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T Nemoto

Publications and source records attributed to T Nemoto.

At least 217 records · Page 12Linked to original sources

Ribonucleic acid association with androgen receptor from rat submandibular gland.

The transformed androgen receptor from rat submandibular gland converts to a faster sedimenting form (6-8S) on a glycerol gradient centrifugation after withdrawal of a transformation-inducing reagent (KCl or ATP). In this report, the association of cytosolic RNA with the transformed androgen receptor was investigated as a possible mechanism of molecular conversion of the androgen receptor. When the transformed and converted androgen receptors were treated with RNase A, these receptors sedimented at 4.5S in a low-salt glycerol gradient. Addition of RNA from rat submandibular gland to the RNase-Sepharose-treated transformed receptor caused a shift of receptor peak from 4.5S to 5.8S. RNA from rat submandibular gland, yeast RNA and E. coli rRNA inhibited DNA-cellulose binding of a RNase-treated transformed receptor in the absence of molybdate. These observations suggest that conversion from the transformed 4S androgen receptor to a 6-8S form resulted from the association of RNA(s) with the transformed receptor.

Animals↗

Interaction of glucocorticoid receptor from rat liver with protamine and arginine.

The nontransformed glucocorticoid receptor (GR) from rat liver was found to bind to protamine-Sepharose and could be recovered by a salt gradient without a change in molecular configuration. The nontransformed GR also bound to arginine-Sepharose, but the transformed GR did not bind to either resin. Ligand-free GR interacted with both resins and was eluted without loss of its steroid binding ability. The bindings of GR to protamine- and arginine-Sepharose were saturable. The apparent dissociation constants of GR on protamine-Sepharose varied from 0.34 nM (-molybdate) to 0.68 nM (+ 10 mM molybdate) and those on arginine-Sepharose were 1.99 nM (-molybdate) and 0.65 nM (+ 10mM molybdate), respectively. The maximum binding capacity was achieved by arginine-Sepharose in the absence of molybdate. Higher salt concentrations (0.5 M NaCl) were required to elute GR from protamine-Sepharose than from arginine-Sepharose (approx 0.03 M NaCl). However, the effectiveness of several salts for the elution of GR was consistent in both resins as follows; MgCl2 = CaCl2 = Na2WO4 greater than (NH4)2SO4 = Na2MoO4 greater than arginine-HCl greater than lysine-HCl greater than KCI = NaCl. These results suggest that GR interacts with arginine residues in protamine. Chromatography using these resins resulted in 7-10-fold purification of occupied and unoccupied nontransformed GRs.

Animals↗

Stability and transformation of the glucocorticoid receptor under acidic conditions.

The [3H]triamcinolone acetonide ([3H]TA)-binding ability of the rat liver glucocorticoid receptor (GR) was investigated under acidic conditions, ranging from pH 2 to 7.3. Both in the presence and absence of 10 mM molybdate, the [3H]TA-binding ability decreased below pH 6.5 and was almost completely lost below pH 5, pH 5.9 +/- 0.1 giving 50% [3H]TA-binding. The binding ability was recovered when the pH of the cytosol was reversed to 7.3 or the precipitate obtained on acidification was dissolved in a buffer of pH 7.3. Moreover, in the absence of molybdate, the [3H]TA-GR complexes formed at pH 7.3 remained unchanged until pH 5. Then they decreased, pH 3.9 +/- 0.1 giving 50% binding, and completely disappeared at pH 3. [3H]TA-binding activity recovered from the precipitate also decreased in a similar pH region (a 50% decrease in binding being observed at pH 4.2 +/- 0.04). These results suggest that rat liver GR is rather resistant under acidic conditions and that it exists in a peculiar state below pH 5.9 to approximately 4 as to its ligand binding property: unoccupied GR has no [3H]TA-binding ability but [3H]TA-GR complexes once formed at neutral pH do not dissociate. [3H]TA-GR complexes recovered from the precipitate at pH 5 had a Stokes radius of 7.5 nm, little DNA-cellulose-binding ability and sedimented at 8.6S on glycerol gradient centrifugation, indicating that the receptor existed in a nontransformed state. In addition, both occupied and unoccupied GR were transformed at about pH 4, their being 50% transformation. This transformation was accompanied by irreversible denaturation of the receptor.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Clinical experiences using pneumatic massage therapy for edematous limbs over the last 10 years.

