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Biomedical subjects

M Shinohara

Publications and source records attributed to M Shinohara.

At least 145 records · Page 8Linked to original sources

Ultrasonographic texture characterization of salivary and neck masses using two-dimensional gray-scale clustering.

OBJECTIVES: To evaluate the clinical usefulness of texture analysis in ultrasonography (US) of head and neck masses. METHODS: Sixty-one cases with salivary or neck masses were examined by US with a linear 7.5MHz array. Images were digitized and quantitatively evaluated by two-dimensional gray-scale clustering using two texture indices, M and SD, as measures of coarseness and homogeneity respectively. RESULTS: Coarseness of Warthin's tumors was significantly higher than that of the other salivary tumors. Malignant lymphomas had finer texture values than other lymph node lesions. CONCLUSIONS: Quantitative texture analysis appears to be a useful complement in diagnostic US.

Diagnosis, Differential↗

Characterization of the roles of the Saccharomyces cerevisiae RAD54 gene and a homologue of RAD54, RDH54/TID1, in mitosis and meiosis.

The RAD54 gene, which encodes a protein in the SWI2/SNF2 family, plays an important role in recombination and DNA repair in Saccharomyces cerevisiae. The yeast genome project revealed a homologue of RAD54, RDH54/TID1. Properties of the rdh54/tid1 mutant and the rad54 rdh54/tid1 double mutant are shown for mitosis and meiosis. The rad54 mutant is sensitive to the alkylating agent, methyl methanesulfonate (MMS), and is defective in interchromosomal and intrachromosomal gene conversion. The rdh54/tid1 single mutant, on the other hand, does not show any significant deficiency in mitosis. However, the rad54 rdh54/tid1 mutant is more sensitive to MMS and more defective in interchromosomal gene conversion than is the rad54 mutant, but shows the same frequency of intrachromosomal gene conversion as the rad54 mutant. These results suggest that RDH54/TID1 is involved in a minor pathway of mitotic recombination in the absence of R4D54. In meiosis, both single mutants produce viable spores at slightly reduced frequency. However, only the rdh54/tid1 mutant, but not the rad54 mutant, shows significant defects in recombination: retardation of the repair of meiosis-specific double-strand breaks (DSBs) and delayed formation of physical recombinants. Furthermore, the rad54 rdh54/tid1 double mutant is completely defective in meiosis, accumulating DSBs with more recessed ends than the wild type and producing fewer physical recombinants than the wild type. These results suggest that one of the differences between the late stages of mitotic recombination and meiotic recombination might be specified by differential dependency on the Rad54 and Rdh54/Tid1 proteins.

Amino Acid Sequence↗

An association between salivary gland disease and serological abnormalities in Sjögren's syndrome.

In the labial salivary glands (LSGs) of 16 primary and 18 secondary Sjögren's syndrome (SS) patients, infiltrating lymphocytes were histologically and immunohistochemically examined; also, the serum levels of rheumatoid factor, antinuclear antibodies, anti-DNA antibodies, anti-SS-A and anti-SS-B antibodies, and immunoglobulins (including IgG, IgM and IgA) were all assayed. An immunohistochemical analysis of the lymphocyte subsets in LSGs revealed that severe lymphocytic infiltration was frequently accompanied by marked B cell accumulation both in primary and secondary SS patients. Furthermore, local B cell accumulation was also closely associated with elevated levels of anti-SS-A and anti-SS-B antibodies and IgG, and this association was statistically significant in the group with primary SS but not in the group with secondary SS. Thus, local lymphocytic infiltration, especially B cell accumulation, in the salivary glands is suggested to be involved in serological abnormalities in primary SS, while complicated autoimmune diseases other than SS may also be involved in serological abnormalities in secondary SS.

Adolescent↗

[The clinical study of radical cystectomy for lymph node positive bladder cancer].

