[Function of membrane phospholipids in Saccharomyces cerevisiae].
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Membrane Lipids↗
Biomedical subjects
A Ohta
Publications and source records attributed to A Ohta.
Localization of nitric oxide synthase-immunoreactive neurons in the solitary nucleus and ventrolateral medulla oblongata of the rat: their relation to catecholaminergic neurons.
The morphological relationship between nitric oxide (NO) and catecholamines in the solitary nucleus (SOL) and ventrolateral medulla oblongata (VLM) was studied by a double immunostaining method with antibodies against NO synthase (NOS), an NO-synthesizing enzyme, and tyrosine hydroxylase (TH), a catecholamine-synthesizing enzyme. Although NOS- and TH-immunoreactive neurons were widely distributed in the SOL and VLM, these immunoreactivities did not coexist in any single neurons. NOS-immunoreactive neurons formed clusters in some restricted regions, i.e. in the medial subnucleus of the SOL, where both NOS- and TH-immunoreactive neurons showed a complementary distribution. These findings suggest that NO-producing neurons constitute a subclass that is distinct from that of catecholaminergic neurons.
Amino Acid Oxidoreductases↗
Age-related deterioration in conditional avoidance task in the SAM-P/10 mouse, an animal model of spontaneous brain atrophy.
A novel inbred strain of mouse 'SAM-P/10' (Senescence Accelerated Mouse) is a model of age-related brain atrophy characterized by age-related loss and shrinkage of neurons in the cerebral neocortex. Age-related changes in learning and memory skills of SAM-P/10 mice were investigated using a newly developed conditional avoidance task in a T-maze. Comparisons were made with findings in the SAM-R/1 strain which shows a little loss and no shrinkage of neocortical neurons. Four-month-old SAM-R/1 and SAM-P/10 performed well during a 10-day training schedule of the conditional avoidance task. SAM-R/1 mice over 17 months of age were slower learners than younger SAM-R/1 mice but reached nearly the same high percentage avoidance as seen in the 4-month-old mice during the last 4 days of the schedule. Performance of the SAM-P/10 mice gradually worsened with aging and 10- to 12-month-old SAM-P/10 mice could not reach the percentage avoidance seen with the 4-month-old mice, even after the 10-day training. When the mean percentage of successful avoidance or escape behavior on every training day was plotted, the curves were much the same for both SAM-R/1 and SAM-P/10 mice, of any age. These results show that aged SAM-P/10 mice retained the left-right turning discrimination in the T-maze and lost the ability to predict the forthcoming aversive shock by associating conditioned stimulus and unconditioned stimulus.
Aging↗
Improved or fatal acute disseminated intravascular coagulation in systemic lupus erythematosus.
To analyze the outcome of systemic lupus erythematosus (SLE) associated with acute disseminated intravascular coagulation (DIC) and also to clarify the clinical factor(s) contributing to the outcome, we retrospectively investigated 120 SLE patients treated between 1981 and 1991. Eight of these patients (6.7%) developed acute DIC; four recovered and the other four died within 2 weeks of onset. Infection preceded acute DIC in all these patients. Acute DIC associated with atypical pneumonia was always fatal, while the patients with pharyngitis or urinary tract infection survived when they were treated adequately. Comparison of the dead and surviving groups revealed that the activity of SLE before the onset of DIC, the severity of DIC, and the treatment given for DIC and the coexistent infection were not significantly related to a fatal outcome. However, severe infection such as atypical pneumonia in patients with secondary immunodeficiency was likely to be fatal irrespective of the presence of DIC.
Acute Disease↗
Cloning of the chitin synthase 3 gene from Candida albicans and its expression during yeast-hyphal transition.
The chitin synthase 3 gene (CACHS3) has been cloned from Candida albicans. The yeast CAL1 gene encoding the chitin synthase 3 of Saccharomyces cerevisiae was used as a probe for the isolation of the gene from C. albicans. The CAL1 homolog was identified in Southern blots of C. albicans genomic DNA and cloned from a C. albicans genomic DNA library. The nucleotide sequences of two partial clones were determined and combined giving a total length of 4610 bp. A continuous open reading frame of 3525 bp encoding a predicted protein of 1175 amino acids and molecular mass of 131 850 daltons was identified. A comparison of the deduced amino acid sequences of CAL1 and the Candida chitin synthase 3 protein showed 59.3% identity. Southern blot analysis indicates that the CACHS3 gene is present in a single copy in the genome and maps to chromosome I. Northern blot analysis shows that expression of chitin synthase 3 gene is dramatically increased during the transition from the yeast to the hyphal form of C. albicans. This change in transcription level strongly suggests that CACHS3 may play a role in Candida morphogenesis.
