheniramine (50 mg/kg) was administered orally immediately after, 5 h, or 9 h after antigen challenge, it did not show any effect on sheep red blood cells (SRBC)-induced delayed footpad reaction (FPR) in mice. On the other hand, when chlorpheniramine was administered orally 16 h after antigen challenge, it significantly inhibited SRBC-induced delayed FPR (p < 0.01). When repirinast (30, 100 mg/ kg) was administered orally 16 h after antigen challenge, it significantly inhibited delayed FPR in a dose-dependent manner. BW-755C (50 mg/kg) showed no significant effect on delayed FPR when it was administered 1 h, 7 h, or 13 h after challenge. However, it significantly inhibited delayed FPR when it was administered 16 h after antigen challenge (p < 0.01). Ibuprofen, diclofenac, or phenylbutazone showed no significant effect on delayed FPR even if it was administered 16 h after antigen challenge. These results demonstrate that mast cells could be involved in DTH reactions and that histamine and leukotrienes may play an important role in DTH reactions in mice.

Animals↗
PubMed1995

Expression and functional analyses of the Dxpa gene, the Drosophila homolog of the human excision repair gene XPA.

Xeroderma pigmentosum (XP) is a human hereditary disease characterized by a defect in DNA repair after exposure to ultraviolet light. Among the seven groups of XP, group A (XP-A) patients show the most severe deficiency in excision repair and a wide variety of cutaneous and neurological disorders. We have cloned homologs of the human XPA gene from chicken, Xenopus, and Drosophila, and sequence analysis revealed that these genes are highly conserved throughout evolution. Here, we report characterization of the Drosophila homolog of the human XPA gene (Dxpa). The Dxpa gene product shows DNA repair activities in an in vitro repair system, and Dxpa cDNA has been shown to complement a mutant allele of human XP-A cells by transfection. Polytene chromosome in situ hybridization mapped Dxpa to 3F6-8 on the X chromosome, where no mutant defective in excision repair was reported. Northern blot analysis showed that the gene is continuously expressed in all stages of fly development. Interestingly, the Dxpa protein is strongly expressed in the central nervous system and muscles as revealed by immunohistochemical analysis using anti-Dxpa antibodies, consistent with the results obtained in transgenic flies expressing a Dxpa-beta-galactosidase fusion gene driven by the Dxpa promoter.

Amino Acid Sequence↗
PubMed1995

DNA repair protein XPA binds replication protein A (RPA).

XPA is a zinc finger DNA-binding protein, which is missing or altered in group A xeroderma pigmentosum cells and known to be involved in the damage-recognition step of the nucleotide excision repair (NER) processes. Using the yeast two-hybrid system to search for proteins that interact with XPA, we obtained the 34-kDa subunit of replication protein A (RPA, also known as HSSB and RFA). RPA is a stable complex of three polypeptides of 70, 34, 11 kDa and has been shown to be essential in the early steps of NER as well as in replication and recombination. We also demonstrate here that the RPA complex associates with XPA. These results suggest that RPA may cooperate with XPA in the early steps of the NER processes.

Base Sequence↗
PubMed1994

Purification and cloning of a nucleotide excision repair complex involving the xeroderma pigmentosum group C protein and a human homologue of yeast RAD23.

Complementation group C of xeroderma pigmentosum (XP) represents one of the most common forms of this cancer-prone DNA repair syndrome. The primary defect is located in the subpathway of the nucleotide excision repair system, dealing with the removal of lesions from the non-transcribing sequences ('genome-overall' repair). Here we report the purification to homogeneity and subsequent cDNA cloning of a repair complex by in vitro complementation of the XP-C defect in a cell-free repair system containing UV-damaged SV40 minichromosomes. The complex has a high affinity for ssDNA and consists of two tightly associated proteins of 125 and 58 kDa. The 125 kDa subunit is an N-terminally extended version of previously reported XPCC gene product which is thought to represent the human homologue of the Saccharomyces cerevisiae repair gene RAD4. The 58 kDa species turned out to be a human homologue of yeast RAD23. Unexpectedly, a second human counterpart of RAD23 was identified. All RAD23 derivatives share a ubiquitin-like N-terminus. The nature of the XP-C defect implies that the complex exerts a unique function in the genome-overall repair pathway which is important for prevention of skin cancer.

