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Biomedical subjects

Y Kondo

Publications and source records attributed to Y Kondo.

At least 91 records · Page 5Linked to original sources

"Avian-type" renal medullary tubule organization causes immaturity of urine-concentrating ability in neonates.

BACKGROUND: While neonatal kidneys are not powerful in concentrating urine, they already dilute urine as efficiently as adult kidneys. To elucidate the basis for this paradoxical immaturity in urine-concentrating ability, we investigated the function of Henle's loop and collecting ducts (IMCDs) in the inner medulla of neonatal rat kidneys. METHODS: Analyses of individual renal tubules in the inner medulla of neonatal and adult rat kidneys were performed by measuring mRNA expression of membrane transporters, transepithelial voltages, and isotopic water and ion fluxes. Immunofluorescent identification of the rCCC2 and rCLC-K1 using polyclonal antibodies was also performed in neonatal and adult kidney slices. RESULTS: On day 1, the transepithelial voltages (V(Ts)) in the thin ascending limbs (tALs) and IMCDs were 14.6 +/- 1.1 mV (N = 27) and -42.7 +/- 6.1 mV (N = 14), respectively. The V(Ts) in the thin descending limbs (tDLs) were zero on day 1. The V(Ts) in the tALs were strongly inhibited by luminal bumetanide or basolateral ouabain, suggesting the presence of a NaCl reabsorption mechanism similar to that in the thick ascending limb (TAL). The diffusional voltage (V(D)) of the tAL due to transepithelial NaCl gradient was almost insensitive to a chloride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB). The V(Ts) in the IMCDs were strongly inhibited by luminal amiloride. On day 1, both the tDL and tAL were impermeable to water, indicating the water impermeability of the entire loop. Diffusional water permeability (P(dw)) and urea permeabilities (P(urea)) in the IMCDs indicated virtual impermeability to water and urea on day 1. Stimulation by vasopressin (1 nmol/L) revealed that only P(dw) was sensitive to vasopressin by day 14. A partial isoosmolar replacement of luminal urea by NaCl evoked negligible water flux across the neonatal IMCDs, indicating the absence of urea-dependent volume flux in the neonatal IMCD. These transport characteristics in each neonatal tubule are similar to those in quail kidneys. Identification of mRNAs and immunofluorescent studies for specific transporters, including rAQP-1, rCCC2, rCLC-K1, rENaC beta subunit, rAQP-2, and rUT-A1, supported these findings. CONCLUSION: We hypothesize that the renal medullary tubule organization of neonatal rats shares a tremendous similarity with avian renal medulla. The qualitative changes in the organization of medullary tubules may be primarily responsible for the immature urine-concentrating ability in mammalian neonates.

Age Factors↗

LST-2, a human liver-specific organic anion transporter, determines methotrexate sensitivity in gastrointestinal cancers.

BACKGROUND & AIMS: One approach to the development of targeted cancer chemotherapy exploits increased uptake of the agent into neoplastic cells. In this scenario, higher concentrations of the agent in cancer cells are responsible for differential killing, whereas the low concentration in normal human cells decreases side effects. The aim of this study was to isolate an organic anion transporter that is weak in normal cells, but abundantly expressed in cancer cells, to deliver the anticancer drugs to the cells. METHODS: A human liver complementary DNA (cDNA) library was screened with liver-specific transporter (LST)-1 cDNA as a probe. Northern blot analyses were performed using the isolated cDNA (termed LST-2). An LST-2-specific antibody was raised, and immunohistochemical analyses including immunoelectron microscopy were performed. Xenopus oocyte expression system was used for functional analysis. We also established a permanent cell line that consistently expresses LST-2 to examine the relationship between methotrexate uptake and sensitivity. RESULTS: The isolated cDNA, LST-2, has 79.7% of overall homology with human LST-1. LST-2 exclusively expressed in the liver under normal conditions and its immunoreactivity was highest at the basolateral membrane of the hepatocytes around the central vein. Although its weak expression in the liver, LST-2 is abundantly expressed in the gastric, colon, and pancreatic cancers. On the other hand, the LST-1 was only detected in a hepatic cell line. LST-2 transports methotrexate in a saturable and dose-dependent manner. Furthermore, introduction of the LST-2 gene into mammalian cells potentiates sensitivity to methotrexate. CONCLUSIONS: LST-2 is one of the prime candidate molecules for determining methotrexate sensitivity and may be a good target to deliver anticancer drugs to the gastrointestinal cancers.

