Tumor-specific autologous cytotoxic T lymphocytes from tissue sections.
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Aged↗
Biomedical subjects
M Nishida
Publications and source records attributed to M Nishida.
Lung granulomatous response induced by infection with the intestinal nematode Nippostrongylus brasiliensis is suppressed in mast cell-deficient Ws/Ws rats.
Certain nematode infections induce eosinophil infiltration and granulomatous responses in the lungs. To examine the role of mast cells in the development of lung lesions, normal +/+ and genetically mast cell-deficient Ws/Ws rats were infected with the nematode Nippostrongylus brasiliensis. In +/+ rats, numbers of eosinophils in bronchoalveolar lavage fluid (BALF) increased significantly 3-7 days after infection, and granulomatous responses composed of histiocytes/ macrophages and multinucleate giant cells were triggered in the lungs 3-14 days after infection. Challenge infection, which was carried out on day 28 after primary infection, induced much higher levels of granulomatous response than after primary infection, suggesting that the response is mediated at least in part by an immunological mechanism. In Ws/Ws rats, both the eosinophil percentage in BALF and the size of the granulomas in the lungs were significantly smaller than in +/+ rats after primary as well as after challenge infection. The amount of rat mast cell protease (RMCP) II in +/+ rat BALF was increased 1 day after primary infection and more significantly after challenge infection, suggesting that lung mucosal mast cells were activated more markedly after the challenge infection. In Ws/Ws rats, RMCP II was undetectable throughout the observation period. The time course of nematode migration in the lungs did not differ in +/+ and Ws/Ws rats. These results suggest that mast cell activation might be relevant to eosinophil infiltration and granulomatous response in the lungs, although the responses do not affect lung migration of the nematode.
Animals↗
Mutation analysis of gonadotropin receptor and G protein genes in various types of human ovarian tumors.
The heterotrimeric guanine-nucleotide-binding proteins (G proteins) and G protein-coupled hormone receptors including gonadotropin receptors have been suggested to play a role in ovarian tumorigenesis. However, no functional significance of gonadotropin receptors and G proteins in this process has been demonstrated. To investigate this issue, we examined point mutations in these genes in various types of ovarian tumors by polymerase chain reaction-single strand conformation polymorphism analysis and direct sequencing. Among 37 tumors (20 epitherial, 8 sex cord-stromal, and 9 germ cell tumors) and 5 carcinoma cell lines examined, no mutational sequence of G protein-interaction domains of luteinizing hormone receptor and follicle-stimulating hormone receptor, or "hot spots" of the alpha subunit of adenylyl cyclase-stimulating G protein and -inhibitory G protein was observed. Although this analysis was performed on only a limited number of tumors and cell lines, and on limited gene loci, the results suggest that mutational activation in gonadotropin receptors and G proteins is not crucial for ovarian tumorigenesis.
Female↗
Gene rearrangements in snake mitochondrial genomes: highly concerted evolution of control-region-like sequences duplicated and inserted into a tRNA gene cluster.
Mitochondrial DNA (mtDNA) regions corresponding to two major tRNA gene clusters were amplified and sequenced for the Japanese pit viper, himehabu. In one of these clusters, which in most vertebrates characterized to date contains three tightly connected genes for tRNA(Ile), and tRNA(Gln), and tRNA(Met), a sequence of approximately 1.3 kb was found to be inserted between the genes for tRNA(Ile) and tRNA(Gln). The insert consists of a control-region-like sequence possessing some conserved sequence blocks, and short flanking sequences which may be folded into tRNA(Pro), tRNA(Phe), and tRNA(Leu) genes. Several other snakes belonging to different families were also found to possess a control-region-like sequence and tRNA(Leu) gene between the tRNA(Ile)and tRNA(Gln) genes. We also sequenced a region surrounded by genes for cytochrome b and 12S rRNA, where the control region and genes for tRNA(Pro) and tRNA(Phe) are normally located in the mtDNAs of most vertebrates. In this region of three examined snakes, a control-region-like sequence exists that is almost completely identical to the one found between the tRNA(Ile) and tRNA(Gln) genes. The mtDNAs of these snakes thus possess two nearly identical control-region-like sequences which are otherwise divergent to a large extent between the species. These results suggest that the duplicate state of the control-region-like sequences has long persisted in snake mtDNAs, possibly since the original insertion of the control-region-like sequence and tRNA(Leu) gene into the tRNA gene cluster, which occurred in the early stage of the divergence of snakes. It is also suggested that the duplicated control-region-like sequences at two distant locations of mtDNA have evolved concertedly by a mechanism such as frequent gene conversion. The secondary structures of the determined tRNA genes point to the operation of simplification pressure on the T psi C arm of snake mitochondrial tRNAs.
