Biomedical subjects
Publications and source records attributed to T Matsubayashi.
A physical map of the unicellular cyanobacterium Synechococcus sp. strain PCC6301 genome has been constructed with restriction endonucleases PmeI, SwaI, and an intron-encoded endonuclease I-CeuI. The estimated size of the genome is 2.7 Mb. On the genome 49 genes or operons have been mapped. Two rRNA operons are separated by 600 kb and transcribed oppositely.
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A part of the tRNA(Leu)(UAA) gene containing a 240-nucleotide group I intron was amplified by PCR from cyanobacterium Synechococcus PCC 6301 genomic DNA. The pre-tRNA synthesized from the cloned PCR product was efficiently self-spliced in vitro under physiological conditions. The gene encoding the tRNA(Leu)(UAA), trnL-UAA, was isolated from a Synechococcus PCC 6301 genomic library and the nucleotide sequence of a 2,167-bp portion was determined. The trnL-UAA consists of a 34-bp 5' exon, a 240-bp group I intron and a 50-bp 3' exon. In addition, three open reading frames (ORF1, ORF2 and ORF3) were found in the 5' and 3' flanking regions of trnL-UAA. The predicted protein sequence of ORF3, which is located 74-bp upstream from trnL-UAA on the opposite strand, shows 66.2% amino acid identity to that of the Synechocystis PCC 6803 gene encoding subunit L of NADH dehydrogenase (ndhL).
An unrelated donor bone marrow transplant (UD-BMT) was carried out on a 10-year-old patient with metachromatic leukodystrophy (MLD). We collected cerebrospinal fluid (CSF) on day +168 and cultured it with recombinant IL-2 and PHA-P to examine the origin of cells in the CSF. Analysis on variable number of tandem repeat (VNTR) of lymphocytes in the CSF amplified by PCR revealed that lymphocytes in the CSF were of donor origin. These data support that BMT at an early stage may prevent deterioration in MLD. Although the patient developed grade III acute GVHD with rash and diarrhea, we successfully treated acute GVHD using rabbit anti-human thymocyte immunoglobulin (ATG). UD-BMT may be an alternative treatment for patients with MLD in the absence of an HLA matched family donor.
An accurate physical map of the genome of a cyanobacterium, Synechocystis sp. strain PCC6803, was constructed on the basis of restriction and linking clone analysis. The genome contained 6 recognition sites for AscI, 25 sites for MluI, and 31 sites for SplI, and the entire genome size was estimated to be 3.6 Mb. Sixteen genes or gene clusters, including those involved in the photosynthetic systems, were localized on the physical map of the genome by hybridization. In the course of the above analysis, two extra chromosomal units with approximate sizes of 110 kb and 125 kb were identified.
From September 1986 to December 1993 31059 anti-HIV antibody tests were performed on the samples from our clinic, from 29 health centers and their branches of Osaka Prefecture, from a hospital and from high risk groups. Enzyme immune assay (EIA) was used up to 1988 and from 1989 particle agglutination (PA) has been employed. The indeterminates of Western blot (WB) were seen in 5 EIA positives and in 2 PA positives. False positive rate of EIA was 0.235% (11/467) and that of PA was 0.011% (2/17922). Two false negative cases of anti-HIV-1 antibody test due to window period were documented and the importance of co-use of antigen test at the time of confirmative antibody tests was discussed.
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We have purified a chloroplast elongation factor Tu (EF-Tu) from tobacco (Nicotiana tabacum) and determined its N-terminal amino acid sequence. Two distinct cDNAs encoding EF-Tu were isolated from a leaf cDNA library of N. sylvestris (the female progenitor of N. tabacum) using an oligonucleotide probe based on the EF-Tu protein sequence. The cDNA sequence and genomic Southern analyses revealed that tobacco chloroplast EF-Tu is encoded by two distinct genes in the nuclear genome of N. sylvestris. We designated the corresponding gene products EF-Tu A and B. The mature polypeptides of EF-Tu A and B are 408 amino acids long and share 95.3% amino acid identity. They show 75-78% amino acid identity with cyanobacterial and chloroplast-encoded EF-Tu species.
We have developed an automated latex agglutination test for Treponema pallidum (TPLA) for measurement of the concentration of treponemal antibodies in syphilitic sera. The assay needs neither a complicated pretreatment of the sera nor special techniques. Intra- and interassay precision studies showed high reproducibility. Potentially interfering substances, such as bilirubin, hemoglobin, triglycerides, and rheumatoid factor, did not affect the results. The results obtained with the TPLA test showed a strong correlation with those from conventional methods. This test for antibodies to T. pallidum may be useful for the mass screening of syphilitic infections.
Functional hepatic imaging was performed using 99mTc-DTPA-galactosyl human serum albumin (99mTc-GSA), a radiolabeled ligand that reacts specifically to the asialoglycoprotein receptor that resides at the plasma membrane of hepatocytes, in 21 patients: five with chronic active hepatitis (CAH), 14 with compensative liver cirrhosis (LC), one with chronic inactive hepatitis and one with acute hepatitis. The former two diseases were mainly investigated. Serial liver images were acquired at the rates of 10 sec/frame for 0-5 min and 2 min/frame for 6-30 min after the injection of 99mTc-GSA, and the images were compared with 99mTc-phytate images in 2 patients with CAH and 11 with LC, and those with portal scintigrams using 123I-iodoamphetamine (IMP) in 3 patients with LC. The images using 99mTc-GSA were in better agreement with hepatic function than those using 99mTc-phytate, and with the findings of portal scintigraphy using 123I-IMP. LHL15 (liver/liver and heart radioactivities at 15 min after the injection of 99mTc-GSA) correlated with the hepaplastin test (r = 0.978 in CAH, and r = 0.544 in LC), indicators of hepatic reserve. These results suggest that liver scintigraphy using 99mTc-GSA might be a useful method for evaluating liver function.