BACKGROUND: The impact of pelvic lymph node dissection on the survival of patients with lymph node positive bladder cancer is controversial. We analyzed the records of the bladder cancer patients treated with radical cystectomy. METHODS: Between 1976 and 1993, 84 patients with transitional cell carcinoma of the bladder were treated with radical cystectomy at Tokyo Metropolitan Komagome General Hospital. Sixteen patients (19.0%) had pathologically proved nodal metastases. These patients' records were reviewed with regard to pathological status, and the outcome compared with negative lymph node patients. The two patients with pN3, which we could not completely undergo lymph node dissection of, were excluded from the analyses for recurrence and survival. RESULTS: There were 10 males and 6 females, aged between 47 and 83 with a mean of 68.3 years. There were significant differences between the patients with positive lymph node and negative in the rate of grade 3 and the rate of infiltration of cancer to the vein and lymphatics of the bladder walls. The frequency of the patients of positive lymph node was 5.3% in pT1, 10.0% in pT2, 28.6% in pT3a, 32.0% in pT3b, 50.0% in pT4, and none in pTis, pTa and pT4is. The five-year survival rates estimated by the Kaplan-Meier method were 68.6% over all 82 patients, 75.7% for the patients with negative lymph node and 30.7% for positive. Of the patients with positive lymph node, two-year survival was 80.0% for pT2-3a, versus 0% for pT3b-4. This difference was highly significant (p < 0.01). But survival analysis for pT2-3a showed no significant difference between the lymph node negative patients and positive. The recurrence rate was 57.1% (8 of 14) in patients with positive lymph node and seven patients suffered from distant metastases. All the patients which had recurrent tumors died by bladder cancer. CONCLUSION: Radical cystectomy with lymph node dissection can provide favorable outcome in lymph node positive patients with low stage (pT2-3a). But the patients with high stage (> or = pT3b) were poor prognosis. Most of the patients died by distant metastases.

Aged↗

[Decreasing heart rate and shortening of the arterial pulse propagation time by acupuncture in the spectral analyses].

By utilizing NAITO's analytical indices of the high frequency energy (HFEI) [sigma p x f (0.15 Hz < frequency < 0.45 Hz)] and the low frequency energy (LFEI) [sigma p x f (0.025 Hz < f < 0.15 Hz)] of R-R intervals combined with UEDA's index of fluctuation in propagation time of radial arterial pulses (i.e. R-P intervals), the author investigated relation between elongation in R-R and shortening in R-P intervals by application of electrical acupuncture (1 Hz) on traditional acupoints (HT7:SHENMEN and PC4: XIMEN). The spectral heart rate analyses have been performed in 16 males and 14 females (total 30, i.e. 18 physically healthy volunteers and 12 patients). Those who were suffering from any cardiovascular diseases were excluded. Results obtained are as follows; (1) The mean R-R intervals were significantly prolonged from 10 minutes after needle insertion to the end, i.e. 10 minutes after removal of the needles. (2) The R-P intervals were significantly shortened after 10 minutes of needle insertion. It is assumed that the decrease in heart rate by acupuncture was due to relative superiority of the vagal tone and the shortened R-P intervals, and was due to sympathetic nerve activities stimulated by acupuncture, so as to expedite arterial pulse wave propagation by the arterial contraction. Thus, increased activities of both sympathetic and vagal nerves can coexist simultaneously during acupuncture.

Acupuncture Therapy↗

[Responses to surgical stress in blood coagulation and fibrinolysis, platelet counts and thromboxane B2 after esophageal cancer operation].