Animals↗
[Development of multi-drug resistant mutants of clinical isolates of Pseudomonas aeruginosa after exposure to fluoroquinolone].
Eight multi-drug resistant mutants (4.94%) were found in 162 clinical isolates of P. aeruginosa after exposure to norfloxacin at different concentrations (1/4, 1/2, 1, 2 and 4 MICs) and were investigated for the mechanisms of drug resistance. All the mutants were eight-times more resistant to norfloxacin than the respective parents, and showed the cross-resistance to the other fluoroquinolones. However, these mutants differed in the drug-resistant patterns; the 94-74 mutant was resistant to carbenicillin, ceftazidime and chloramphenicol, TA-15 mutant was resistant to imipenem, and the 93-183 was resistant to carbenicillin, ceftazidime and gentamicin. TA-16 mutant only showed a marked decrease in the bacterial uptake of norfloxacin. Profiles of the outer membrane proteins of the mutants were analyzed by SDS-PAGE method. The six mutants, except for TA-52 and 93-183 mutants, increased the intensity of bands in the 46 KD region. Three mutants (TA-15, TA-16 and 93-183) decreased the intensity of 44 KD (OMPE) and also the former two decreased the intensity of 22 KD (OMP G). The gryA mutations associated with fluoroquinolone-resistance were investigated for the eight mutants, and the 93-183 mutant showed to have the gyrA mutation caused by the alteration in the amino acid sequence of gyrA; Thr-83 (ACC) to Ile (ATC). The result of the present study indicate that multi-drug resistance of clinical isolates of P. aeruginosa develops through different types of mechanisms, and is not easily explained fully by the present results, and suggest that many factors attributed to the development of the resistances in those mutants remains to be clarified.
Anti-Infective Agents↗
[Drug-resistance patterns and biological properties of MRSA isolated from different geographical areas of Japan].
One hundred isolates of methicillin-resistant Staphylococcus aureus (MRSA) collected from hospitals located in different geographical areas of Japan were used in the present study. The susceptibilities of the strains to gentamicin (GM), erythromycin (EM), tetracycline (TC) and ofloxacin (OFLX) were determined and classified into twelve groups according to their differences in the patterns of the resistance to the four drugs. Of the 100 strains tested, 75 belonging to the main four groups were investigated for the relationships between their patterns of the drug-resistance and biological properties such as coagulase, enterotoxin and phage types, TSST-1 and beta-lactamase producibilities, and percentages of the strains carrying plasmids of large size (> or = 20,000 bp). The main four patterns of drug resistance of the 75 strains were as follows: the first group (33 strains resistant to GM, EM, TC and OFLX), the second group (15 strains resistant to EM, TC and OFLX), the third group (11 strains resistant to GM, EM and OFLX) and the fourth group (16 strains resistant to EM and OFLX) and the remaining 25 strains were divided into further groups. A considerable number of the strains in the third group differed markedly in some biological properties from those in the other groups; coagulase typing (III type-50%: the other groups-II type), enterotoxin typing (64%-non-production: the other groups-C type), TSST-1 producibility (36%-production: the other groups-76 to 88%-production) and phage typing (55%-non-typable: the other groups-80 to 97%-non-typable). In beta-lactamase and plasmids of large size, the first group consisted of the strains with beta-lactamase producibility at the ratio of 50:50, but the other groups consisted of most of the strains with the beta-lactamase producibility. Most of the strains belonging to the second and fourth groups carried large size of plasmids, but 36 and 46% of those belonging to the first and third groups did not carry them. On the other hand, our results of the susceptibility test for the 100 strains showed that a considerable number of strains were susceptible to GM (35%) and TC (42%) at therapeutically significant concentrations.
