Biomedical subjects

Taketoshi Wakabayashi

Publications and source records attributed to Taketoshi Wakabayashi.

11 recordsLinked to original sources

PubMed2005

JNK inhibitory kinase is up-regulated in retinal ganglion cells after axotomy and enhances BimEL expression level in neuronal cells.

Optic nerve transection results in retinal ganglion cell (RGC) death in adult mammals, after the alteration of gene expression of RGCs. To elucidate the molecular mechanism by which axotomy induces RGC death, we isolated the molecules up-regulated after optic nerve transection. One of these, axotomy-related [corrected] gene (ARG)357, an 898-amino-acid [corrected] protein containing a complete serine-threonine kinase domain, was isolated from a subtraction library of the rat retina. The sequence showed that this gene was a rat homolog of human c-Jun N-terminal kinase (JNK) inhibitory kinase and so belonged to the germinal center kinase-VIII subfamily of Sterile20s protein kinase. We designated ARG357 as rat JNK inhibitory kinase (JIK). Rat JIK was expressed ubiquitously in various tissues and was highly expressed in the retina, with selective expression in RGCs. After axotomy, BimEL and Hrk, which are BH3-only proteins, and rat JIK were up-regulated in RGCs. Overexpression of rat JIK in neuronal cells up-regulated the expression of BimEL, but not that of Hrk. These results indicate that JIK may contribute to axotomy-induced RGC death by up-regulating the expression of BH3-only protein.

Adaptor Proteins, Signal Transducing↗

PubMed2005

Toxicological evaluation of micafungin ophthalmic solution in rabbit eyes.

There have been no reports of the topical application of micafungin to the eye. The aim of this study was to evaluate the safety of topical instillation of 0.1% micafungin ophthalmic solution in rabbit eyes. In New Zealand white rabbits (n = 6), 50 microL of 0.1% micafungin solution was topically instilled to 1 eye, and 50 microL of sterile saline was applied to the other eye. Both eyedrops were administered hourly from 7 A.M. for 7 days. Measurements were conducted on corneal thickness, intraocular pressure, endothelial cell density, and lactate dehydrogenase (LDH) activity of tear samples. The eyes were examined slit-lamp biomicroscopically and histopathologically. Topical micafungin application for 1 week did not induce any changes in intraocular pressure, endothelial cell density, and tear LDH. Corneal thickness after instillation was slightly, but significantly, smaller in the micafungin group than in the control group (P = 0.0156, paired t test), but this difference disappeared within 24 hours after the final instillation. Biomicroscopy and histopathology revealed no significant toxic influence of micafungin application on the cornea. Topical instillation of micafungin solution had no apparent toxicity to the cornea. These results warrant future studies on the efficacy of micafungin ophthalmic solution against corneal fungal infection.

Animals↗

PubMed2004

Rapid and sensitive diagnosis of adenoviral keratoconjunctivitis by loop-mediated isothermal amplification (LAMP) method.

PURPOSE: To develop a new method to detect and type adenoviruses directly from conjunctival scrapings using loop-mediated isothermal amplification (LAMP) with adenovirus (ad) type specific primer. METHODS: Using primers specific for the gene of ad1, ad3, ad4, ad8, ad19 and ad37, heat denatured adenovirus DNA was amplified by the LAMP and polymerase chain reaction (PCR). Alkaline lysed adenovirus prototype and conjunctival scrapings were also used directly as templates. RESULTS: Type specific primers amplified ad genes of the corresponding ad prototype specifically. The specific amplification was observed in both heat denatured and alkaline lysed samples. The amplified product was first detected within 45 min. Ad genotypes of clinical samples determined by the LAMP method were almost identical to those determined using the PCR-sequencing method. CONCLUSIONS: LAMP based isothermal amplification of adenovirus genome for detection and typing of adenoviruses is faster than PCR based methods. This new method will be useful for rapid diagnosis and typing of adenoviral conjunctivitis.

Adenovirus Infections, Human↗

PubMed2004

Long-term changes in rabbit cornea after ionizing radiation.

PURPOSE: To investigate long-term changes in the cornea after ionizing irradiation in rabbits. METHODS: Mature albino rabbits (n = 4) were unilaterally irradiated with 20 Gy of x-rays. The contralateral eye served as a control. The rabbits were examined with slit-lamp biomicroscopy for 5 years. The eyes were then enucleated for histopathologic examinations with light microscopy and transmission electron microscopy. RESULTS: On biomicroscopy, there was no corneal abnormality during the 5-year follow-up period. On histopathology, no abnormality was observed in the corneal epithelium. In the endothelium, a vacuole-like structure was recognized by light microscopy, and transmission electron microscopy revealed marked enlargement of intercellular space and anamorphosis of the nuclei. CONCLUSION: Moderate x-ray irradiation to rabbit eye did not induce any long-term damage to the corneal epithelium, but the endothelium demonstrated persistent and irreversible damage, which was observed even 5 years after irradiation.

Animals↗

PubMed2002

Up-regulation of Hrk, a regulator of cell death, in retinal ganglion cells of axotomized rat retina.

Hrk, a regulator of cell death, belongs to the family of pro-apoptotic BH3-only proteins and is known to induce apoptosis in nerve tissue. We examined Hrk gene expression to clarify the role of BH3-only proteins in apoptosis of retinal ganglion cells (RGCs) after optic nerve transection in the adult rat. Reverse transcription-polymerase chain reaction showed that Hrk was up-regulated in retina from 12 h after axotomy, and continued to be elevated for 1 week. In situ hybridization histochemistry revealed that Hrk was expressed in a sub-population of axotomized RGCs. These results suggest that Hrk is involved in the induction of apoptosis in RGCs after optic nerve transection.