Studies on responses to surgical stress in blood coagulation and fibrinolysis, platelet counts and thromboxane B2 (TXB2) were carried out with 18 esophageal cancer patients who had undergone radical esophagectomy through right thoracotomy and reconstruction with gastric tube. Plasma levels were measured for the following for coagulation assessment: thrombin.antithrombin III complex (TAT), soluble fibrin monomer complex(SFMC), fibrinogen, antithrombin III, protein C and thrombomodulin. Selected fibrinolytic markers are: tissue plasminogen activator.plasminogen activator inhibitor 1 complex (tPA.PAI1C), plasminogen, alpha 2 plasmin inhibitor, plasmin. alpha 2 plasmin inhibitor complex(PIC), FDP and D-dimer. Peripheral venous blood samples were taken from the patients before the operation, immediately after the operation and on each of the first, second, third, seventh and fourteenth day after the operation. It was observed that TAT, SFMC, tPA.PAI-1C and TXB2 were remarkably altered immediately after the operation. This indicates that the major surgical stress significantly activated coagulation, fibrinolysis and platelets. Higher plasma levels of TAT compared to the pre-operation level was recorded for two weeks after the operation. Furthermore, in four cases, SFMC became positive during three to seven days after operation. These facts indicate that the activation of coagulation persisted during the days after operation. PIC began to increase from the 2nd to 3rd days after operation, reaching the maximum on the 7th day. Biphasic changes which peaked twice on the 1st and 7th days after operation were shown in plasma levels of FDP and D-dimer. These results indicate that the activation of fibrinolysis also persisted during the days after operation. The activation of coagulation and fibrinolysis may persist at least for two weeks after major surgical operation. Careful observation for the states of these systems was thought to be needed during the post-operative days, and the molecular markers could be useful to assess subclinical changes of these systems.

Adult↗

Correlation between HLA-DR expression and level of poly(ADP-ribose) synthetase in human thyroid carcinoma cells.

The expression of MHC class II molecules is normally restricted to antigen presenting cells. Aberrant expression of class II molecules, however, was detected in the thyrocytes of autoimmune thyroid diseases. We attempted to regulate the expression of HLA-DR molecules in thyroid carcinoma cells by expressing the exogenous poly(ADP-ribose) synthetase gene. We transfected a metal inducible expression plasmid capable of expressing poly(ADP-ribose) synthetase gene into thyroid carcinoma 8505C cells and the transformants, treated with metal and IFN-gamma, were separated by Magnetic Cell Separation. The activity of the synthetase was increased in the HLA-DR-enriched transformants as compared with that in control or the HLA-DR+ transformants. RNA blot analysis and flow cytometric analysis revealed that the IFN-gamma-inducible expression of HLA-DR molecules was depressed by the induction of exogenous poly(ADP-ribose) synthetase gene. This result indicates that HLA-DR expression was correlated with the level of poly(ADP-ribose) synthetase in human thyroid carcinoma cells. Furthermore we examined the level of poly(ADP-ribose) synthetase in patients with autoimmune thyroid diseases. We observed a significant decrease in poly(ADP-ribose) synthetase in the patients. Taken together with the previous observation, the decrease in poly(ADP-ribose) synthetase is closely linked to the aberrant expression of HLA-DR molecules in some autoimmune thyroid diseases.

Autoimmune Diseases↗

The effect of leukocyte function of streptozotocin-induced diabetes in naturally occurring gingivitis rat.

Although diabetes mellitus is known to aggravate periodontal disease, the precise relationship between these two entities is far from being completely understood. Further study of this relationship was therefore undertaken in the form of observation of both naturally occurring gingivitis in rats (ODUS/Odu) and effects produced by induction of experimental diabetes mellitus by injection of streptozotocin (STZ: 65 mg/kg, i.v.). At one and 3 mon after STZ injection, liquid paraffin was injected intraperitoneally. Four days thereafter, pocket probing depths of rats were measured and blood samples as well as peritoneal macrophages were collected from both experimental animals and non diabetic controls. Both chemotaxis and phagocytosis of macrophages were studied. At one and 3 mon after STZ injection, pocket probing depths of diabetic animals were significantly deeper than those of controls (p < 0.001). Pocket probing depths were deeper at 3 mon after STZ injection than after 1 mon in diabetic animals. At three months after STZ injection, there was a high degree of positive correlation between pocket probing depths, blood glucose levels, triglyceride, and hemoglobin A1c levels (p < 0.01). Also, macrophage chemotaxis was more suppressed in diabetic rats than it was in controls. Additionally, both phagocytosis ratios and phagocytosis indices of macrophages in the diabetes group were significantly more suppressed than those in controls in both experimental periods (p < 0.001). Findings suggest that both chemotaxis and phagocytosis are compromised in macrophages from rats rendered diabetic by STZ injection. Thus as host defense mechanisms become weakened, there is a corresponding progression of periodontal disease.

