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Biomedical subjects

M Imanishi

Publications and source records attributed to M Imanishi.

At least 55 records · Page 3Linked to original sources

Artificial nine zinc-finger peptide with 30 base pair binding sites.

It is well-known that DNA binding of native nine zinc-finger protein TEIIIA is dominated by interaction of select few fingers. Newly designed zinc-finger peptide Sp1ZF9 containing nine Cys2-His2 type motifs has been manipulated. The DNA-binding property of Sp1ZF9 was compared with those of native three zinc-finger Sp1(530-623) and artificial six zinc-finger Sp1ZF6 peptides. Although the equilibrium time was less than 0.5 h for Sp1(530-623)-DNA complex, Sp1ZF6 and Sp1ZF9 required approximately 48 and 72 h, respectively, for full complex formation. Evidently, the footprinting analysis demonstrated that Sp1ZF9 and Sp1ZF6 bind at least 27 and 18 contiguous base pairs of DNA sequence, respectively. Sp1ZF9 showed two step bindings to DNA, namely first the recognition of GC (5'-GGG-GCG-GGGCC-3') sequence by the N-terminal Sp1 domain and next the recognition of the corresponding target sequences by the middle and C-terminal Sp1 domains. In contrast with unimolecular binding of Sp1ZF9 and Sp1ZF6, two Sp1(530-623) molecules bind to one GCIII (5'-GGG-GCG-GGG-GGG-GCG-GGG-GGG++ +-GCG-GGGCC-3') site region. Semispecific complex formed at the beginning of Sp1ZF9-DNA interaction has also been characterized by kinetic analysis using surface plasmon resonance. Interestingly, the association rate constants for GC and GCIII complexes of Sp1ZF9 are smaller than those of the corresponding Sp1(530-623) complexes. Of special interest is the fact that new nine zinc-finger peptide Sp1ZF9 can bind to DNA sequence of approximately 30 base pairs. Such multi zinc-finger peptides may be useful as genome-specific transcriptional switches in future.

Base Pairing↗

Intracranial gas on CT after cardiopulmonary resuscitation: 4 cases.

We report four patients in whom gas was seen in the head on CT shortly after cardiopulmonary resuscitation. The gas was in the posterior cranial fossa, presumably within veins, or in the cavernous sinus. The cause of the cardiac arrest was myocardial infarction in three patients and hanging in one. All had peripheral or central venous lines. The mechanism by which gas appeared in the intracranial veins is discussed.

Cardiopulmonary Resuscitation↗

A modified microsyringe for extracellular recording of neuronal activity.

We describe a modified Hamilton microsyringe that allows extracellular recording of neuronal activity and subsequent injections. It is assembled with a Hamilton removable needle and a syringe for injection, a Teflon-coated tungsten wire for recording, and polyimide tubing as a sheath. The device is inexpensive and easy to handle in anatomical and physiological experiments in awake monkeys.

Animals↗

Post-transplant diabetes with anti-glutamic acid decarboxylase antibody during tacrolimus therapy.

A 54-year-old man undergoing hemodialysis because of end-stage renal failure was transplanted with a cadaver kidney in November 1997. He had no history of diabetes. Tacrolimus was used as the primary immunosuppressant. Three weeks after transplantation, he developed insulin-requiring diabetes mellitus. Anti-glutamic acid decarboxylase antibody was not detected on the third post-operative day, but appeared 4 weeks after transplantation. The recipient had DNA haplotypes that indicated susceptibility to Type 1 diabetes in Japanese subjects. Immunosuppressive therapy was changed from tacrolimus to cyclosporin. Thereafter, titer of anti-glutamic acid decarboxylase antibody disappeared and the patient's insulin requirement was notably reduced. The mechanism underlying the development of diabetes in this case appears to be, in part, direct beta-cell toxicity due to tacrolimus therapy, resulting in secondary beta-cell autoimmunity. This case suggests that tacrolimus therapy after transplantation should be used with caution in patients with genetic susceptibility to Type 1 diabetes.

Antibodies↗

Oxidation products of uric acid and ascorbic acid in preterm infants with chronic lung disease.