Amino Acid Sequence↗
Cloning of the C-URA3 gene and construction of a triple auxotroph (his5, ade1, ura3) as a useful host for the genetic engineering of Candida maltosa.
The C-URA3 gene of the n-alkane assimilating-yeast Candida maltosa was cloned by complementation of the ura3 mutation of Saccharomyces cerevisiae. The nucleotide sequence of C-URA3 and its deduced amino-acid sequence showed significant homology to those of the orotidine 5'-phosphate decarboxylases of other fungal species. To construct a useful host for genetic engineering of C. maltosa using C-URA3 as a marker, one allele of C-URA3 in a double auxotroph (his5, ade1) was disrupted by C-ADE1, and subsequently two kinds of ura3 mutants were isolated by selecting for spontaneous 5-fluoro-orotic acid (5FOA) resistance. One of the mutants was homozygous for the disruption (ura3::C-ADE1/ura3::C-ADE1); the other was heterozygous (ura3::C-ADE1/ura3). The ura3::C-ADE1 allele in the latter strain was re-substituted by C-URA3 to rescue the adenine auxotroph (his5, ade1, C-URA3/ura3). Finally, by selecting a 5FOA-resistant mutant, a triple auxotroph (his5, ade1, ura3/ura3) was isolated.
Amino Acid Sequence↗
Successful treatment of autoimmune thrombocytopenia and rapidly progressive pneumonitis with recurrent pneumothoraces in a patient with rheumatoid arthritis.
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Arthritis, Rheumatoid↗
Elevation of plasma angiotensinogen in rats with experimentally induced nephrosis.
In order to determine the activity of the renin-angiotensin system in the nephrotic syndrome, the plasma concentration of angiotensinogen was measured in rats with puromycin aminonucleoside (PA)-induced nephrosis using two different methods: a direct radioimmunoassay, which measures both angiotensinogen and des-angiotensin I-angiotensinogen, and an indirect assay, which measures angiotensin I liberated from angiotensinogen by excess renin. The plasma concentration of angiotensinogen as measured by the direct assay increased before the appearance of PA-induced hypoproteinemia or proteinuria and subsequently decreased to normal levels simultaneously with the appearance of proteinuria. The indirect assay of angiotensinogen also demonstrated an increased concentration of plasma angiotensinogen before the development of nephrosis, but the level decreased to below normal after the appearance of proteinuria. Both plasma renin concentration and renin activity also increased simultaneously with the increase in plasma angiotensinogen. The difference between the concentrations of plasma angiotensinogen determined by these methods increased before and during the early phase of PA-induced nephrosis, suggesting the increased consumption of angiotensinogen by renin during this period. Measurement of plasma corticosterone and serum interleukin-6 revealed that these circulating factors were not involved in the elevation of plasma angiotensinogen in rats with PA-induced nephrosis. These results indicate that the renin-angiotensin system is activated before the appearance of PA-induced nephrotic syndrome.
Angiotensinogen↗
Effects of dimethylpyrazine isomers on reproductive and accessory reproductive organs in male rats.
The effects of dimethylpyrazine isomers on reproductive and accessory reproductive organs in male rats were studied. Following the administration of 2,5-dimethylpyrazine (100 mg/kg, s.c.) the weight of prostate and seminal vesicles, as well as testosterone levels in plasma were significantly decreased. One isomer of dimethylpyrazine, 2,6-dimethylpyrazine (100 mg/kg, s.c.), affected only the seminal vesicles, while 2,3-dimethylpyrazine had no influence on accessory reproductive organs. Following administration of 100 mg/kg of 2,5-dimethylpyrazine, acid phosphatase activity in the prostate and fructose content in the seminal vesicles were also significantly decreased compared with tissues from vehicle-treated animals. Weight of the testes and acid phosphatase activity therein were not affected following the administration of 2,5-dimethylpyrazine, nor were numbers of spermatozoa in the epididymis. These results showed that 2,5-dimethylpyrazine induced decrease in prostate and seminal vesicle weight by inhibiting testosterone uptake and reducing plasma testosterone levels.
Acid Phosphatase↗
Identification of cDNA clones coding for the style specific S11a-glycoprotein gene associated with gametophytic self-incompatibility in tomato (Lycopersicon peruvianum).
We isolated 18 cDNA candidates that were expected to encode parts of the self-incompatibility-associated RNase of Lycopersicon peruvianum PI 126441 (S11a-plant) from a style cDNA library. Polymerase chain reaction was used with oligonucleotides derived from conserved amino acid sequences of RNases from fungi and S-associated RNases in other species of Solanaceae. Two of these clones (both do not have full length sequences) gave 826-bp and 730-bp sequences, and they had the same open reading frame, named ORF-1. Comparison of the deduced amino acid sequence of ORF-1 with S-associated RNase of other Solanaceous plants showed a high degree of similarity. mRNA encoding this ORF-1 was about 1000 bases long and detected only in the style tissue by Northern analysis. Using one of the cDNA clones as a probe, we detected sequence variability among three different S-genotypes of randomly chosen wild-type tomatoes by Southern analysis. From these results, it was concluded that ORF-1 encodes the self-incompatibility associated S-glycoprotein of L. peruvianum PI 126441 (S11a-plant).