Anti-Bacterial Agents↗
[Drug-resistance patterns of clinical isolates of Pseudomonas aeruginosa in regard to their lipopolysaccharide-chain sizes].
Pseudomonas aeruginosa strains isolated from patients with different types of the infections each consisted of LPSs different in chain sizes. The drug-susceptibility patterns of these strains of P. aeruginosa were investigated to clarify the relationship between the LPS-compositions and susceptibility to some kinds of anti-pseudomonal drugs. The susceptibilities of nineteen strains (seven long-LPS strains, four short-LPS strains and eight LPS-deficient strains) to piperacillin, ceftazidime, gentamicin, norfloxacin and polymyxin-B were determined and these strains were classified into six types (Types I-VI) according to their drug-resistance patterns. Six of the eight LPS-deficient strains were found to be highly resistant to gentamicin alone (Type IV). Four of the seven strains with the long-LPS and one strain with the short-LPS were resistant to three drugs such as piperacillin, ceftazidime and norfloxacin, and classified into Type I. These results indicated that the major part of the LPS-deficient strains and the considerable part of the long-chain LPS strains of P. aeruginosa tested had each characteristic profile in the drug resistance. The outer membrane proteins of thirteen strains, consisting of different types of LPS compositions, were analyzed by SDS-PAGE. The strains belonging to the same types of the drug-resistance patterns were found to have similar OMP-profiles, although a few exceptions were found. beta-lactamase and gentamicin-inactivating activities were determined for piperacillin-resistant and gentamicin-resistant strains, respectively. Of the piperacillin-resistant strains tested, the activity of beta-lactamase was high in one (No. 8) only, low in four and not found in four. The results showed that degrees of resistance of P. aeruginosa strains tested to piperacillin did not correlate to their producibility of beta-lactamase except one strain. Of the nine gentamicin-resistant strains tested, the gentamicin inactivating activity was high in one (No. 30) only, moderate in six and low in two. These results suggested that the significant levels of piperacillin- or gentamicin-resistance of P. aeruginosa isolated tested might be expressed each due to their decreased abilities for drug-permeabilities in addition to drug-inactivating activities such as beta-lactamase or gentamicin-modifying enzyme. In the case of some resistant strains, the resistance to piperacillin or gentamicin was not explained by the results of the present study. Therefore, we must investigate the possibility that other mechanisms participate in the resistance of these strains.
Drug Resistance, Microbial↗
Alpha 1-adrenergic stimulation induces cardiac tolerance to hypoxia via induction and activation of Mn-SOD.
We examined whether or not alpha 1-adrenergic stimulation increases the tolerance of the heart to ischemia using a hypoxia-reoxygenation model of cardiac myocytes. After exposure to norepinephrine (NE; 0.2 microM) for 24 h, the manganese superoxide dismutase (Mn-SOD) content and activity in the cells were increased from 0.61 +/- 0.03 to 0.87 +/- 0.04 microgram/dish and 22 +/- 1 to 55 +/- 4 U/dish, respectively. The specific activity of Mn-SOD was also increased from 36 to 63 U/microgram Mn-SOD protein after the stimulation with NE. Prazosin (2 microM) abolished the increase in Mn-SOD activity (U/mg total protein). Creatine kinase (CK) release after hypoxia (PO2 7 mmHg; 3 h)-reoxygenation (1 h) from cells pretreated with NE in the presence of propranolol and yohimbine for 24 h was attenuated by 48% compared with that from cells without NE stimulation. When antisense oligodeoxyribonucleotides to Mn-SOD were added to myocyte cultures, the increase in Mn-SOD activity (U/mg total protein) and the attenuation of CK release after the addition of NE in the presence of propranolol and yohimbine were not observed. These results suggest that alpha 1-adrenergic stimulation increases the tolerance of myocytes to hypoxia through induction and activation of Mn-SOD.
