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Biomedical subjects

J Imai

Publications and source records attributed to J Imai.

At least 37 records · Page 2Linked to original sources

cDNA cloning of myosin heavy chain isoforms from carp fast skeletal muscle and their gene expression associated with temperature acclimation.

We have isolated cDNA clones encoding fast skeletal muscle myosin heavy chains of carp acclimated to 10, 20 and 30 degrees C for over 5 weeks. All clones covered at least the full length of L-meromyosin, the C-terminal part of the myosin molecule. Nucleotide sequence analysis on cDNA clones showed three types of 3' untranslated sequences, demonstrating that carp expresses at least three myosin heavy chain isoforms in fast skeletal muscle in an acclimation-temperature-dependent manner. cDNAs were identified which were the predominant types expressed in 10 degrees C- and 30 degrees C-acclimated fish, as well as an intermediate type present at all acclimation temperatures. Northern blot analysis using probes of three kinds of DNA fragments from the 3' untranslated region of carp acclimated to 10, 20 and 30 degrees C further confirmed the presence of acclimation-temperature-specific isoforms. In addition, it was found that mRNA levels of three isoforms were altered in an acclimation-temperature-dependent manner. When the deduced amino acid sequences of three types of carp L-meromyosin were compared with those of homoiotherms, the 30 degrees C-acclimated type was more similar to those of homoiotherms than was the 10 degrees C-acclimated type.

Acclimatization↗

99Tcm-MAG3: a sensitive indicator for evaluating perfusion and rejection of renal transplants.

Radionuclide renography has a role in evaluating perfusion of transplanted kidneys. In the course of rejection, cortical perfusion decreases before urinary excretion changes. Based on the facts that 99Tcm-MAG3 has different pharmacokinetics and shows a higher kidney-to-background count ratio than 99Tcm-DTPA, we postulated that 99Tcm-MAG3 was a sensitive and reproducible agent to measure cortical perfusion of transplanted kidneys. To clarify the feasibility of using 99Tcm-MAG3 to measure the cortical perfusion index (CPI), sequential renography was performed using 99Tcm-DTPA and 99Tcm-MAG3 in 14 patients with stable renal transplants, who had changes in serum creatinine concentration of less than 50% between the two studies. The CPI was calculated with 99Tcm-DTPA and 99Tcm-MAG3 and these were then compared and correlated with concurrent serum creatinine concentration. The CPI with 99Tcm-MAG3 was 1.43 times that with 99Tcm-DTPA in patients with changes in serum creatinine concentration equal to or less than 20%, and regression analysis revealed that the difference in CPI was larger in patients with more severely decreased renal perfusion than in patients with normal or mildly decreased renal perfusion. This preliminary study has indicated that the CPI with 99Tcm-MAG3 is a sensitive index for detecting changes in renal function, and thus is a feasible indicator of cortical perfusion when evaluating the rejection of transplanted kidneys.

Adult↗

Determination of the 8-methyl ether of xanthurenic acid in human serum by high-performance liquid chromatography with fluorescence detection.

We developed a simple and sensitive assay for the 8-methyl ether of xanthurenic acid in serum by high-performance liquid chromatography with fluorescence detection (excitation at 340 nm, emission at 450 nm). The compound under study in serum samples was extracted with Sep-Pak C18 cartridge and the extract was applied to an octadecylsilane-bonded column (Nucleosil 5C18, 150 x 4 mm I.D.). The mobile phase used was a mixture of 0.05 M sodium acetate buffer (pH 6.0), containing 5 mM sodium 1-octanesulfonate and 0.1 mM Na2EDTA, and acetonitrile (93:7, v/v). The 8-methyl ether of xanthurenic acid from serum samples was sufficiently separated to be clearly distinguishable. The quantification limit was 2 x 10(-14) mol, which was sensitive enough to detect 8-methyl ether of xanthurenic acid in serum from normal subjects. The method was applied to samples from patients with deficiency in tryptophan metabolism, xanthurenic acid/3-hydroxy kynurenineuria and showed a striking elevation in serum 8-methyl ether of xanthurenic acid.

Amino Acid Metabolism, Inborn Errors↗

Presence of antibodies to p21X and/or p27rex proteins in sera from human T-cell leukemia virus type I-infected individuals.