Amino Acid Sequence↗

New method of on-line quantification of regional wall motion with automated segmental motion analysis.

We have recently developed an automated segmental motion analysis (A-SMA) system, based on an automatic "blood-tissue interface" detection technique, to provide real-time and on-line objective echocardiographic segmental wall motion analysis. To assess the feasibility of A-SMA in detecting regional left ventricular (LV) wall motion abnormalities, we performed 2-dimensional echocardiography with A-SMA in 13 healthy subjects, 22 patients with prior myocardial infarction (MI), and 9 with dilated cardiomyopathy (DCM). Midpapillary parasternal short-axis and apical 2- and 4-chamber views were obtained to clearly trace the blood-tissue interface. The LV cavity was then divided into 6 wedge-shaped segments by A-SMA. The area of each segment was calculated automatically throughout a cardiac cycle, and the area changes of each segment were displayed as bar graphs or time-area curves. The systolic fractional area change (FAC), peak ejection rate (PER), and filling rate (PFR) were also calculated with the use of A-SMA. In the control group, a uniform FAC was observed in real time among 6 segments in the short-axis view (60% +/- 10% to 78% +/- 9%), or among 5 segments in either the 2-chamber (59% +/- 12% to 75% +/- 16%) or 4-chamber view (58% +/- 13% to 72% +/- 12%). The variations of FAC, PER, and PFR were obviously decreased in infarct-related regions in the MI group and were globally decreased in the DCM group. We conclude that A-SMA is an objective and time-saving method for assessing regional wall motion abnormalities in real time. This method is a reliable new tool that provides on-line quantification of regional wall motion.

Adult↗

Neonatal periventricular-intraventricular hemorrhage. Subacute and chronic MR findings.

PURPOSE: To evaluate MR findings at subacute and chronic stages after neonatal periventricular-intraventricular hemorrhage (PIVH), and to determine the clinical significance of follow-up MR. MATERIAL AND METHODS: Twenty-six children (estimated gestational age, 23-39 weeks; mean 29 weeks) with a history of previous PIVH underwent MR examination during their subacute and chronic clinical courses. PIVHs were initially detected with ultrasound examination in all cases. PIVH was divided into three grades (mild, moderate, and severe) according to the findings at the initial US studies, which were correlated to the MR findings. RESULTS: Abnormal signal intensities related to hemorrhage were demonstrated in 10 of 21 patients (47.6%) on initial MR studies; the very low signal intensities on T2-weighted images found on the periventricular wall with a linear or a spotty shape had disappeared at one year after the initial US. The children with ventriculomegaly and periventricular leukomalacia (PVL) were increased in number on the follow-up studies. CONCLUSION: MR imaging provided valuable information about the consequences of neonatal PIVH such as developments of ventriculomegaly or PVL. It is noteworthy that hemorrhagic lesions could not be detected in half of the cases at the subacute or chronic stage of PIVH.

Brain↗

Scrotal reconstruction with a thinned flap based on both inferior epigastric arteries. Case report.

A 62-year-old man presented with an extensive defect of the scrotum caused by Fournier's gangrene. We reconstructed the scrotum using a thinned flap based on both inferior epigastric arteries one month after the first debridement. This method has the advantage of achieving adequate mobility of the reconstructed scrotum. The results were satisfactory both functionally and cosmetically, although we had to repair a hernia at the donor site a year later.

Debridement↗

Protective effects of fluvastatin against reactive oxygen species induced DNA damage and mutagenesis.

Oxidative stress may be an important factor in the development of diabetic complications. Advanced glycation end-products have drown attention as potential sources of oxidative stress in diabetes. We investigated the protective effects of fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, on oxidative DNA damage from reactive oxygen species or advanced glycation end-products in vitro, as well as effects of main fluvastatin metabolites and other inhibitors of the same enzyme, pravastatin and simvastatin. Protective effects were assessed in terms of the DNA breakage rate in a single-stranded phage DNA system in vitro. DNA was exposed to either reactive oxygen species or advanced glycation end-products. Fluvastatin and its metabolites showed a strong protective effect comparable to those seen with thiourea and mannitol, though pravastatin and simvastatin did not exert clear protective effects. Furthermore, fluvastatin reduced the mutagenesis by reactive oxygen species or advanced glycation end-products in Salmonella typhimurium test strains. Both pravastatin and simvastatin still lacked protective activity. Fluvastatin and its metabolites protect against oxidative DNA damage and may reduce risk of consequent diabetic complications.