Animals↗

PubMed2002

Posterior capsule staining and posterior continuous curvilinear capsulorhexis in congenital cataract.

We report 2 cases of indocyanine green (ICG) staining used for posterior continuous curvilinear capsulorhexis (PCCC) in congenital cataract combined with anterior vitrectomy. In the first case, because of corneal opacity, the visibility of the posterior capsule was poor without staining. After the extraction of the cataract, a PCCC was performed after ICG staining of the posterior capsule. In the second case, after cataract removal, ICG staining was used to better visualize the posterior capsule. In both cases, the PCCC was successfully completed because of better visualization of the stained posterior capsule flap against the transparent anterior hyaloid face of the vitreous. Clear visual axes have been maintained.

Capsulorhexis↗

PubMed2002

Detection of CMV in plasma and aqueous humor specimens from AIDS patients with CMV retinitis using the amplicor CMV test.

A DNA hybridization test (the Amplicor CMV test) for CMV DNA was used to detect CMV DNA in paired plasma and aqueous humor specimens from 37 AIDS patients with ophthalmoscopically diagnosed CMV retinitis. CMV was detected in 25/37 (67.6%) plasma specimens and in 32/37 (86.5%) aqueous humor specimens. Thus, the Amplicor CMV test using aqueous humor specimens seems to be a useful diagnostic test for CMV retinitis and one that is more sensitive than determination of CMV DNA in plasma specimens.

AIDS-Related Opportunistic Infections↗

PubMed2002

Intraocular and plasma HIV-1 RNA in HIV-infected patients with CMV retinitis and HIV-infected controls.

HIV-1 RNA was quantified in intraocular specimens and plasma from AIDS patients with CMV retinitis undergoing therapeutic vitrectomy and in 8 control subjects undergoing cataract surgery using the Amplicor RT-PCR-based assay. The HIV-1 RNA concentration in the intraocular specimens was significantly lower than in plasma. Patients with CMV retinitis and plasma HIV-1 RNA concentrations > 100,000 copies/ml generally had detectable HIV-1 RNA in their intraocular specimens. These findings point to the risk of percutaneous injury from a sharp instrument used in HIV-infected patients during ophthalmic surgery and possible transmission of HIV during such surgery. Prevention of percutaneous contact will require the use of surgical instruments that reduce the likelihood of injury, as well as the wearing of double gloves during surgery.

AIDS Serodiagnosis↗

PubMedDate not supplied

Rapid and sensitive diagnosis of adenoviral keratoconjunctivitis by loop-mediated isothermal amplification (LAMP) method.

Adenovirus Infections, Human↗

PubMedDate not supplied

Detection of CMV DNA in the aqueous humor of AIDS patients with CMV retinitis by AMPLICOR CMV test.

The present study was performed to detect the cytomegalovirus (CMV) DNA in the aqueous humor from the eyes of acquired immunodeficiency syndrome (AIDS) patients with CMV retinitis. Detection of CMV DNA in the aqueous humor in the eyes with active CMV retinitis was compared with detection of CMV DNA in inactive retinitis. CMV DNA in the aqueous humor was evaluated before and after treatment with intravitreal injection of ganciclovir. CMV DNA in the aqueous humor was measured by AMPLICOR CMV test. Forty-two eyes of 35 AIDS patients were diagnosed ophthalmoscopically as having CMV retinitis that was subclassified as either active or inactive. The active and inactive CMV retinitis cases were distinguished based on clinical evaluations and fundus photographs. The results showed that 37 of the 42 eyes (88.1%) were positive for CMV DNA prior to treatment, while in 29 of these 37 eyes (78.4%), the aqueous humor became CMV DNA-negative after the treatment. Successful treatment with the intravitreal injection of ganciclovir was associated with a reduction in the detection of CMV DNA in the aqueous humor. CMV DNA was not detected in the aqueous humor of patients with quies cent CMV retinitis. In conclusion, the AMPLICOR CMV test was found to be a reliable tool for differentiating active and inactive CMV retinitis, and is useful for helping to select the optimal treatment regimen. The intravitreal injection of ganciclovir is highly effective in reducing detectable CMV DNA in the aqueous humor.

AIDS-Related Opportunistic Infections↗

PubMedDate not supplied

Heat treatment enhances healing process of experimental pseudomonas corneal ulcer.

We investigated the effects of hyperthermia on the healing process of experimental Pseudomonas corneal ulceration (PCU). Hartley guinea pigs were used to develop animal models of PCU. As a heat source, disposable chemical pocket warmers were applied. The healing process of PCU was compared between the heat-treated corneas and the control corneas. The severity of infection and the degree of angiogenesis were classified by a clinical scoring system. The animals were euthanized 14 days after infection and the corneas were submitted for histopathological examination. The expression of vascular endothelial growth factor (VEGF) was examined immunohistochemically. Comparative reverse transcription polymerase chain reaction was performed to measure the expression level of VEGF in the cornea. Hyperthermia significantly promoted corneal epithelization and neovascularization in the PCU model. Heat treatment significantly decreased the number of viable Pseudomonas organisms present in PCU. On immunohistochemistry, the heated cornea demonstrated more intense staining for VEGF. Comparative reverse transcription polymerase chain reaction showed upregulation of the expression level of VEGF mRNA in the heat-treated cornea. Hyperthermia accelerated the healing process of PCU with increased corneal neovascularization. Angiogenesis may play an important role in the PCU healing process, which is enhanced by the heat treatment.

Animals↗