The human T-cell leukemia virus type I (HTLV-I) pX gene encodes three nonstructural proteins, p40tax, p27rex and p21X. So far, natural antibodies to p27rex and/or p21X have not been found in sera from HTLV-I-infected individuals, although antibodies to p40tax have been found. Recently, the viral transcripts specific for these proteins were detected in fresh peripheral blood mononuclear cells from HTLV-I-infected individuals by the polymerase chain reaction coupled to reverse transcription, showing the in vivo expression of these proteins. We detected antibodies to p21X and p27rex by an enzyme-linked immunosorbent assay (ELISA) system using a recombinantly produced p21X protein as a common antigen, because p21X is identical to the C-terminal portion of p27rex. The sensitivity of the ELISA was determined to be approximately 100 times greater than that of Western blotting. From the analyzed sera of 31 ATL patients, 30 asymptomatic carriers, 18 HAM patients and 100 healthy donors, three specimens from one ATL patient and two carriers were found to be positive for anti-p21X/p27rex antibodies. The specificity of the ELISA reaction was confirmed by the competitive ELISA test with the highly purified recombinant p21X protein. As of result, we first determined the presence of anti-p21X/p27rex antibodies in a small percentage (3.8%) of the sera from HTLV-I-infected individuals. Even sera from the ATL patients, whose fresh PBMCs contained the transcripts for these proteins, were not found to contain these antibodies, suggesting that the immune response to these proteins is low in HTLV-I-infected humans.

Animals↗

Genetic analysis of the Saccharomyces cerevisiae RHO3 gene, encoding a rho-type small GTPase, provides evidence for a role in bud formation.

RHO3 encodes a Rho-type small GTPase of the yeast Saccharomyces cerevisiae. We isolated temperature-sensitive alleles and a dominant active allele of RHO3. Ts- rho3 cells lost cell polarity during bud formation and grew more isotropically than wild-type cells at nonpermissive temperatures. In contrast, cells carrying a dominant active mutant RHO3 displayed cold sensitivity, and the cells became elongated and bent, often at the position where actin patches were concentrated. These phenotypes of the rho3 mutants strongly suggest that RHO3 is involved in directing the growing points during bud formation. In addition, we found that SRO6, previously isolated as a multicopy suppressor of rho3, is the same as SEC4. The sec4-2 mutation was synthetic lethal with temperature-sensitive rho3 mutations and suppressed the cold sensitivity caused by a dominant active mutant RHO3. The genetic interactions between RHO3 and SEC4, taken together with the fact that the Rab-type GTPase Sec4p is required to fuse secretory vesicles together with plasma membrane for exocytosis, support a model in which the Rho3p pathway modulates morphogenesis during bud growth via directing organization of the actin cytoskeleton and the position of the secretory machinery for exocytosis.

Amino Acid Sequence↗

[Renal cell carcinoma in childhood: a case report].

A case of renal cell carcinoma (RCC) in a teenage patient is reported. A 13-year-old girl visited our hospital with the complaint of dyspnea and right abdominal mass. CT scan and chest X-P revealed a right renal tumor with multiple lung metastasis. Wilms' tumor was suspected and a combination chemotherapy consisting of actinomycin D and vincristin was performed. Since the tumor was insensitive to these agents, open biopsy was done. The pathological findings showed clear cell renal cell carcinoma. Despite the administration of alpha interferon, 5-FU and cimetidine, the disease was progressive and the patient died 5 months after diagnosis. It is important to consider the possibility of renal cell carcinoma in teenage patients with a renal mass, since more than 40% of the renal tumors in this age group are renal cell carcinomas.

Adolescent↗

Temperature acclimation induces light meromyosin isoforms with different primary structures in carp fast skeletal muscle.

Carp acclimated to 10 degrees C gave 69k, 66k, and 62kDa light meromyosin (LMM) fragments in SDS-PAGE, while fish acclimated to 30 degrees C gave 74k, 69k, 66k, and 62kDa fragments. The microsequence analysis revealed that the 69k and 66kDa components from the 10 degrees C-acclimated carp contained an N-terminal amino acid sequence different from that of 62kDa. The four fragments from the 30 degrees C-acclimated carp showed the same sequence as that of the 69k and 66kDa components from the 10 degrees C-acclimated carp, except that the 2nd amino acid, Ala, of the 10 degrees C-acclimated LMM was replaced by Thr. DNA fragments encoding an N-terminal region of LMM were amplified by PCR or reverse transcriptase-PCR, demonstrating that the two acclimated groups further contained several amino acids substituted.

