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Biomedical subjects

M Fukuda

Publications and source records attributed to M Fukuda.

At least 37 records · Page 2Linked to original sources

Does corpus luteum locally affect follicular growth negatively?

In this study bilateral ovarian follicular growth during the luteal phase was investigated in relation to the ovary where ovulation occurred. The diameter of the largest follicle in the contralateral ovary without corpus luteum and in the ipsilateral ovary with corpus luteum was measured using vaginosonography in a total of 66 natural cycles of 27 normally cycling women undergoing treatment with intrauterine insemination (IUI). None of the women received ovarian stimulation or luteal support. Follicles from 2 to 11 mm in diameter were measured in early luteal phase (day +1 to +4), mid-luteal phase (day +5 to +9) and late luteal phase (day +10 onwards). The mean diameters of the largest follicle in the contralateral ovary without corpus luteum during the early, mid- and late luteal phases were 6.81 +/- 1.33 (mean +/- SD), 6.14 +/- 1.29 and 5.71 +/- 1.17 mm respectively, while those of the ipsilateral ovary with corpus luteum were 6.48 +/- 1.40, 5.65 +/- 1.47 and 4.98 +/- 1.19 mm respectively. While there was no significant difference during the early luteal phase, the mean diameter of the largest follicle in the ipsilateral ovary with corpus luteum was significantly smaller than that of the contralateral ovary without corpus luteum during the mid-luteal phase (P < 0.004) and the late luteal phase (P < 0.0005). These results indicate that the corpus luteum locally affects neighbouring follicular growth negatively during the luteal phase of the menstrual cycle, with the most pronounced effect expressed in the mid- and late luteal phases.

Adult

Increased cell proliferation of urothelium in rat bladder following unilateral nephrectomy.

PURPOSE: The aim of this study is to investigate the effect of unilateral nephrectomy on cell proliferation of normal urinary bladder epithelium in adult rats. MATERIALS AND METHODS: Following unilateral nephrectomy or sham operation 5 rats for each time point were sacrificed and their bladders were removed at 6 and 12 hours and also on 1, 2, 4, 7, 14, 28, and 56 days. Another 5 rats were not operated and served as the control. The percent number of proliferating cell nuclear antigen (PCNA) positive cells and the number of argyrophilic nucleolar organizer regions (AgNORs) were determined for each bladder. RESULTS: In the nephrectomized groups, a significant increase of PCNA expression in urothelial cells of the bladder was found at each sacrifice time with the exception of 28 and 56 days when compared with that of the control and corresponding sham-operated groups. Percent PCNA labeling index (PCNA-LI) reached a peak value of approximately 9.1% on day 2 post operation. From day 4 post nephrectomy, however, PCNA-LI decreased gradually reaching control levels by day 28. No significant difference was found between the control and sham-operated groups. The mean number of AgNOR dots in nephrectomized rats sacrificed on day 1, 2, 4, 7 and 14 post operation was significantly larger than that of the control and corresponding sham-operated groups. CONCLUSION: The results suggest that unilateral nephrectomy temporarily enhances normal urothelial cell proliferation in adult rat bladder.

Animals

The entire nucleotide sequences of two GB virus C/hepatitis G virus isolates of distinct genotypes from Japan.

Recently, putative viral agents responsible for human non-A to E hepatitis have been independently reported by two groups of investigators and designated GB virus C (GBV-C) and hepatitis G virus (HGV), respectively. The entire nucleotide sequences were determined for two viral genomes isolated from Japanese blood donors with GBV-C RNA. One of them (GT230) had a total genomic length of 9390 nucleotides (nt) with 5' and 3' untranslated regions of 551 and 313 nt, while the other (GT110) had genomic lengths of 9395, 281 and 315 nt, respectively. They both had a single long open reading frame, encoding 2842 amino acids (aa) in GT230 and 2933 aa in GT110. Surprisingly, they both lacked a clearly identifiable core gene, and possessed the E1/E2 gene with only four potential N-linked glycosylation sites. Pairwise comparison and phylogenetic analysis of the entire sequence indicated that the prototype GBV-C and two HGV isolates reported, as well as GT230 and GT110, are the same virus possibly of different genotypes. The five GBV-C/HGV isolates were variable up to 13.8% in the genomic nucleotide sequence, and contained deletions and insertions within the 5'-terminal 518-593 nt, which resulted in four different sizes of predicted polyproteins encoded by genomes of individual isolates. By contrast, the 3' untranslated region was well conserved. The high degree of sequence conservation within this region would favour it as a target for sensitive detection of GBV-C/HGV RNA.