Adrenergic alpha-Agonists↗
Preparation of anti-human thymidine phosphorylase monoclonal antibodies useful for detecting the enzyme levels in tumor tissues.
The antitumor activity of cytostatic 5'-deoxy-5-fluorouridine (5'-dFUrd) depends on its being converted to 5-fluorouracil (5-FUra) by the enzyme thymidine phosphorylase (dThdPase, EC 2.42.4). We prepared mouse anti-human dThdPase monoclonal antibodies to serve as tools for clinical studies with this drug. Partially purified dThdPase obtained form HCT116 human colon cancer cells grown in athymic mice was used as and antigen for the immunization of mice. Six hybridomas were cloned which produced anti-human dThdPase antibodies, as detected by Western blot analysis with human dThdPase. With these antibodies, we developed an ELISA method sensitive enough to measure dThdPase levels, even in tumor tissue samples weighing as little as 10 mg. In addition, one monoclonal antibody was suitable for immunologically staining the enzyme in tumor tissues. Thus, these anti-human dThdPase monoclonal antibodies could be used to measure levels of the enzyme in tumor cells, which is essential for the activation of 5'-dFUrd.
Animals↗
Insulin resistance and body fat distribution.
Body fat distribution can be assessed by computed tomography (CT). The ratio of umbilicus was used to classify obese subjects as having visceral fat obesity (VFO) or subcutaneous fat obesity (SFO). Serum triglyceride and total cholesterol levels and plasma glucose area in an oral glucose tolerance test were higher in patients with VFO than in those with SFO. Significant positive correlations were demonstrated between V/S ratio and plasma glucose area, serum triglyceride level, and total cholesterol level as well as systolic or diastolic blood pressure. VFO was more frequently associated with coronary artery disease. Moreover, VFO was more often accompanied by multiple risk factors than was SFO. Steady-state plasma glucose (SSPG) level was significantly higher in patients with VFO than with SFO, suggesting that insulin resistance may be more remarkable in VFO than in SFO. Furthermore, visceral fat accumulation was also associated with these complications even in nonobese subjects. Visceral fat area (VFA) was significantly correlated with fasting plasma glucose, serum triglyceride, and total cholesterol levels. Animal models such as Goto-Kakizaki (GK) rats with ventromedial hypothalamus (VMH) lesions and Otsuka-Long-Evans-Tokushima-Fatty (OLETF) rats were accompanied by visceral fat accumulation and an early stage of aortic atherosclerosis. Aging, sex hormone, genetic, and dietary factors and physical inactivity may induce visceral fat accumulation. Visceral fat is characterized by its high lipogenic activity as well as its accelerated lipolytic activity. High levels of portal free fatty acids (FFAs) may eventually result in an enhancement of hepatic triglyceride synthesis, causing hyperlipidemia. High portal FFA levels would also induce insulin resistance, thereby causing glucose intolerance, hypertension, and finally atherosclerosis. We propose a term, "visceral fat syndrome," as a highly atherogenic state, which includes visceral fat accumulation, glucose intolerance (insulin resistance), hyperlipidemia, and hypertension.
Adipose Tissue↗
[Adjuvant chemotherapy with cyclophosphamide, adriamycin, and CDDP (CAP) for high risk endometrial cancer after complete surgery].