Acclimatization↗

Incidence of human T lymphotropic virus type I seroconversion after age 40 among Japanese residents in an area where the virus is endemic.

To clarify how seroconversion after age 40 years contributes to the seroprevalence of human T lymphotropic virus type I (HTLV-I), Japanese residents were studied in an area of Kyushu in which HTLV-I is endemic. Cross-sectional data showed higher seroprevalences among subjects > or = 40 than in those < 40 years old (27.0% vs. 7.9%). The prevalence was also greater in women than men (25.5% vs. 17.6%). Six hundred subjects > or = 40 years old were tested during 1976-1993. Among 8 who seroconverted, 5 had an HTLV-seropositive spouse and 2 seroconverted after blood transfusions. The incidences per 1000 person-years of seroconversion in age groups 40-49, 50-59, and > or = 60 years were, respectively, 0.0, 0.0, and 3.3 (range, 1.38-5.13) in men and 0.0, 5.3 (range, 0.11-7.99), and 1.4 (range, 0.00-1.60) in women. The seroconversion incidence was insufficient to account for the rise of seroprevalence in those > or = 40 years old, especially among women. These results suggest that the high rate and sex discordance in HTLV-I seroprevalence after age 40 may depend mainly on the birth cohort effect for vertical or horizontal transmission.

Adult↗

Serum cryoglobulin and chronic hepatitis C virus disease among Japanese patients.

OBJECTIVES: Hepatitis C virus (HCV)-associated mixed cryoglobulins appear to be detected often in hepatitis C-related chronic liver disease. The association of the two phenomenon among Japanese patients is the subject of the present study. METHODS: Serum levels of total hemolytic complement (CH50) and anti-C3d-binding immune complex, as well as the prevalence of cryoglobulins, were studied in 213 patients with chronic liver disease (hepatitis C, 155; hepatitis B, 58). Cryoprecipitates were tested for anti-HCV Ab and HCV RNA. RESULTS: CH50 activity was significantly lower in patients with hepatitis C than in those with hepatitis B except in responders to interferon who showed a sustained loss of HCV RNA. Cryoglobulins were detected in 24 (37%) of 65 patients with hepatitis C; they generally consisted of polyclonal immunoglobulins but one case. Cryoglobulins were more frequently observed in cirrhotic patients and in those with a longer duration of disease. Cryoglobulinemia-related clinical signs such as vasculitis occurred in only three cases. Patients with cryoglobulins had lower CH50 activity and higher immune complex values than those without cryoglobulins. Anti-HCV Ab and HCV RNA were detected in all cryoprecipitates tested. CONCLUSIONS: These findings suggest that HCV is a major cause of cryoglobulins and advanced liver damage. However, serum cryoglobulins with polyclonal immunoglobulins appear to be less frequent among Japanese patients than among those studied in Western countries.

Adult↗

[Effects of 5-fluorouracil in shrinking residual gastric cancer: a case report].

The patient was a 79-year-old man who underwent extensive gastrectomy and Billroth-II reconstruction for gastric ulcer in 1974. He complained of heartburn in November, 1992, and endoscopy for the residual stomach revealed IIb + IIa-like lesion mainly in the posterior wall of the anastomosed site, which was diagnosed as well differentiated adenocarcinoma. As the patient refused to be operated, 150 mg/day of 5-FU was administered. For 4 months before he decided to undergo operation, 12.9 g of 5-FU was administered in total. After rehospitalization, endoscopy of the residual stomach showed shrinkage of the lesion, and biopsy revealed no malignant findings. The pathological examination after total resection of the residual stomach showed atypism, but no evidence to support malignancy. He has continued to be well without sign of recurrence for 1 year since the operation. The administration of 5-FU is widely used as preoperative adjuvant chemotherapy. This case it is rare for 5-FU to cause the disappearance or falling off of malignant cells.

Adenocarcinoma↗

Determination of 8-methyl ether of xanthurenic acid in human urine by high-performance liquid chromatography.