Allantoin, the oxidation product of uric acid (UA), can be used as an in vivo marker of free radical generation. The aims of the present study were to evaluate the allantoin changes in plasma and bronchoalveolar lavage fluid (BALF) as well as to examine plasma levels of ascorbic acid (AA) and its oxidation product, dehydroascorbic acid (DHAA), in infants with or without chronic lung disease (CLD) during the first week of life. The study population was 20 infants of 24-30 weeks gestation, comprising 10 who subsequently developed CLD and 10 without CLD. In the CLD infants, the plasma allantoin/UA ratio showed a significant increase after day 1 and continued to increase gradually to reach a peak on day 6 (6.5 +/- 4.1% for CLD and 2.1 +/- 0.9% for non-CLD infants). The allantoin/UA ratio in BALF was also higher in CLD infants and the difference reached significance on days 4-6 (41.2 +/- 15.8% for CLD and 11.7 +/- 9.9% for non-CLD infants). In contrast to allantoin, the plasma DHAA/AA ratio did not differ between the 2 groups throughout the study period. Our findings that the allantoin/UA ratios were significantly higher in CLD than non-CLD infants not only in plasma but also in BALF, and that the intergroup differences of this ratio in both plasma and BALF was more prominent in the latter half of the first week of life further confirm our previous speculation that oxygen radicals are involved in the development of neonatal CLD.

Allantoin↗

Aspirin renography in diagnosis of unilateral renovascular hypertension.

We previously proposed that aspirin can enhance the diagnostic sensitivity of renography with [123I] orthoiodohippurate (OIH) in patients with suspected unilateral renovascular hypertension (RVH). In this study we assessed the sensitivity and specificity of this method. Twenty-four patients, 14 with unilateral RVH and 10 with essential hypertension, were studied. For all patients with RVH, definitive diagnosis was based on the results of angiography and the response to renal arterial angioplasty after this study. For all patients with essential hypertension, we confirmed that there was little if any stenosis of the renal artery by digital subtraction angiography or Doppler sonography. Renography with [123I]OIH or 99mTc-mercaptoacetyltriglycine (MAG3) was done once before and once after the oral administration of aspirin (20 mg/kg). We considered renal blood flow to be decreased if the time to the peak in renography was 5 min or more, and defined the peak times of the kidneys as different if separated by 2 min or more. Unilateral RVH was diagnosed when both criteria were met. Renography before aspirin administration indicated RVH in 7 of the 14 patients with RVH, and renography after aspirin indicated RVH in 13 of the 14 patients. Of the 10 patients with essential hypertension, renography before and after aspirin administration showed no difference between the kidneys in 8 patients. Thus, aspirin renography increased the sensitivity from 50% to 93%, but did not change the specificity (80%) in the diagnosis of RVH. These results suggest that renography with [123I]OIH or 99mTc-MAG3 for the diagnosis of unilateral RVH is more sensitive when aspirin is used.

Adult↗

Evaluation of rat sperm by flow cytometry: simultaneous analysis of sperm count and sperm viability.

In this study, we conducted a simultaneous analysis of sperm count and viability in rats by flow cytometry (FCM). Epididymal fluids were taken from the caudal epididymis of 12 to 13 week-old Sprague-Dawley rats. The fluids were weighed and mixed with Dulbecco's phosphate buffered saline (D-PBS). Propidium iodide, which can stain only dead sperm, was used to distinguish viable and dead sperm. The sperm count and viability analyzed by FCM were 1.28 x 10(6)/mg and 78.0%, respectively. These values were consistent with the corresponding values (1.39 x 10(6)/mg and 81.0%) that were directly determined microscopically in the fluids of the same sample. In addition, when the original mixture containing sperm was diluted two times and four times with D-PBS, or was diluted two times with D-PBS containing only killed sperm, the sperm count and viability determined by FCM also correlated well with the sperm count (r = 0.96, P < 0.01) and sperm motility (r = 0.99, P < 0.01) by direct microscopic observation, respectively. In conclusion, the present flow cytometric analysis would be practical for the simultaneous determination of sperm count and viability in rat epididymal fluids.

Animals↗

Glomerular charge and size selectivity assessed by changes in salt intake in type 2 diabetic patients.

OBJECTIVE: To evaluate glomerular charge selectivity in patients with type 2 diabetes, we studied changes in fractional clearance of proteins with different sizes and charges when patients were placed on two diets with different salt contents. RESEARCH DESIGN AND METHODS: Nineteen patients with type 2 diabetes and normoalbuminuria (< 20 micrograms/min, n = 8), microalbuminuria (20-100 micrograms/min, n = 7), or advanced albuminuria (> 100 micrograms/min, n = 4) were placed on a low-salt diet (85 mEq of sodium daily) or a high-salt diet (255 mEq of sodium daily) for 1 week, and then on the other diet, in random order. Fractional clearances of albumin and immunoglobulin G (IgG) were calculated on the last 3 days of each diet. RESULTS: In patients with normoalbuminuria, the high-salt diet increased the fractional clearance of IgG, which is electrically neutral, but the fractional clearance of albumin, which is anionic, was unaltered, suggesting that the pore charge of the glomerular barrier was unaffected. However, in patients with microalbuminuria, the high-salt diet increased the fractional clearances of IgG and albumin equally, indicating some neutralization of the pore charge. Fractional clearance of IgG in these first two groups was similar when salt intake was low, so pore size was the same in these groups. In patients with advanced albuminuria, fractional clearance of IgG was higher than in the other groups, indicating that size selectivity had worsened. CONCLUSIONS: In type 2 diabetic patients, charge selectivity is lost before size selectivity as diabetic nephropathy progresses.