One hundred and six patients with endometrial cancer were treated at Tsukuba University Hospital between 1983 and 1992. Sixteen patients who underwent complete resection were at high risk for recurrence and were given adjuvant platinum-based, multiagent chemotherapy instead of adjuvant radiotherapy. Eligibility criteria included outer-third myometrial invasion (group 1; n = 5), pelvic lymph node metastasis (group 2; n = 3) or both myometrial invasion and lymph node metastasis (group 3; n = 8). Of these three groups at high risk for recurrence, each patient in groups 1 and 2 developed recurrence and died. In group 3 two patients developed recurrence and one patient has died. There was one treatment death due to neutropenic sepsis in group 3. The recurrence sites in all four patients were local and no distant recurrence was noted. In other words, of 15 patients (excluding one treatment death) at high risk for recurrence, three of 12 patients (25%) with deep myometrial invasion and three of 10 patients (30%) with positive pelvic lymph nodes developed recurrence and died. The other 12 patients (include one patient with recurrence; 100 months) have survived for a long interval (range, 45-131 months). The survival rates for stage I, II, III, and IV are 92.6, 89.5, 60.0%, respectively. Patients with stage I in this study had a better survival than those with stage I in the 1984 annual report of the Japan Society of Obstetrics and Gynecology (p < 0.05). The findings of this prospective clinical trial supported those of the randomized trial comparing adjuvant CAP with radiation therapy.
Antineoplastic Combined Chemotherapy Protocols↗
Effects of hepatic denervation on ischemia-reperfusion injury in dogs.
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Alanine Transaminase↗
[An immunohistological study on expression of glutathione S-transferase pi (form) in human endometrial carcinoma].
Ninety-five specimens from patients with endometrial carcinoma (82 of endometrial type, 6 of adenoacanthoma, 4 of adenosquamous carcinoma, 3 of atypical endometrial hyperplasia) and 13 with ovarian endometrioid carcinoma were stained immunohistochemically with a rabbit polyclonal antibody prepared against the placental form of the enzyme glutathione S-transferase pi (GST-pi). Histological studies showed that the degree of staining decreased as the tumor lost its differentiation in endometrial carcinoma, but the degree of staining was independent of the differentiation in the case of ovarian endometrioid carcinoma. A comparison between the grade of staining of GST-pi in 82 cases of the endometrial type of endometrial carcinoma and 13 cases of ovarian endometrioid carcinoma revealed a stronger stain for the endometrial carcinoma than for ovarian endometrioid carcinoma (p < 0.01, Mann-Whitney's U test). Therefore, the GST-pi value for endometrial carcinoma was different from that for endometrioid carcinoma. In general, as compared with ovarian endometrioid carcinoma, endometrial carcinoma is considered to be resistant to chemotherapeutic agents. In conclusion, these results suggest that there is an apparent correlation between the GST-pi value and chemoresistance of the tumor.
Adenocarcinoma↗
[Usefulness of reaction times in high-cortical recognition tasks].
We compared reaction times for high-cortical recognition tasks with kana-searching test in a group of 46 normal individuals aged 21 to 83 who gain a full score in Mini-Mental State. The following tasks of reaction time were used; simple reaction time (SRT), choice reaction time (CRT), memory scanning time (MST), and visual searching time (VST). The results were consistent and demonstrated that mean reaction time of 46 individuals increased in difficult tasks such as MST or VST. All reaction times increased depending on aging, which became longer in difficult tasks. Score of kana-searching test was correlated with MST and VST, but did not correlate with SRT and CRT. These findings suggest that reaction times may be useful for mass-screening of high-cortical functions in different ages as well as the kana-searching test.
Adult↗
[A case of bilateral pneumothorax in the patient with Marfan syndrome].
A case of bilateral pneumothorax in the patient with Marfan Syndrome was reported 13-year-old male was admitted to our hospital with diagnosis of left pneumothorax. He was tall and thin with long tapered extremities and echography revealed annulo-aortic-ectasia. A diagnosis of Marfan Syndrome was established by these characteristic of skeletal and cardiovascular findings. Air leakage continued inspite of the drainage of left chest cavity. Therefore, the resection of bullae was surgically undertaken. Post-operative course was uneventful and he discharged on 18th post-operative day. The pneumothorax of the right side occurred six months after first operation. After chest drainage, the surgical treatment was performed. However, he had recurrence of right pneumothorax six months after second operation. He was treated successfully with drainage tube and minocycline injection into the right pleural cavity. He should be carefully followed up, because the patient of Marfan Syndrome often dies due to cardiovascular abnormalities.
Adolescent↗
[An immunohistological study on expression of glutathione S-transferase pi (form) in dysplastic and neoplastic human uterine cervix lesions].