We developed a simple and sensitive assay for the urinary 8-methyl ether of xanthurenic acid (XA-OMe) by high-performance liquid chromatography with fluorescence detection (excitation at 340 nm; emission at 450 nm). Urine samples were diluted with 0.03 M potassium phosphate buffer (pH 6.0) and applied to an octadecylsilane-bonded column (Nucleosil 5C18, 150 x 4 mm I.D.). The mobile phase used was a mixture of this same buffer and acetonitrile (1000:140, v/v). Both direct injection of urine and solvent extraction prior to HPLC were tested and showed a good correlation and sensitivity, although the peak of XA-OMe was occasionally less distinguishable from close peaks in urine from normal controls by the direct injection method. The quantification limit was 5 x 10(-14) mol which was sensitive enough to detect XA-OMe in urine from normal subjects. The method was applied to samples from patients with a deficiency in tryptophan catabolism, xanthurenic acid/3-hydroxykynurenine-uria and showed a striking elevation in urinary XA-OMe excretion.

Child, Preschool↗

Pharmacokinetics of the garlic compound S-allylcysteine.

The pharmacokinetic behavior of S-allylcysteine (SAC), one of the biologically active transformation products from garlic, was investigated after oral administration to rats, mice, and dogs. SAC was rapidly and easily absorbed in the gastrointestinal tract and distributed mainly in plasma, liver, and kidney. The bioavailability was 98.2, 103.0, and 87.2% in rats, mice, and dogs, respectively. SAC was mainly excreted into urine in the N-acetyl form in rats; however, mice excreted both SAC and the N-acetyl form. The half-life of SAC was longer in dogs than in rats and mice.

Animals↗

Antioxidant and radical scavenging effects of aged garlic extract and its constituents.

The antioxidant properties of three garlic preparations and organosulfur compounds in garlic have been determined. Aged garlic extract inhibited the emission of low level chemiluminescence and the early formation of thiobarbituric acid-reactive substances (TBA-RS) in liver microsomal fraction initiated by t-butyl hydroperoxide. However, the water extracts of raw and heat-treated garlic enhanced the emission of low level chemiluminescence. Among the variety of organosulfur compounds, S-allylcysteine (SAC) and S-allylmercaptocysteine (SAMC), the major organosulfur compounds found in aged garlic extract, showed radical scavenging activity in both chemiluminescence and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays, indicating that these compounds may play an important role in the antioxidative activity of aged garlic extract.

Animals↗

[Usefulness of the measurement of serum soluble IL-2 receptor alpha chain levels in clinical monitoring of non-Hodgkin lymphoma].

We measured the serum concentrations of the soluble form of interleukin-2 receptor alpha chain (sIL-2R alpha) in 25 patients with non-Hodgkin lymphoma (NHL) and 1 patient with virus-associated hemophagocytic syndrome (VHAS) by an enzyme-linked immunosorbent assay (ELISA) using two anti-IL-2R alpha monoclonal antibodies which recognize different epitopes. The sIL-2R alpha levels were markedly higher (n = 12, range = 469.2-11020 U/ml, mean +/- SD = 6153.0 +/- 1687.2 U/ml) in the sera of patients with NHL than in healthy individuals (n = 46, range = 290.1-849.3 U/ml, mean +/- SD = 459.8 +/- 126.9 U/ml, p < 0.01). The serum sIL-2R alpha levels in the NHL patients were closely associated with the stage of disease. The mean value of the serum sIL-2R alpha in each stage of NHL was as follows: stage I (469.2 U/ml, n = 1), stage II (4879.0 U/ml, n = 3), stage III (7364.8 U/ml, n = 8), stage IV (13796.2 U/ml, n = 10). However, there was no clear correlation between the elevation of serum sIL-2R alpha level and the pathological (LSG) classification of NHL. The serum sIL-2R alpha levels serially measured during the clinical course of 2 NHL patients were closely associated with the severity and the progression of the disease. Thus, the measurement of serum sIL-2R alpha levels is very useful not only for the diagnosis but also to monitor the clinical course of NHL.

Aged↗

A genetic system controlling mitochondrial fusion in the slime mould, Physarum polycephalum.