Amino Acid Sequence

Gene mutation analysis and quantitation of DNA topoisomerase I in previously untreated non-small cell lung carcinomas.

To elucidate whether gene alterations of topoisomerase I (topo I) exist in untreated non-small cell lung carcinomas (NSCLC), polymerase chain reaction-single strand conformation polymorphism analysis was performed in forty-four NSCLC tissue samples. Gene alterations of topo I were sought in three regions, near codons 361 and 363, 533, and 722 and 729, where point mutations have been found in resistant tumor cell lines selected by chronic camptothecin exposure. In addition, nuclear topo I contents were determined by immunoblotting. No mobility shifts were observed compared to the pattern observed in a normal control at any of the three regions in any sample, whereas topo I levels showed an approximately 12-fold variation. The variation is remarkably large compared to those seen in previous in vitro and in vivo studies. The results suggest that mutations of topo I may not contribute to intrinsic resistance of NSCLC to camptothecins, but low topo I levels may account, at least in part, for the resistance.

Adenocarcinoma

Analysis of urinary red blood cells of healthy individuals by an automated urinary flow cytometer.

A newly developed automated urinary flow cytometer allows clear discrimination of erythrocytes from other solid components of urine. In this study, the normal range of the urinary erythrocyte count and the source of urinary erythrocytes in healthy individuals were investigated using this analyzer. For the diagnosis of the source of the urinary erythrocytes, Kitasato University Kidney Center criteria for this analyzer were applied. The subjects were 133 regularly employed volunteers (age range 20-48 years, mean 30.5) who noted no urinary tract symptoms and showed normal blood pressure, consisting of 41 females not in their menstrual period (age range 20-39 years, mean 24.8) and 92 males (age range 20-48 years, mean 33.1). Mid-stream voided urine was collected from these subjects using urine sampling cups, immediately transferred to 50-ml sterilized Spitz tubes, and analyzed within 30 min using the automated urinary flow cytometer. Urinary erythrocytes were derived from glomeruli in all samples of healthy subjects. The urinary erythrocyte count showed a logarithmic normal distribution. Values 2 SD of the urinary erythrocyte count in healthy individuals or higher were regarded as abnormal, and hematuria was considered to be positive when 11.0/microliter or more erythrocytes were observed by this analyzer. The finding by this analyzer corresponded to the report of Birch et al.

Adult

Use of an osseointegrated implant for dental rehabilitation after cleft repair by periosteoplasty: a case report.

OBJECTIVE: We report here on a patient with unilateral cleft lip and alveolus who underwent dental rehabilitation of cleft alveolus using an osseointegrated implant after cleft repair by periosteoplasty. The patient, whose lateral incisor was congenitally missing, had periosteoplasty, followed by excellent bone formation at the cleft alveolus within 2 years. After the completion of orthodontic alignment of the maxillary dental arch, a Brånemark single-tooth implant was placed into the bone-formed alveolus. CONCLUSION: This treatment procedure offers an option of dental rehabilitation for the alveolar clefts of patients with cleft lip and/or palate.

Alveolar Process

[Intraoperative monitoring for functional neurosurgery during intravenous anesthesia with propofol].

We studied the effects of propofol on electrophysiologic monitoring for functional neurosurgery. In six patients with intractable epilepsy, electrocorticograms (ECoGs) were monitored for epilepsy surgery, and in two of them, somatosensory evoked potentials (SEPs) were monitored because of the focus adjacent to the central sulcus. In four patients with hemifacial spasm, brain stem auditory evoked potentials (BAEPs) and abnormal muscle responses (AMRs) were monitored during microvascular decompression (MVD). In two patients with Parkinson's disease and in one patient with post-traumatic tremor, neural noise levels were recorded from microelectrodes during posteroventral pallidotomy and Vim thalamotomy. In each case of epilepsy surgery, during intravenous anesthesia with propofol, spike activity was recordable enough to identify the resective area and the residual spikes. SEP phase reversal was obtained in two patients and an exact determination of the central sulcus was possible. BAEPs and AMRs were obtained in all MVDs. To record neural noise levels, the infusion of propofol was decreased in two cases of posteroventral pallidotomy, and it was stopped in one case of Vim thalamotomy. In these patients, neural noise levels were recorded and were useful for identifying the target. Propofol is a potentially useful anesthetic agent for electrophysiologic monitoring during functional neurosurgery.