The glutathione S-transferase (GST) pi has been studied in association with the mechanisms of multidrug resistance and as a marker for malignant tumors. In this study, specimens from 92 cases of cervical neoplasms and 10 cases of normal squamous epithelium adhering to myoma were stained immunohistochemically with a rabbit polyclonal antibody to GST-pi. In 6 cases of normal squamous epithelium, the intermediate layer was positively stained with the GST-pi antibody. In all 20 cases of dysplasia, the cells with koilocytotic atypia were stained positively. In all 10 cases of carcinoma in situ and all 16 cases of stage Ia squamous cell carcinoma, various intensities of GST-pi staining were demonstrated. Forty-six specimens of stage Ib or more squamous cell carcinoma were positive for GST-pi binding except only one case. In general, squamous cell carcinoma of the uterine cervix is resistant to chemotherapeutic agents. GST-pi is most frequently stained in cervical squamous cell carcinoma as compared with ovarian or endometrial carcinoma. In conclusion, these results suggest that GST-pi may be a marker for cervical squamous cell carcinoma.
Adult↗
A nitric oxide donor reverses myocardial injury in rabbits with acute hypercholesterolemia.
We evaluated the propagation of myocardial injury in a model of coronary artery occlusion and reperfusion in control and hypercholesterolemic rabbits. This was done by examining the differences in the infarct size and in the extent of leukocyte accumulation resulting from coronary artery occlusion (30 min) followed by reperfusion (2 or 48 hr) in rabbits fed 1% cholesterol for 4 days vs. controls not fed cholesterol. There was no significant difference in the infarct size in the 2-hr (45.7 +/- 6.7%, n = 8) vs. 48-hr (48.8 +/- 5.8%, n = 9) models of reperfusion in control rabbits. However, infarct size in the cholesterol-fed rabbits at 2 hr (64.0 +/- 4.1%, n = 6) or 48 hr (72.3 +/- 3.0%, n = 8) of reperfusion significantly exceeded that in the corresponding controls (P < .05). The infarct in cholesterol-fed rabbits at 2 hr of reperfusion was smaller than that at 48 hr of reperfusion, but not significantly. Treatment with S-nitroso-N-acetylpenicillamine, a nitric oxide donor, effectively reduced the size of infarct in the cholesterol-fed rabbits. However, treatment with N-acetylpenicillamine had no infarct-limiting effect. When we evaluated the extent of leukocyte accumulation in the ischemic myocardium, as assessed by myeloperoxidase activity, a positive correlation was observed between myeloperoxidase activity and infarct size at 48 hr of reperfusion. Results indicate that the propagation of myocardial ischemia and reperfusion injury in acutely hypercholesterolemic rabbits differed from that in controls. An exogenous nitric oxide donor effectively reduced the size of infarct associated with the reduction in the accumulation of leukocytes 48 hr after reperfusion, which suggests that a reduction in the production of endogenous nitric oxide during ischemia and reperfusion may aggravate the severity of myocardial injury in acutely hypercholesterolemic rabbits.
Animals↗
A novel nonsense mutation associated with an exon skipping in a patient with hereditary protein S deficiency type I.
The genetic defect in a patient with hereditary type I protein S (PS) deficiency was investigated. All the exons and intron-exon junctions of the patient's PS gene were amplified by PCR and subjected to heteroduplex screening. Only the PCR product of exon 4 revealed heteroduplex bands. A novel nonsense mutation, Ser62 (TCA) to Stop (TGA) was found in exon 4. RT-PCR detected the aberrant mRNA in the patient's platelets, which was markedly reduced in amount and lacked the region of exon 4, suggesting that the nonsense mutation affected the mutated mRNA metabolism and induced exon skipping. The skipping of exon 4 causes an in-frame deletion of 29 amino acids which just construct the thrombin-sensitive region of the PS molecule. The loss of such an important domain as well as the quantitative decrease in the mutated mRNA appear to be responsible for the type I PS deficiency in this patient.
Exons↗