We have identified two distinct mitochondrial phenotypes, namely, Mif+ (mitochondrial fusion) and Mif- (mitochondrial fusion-deficient), and have studied the genetic system that controls mitochondrial fusion in the slime mould, Physarum polycephalum. A mitochondrial plasmid of approximately 16 kbp was identified in all Mif+ plasmodial strains. This plasmid is apparently responsible for promoting mitochondrial fusion, and it is inserted into the mitochondrial DNA (mtDNA) in successive sexual crossing with Mif- strains. This recombinant mtDNA and the unchanged free plasmid spread through the mitochondrial population via the promotion of mitochondrial fusion. The Mif+ strains with the plasmid were further classified as being two types: high frequency and low frequency mitochondrial fusion. Restriction analysis of the mtDNA suggested that the high frequency mitochondrial fusion type was more often heteroplasmic; within each plasmodium, mtDNAs of both parental types were usually present, in addition to the presence of the plasmid. Genetic analysis with the progeny obtained from crossing myxamoebae derived from three different isolates suggested that these progeny carried different alleles at a nuclear locus that controlled the frequency of mitochondrial fusion. These alleles (mitochondrial mating-type alleles, mitA1, 2 and 3) appear to function like the mating type of the myxamoebae; mitochondrial fusion occurs at high frequency with the combination of unlike alleles, but at low frequency with the combination of like alleles.

Alleles↗

Southernmost carriers of HTLV-I/II in the world.

To clarify the real distribution of HTLV-I and -II carriers among indigenous people in central and South America, blood samples collected from indigenous people in isolated regions of Southern Chile were examined. Among 199 inhabitants from Chiloe Island and Pitrufquen town, three cases (1.5%) showed positive anti-HTLV-I antibodies. Two out of the three (82-year-old male and 58-year-old female) reacted to HTLV-II-specific Gag and/or Env proteins but not to HTLV-I-specific ones. The latter case was confirmed as an HTLV-II carrier by polymerase chain reaction test.

Aged↗

[Preliminary study of HTLV-I seroprevalence in Chilean Indian populations].

Aiming to seek the origin and define the prevalence of HTLV-1 infections, 464 blood samples from aboriginal populations proceeding from isolated regions of the north and south of Chile were studied. Antibodies against HTLV were measured with agglutination tests and confirmed with immuno-fluorescence and Western Blotting. Seven out of 107 (6.5%) blood samples from Atacama indians, 2 out of 202 (1%) from Mapuche indians and 3 out of 155 (1.9%) from Huilliche aborigines were positive. These results highlight an important presence of the virus in indigenous populations, specially in the extremes of the country. These findings could suggest an indigenous (mongoloid) origin of HTLV-1 in Chile, specially in Chiloe, where apparently there has been no contact with african or japanese populations, that could be the origin of the contamination.

Adolescent↗

Herbal medicine use, Epstein-Barr virus, and risk of nasopharyngeal carcinoma.

Herbal medicine use is thought to be linked to nasopharyngeal carcinoma (NPC) either through its ability to reactivate the Epstein-Barr virus (EBV) or through a direct promoting effect on EBV-transformed cells. To investigate this, 104 histologically confirmed NPC cases and 205 matched controls were studied in The Philippines. Blood was collected to assess antibody titers against EBV, and an interview was administered which elicited information concerning herbal medicine use and other risk factors for NPC. Subjects strongly positive for anti-EBV antibodies (Epstein-Barr nuclear antigen [EBNA]) (titers greater than or equal to 1:80) were at a 21-fold excess risk of disease (95% confidence interval, 8.4, 51.8). Herbal medicine use was also associated with NPC (relative risk, 2.5; 95% confidence interval, 1.4, 4.5). Associations persisted after adjustment for education, smoking, Chinese ancestry, and consumption of salted fish. Exposure to herbal medicines among subjects testing negative/weakly positive for anti-EBNA antibodies was not associated with an elevation in risk (relative risk, 0.6), strong positivity to anti-EBNA antibodies in the absence of herbal medicine use was associated with a significant 16-fold excess risk of disease, and exposure to herbal medicines among subjects testing strongly positive for anti-EBNA antibodies was associated with a significant 49-fold excess risk of NPC when cases were compared to controls. Similar results were obtained when other serological measures of EBV exposure were used. Anti-EBV antibody titers were elevated in herbal medicine users compared to nonusers among cases but not among control subjects. This suggests that, if herbal medicines interact with EBV in the development of NPC, they do not do so by reactivating EBV infection but rather through a direct proliferative effect on EBV-transformed cells. Although the interaction between EBV and herbal medicines is biologically plausible, larger, more detailed studies need to be conducted to validate this preliminary finding.

Antibodies, Viral↗