Adult

Infection with GB virus C in patients with chronic liver disease.

Infection with putative non-A to E hepatitis virus, designated GB virus C (GBV-C), was surveyed in 286 patients with chronic liver disease in Japan. RNA of GBV-C was detected, by reverse-transcription polymerase chain reaction with nested primers from the 5'-noncoding region, in 19 patients (6.6%) at a frequency higher (P < 0.001) than in three of 275 (1.1%) normal controls. It was detected in three of 83 (4%) patients with hepatitis B virus infection, 15 of 188 (8%) patients with hepatitis C virus infection, and one of 12 (8%) patients without evidence of ongoing infection with hepatitis B or C virus. GBV-C RNA was detected in nine of 186 (5%) patients with chronic hepatitis aged 51.2 +/- 13.3 years, six of 64 (9%) with liver cirrhosis aged 62.9 +/- 11.4 years, and four of 36 (11%) with hepatocellular carcinoma aged 62.0 +/- 11.1 years. Nucleotide sequences of 100 base pairs in the helicase region of GBV-C isolates from the 19 patients varied up to 21%, while sequences of 33 deduced amino acids were conserved and differed only by up to 6%. These results indicate that infection with GBV-C in patients with non-B, non-C chronic liver disease would not be frequent, although the sensitivity of the detection method could be improved. Coinfection of GBV-C with hepatitis B or C virus, as well as the duration of infection, might accelerate the progression of chronic liver disease.

Amino Acid Sequence

Duplex PCR of the Y-27H39 and HPRT loci with reference to Japanese population data on the HPRT locus.

Using a duplex polymerase chain reaction (PCR) system, we successfully amplified GATA microsatellite loci Y-27H39 and HPRT on the Y and X chromosomes, respectively, and detected their alleles with a digoxigenin-labeled (GATA)6 probe. Among 450 unrelated Japanese including 225 females, we found 6 HPRT alleles ranging from 155 to 175 nucleotides with a simple repeat structure comprising 11 to 16 repeats. The duplex PCR is highly informative in Japanese.

Alleles

[Metastatic renal tumor originating from esophageal cancer: report of 2 cases].

We report 2 cases of esophageal cancer metastatic to the kidney. The first case was in a 57-year-old man who complained of severe right flank pain. He had underwent an operation for esophageal cancer 2 months previously. A computerized tomography (CT) scan revealed a wedge-shaped, low-density mass in the right kidney. Right nephrectomy revealed squamous cell carcinoma. He has remained free of recurrence 3 months postoperatively. The second case was in a 57-year-old man with esophageal cancer treated by radiation therapy. Severe right flank pain and gross hematuria appeared after 1 year. A CT scan showed a wedge-shaped, low-density tumor in the right kidney accompanied with a tumor thrombus in the inferior vena cava. Right nephrectomy as well as resection of the thrombus were performed. Pathological diagnosis was squamous cell carcinoma. He died of cancer 2 months postoperatively.

Carcinoma, Squamous Cell

An inhibitory effect of actin on casein kinase II activity in vitro.

The inhibitory effect of actin on protein phosphorylation by three distinct protein kinases (CK-II, A-kinase and MAP-kinase) was examined in vitro. It was found that: (i) actin inhibits the activities of alpha-monomeric CK-II (CK-IIalpha) as well as oligomeric CK-II (alpha2beta2) in a dose-dependent manner, but has no effect on the activities of the two other kinases; and (ii) actin-induced inhibition of CK-II activity is due to the binding of actin to the alpha-subunit of CK-II and is non-competitive with its phosphate acceptors. In addition, it is demonstrated that actin binds directly to CK-II: both actin and CK-II are coprecipitated by anti-serum against Drosophila CK-IIbeta or by specific IgG against Ascaris suum muscle actin. The results presented here suggest that actin can suppress CK-II-mediated signal transduction.

