The C57BL/6J-fe/fe mouse is a coat color mutant. The coat color of the homozygote mouse becomes progressively lighter with advancing age. The faded gene (fe) of C57BL/6J-fe/fe was mapped in a 2.0 cM distal to D10mit191 by our group. To make a high-resolution map, we used the Korean wild mouse (KWHM) for a backcross panel, which was captured in 1995 and has been maintained as an inbred line by our laboratory. In the inter-specific backcross panel (N=400), the fe gene was mapped to 1.0 cM distal to D10mit156. The gene order was defined: centromere -D10mit3/85 (1.3+/-0.6 cM)-D10mit155 (1.3+/-0.6 cM)-D10mit191 (2.0+/-0.7 cM)-D10mit156 (1.0+/-0.5 cM)-fe-D10mit193 (1.3+/-0.6 cM)-D10mit54 (1.0+/-0.5 cM)-D10mit44 (8.5+/-1.4 cM)-D10mit42 (10.0+/-1.5 cM). The measured distance between D10mit191 and D10mit 44 differed in both inter-specific (DBA/2) and intra-specific (KWHM) backcross panels (14.2 vs 13.8 cM). Taken together, our high-resolution linkage map of the fe locus from an intra-specific backcross panel will provide a good entry point to isolate the fe gene.
Animals ; Centromere ; Chromosomes, Human, Pair 10 ; Gene Order ; Hair Color ; Homozygote ; Mice

Loss-of function mutations in the transmembrane inner ear expressed (Tmie/TMIE) gene have been shown to cause deafness in mice and humans (DFNB6). Previous studies report that the circling mouse can be an animal model for DFNB6. However, the expression pattern of Tmie protein in postnatal developmental stages has not been clearly revealed. In this study we tried to investigate the expression of Tmie protein in the liver, spleen, kidney, and lung, as well as in the cochlea. We examined various tissue samples from five different age groups of C57BL/6J animals. Using western blotting analysis, the expression of Tmie protein in these organs has been identified. The results show that Tmie protein expression in the cochlea has been increased in postnatal developmental stages, indicating that Tmie plays an important role in not only the development and also in the function of the cochlea. The expression pattern of Tmie in adult mouse organs such as the liver, spleen, kidney, and spleen significantly vary in adult rats. The order of Tmie expression level in mice (63 days after birth) was spleen, liver, lung, cochlea, and kidney, whereas in the adult rat it was liver, cochlea, lung, spleen, and kidney.
Adult ; Animals ; Blotting, Western ; Cochlea ; Deafness ; Ear, Inner ; Humans ; Kidney ; Liver ; Lung ; Mice ; Models, Animal ; Rats ; Spleen

The faded mouse is a coat color mutant that shows faded coat color and age-related loss of pigmentation. This mutation is transmitted by an autosomal recessive gene with 100% penetrance. In the present study, we carried out linkage analysis of the faded (fe) gene using intra-specific backcross panels. Affected faded mice were carefully confirmed by their faded coat color at about 4 weeks of age. In the intra-specific backcross between faded and CBA mice (n=198), the fe gene was mapped to a region 2.1 cM distal to D10mit191. Therefore, the gene order was defined as follows: centromere-D10mit51 (12.4+/-2.4 cM)-D10mit191 (2.1+/-1.0 cM)-fe-D10mit44 (13.3+/-2.4 cM)-D10mit42 (14.4+/-2.5 cM). This linkage map of the fe locus will provide a good entry point to isolate the fe gene. Since the faded mouse has pigmentary abnormalities, this mutant may be a useful model for studies of pigmentary abnormalities in humans.
Animals ; Chromosomes, Human, Pair 10 ; Gene Order ; Genes, Recessive ; Humans ; Mice ; Mice, Inbred CBA ; Penetrance ; Pigmentation

Radical nephrectomy with tumor thrombectomy for advanced renal cell carcinoma is an oncologically relevant approach that can achieve long-term survival even in the presence of distant metastases. However, the surgical techniques pose significant challenges. The objective of this clinical review was to present technical recommendations for tumor thrombectomy in the vena cava to facilitate surgical treatment. Transesophageal echocardiography is required to prepare for this procedure. Cardiopulmonary bypass should be considered when the tumor thrombus has invaded the cardiac chamber and clamping is not feasible because of the inability to milk the intracardiac chamber thrombus in the caudal direction. Prior to performing a cavotomy, it is crucial to clamp the contralateral renal vein and infrarenal and suprahepatic inferior vena cava (IVC). If the suprahepatic IVC is separated from the surrounding tissue, it can be gently pulled down toward the patient’s leg until the lower margin of the atrium becomes visible. Subsequently, the tumor thrombus should be carefully pulled downward to a position where it can be clamped. Implementing the Pringle maneuver to reduce blood flow from the hepatic veins to the IVC during IVC cavotomy is simpler than clamping the hepatic veins. Sequential clamping is a two-stage method of dividing thrombectomy by clamping the IVC twice, first suprahepatically and then midretrohepatically. This sequential clamping technique helps minimize hypotension status and the Pringle maneuver time compared to single clamping. Additionally, a spiral cavotomy can decrease the degree of primary closure narrowing. The oncological prognoses of patients can be improved by incorporating these technical recommendations.