Animals↗

Specificity determinants in interaction of the initiator (Rep) proteins with the origins in the plasmids ColE2-P9 and ColE3-CA38 identified by chimera analysis.

The ColE2-P9 rep protein specifically binds to the orgin and initiates DNA synthesis. Interaction of the Rep protein with the origins of plasmids ColE2-P9 and ColE-3-CA38 (one of the close relatives of ColE2-P9) is plasmid-specific. By using chimeric rep genes and chimeric origins we showed that the two region, A and B, in the C-terminal regions of the Rep proteins and the two sites alpha and beta, in the origins are important for the determination of specificity. When each of the A/alpha and B/beta pairs is from the same plasmids, the plasmid replication is efficient. On the other hand, if only the A/alpha pair is from the same plasmids, the plasmid replication is inefficient. For the region A, the plasmid-specificity is mainly determined by the presence or absence of a nine-amino acid sequence. For the region B, the specificity is probably determined by several amino acids. The region B, contains a segment of amino acid sequence which shows significant homology with the DNA recognition helices of various DNA binding proteins. At the site alpha, the single additional base-pair in the ColE3-CA38 origin can be either A/T or T/A. At the site beta, however, the single additional base-pair in the ColE2-P9 origin must be G/C. Among other possibilities we propose that the region A is a linker connecting the two domains in the Rep protein involved in DNA-binding and that the region B is a part of the sequence-specific DNA-binding domain.

Amino Acid Sequence↗

Size-dependent release of carboxyfluorescein from cetylmannoside-modified liposomes in human plasma.

The interaction of liposomes with human plasma was investigated using 6(5)-carboxyfluorescein (CF) as an aqueous phase marker of cetylmannoside-modified multilamellar vesicles (Man-MLVs) of various sizes. The release of CF decreased with increasing liposome concentration. The time courses of the CF release from Man-MLVs were monitored continuously and were analysed kinetically. The curves were characterized by two phases, the first-order release process and the maximum release, which represent the rate and the extent of CF release, respectively. The increase of liposome size increased the rate of release by 42% and the extent of release by 121%, respectively. These effects of liposome size on the release processes were suggested to result from the size-dependent affinities of liposomes to the human complement system. The assay system of liposomally bound fragments of complement component 3 (C3), such as C3b and/or iC3b, was developed by applying a sandwich enzyme-linked immunospecific assay. The percentage of C3 fragments to total proteins bound to liposomes increased with the size of liposomes and there was a good correlation between the extent of CF release and the percentage of C3 fragments bound. These results indicated that Man-MLVs released entrapped CF via activating the human complement system and the affinity of Man-MLV to complement increased with the size of Man-MLVs in human plasma. These in vitro results suggest the role of complement as an opsonin in the disposition of Man-MLVs in humans.

Drug Carriers↗

Cytokine messenger RNA expression in the labial salivary glands of patients with Sjögren's syndrome.

OBJECTIVE: To characterize the initiation and progress of localized autoimmune damage in Sjögren's syndrome (SS), an autoimmune disease that is also considered to be a lymphoaggressive disorder, by examining the pattern of cytokine production at the site of autoimmune damage. METHODS: Using a polymerase chain reaction-based method, cytokine messenger RNA (mRNA) expression in the labial salivary glands of 15 patients with SS was investigated. In addition, the infiltrating lymphocytes in the labial salivary glands were examined immunohistochemically. RESULTS: Messenger RNAs of Th1 cytokines, such as interleukin-2 (IL-2) and interferon-gamma, were consistently detected in all patients, while Th2 cytokine mRNAs, such as IL-4 and IL-5, were detected in some cases, in association with strong B cell accumulation in the labial salivary glands. Other cytokine mRNAs produced by a variety of cell types, including IL-10, IL-6, and transforming growth factor beta (TGF beta), were also consistently detected in all patients, while IL-12 mRNA was detected in some of the patients. CONCLUSION: These results suggest that Th1 cytokines, as well as IL-10, IL-6, and TGF beta, are essential in the induction and/or maintenance of SS, while Th2 cytokines are involved in the progression of the disease process, especially local B cell activation.