Actins

Mutation of the pleckstrin homology domain of Bruton's tyrosine kinase in immunodeficiency impaired inositol 1,3,4,5-tetrakisphosphate binding capacity.

Bruton's tyrosine kinase (Btk), a cytoplasmic protein-tyrosine kinase, plays a pivotal role in B cell activation and development. Mutations in the pleckstrin homology (PH) domain of the Btk gene cause human X-linked agammaglobulinemia (XLA) and murine X-linked immunodeficiency (Xid). In this paper, we report that the PH domain of Btk functions as an inositol 1,3,4,5-tetrakisphosphate (IP4), inositol 1,3,4,5,6-pentakisphosphate, and inositol 1,2,3,4,5,6-hexakisphosphate (IP6) binding domain (Kd of approximately 40 nM for IP4), and that all of the XLA (Phe replaced by Ser at position 25 (F25S), R28H, T33P, V64F, and V113D) and Xid mutations (R28C) found in the PH domain result in a dramatic reduction of IP4 binding activity. Furthermore, the rare alternative splicing variant, with 33 amino acids deleted in the PH domain, corresponding to exon 3 of the Btk gene, also impaired IP4 binding capacity. In contrast, a gain-of-function mutant called Btk*, which carries a E41K mutation in the PH domain, binds IP6 with two times higher affinity than the wild type. Our data suggest that B cell differentiation is closely correlated with the IP4 binding capacity of the PH domain of Btk.

Agammaglobulinaemia Tyrosine Kinase

6'-Sulfo sialyl Lex but not 6-sulfo sialyl Lex expressed on the cell surface supports L-selectin-mediated adhesion.

In order to determine if a sulfated oligosaccharide on the cell surface can function as an L-selectin ligand, a novel approach for in vitro transfer of oligosaccharides was utilized (Srivastava, G., Kaun, K. J., Hindsgaul, O., and Palcic, M. M. (1992) J. Biol. Chem. 267, 22356-22361). CHO cells were incubated with synthetic 6'-sulfo sialyl Lex, NeuNAcalpha2-->3(sulfate-6)Galbeta1-->4(Fucalpha1-->3) GlcNAc or 6-sulfo sialyl Lex, NeuNAcalpha2-->3Galbeta1-->4[(Fucalpha1-->3)sulfate--> 6GlcNAc] oligosaccharide linked to C-6 of a fucose residue in GDP-fucose and a milk fucosyltransferase. The resultant CHO cells expressing 6'-sulfo sialyl Lex or 6-sulfo sialyl Lex on their cell surface were tested for adhesion to E-selectin and L-selectin chimeric proteins coated on plates. The results indicate that 6'-sulfo sialyl Lex supports L-selectin-mediated adhesion much better than sialyl Lex similarly tagged on the cell surface. In contrast, 6-sulfo sialyl Lex containing a sulfate group on the N-acetylglucosamine residue did not support adhesion with either selectin. These combined results suggest that 6'-sulfo sialyl Lex is a much better ligand than sialyl Lex oligosaccharide for L-selectin.

Animals

Cytoplasmic localization of mitogen-activated protein kinase kinase directed by its NH2-terminal, leucine-rich short amino acid sequence, which acts as a nuclear export signal.

Mitogen-activated protein kinase (MAPK) is activated in cytoplasm in response to extracellular signals and then is translocated to nucleus. A directed activator for MAPK, MAPK kinase (MAPKK), stays in cytoplasm to transmit the signal from the plasma membrane to MAPK. Here we show that MAPKK contains a short amino acid sequence in the N-terminal region (residues 32-44), which acts as a nuclear export signal (NES) and thus is required for cytoplasmic localization of MAPKK. This NES sequence of MAPKK, like that of protein kinase inhibitor of cAMP-dependent protein kinase or Rev, is rich in leucine residues, which are crucial for the NES activity. Furthermore, the NES peptide of protein kinase inhibitor, as well as the NES peptide of MAPKK, inhibited the nuclear export of ovalbumin conjugated to the NES peptide of MAPKK. These results may suggest a common mechanism of nuclear export using a general leucine-rich NES.

Amino Acid Sequence