Amino Acid Sequence↗
PubMed1994

Chromosomal localization of three repair genes: the xeroderma pigmentosum group C gene and two human homologs of yeast RAD23.

The nucleotide excision repair (NER) disorder xeroderma pigmentosum (XP) is characterized by sun (UV) sensitivity, predisposition to skin cancer, and extensive genetic heterogeneity. Recently, we reported the cloning and analysis of three human NER genes, XPC, HHR23A, and HHR23B. The previously cloned XPC gene is involved in the common XP complementation group C, which is defective in excision repair of non-transcribed sequences in the genome. The XPC protein was found to be complexed with the product of HHR23B, one of the two human homologs of the Saccharomyces cerevisiae NER gene RAD23. Here we present the chromosomal localization by in situ hybridization using haptenized probes of all three genes. The HHR23A gene was assigned to chromosome 19p13.2. Interestingly, the HHR23B and XPC genes, the product of which forms a tight complex, were found to colocalize on band 3p25.1. Pulsed-field gel electrophoresis revealed that the HHR23B and XPC genes possibly share a MluI restriction fragment of about 625 kb. Potential involvement of the HHR23 genes in human genetic disorders is discussed.

Animals↗
PubMed1994

Immunocytochemical analyses of distributions of Na, K-ATPase and GLUT1, insulin and transferrin receptors in the developing retinal pigment epithelial cells.

The retinal pigment epithelium (RPE) is unique in that Na,K-ATPase is predominantly localized on its apical surface. We studied the distributions of Na,K-ATPase and glucose transporter GLUT1, insulin and transferrin receptors in developing rat RPE cells immunocytochemically. Na,K-ATPase, first detected in 17-day-old embryonic eyes, was already distributed predominantly on the apical surface. This reversed distribution of Na,K-ATPase was maintained throughout their life. Insulin receptor and transferrin receptor were distributed exclusively on the basolateral surface. By quantitative immunogold electron microscopic technique we found that glucose transporter GLUT1 is distributed almost equal in amount on both the apical and basolateral surfaces of RPE cells, thus presumably constructing an efficient pathway for glucose transport from the choriocapillaries to the neural retina through the blood-retinal barrier. These results suggest that in the RPE cells the intrinsic basolateral plasma membrane proteins are sorted out at least in three different ways.

Animals↗
PubMed1994

[Interferon-associated retinopathy].

During interferon (IFN) treatment for chronic hepatitis C, retinopathy develops in about one half of the patients. This "IFN-associated retinopathy" is characterized by cotton-wool spots (ischemic lesions) and superficial hemorrhage around the optic disc. It usually occurs 1 to 3 months after initiation of IFN therapy. Most of the patients with retinopathy have no symptoms, while some complain of impairment of visual acuity or flying specks. IFN-associated retinopathy tends to improve without any ophthalmic treatment and IFN therapy can be continued under careful observation. A complication of diabetes mellitus or a decrease in platelet count during IFN treatment may aggravate IFN-associated retinopathy. We emphasize that careful ophthalmologic follow-up is needed for all patients under IFN therapy.

Adult↗
PubMed1994

[Immunohistological study in Bruch's membrane of senescence accelerated mouse].

Age-related macular degeneration is one of the major causes of severe visual loss is elderly individuals. However, relatively little is known about its etiology. The disease may be associated with senescence. Ultrastructural and immunohistochemical studies on SAM (senescence accelerated mouse) eyes were carried out to learn details of aging changes in the retinal pigment epithelium (RPE) and Bruch's membrane. SAM P 1 mice aged 2, 10, 14 months were examined in this study. The eyes were analysed for type IV collagen and heparan sulfate proteoglycan (HSPG) by the avidin-biotin-peroxidase complexes (ABC) method and post-embeddig immunolocalization with colloidal gold. With the ABC method, the basement membranes of both the RPE and the choriocapillaris showed markedly positive staining when treated with anti-type IV collagen antibody and moderately positive staining when treated with anti-HSPG antibody. In ultrastructural immunolocalization, both basement membranes showed fairly heavy labeling in response to the antibodies to type IV collagen, and moderate labeling in response to the antibodies to HSPG. With aging, the thickness of the basement membrane of the choriocapillaris and gold particle labeling by the antibodies to type IV collagen increased. The gold particle labeling by the antibodies to HSPG increased slightly, but was distributed sparsely. These results showed the advancing process of senescence changes in Bruch's membrane.

Aging↗