Adult↗

[Myxoid liposarcoma surrounding non-functioning transplanted kidney after living renal transplantation: a case report].

A 19-year-old male visited our clinic with the complaint of lower abdominal distention, January 1992. He had undergone living renal transplantation from his mother as a donor on June 15, 1988. But the transplanted kidney had become functional loss by chronic rejection 3 years after transplantation. The computed tomography showed huge low density mass around the non-functioning transplanted kidney. En block resection of the tumor and the transplanted kidney was performed. The tumor was pathologically diagnosed as mixoid liposarcoma. It was proved that the tumor cells were derived from recipient cells by the investigation of HLA-DRB1 DNA typing.

Adult↗

[Is tacrolimus effective for ongoing renal allograft rejection?].

Tacrolimus has already gained a high reputation as an induction-maintenance immunosuppressive therapy after kidney transplantation. Recently, it is being used as rescue therapy against rejection, and its effectiveness also appears to have been established to some extent. In this study, we evaluated the efficacy of Tacrolimus rescue therapy at 4 institutions in the Kinki District. The subjects were 19 patients treated with Tacrolimus against rejection observed during immunosuppressive therapy using cyclosporin. Evaluation was made by classifying the patients into 6 with acute rejection that occurred within 3 months after transplantation (AR), 4 with late onset acute rejection that developed more than 3 months after operation (LAR), and 9 patients with chronic rejection (CR). In the AR group, many patients received combination therapy at the introduction of Tacrolimus, and the long-term outcome was satisfactory. Tacrolimus was effective in 2 (50%) of the 4 patients in the LAR group. The trough levels of Tacrolimus at its introduction were 10-15 ng/ml in the AR and LAR groups. Deterioration of the transplanted kidney function was prevented in 3 (50%) out of 6 patients in the CR group observed for less than 1 year, but it deteriorated in all 3 patients observed for 1 year or longer. The trough levels of tacrolimus at its introduction were 5-10 ng/ml in many patients in the CR group. The rescue therapy using Tacrolimus was effective against acute rejection but further follow-up is considered to be needed to evaluate its efficacy against chronic rejection.

Graft Rejection↗

[Postoperative complications after Mainz pouch construction--with emphasis on measures to deal with narrowing of the duct aperture].

Thirty patients underwent Mainz pouch construction between December 1992 and January 1998. The post-operative observation period varied from 6 to 57 months (average, 38.5 months). A mechanism to ensure urinary continence was provided in 24 cases, while formation of a nipple valve by intussusception of the ileum with seromuscular stripping and submucosally embedded in situ appendix was performed in 6 cases. An umbilical stoma was adopted in all cases. Late-phase complications included 3 cases of hydronephrosis requiring treatment. Efferent limb of the stoma was observed in 8 cases, 6 of which were attributable to mild constriction of the stoma at the site of anastomosis of the navel cavity. In those cases in which stomal stricture occurred, difficulty in withdrawing urine was eliminated by providing a fistula stopper. Although few clinical methods to deal with stomal stricture have been reported, we found the fistula stopper to be useful providing good external appearance and patients' quality of life.

Adult↗

[Application of a semi-automatic ROI setting system for brain PET images to animal PET studies].

ProASSIST, a semi-automatic ROI (region of interest) setting system for human brain PET images, has been modified for use with the canine brain, and the performance of the obtained system was evaluated by comparing the operational simplicity for ROI setting and the consistency of ROI values obtained with those by a conventional manual procedure. Namely, we created segment maps for the canine brain by making reference to the coronal section atlas of the canine brain by Lim et al., and incorporated them into the ProASSIST system. For the performance test, CBF (cerebral blood flow) and CMRglc (cerebral metabolic rate in glucose) images in dogs with or without focal cerebral ischemia were used. In ProASSIST, brain contours were defined semiautomatically. In the ROI analysis of the test image, manual modification of the contour was necessary in half cases examined (8/16). However, the operation was rather simple so that the operation time per one brain section was significantly shorter than that in the manual operation. The ROI values determined by the system were comparable with those by the manual procedure, confirming the applicability of the system to these animal studies. The use of the system like the present one would also merit the more objective data acquisition for the quantitative ROI analysis, because no manual procedure except for some specifications of the anatomical features is required for ROI setting.

Animals↗

Cancer risk after renal transplantation in Japan.