Bacteriophages↗

Antioxidative effects of fluvastatin and its metabolites against oxidative DNA damage in mammalian cultured cells.

We investigated the effects of fluvastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, on reactive oxygen species (ROS) and on oxidative DNA damage in vitro, as well as the effects of the main fluvastatin metabolites (M2, M3, and M4) and other inhibitors of the same enzyme, pravastatin and simvastatin. The hydroxyl radical and the superoxide anion scavenging activities of fluvastatin and its metabolites were evaluated using an electron spin resonance spectrometer. Fluvastatin and its metabolites showed superoxide anion scavenging activity in the hypoxanthine-xanthine oxidase system and a strong scavenging effect on the hydroxyl radical produced from Fenton's reaction. Protective effects of fluvastatin on ROS-induced DNA damage of CHL/IU cells were assessed using the single-cell gel electrophoresis assay. CHL/IU cells were exposed to either hydrogen peroxide or t-butylhydroperoxide. Fluvastatin and its metabolites showed protective effects on DNA damage as potent as the reference antioxidants, ascorbic acid, trolox, and probucol, though pravastatin and simvastatin did not exert clear protective effects. These observations suggest that fluvastatin and its metabolites may have radical scavenging activity and the potential to protect cells against oxidative DNA damage. Furthermore, ROS are thought to play a major role in the etiology of a wide variety of diseases such as cellular aging, inflammation, diabetes, and cancer development, so fluvastatin might reduce these risks.

Animals↗

Pancreaticobiliary arteriovenous malformation with common bile duct dilation in a patient with hemobilia.

Arteriovenous malformation of the pancreas is a very rare entity. We describe a case involving the head of the pancreas associated with progressive hemobilia bleeding from the lower part of the common bile duct. The patient was a 50-year-old man with acute epigastralgia. Endoscopic retrograde cholangiopancreatography revealed hemobilia and cystic dilation of the common bile duct. Angiography demonstrated increased blood volume in the head of the pancreas and early filling of the superior mesenteric and portal veins. Abdominal pain and progressive anemia caused by hemobilia required surgical treatment. Histologic examination of the resected specimen revealed marked proliferation of the blood vessels in the pericholedochal area and the exact point of bleeding from the pancreaticobiliary arteriovenous malformation.

Angiography↗

Transvaginal ultrasound of urethral sphincter at the mid urethra in continent and incontinent women.

PURPOSE: Few studies are available on imaging the urethral sphincter. We performed transvaginal ultrasonography to examine the muscle in continent and incontinent women. MATERIALS AND METHODS: Transvaginal ultrasonography was performed using a 7.5 MHz. transrectal probe in 19 continent and 69 incontinent supine women. Incontinent cases were classified as urge (14) and stress (55), and included urethral hypermobility (22) and intrinsic sphincter deficiency (33). Cadaver specimens were also examined to confirm the anatomy of ultrasound images. RESULTS: Transvaginal ultrasonography showed the urethra as a round structure consisting of a relatively high echogenic central zone and a relatively low echogenic peripheral zone. Matching ultrasound images with the anatomy of cadaver specimens indicated that the peripheral zone represents rhabdosphincter most reliably at the lateral aspects. Thus, its thickness was measured at the 9 o'clock position of the mid urethra. Mean thickness plus or minus standard error of mean was significantly decreased in stress incontinence compared with continence and urge incontinence (2.14 +/- 0.04, 2.78 +/- 0.08 and 2.87 +/- 0.11 mm., respectively, p <0. 001), while in intrinsic sphincter deficiency it was thinner than in hypermobility (2.00 +/- 0.05 versus 2.35 +/- 0.06 mm., p <0.01). There was no difference in central zone thickness according to continence status. CONCLUSIONS: Urethral structures may be visualized by transvaginal ultrasonography. The rhabdosphincter image was thinner in stress incontinence, especially in intrinsic sphincter deficiency. Transvaginal ultrasonography may be a useful clinical test for examining the morphology of the urethral sphincter muscle in women.

Cadaver↗

Identification and characterization of the allergens in the tomato fruit by immunoblotting.