Amino Acid Sequence↗
Evolutionary position of n-alkane-assimilating yeast Candida maltosa shown by nucleotide sequence of small-subunit ribosomal RNA gene.
We clarified the evolutionary position of Candida maltosa, an n-alkane-assimilating yeast, by sequencing the nucleotides of the small-subunit ribosomal RNA gene. Phylogenetic analyses showed the close evolutionary relationships of C. maltosa with C. tropicalis, C. viswanathii, C. albicans, C. parapsilosis, and C. guilliermondii, forming a sub-group within this genus.
Alkanes↗
Effects of the novel thyrotropin-releasing hormone analogue Na -((1S,2R)-2-methyl-4-oxocyclopentylcarbonyl)-L-histidyl-L-prol ina mide monohydrate on the central nervous system in mice and rats.
The central nervous system (CNS) effects of a novel thyrotropin-releasing hormone (TRH) analogue, JTP-2942 (Na-alpha- ((1S,2R)-2-methyl-4-oxocyclopentylcarbonyl)-L-histidyl-L-pro linamide monohydrate, CAS 131404-34-7) were investigated and compared with those of TRH. When administrated subcutaneously, JTP-2942 was about 80 times more potent than TRH in the antagonization of reserpine-induced hypothermia, and when administrated intravenously it was about 16 times more potent than TRH in the potentiation of flexor reflexes. Furthermore, oral administration of JTP-2942 was able to antagonize a chlorpromazine-induced reduction in locomotor activity, while TRH was far less effective. In tests on the recovery from pentobarbital-induced sleep and disturbance of consciousness induced by concussive head trauma, JTP-2942 was about 30 and 3 times more potent than TRH, respectively. Thus, JTP-2942 had a far stronger and more persistent action compared with TRH in regard to these effects. However, JTP-2942 had a 3-fold lower effect on thyroid stimulating hormone (TSH) release than TRH. These results suggest that the stimulatory effects of JTP-2942 are selective for the CNS and that this TRH analogue may be applicable to clinical use.
Animals↗
[A case report of cor triatriatum benefit of MRI for preoperative diagnosis and surgical method].
Cor triatriatum is uncommon in all congenital heart diseases. It is a malformation resulting in a separation of the left atrium or right atrium into two chambers due to a congenitally abnormal diaphragm. We wish to present a case of cor triatriatum in which MRI was found most useful for preoperative diagnosis and surgical procedure. A 2-year-old girl was transferred to us for severe pulmonary congestion as shown on chest X-ray. Echocardiography showed abnormal diaphragm in the left atrium. MRI demonstrated clearly the relationship between left pulmonary vein and the abnormal diaphragm. Therefore we should preoperatively determine type I A according to the Lucas and Schmidt's classification. Cardiac catheterization showed moderate pulmonary hypertension and confirmed cor triatriatum. The resection of the abnormal diaphragm was performed under extracorporeal circulation with moderate hypothermia. The postoperative course was uneventful. MRI is a very useful non-invasive technique in making a diagnosis and in choosing the appropriate surgical procedure for cor triatriatum.
Child, Preschool↗
[Clinical study of Still's disease as a distinct disease entity].
There seems to be yet unresolved questions as to whether child-onset and adult-onset Still's disease are truly the same disease and whether adult-onset Still's disease is a disease entity independent of other rheumatic diseases in the adult. In order to clarify these issues, we have analyzed statistically the clinical features of Still's disease of various age-onset and also compared the features of adult-onset Still's disease with those of other rheumatic diseases having similar manifestations. Clinical data used for the study were those collected from 32 institutions in Japan through questionnaire method by Adult Still's Disease Research Committee, and the patients with adult Still's disease and with other rheumatic diseases were definitely diagnosed by the Committee. When the sensitivity of each of total 90 clinical items was compared between child-onset adult Still's disease (11 cases) and adult-onset Still's disease (77 cases), and between young adult-onset (16-35 years) Still's disease (48 cases) and aged adult-onset (45 years or older) Still's disease (15 cases), only 7-9% of the items showed significant difference by chi 2-test. This was the same range as when the sensitivity of the clinical items obtained from the literature of child-onset Still's disease (26 cases) was compared with that of adult-onset Still's disease (77 cases). On contrary to this range, over half of the items showed the significant difference in sensitivity when adult-onset Still's disease was compared with other rheumatic diseases such as seronegative rheumatoid arthritis (31 cases), malignant rheumatoid arthritis (25 cases), and polyarteritis nodosa (31 cases).(ABSTRACT TRUNCATED AT 250 WORDS)
Adolescent↗
[A case of malignant rheumatoid arthritis with long-term improvement by regular use of cryofiltration plasmapheresis].