Damaged DNA binding (DDB) protein is an important gene in the repair of damaged DNA. DDB is a heterodimer (DDB1 and DDB2) protein, murine DDB2 has 10 exons about 1.5kb in size (Genbank Accession No. AY027937). Here we identified five DDB2 variants (M1-M5) from various mouse tissues that are generated by alternative splicing. We used reverse transcription-PCR (RT-PCR) to identify splicing variants and isolated PCR products using an agarose-gel PCR purification kit. All isolated PCR products were cloned and the structure of splicing variants was confirmed by sequencing. The first splicing variant M1 was generated by omission of exon 4. The second splicing variant M2, by omission of exons 4-5. The third variants M3 was generated by omission from the middle of exon 1 to exon 6 and was expressed in the heart. Fourth variants M4 was generated by omission of exon 2 and exons 4-7. M5, the last splicing variant was generated by omission of exons 4-7. M4 and M5 were expressed in the spleen. Analysis of tissue distribution by RT-PCR indicates that M1 is most highly expressed in the mouse brain. These results indicated that murine DDB2 has five splicing variants and splicing variants expression patterns were different depending on mouse tissue. Further functional studies of each splicing variants will provide more information about the molecular mechanism of DDB2 function and DDB2 gene expression regulation.
Alternative Splicing ; Animals ; Brain ; Clone Cells ; DNA ; Exons ; Gene Expression Regulation ; Heart ; Hydrocarbons, Chlorinated ; Mice ; Polymerase Chain Reaction ; Spleen ; Tissue Distribution

Circling mouse (C57BL/6J-cir/cir) deleted the transmembrane inner ear (Tmie) gene is an animal model for human non-syndromic recessive deafness, DFNB6. In circling mouse, hair cells in the cochlea have degenerated and hair bundles have become irregularity as time goes on. Tmie protein carries out a function of the mechanoelectrical transduction channel in cochlear hair cells. Myosin7a (MYO7A) protein has key roles in development of the cochlear hair bundles as well as in the function of cochlear hair cells. To find whether Tmie protein interacts with MYO7A proteins in the cochlea postnatal developmental stage, we investigated expression of the MYO7A proteins in the cochlear hair cells of circling mice by western blot analysis and whole mount immunofluorescence at postnatal day 5 (P5). The expression of MYO7A showed statistically significant increase in the cochlea of C57BL/6J-+/cir and C57BL/6J-cir/cir mice than that of C57BL/6J-+/+ mice. The MYO7A intensity of the cochlear hair cells also increased in C57BL/6J-+/cir and C57BL/6J-cir/cir mice compared with those of C57BL/6J-+/+ mice. Taken together, the results indicate that Tmie protein may have an important role with MYO7A protein in the development and maintenance of the stereociliary bundles during postnatal developmental stage of the cochlea.
Animals ; Blotting, Western ; Cochlea ; Deafness ; Ear, Inner ; Fluorescent Antibody Technique ; Hair ; Hair Cells, Auditory* ; Humans ; Mice* ; Models, Animal

Mutations in the transmembrane inner ear (Tmie) gene, which encodes the Tmie protein, have been attributed to deafness autosomal recessive 6 (DFNB6), an autosomal nonsyndromic recessive hearing loss disorder. Although the Tmie gene was identified a few years ago, little is known about subcellular localization of the Tmie protein. In order to address this, we developed a stable cell line expressing Tmie protein. The expression of Myc-tagged Tmie protein was confirmed by Western blot analysis using an anti-Myc antibody and localization of the Tmie protein was confirmed by immunostaining, using the anti-Myc antibody as well as the anti-tmie antibody. Our study demonstrates that the Tmie protein is localized mostly in the cellular membrane and to a lesser extent in cytoplasm. These results suggest that our Tmie expressing stable cell line provides a suitable in vitro model to explore Tmie synthesis and functions.
Blotting, Western ; Cell Line ; Cytoplasm ; Deafness ; Ear, Inner ; Hearing Loss ; Membranes