BACKGROUND: Although the tomato fruit (Lycopersicon esculentum) has been widely investigated for breeding purposes, there have been few studies on tomato allergenicity. We attempted to identify the tomato fruit allergens and to compare the concentrations of IgE-binding proteins among the different growth stages with sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. METHODS: An immunoblot experiment on tomato fruit extracts was performed using sera from 11 patients with oral allergy syndrome (OAS) to tomatoes. Bands reacting with IgE from more than half of the OAS patients' sera were excised and subjected to determination of N-terminal amino acid sequences using the automated Edman degradation method. Moreover, we compared the concentrations of these proteins at each growth stage of the tomato fruit with SDS-PAGE and immunoblotting. RESULTS: Four proteins binding with IgE from more than half of the OAS patients' sera were determined to be polygalacturonase 2A (PG2A), beta-fructofuranosidase, superoxide dismutase (SOD) and pectinesterase (PE). The concentrations of PG2A, beta-fructofuranosidase and PE were highest in the red ripening stage with both SDS-PAGE and immunoblotting. CONCLUSION: The concentrations of 3 of 4 tomato allergens increased during ripening.

Adolescent↗

Electromyography of male rat perineal musculature during copulatory behavior.

Electromyogram (EMG) recordings were made from the bulbospongiosus (BS) and ischiocavernosus (IC) muscles of the awake male rat in order to evaluate the roles played by these muscles in the implementation of penile intromission during a copulatory encounter. Male rats with bipolar wire electrodes implanted chronically in the BS and IC muscles were mated with sexually receptive females. Concurrent videotape recordings established a relationship between EMG and components of male rat copulatory behavior, i.e. mount, intromission and ejaculation. On EMGs recorded from the IC muscle, the initial low-amplitude phase at the initiation of each behavioral component was distinguished from the late high-amplitude phase whenever the male accomplished mounting or intromission. In the BS muscle, the late high-amplitude phase was observed only when the male accomplished intromission. Power spectrum analysis by fast Fourier transformation revealed that in the IC muscle the two phases differed at an overall range of 100-1,000 Hz. In the BS muscle, however, the spectra of the two phases differed at a specific range of 400-600 Hz. Recruitment of additional motor units would culminate in such a characteristic rise in the power spectrum. The results suggest that somatosensory inputs contingent on the intromission cause such recruitment in the BS to regulate penile cups and other movements.

Animals↗

Identification of thyroid hormone transporters in humans: different molecules are involved in a tissue-specific manner.

We have recently identified that rat organic anion transporters, polypeptide2 (oatp2) and oatp3, both of which transport thyroid hormones. However, in humans the molecular organization of the organic anion transporters has diverged, and the responsible molecule for thyroid hormone transport has not been clarified, except for human liver-specific transporter (LST-1) identified by us. In this study we isolated and characterized a novel human organic anion transporter, OATP-E from human brain. The isolated complementary DNA encodes a polypeptide of 722 amino acids with 12 transmembrane domains. A rat counterpart, oatp-E, was also identified. Homology analysis and the phylogenetic tree analysis revealed that OATP-E/oatp-E is a subfamily of the organic anion transporter. Human OATP-E transported 3,3',5-triiodo-L-thyronine (K(m), 0.9 microM), thyronine, and rT(3) in a Na(+)-independent manner. Although the clone was isolated from the brain, OATP-E messenger RNA was abundantly expressed in various peripheral tissues. The rat counterpart, oatp-E, also transported 3,3',5-triiodo-L-thyronine. In addition, in this study we revealed that human OATP, which is exclusively expressed in the brain, transported 3,3',5-triiodo-L-thyronine (K(m), 6.5 microM), T(4) (K(m), 8.0 microM), and rT(3). These data suggest that in humans, several different molecules are involved in transporting thyroid hormone: OATP in the brain, LST-1 in the liver, and OATP-E in peripheral tissues.

Amino Acid Sequence↗

Estrogen therapy in a male patient with chronic hepatitis C and irradiation-induced testicular dysfunction.