A Japanese man, 25 years old, suffered from stiffness, swelling, and pain of his joints in May 1983. He was diagnosed as rheumatoid arthritis and was given steroid unsuccessfully. Then, he was admitted to our hospital in February 1984. Because of the presence of high fever and rash on admission, differential diagnosis of RA from adult Stills disease was difficult. However, skin biopsy disclosed apparent vasculitis, leading to the definite diagnosis of malignant RA (MRA). We could not induce any remission by large dose of steroid including pulse therapy, various immuno-suppressants, and anti-rheumatic agents, because of lack of effectiveness and side effects of the drugs. Double filtration plasmapheresis (PA) was performed, but its beneficial effect soon disappeared. On the other hand, cryofiltration PA caused more beneficial and prolonged effect, resulting in improvement. Thereafter, he was successfully followed on regular use of PA for about 6 years. His condition depended on the interval between PA and next PA and on the volume of filtrated plasma. He died of septicemia on March, 1991. We report here the case of MRA with long time improvement by regular use of cryofiltration PA.
Adult↗
Inbred SAM-P/10 as a mouse model of spontaneous, inherited brain atrophy.
We developed a novel inbred strain of mouse with age-related brain atrophy and it was named "Senescence Accelerated Mouse (SAM)-P/10." Macroscopic morphometry indicated that the brains of SAM-P/10 showed age-dependent involutional changes mainly in the frontal portion of the cerebrum. The brain weight decreased by 8.6% throughout the life-span. There were no obvious defects in postnatal development. Semi-macroscopic morphometry revealed a prominent atrophy in the neocortex, olfactory cortex and amygdala. Microscopic morphometry showed that the neocortical neurons were lost with aging, with mostly the large neurons being affected which were lost by 35.6% throughout the life-span. Somata of the neocortical neurons shrank with advancing age. In a control SAM-R/1 strain with only a slight macroscopic involutional change in the brain without weight loss, neither loss of the neocortical large neurons nor shrinkage of the neocortical neurons was evident with aging. Learning and memory skills were evaluated using the one-trial passive avoidance task and conditional avoidance task. Young SAM-P/10 mice performed well in both tasks but older SAM-P/10 showed a poorer performance in both tasks, and this was even poorer than the performance of very old SAM-R/1 mice. Thus, SAM-P/10 can serve as a spontaneous animal model of brain atrophy for a variety of studies of aging of the brain. A better understanding of neurodegenerative diseases with dementia should be forthcoming.
Aging↗
Purification of two chitinases from Rhizopus oligosporus and isolation and sequencing of the encoding genes.
Two chitinases were purified from Rhizopus oligosporus, a filamentous fungus belonging to the class Zygomycetes, and designated chitinase I and chitinase II. Their N-terminal amino acid sequences were determined, and two synthetic oligonucleotide probes corresponding to these amino acid sequences were synthesized. Southern blot analyses of the total genomic DNA from R. oligosporus with these oligonucleotides as probes indicated that one of the two genes encoding these two chitinases was contained in a 2.9-kb EcoRI fragment and in a 3.6-kb HindIII fragment and that the other one was contained in a 2.9-kb EcoRI fragment and in a 11.5-kb HindIII fragment. Two DNA fragments were isolated from the phage bank of R. oligosporus genomic DNA with the synthetic oligonucleotides as probes. The restriction enzyme analyses of these fragments coincided with the Southern blot analyses described above and the amino acid sequences deduced from their nucleotide sequences contained those identical to the determined N-terminal amino acid sequences of the purified chitinases, indicating that each of these fragments contained a gene encoding chitinase (designated chi 1 and chi 2, encoding chitinase I and II, respectively). The deduced amino acid sequences of these two genes had domain structures similar to that of the published sequence of chitinase of Saccharomyces cerevisiae, except that they had an additional C-terminal domain. Furthermore, there were significant differences between the molecular weights experimentally determined with the two purified enzymes and those deduced from the nucleotide sequences for both genes. Analysis of the N- and C-terminal amino acid sequences of both chitinases and comparison of them with the amino acid sequences deduced from the nucleotide sequences revealed posttranslational processing not only at the N-terminal signal sequences but also at the C-terminal domains. It is concluded that these chitinases are synthesized with pre- and prosequences in addition to the mature enzyme sequences and that the prosequences are located at the C terminal.
Amino Acid Sequence↗