Adult↗

Undifferentiated carcinoma of the broad ligament in a 28-year-old woman--a case report and results of immunohistochemical and electron-microscopic studies.

A case of undifferentiated carcinoma of the broad ligament is reported. The patient was a 28-year-old Japanese woman who underwent laparotomy for a cystic and solid tumor of the left broad ligament. The tumor measured 9 cm in diameter and was unattached to the uterus, ipsilateral ovary, or salpinx. Histological examination revealed tumor tissue consisting of solid nests of irregular configuration and rather small to medium-sized undifferentiated epithelial cells. The tumor cells were positive for cytokeratin, but negative for CA125, epithelial membrane antigen, S-100 protein, amylase, carcinoembryonic antigen, chromogranin A, synaptophysin, HHF35, desmin, and vimentin. Electron-microscopic examination revealed gap junctions and primitive cellular junctions, but no intracytoplasmic mucous granules or microvilli on the surface of the tumor cells. This is the first report of undifferentiated carcinoma of the broad ligament as far as we have been able to determine from a search of the literature.

Adnexal Diseases↗

Isolation of a novel lectin from the globiferous pedicellariae of the sea urchin Toxopneustes pileolus.

Sea urchin lectin-I (SUL-I), a 32 kDa lectin was purified from the large globiferous pedicellariae of the sea urchin, Toxopneustes pileolus by using gel permeation chromatography, ion-exchange chromatography and reverse-phase HPLC. SDS-PAGE showed that SUL-I is a monomeric protein with a molecular mass of 32 kDa. Amino acid analysis indicates SUL-I to contain 294 residues. SUL-I was shown to have chemotactic properties for guinea-pig neutrophils at concentrations of 0.625 microgram/ml. These data suggest that a 32 kDa lectin from T. pileolus may be related to defensive role.

Animals↗

T cell receptor V alpha and V beta gene usage by tumour-infiltrating lymphocytes in oral squamous cell carcinoma.

Oral squamous cell carcinomas (SCC) are often infiltrated by a large number of T lymphocytes. To clarify the nature of the tumour-infiltrating lymphocytes (TIL), we examined T cell receptor (TCR) V alpha and V beta gene usage by TIL and peripheral blood mononuclear cells (PBMC) obtained from 10 patients with oral SCC. We obtained RNA from TIL and PBMC, synthesized complementary DNA, and used the polymerase chain reaction (PCR) method with a panel of primers specific for the V gene segment subfamily (V alpha 1-18/V beta 1-20). We thus found that TIL showed more restricted usage of V beta gene families in contrast to PBMC of the same patients while two unique V beta gene (V beta 6 and V beta 5.2) segment transcripts were overexpressed in the TIL of more than half of the patients. On the other hand, no major difference was observed in the V alpha gene usage between the TIL and PBMC of most patients. To characterize these T cell subpopulations with unique V beta gene segment transcripts further, we sequenced the complementarity-determining region 3 in V beta 6-C beta and V beta 5.2-C beta PCR products derived from TIL and PBMC of two selected patients in each case. Although no usage of the conserved amino acid sequence by TIL was detected, the frequent use of V beta 6/J beta 1.1 in one patient and the V beta 6/J beta 2.7 gene segments in another patient was observed. Regarding the V beta 5.2 transcripts, obtained from the other two patients, no preferential usage of specific J beta gene segments by TIL was observed. These results suggest that the unique T cell populations are amplified in patients with oral SCC, possibly as a consequence of an in situ immune reaction.

Adult↗