We report an 18-year-old male patient who developed chronic hepatitis C after blood transfusion and had testicular dysfunction after irradiation for a testicular relapse of childhood acute lymphocytic leukemia after cessation of maintenance therapy, and the initiation of testosterone replacement therapy at puberty. Concomitant administration of estradiol resulted in a reduction in serum alanine aminotransferase and ferritin levels and hepatic iron concentration and staining after 2 years of estrogen therapy, although interferon therapy was withdrawn because of adverse effects. This observation suggests that endogenous estradiol may play a beneficial role in male patients with chronic hepatitis C.

Adolescent↗

Effect of exercise on the metabolism of vitamin B6 and some PLP-dependent enzymes in young rats fed a restricted vitamin B6 diet.

The effect of exercise on vitamin B6 metabolism and PLP-dependent enzymes was studied in rats fed a diet with or without vitamin B6. Metabolism of some amino acids (citrulline, arginine, ornithine and threonine) inhibited in the B6-deficient rats was normalized during exercise. Exercise was also effective in storing vitamin B6 in the body by lowering excretion of vitamin B6, when intake of vitamin B6 was restricted. Aspartatae aminotransferase activity was higher in the red portion of the gastrocnemius muscle than that of the white one, whereas glycogen phosphorylase activity was vice versa and furthermore glycogen content in the white portion was very low in the vitamin B6-deficient rat. From the data obtained, it has been suggested that the red and white portions of the gastrocnemius muscle seemed to be more important in metabolizing amino acids and hydrolyze glycogen, respectively.

Adaptation, Physiological↗

[Method-performance studies of notified analytical method for fenbutatin oxide and cyhexatin].

Method-performance studies were conducted for the notified revised analytical method of fenbutatin oxide (FBTO) and cyhexatin (CHT). FBTO and CHT spiked into rice, soybeans, spinach, orange, tea powder and tea extract at the level of 0.5 microgram/g for FBTO and 0.1 microgram/g for CHT were analyzed in replicate in 6 laboratories. Means recoveries of FBTO were 85.2-96.5% and those of CHT were 83.5-89.2% except from soybeans (46.5%). Repeatability relative standard deviation values of FBTO and CHT in each crop were in the ranges of 2.3-9.4% and 3.2-6.3%, respectively. Reproducibility relative standard deviations were 3.9-12.6% for FBTO and 8.3-12.9% for CHT. Detection limits were 0.015-0.05 microgram/g for FBTO and 0.005-0.02 microgram/g for CHT.

Food Analysis↗

[Method-performance studies of notified analytical method for clofentezine].

Method-performance studies were conducted for the notified revised analytical method of clofentezine. Clofentezine spiked in azuki beans, apple, orange, banana, grape, tea powder and tea extract at the level of 0.2 microgram/g (2 micrograms/g for tea) was analyzed in replicate in 6 laboratories. Mean values of recovery from 7 crops ranged from 78.4 to 85.2%. Repeatability relative standard deviation values ranged from 2.2 to 4.6% and reproducibility standard deviation values ranged from 4.8 to 10.3%. The detection limits were 0.005-0.01 microgram/g. These results show the notified analytical method has good performance.

Chlorobenzenes↗

Combination therapy of 2-5A antisense against telomerase RNA and cisplatin for malignant gliomas.

2',5'-Oligoadenylate (2-5A) linked to an antisense oligonucleotide against human telomerase RNA (2-5A-anti-hTR) is a novel therapeutic modality we have recently developed. We designed the oligonucleotide to target telomerase which maintains chromosome integrity in cancer cells. We have already demonstrated the efficacy of this new therapy in malignant glioma cells with telomerase activity. In the present study, we investigated the effect of 2-5A-anti-hTR in combination with cisplatin, an anti-cancer drug commonly used for malignant glioma patients. Six human malignant glioma cell lines with telomerase activity were treated with 0.5 microM 2-5A-anti-hTR and/or cisplatin (0, 0.1, 1, 5, 10, or 20 microg/ml) for three days, and cell viability was measured using the MTT colorimetric assay. The combination therapy showed synergistic effect at 1 microg/ml cisplatin and additive effect at 5 microg/ml cisplatin. TUNEL staining of the treated cells showed significantly increased apoptotic cells after the combination therapy. Furthermore, tumor growth of subcutaneous xenografts of human malignant glioma cells in nude mice was effectively reduced after the combination therapy for seven days. Our study shows that the tumor-killing effects of the combined treatments are, at least in part, due to induction of apoptosis. 2-5A-anti-hTR is a promising therapy not only when used alone, but also in combination with cisplatin.

Animals↗