A new transcription unit has been identified and characterized in the small single-copy region of tobacco chloroplast DNA. A primary transcript (1550 nucleotides) spanning the entire transcription unit contains no significant open reading frames (ORFs), other than ORF55, recently identified as the gene encoding the ribosomal protein CL32 (rpl32). The leader sequence extends 1101 nucleotides from the rpl32 initiation codon. Primer extension and in vitro capping experiments in combination with ribonuclease protection assays, revealed a promoter situated more than 322 bp inside the coding region of ndhF, which is divergently oriented with respect to rpl32. A canonical Pribnow-box is found just upstream of the transcription start site, but a typical -35 motif was not detected. This is the first internal divergent promoter to be characterized in the chloroplast genome.
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The CT numbers of cortex at the level of 20 cm (CT20) and spongiosa in the lateral condyle at the level of 2 cm (CT20) proximal from the distal end of the femur, and the bone mineral density of spongiosa in the L3 body (BMD), were obtained by QCT. The study included 43 female patients with rheumatoid arthritis (RA), 71 female patients with primary osteoporosis (OP), 20 female nondialyzed patients with chronic renal failure (CRF:nonHD), 37 hemodialyzed patients (CRF:HD), including 13 parathyroidectomized patients (CRF:HD, PTX), and 10 healthy volunteers. CT20 correlated closely with age in RA. CT02 and BMD correlated closely with age in RA and OP. CT20 and CT02 correlated closely with the duration of hemodialysis in CRF:HD, but not with the duration of disease in RA. The values of CT20 and CT02 in the CRF:HD. PTX group were significantly lower than those in the other CRF groups. BMD in the RA groups was not different from that of healthy volunteers. The CT20 values of the one-third of RA patients older than 60 years were extremely low compared with those of the other two-thirds. The results indicated that BMD was useful in assessing bone mineral content in OP, but not in RA. CT02 and CT20 were useful in assessing bone mineral content in these three diseases, CT20 was especially useful for patients in the CRF:HD group and those with RA older than 60 years, but it was not useful in the CRF:nonHD group.
We report two siblings with autoimmune neutropenia of infancy. Patient 1 was hospitalized for viral gastroenteritis at 5 months of age, when his absolute neutrophil count was 684/mm3. Absolute neutrophil counts ranging from 0 to 930/mm3 continued but normalized spontaneously at 25 months of age. Patient 2, a younger brother of patient 1, had prolonged high grade fever at 5 months, when his absolute neutrophil count was 138/mm3. Absolute neutrophil counts ranging from 96 to 1,078/mm3 continued for the following 9 months. Antineutrophil antibodies were detected in the patients' sera. Our cases are siblings and each case was independently affected, since the diagnoses were made at 5 months of age and the interval between the diagnoses was 23 months. Autoimmune neutropenia of infancy is comparatively rare, so that we think that the disease in these siblings was due to a genetic abnormality.
Renoscintigraphy using 123I-orthoiodohipprate (123I-OIH) was conducted, and the dynamic imaging study results and the renograms were analyzed. The results using 123I-OIH were compared with those using 131I-OIH in 22 of 33 patients. Measurement of the effective renal blood flow (ERBF) was also performed using 123I-OIH by the single blood-sample method. 123I is a more suitable nuclide for gamma-camera scintigraphy than 131I in terms of physical characteristics, and a larger dose of 123I-OIH is possible than 131I-OIH because of its lesser radiation. These are the reasons why clearer dynamic images of the blood flow phase, functional phase and excretion phase can be obtained. These make possible diagnoses associated with the organic changes and functions. No essential differences could be found in the functional study results of renogram between the two drugs because on the nuclide of 131I is replaced by 123I, but a renogram with less statistical noise could be obtained using 123I-OIH than 131I-OIH. The radiation dose of 123I-OIH is less than that of 131I-OHI, and clearer dynamic images and functional indexes can be easily obtained in scintigraphy using 123I-OIH. It is concluded that 123I-OIH is a useful radiopharmaceutical for renal examination, and it should replace 131I-OIH.
The CT numbers of cortex at the level of 20 cm (CT20) and of spongiosa in the lateral condyle at 2 cm (CT02) proximally from the distal end of the femur, and the bone mineral density of spongiosa in L3 body (BMD), were obtained by QCT. The study included 48 patients with rheumatoid arthritis or chronic renal failure as well as 10 healthy volunteers. The relationships of CT20 vs BMD in the regions above and below a critical value of BMD were quite different from each other. Similar relationships were observed in the plot of CT20 vs CT02. The results indicated that the demineralization of cortex was much less than that of spongiosa while the mineral content of spongiosa kept higher than a critical value, but the demineralization of cortex surpassed that of spongiosa once the mineral content of spongiosa had become lower than the critical value. It is necessary to assess bone mineral content of cortex especially in patients with lingering imbalance of bone metabolism.
We described a 8-month-old boy of massive hemorrhagic enteritis due to rotavirus complicated by shock. As far as we know, this case is the first patient who developed vascular collapse from the gastrointestinal tract bleeding by rotavirus infection. Clinicians should be alerted to the possibility of the occurrence of severe hemorrhagic enteritis as a life threatening complication in rotavirus induced enteritis.