PURPOSE: Penile circular fasciocutaneous flap urethroplasty is a useful technique for a long anterior urethral stricture due to the flap's hairless nature and ample length. We investigated the surgical outcomes of urethroplasty for a complex anterior urethral stricture, performed using a penile circular fasciocutaneous flap. MATERIALS AND METHODS: Between 2008 and 2013, we performed a retrospective review of 29 patients who underwent urethroplasty using a penile circular fasciocutaneous flap and had at least 6 months of follow-up. A total of 20 cases utilized only a fasciocutaneous flap, while 9 cases combined a fasciocutaneous flap with other surgery. Success was defined as no requirement of additional urethral instrumentation. RESULTS: The overall success rate was 68.9% (20 out of 29 cases) at a median follow-up of 19 months. Furthermore, fasciocutaneous flap urethroplasty rendered the actual stricture-free rate of 79.3%. The location of recurrence was mostly at the junction of the flap. Among 9 surgical failures, 5 cases were treated successfully by using an additional surgical procedure. Fistula repair was needed in 1 case 4 months later. Further, periodic urethral dilation was performed in the remaining 3 cases. The failure rate was significantly higher in patients with suprapubic cystostomy than in patients without suprapubic cystostomy. The most common complication was post-micturition dribbling. CONCLUSIONS: Penile circular fasciocutaneous flap urethroplasty is a useful method for the reconstruction of a long anterior urethral stricture. A sufficient healthy margin should be acquired for better surgical results due to the fact that most recurrence occurs at the junction of the flap.
Cystostomy ; Fistula ; Follow-Up Studies ; Humans ; Male ; Penis ; Recurrence ; Retrospective Studies ; Surgical Flaps ; Urethral Stricture*

PURPOSE: Penile circular fasciocutaneous flap urethroplasty is a useful technique for a long anterior urethral stricture due to the flap's hairless nature and ample length. We investigated the surgical outcomes of urethroplasty for a complex anterior urethral stricture, performed using a penile circular fasciocutaneous flap. MATERIALS AND METHODS: Between 2008 and 2013, we performed a retrospective review of 29 patients who underwent urethroplasty using a penile circular fasciocutaneous flap and had at least 6 months of follow-up. A total of 20 cases utilized only a fasciocutaneous flap, while 9 cases combined a fasciocutaneous flap with other surgery. Success was defined as no requirement of additional urethral instrumentation. RESULTS: The overall success rate was 68.9% (20 out of 29 cases) at a median follow-up of 19 months. Furthermore, fasciocutaneous flap urethroplasty rendered the actual stricture-free rate of 79.3%. The location of recurrence was mostly at the junction of the flap. Among 9 surgical failures, 5 cases were treated successfully by using an additional surgical procedure. Fistula repair was needed in 1 case 4 months later. Further, periodic urethral dilation was performed in the remaining 3 cases. The failure rate was significantly higher in patients with suprapubic cystostomy than in patients without suprapubic cystostomy. The most common complication was post-micturition dribbling. CONCLUSIONS: Penile circular fasciocutaneous flap urethroplasty is a useful method for the reconstruction of a long anterior urethral stricture. A sufficient healthy margin should be acquired for better surgical results due to the fact that most recurrence occurs at the junction of the flap.
Cystostomy ; Fistula ; Follow-Up Studies ; Humans ; Male ; Penis ; Recurrence ; Retrospective Studies ; Surgical Flaps ; Urethral Stricture*

The C57BLKS/J-Lepr(db) mouse has a point mutation in the leptin receptor gene and is one of the most useful animal model for non-insulin dependent diabetes mellitus in human. Since the homozygote of C57BLKS/J-Lepr(db) mouse is infertile, detection of point mutation in the leptin receptor gene is important for efficient maintaining strains as well as mass production of homozygotes. To develop a rapid and efficient genotyping method for C57BLKS/J-Lepr(db) mouse, the tetra-primer amplification-refractory mutation system polymerase chain reaction (ARMS-PCR) was used. The 407 and 199 bp PCR products were amplified from normal (+/+) mice; while the 407 and 268 bp PCR products were amplified from homozygotes (db/db) mice; and the 407, 268, and 199 bp PCR products were amplified from heterozygotes (db/+) mice. This result showed that the tetra-primer ARMS-PCR analysis by us is suitable to detect point mutation of the leptin receptor gene. Taken together, our method will dramatically reduce animal use for maintenance of strains as well as production of homozygote in the C57BLKS/J-Lepr(db) strains.
Animals ; Diabetes Mellitus ; Heterozygote ; Homozygote ; Humans ; Leptin* ; Mice* ; Models, Animal ; Point Mutation ; Polymerase Chain Reaction ; Receptors, Leptin*