Excess of cancer in patients receiving renal transplantation is well-known in Western countries, but information in Japan remains limited. Our study examined whether excess risk is found in patients receiving renal transplantation in Japan. Between 1970 and 1995, 1155 males and 589 females underwent renal transplantation in 6 hospitals, and a total of 12,982 person-years of observation was accumulated. Malignancies developed in 2.6% of patients; O/E ratio was 2.78. Median interval from renal transplantation to tumor development was 58 months. The interval in the patients receiving medication with cyclosporine-A (CyA) (median, 42.5 months) was significantly shorter than that with non-CyA (median, 95.5 months). Median age at the diagnosis of malignancy was 40 years, which is much younger than that in the general population. Relative risk was highest in renal cancer, followed by thyroid cancer, malignant lymphoma and uterine cancer. A distribution of malignancies was different from that reported from Western countries. These findings showed the excess risk of malignancies in Japan with renal transplants, especially in male patients, similar to that observed in Western countries, though the types of malignancy were different.

Adolescent↗

Suppressed alloreactivity and mixed chimerism in rats with accepted cardiac allografts by intrathymic injection of donor bone marrow cells.

Intrathymic injection of donor bone marrow cells (ITBMCs) at the time of transplantation and treatment with antilymphocyte serum (ALS) permitted the indefinite survival of Brown Norway (BN, RT1n) rat heart grafts in 6 out of 8 Lewis (LEW, RT1l) rat recipients, LEW recipients with long-surviving BN heart grafts (LSGs) also accepted additional BN heart grafts without further immunosuppression, though they rejected Piebald Virol Glaxo (PVG, RT1c) rat heart grafts in the usual fashion. In the in vitro study, the proliferative response of the lymphocytes from LEW recipients with LSGs remained suppressed when they were stimulated by BN spleen cells, but not when stimulated by PVG cells. Bone marrow cells (BMCs) from LEW rats with LSGs showed strong, nonspecific, suppressive effects on the proliferative response in the mixed lymphocyte culture reaction, suggesting that one of the possible explanations for tolerance might be the involvement of a suppressor mechanism.

Animals↗

Effects of cicletanine on prostaglandin I2 and E2 levels in patients with essential hypertension.

Cicletanine is a new antihypertensive drug that seems to stimulate the synthesis of prostaglandin (PG) I2. However, there is little evidence that cicletanine increases the level of PGI2 in the systemic blood of human subjects long-term. To investigate the antihypertensive mechanism of cicletanine, we measured serially the systemic blood pressure and the levels of both 6-keto-PGF1alpha (a stable metabolite of PGI2) and PGE2 in plasma and urine after administration of cicletanine. Nine patients with essential hypertension on a diet with sodium intake of 120 mEq/day took 100 mg of the drug orally daily every day for 1 week. Systemic blood pressure was measured hourly for 24 h on day 7 of the control period and on days 1 and 7 of the cicletanine period. The two PGs of interest were extracted, purified by high pressure liquid chromatography, and measured by radioimmunoassay. Cicletanine decreased blood pressure without reflexial tachycardia. The plasma levels of 6-keto-PGF1alpha were slightly, but significantly, higher at 3 h after the administration of cicletanine on both days 1 and 7 of administration (on day 1, 3.88 +/- 1.44 pg/mL and on day 7, 4.07 +/- 0.76, means +/- SD, both P < .05 v before administration on day 1) than before administration on day 1 (3.21 +/- 1.25 pg/mL). Plasma PGE2 was higher before and at 3 h after administration on day 7 than at 12 noon on day 7 of the control period. Cicletanine increased the urinary excretion of the two PGs; the increased PG levels partly account for the increased natriuresis in the first 3 days. The antihypertensive effects of cicletanine taken for 1 week were based on natriuresis caused by increased systemic synthesis of the vasodilator PGI2 and partly by the increased renal synthesis of PGI2 and PGE2.

6-Ketoprostaglandin F1 alpha↗

[A case report of spontaneous hemopneumothorax].

A 55-year-old man was admitted to the private hospital with chief complaints of left chest pain and increasing dyspnea. Chest X ray on admission revealed a collapsed lung and an air fluid line in the left hemothorax. A chest tube drainage was carried out, but hemorrhagic pleural fluid of 1,000 ml was drainaged. So was the patient transferred to our hospital for surgical treatment. Following blood transfusion because of hemorrhagic pleural fluid of 1,500 ml, urgent surgery was carried out. At the operation under thoracoscope, the bleeding from the apex of the thoracic cavity was noted. The bleeding was controlled with endoscopic surgical clip. As it was, however, very difficult to remove massive blood clots under endoscopic surgery, a small thoracotomy was carried out in addition to the thoracoscopic surgery, then the bulla in the apex of left lung was resected and also removed a large amount of blood clots.

Hemopneumothorax↗