UNLABELLED: A pneumatic massage apparatus has been developed for the treatment of peripheral lymphedema. The stagnant lymph and venous blood are displaced toward the heart by this pneumatic massage. It has been applied to more than 650 patients with edematous limbs in the authors' Hadomer clinic over the last ten years. Satisfactory results were obtained, including decrease in swelling, pain, and induration. The authors' clinical experiences and statistical analysis of the patients are reported here. STATISTICAL ANALYSIS: 9 primary (4 males and 5 females) and 646 secondary (28 males and 618 females) patients with lymphatic edema of extremities have been treated. The majority of the patients had postmastectomy edema of the upper extremity. CLINICAL STUDIES: The circumference of the edematous limb was measured before and after pneumatic massage treatment of forty to sixty minutes, daily or every second day. (Formula: see text). The rate of swelling was calculated by the formula described above. Twenty-six cases, treated with pneumatic massage for more than fifteen months, have been investigated. A decrease in the rate of swelling was observed in 14 cases (54%), there was no change in 7 (27%), and an increase was seen in 5 (19%). Electric impedance plethysmographic analysis of peripheral lymph volume and blood flow indicated that pneumatic massage was effective in the treatment of edema of the limbs, and its effectiveness was confirmed also by core temperature measurement of the hand during pneumatic massage. The decreased circumference of the edematous limb was kept constant for a long period of time by daily repeated pneumatic massage and ordinary conservative treatments. In conclusion, most lymphedema is of a secondary type. Pneumatic massage was effective in the treatment of peripheral lymph edema, and the improvement was maintained by repeated daily pneumatic massage and ordinary conservative treatment. Therefore, before it becomes worse, lymphedema of the limb should be treated by use of conservative treatments, including pneumatic massage continued as a long-term schedule.

Adult↗

Ammonium sulfate precipitation of the glucocorticoid receptor from rat liver.

The transformed glucocorticoid receptor (GR) from rat liver precipitated at 30% saturation of ammonium sulfate and sedimented at 4.3 S on glycerol gradient centrifugation, whereas the nontransformed GR precipitated at higher concentrations of ammonium sulfate (40-50% saturation) and sedimented at 8.6 S on a gradient. Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that heat shock protein 90 (hsp 90) precipitated at 40-50% saturation of ammonium sulfate. Moreover, hsp 90 and the nontransformed GR were eluted from DEAE high performance ion-exchange chromatography at similar salt concentrations (0.22-0.23 M NaCl), whereas the transformed GR was eluted at 0.1 M NaCl. Therefore, hsp 90 seems to be responsible for the surface charge characteristics of the nontransformed GR.

Ammonium Sulfate↗

A randomized study of intensive versus moderate chemotherapy programs in metastatic breast cancer.

Two intensive chemotherapy regimens CFPMV (Cytoxan [cyclophosphamide], 5-fluorouracil, prednisone, methotrexate, vincristine) and CA (Cytoxan, Adriamycin [doxorubicin]) were tested against a moderate regimen, CFP, in a prospective three-arm, randomized study with crossover when relapse or failure occurred, in order to assess whether the response, duration of remission, and survival can be altered by using more intensive regimens as first-line or as rescue therapy. All three regimens were equally effective as initial chemotherapy: CFP 26/46 (57%); CFPMV 31/48 (65%) and CA 26/47 (55%) (P = 0.61) with the least toxicity for the CFP regimen. Median duration of remission were 9.5, 11, and 9 months, respectively. Complete responses were almost identical in all three regimens: 4/46 (9%); 6/48 (12%) and 5/47 (11%) (P = 0.94). CFPMV was an effective regimen as second-line therapy: 11/33 (33%) or third-line therapy: 7/21 (33%). The CA regimen was equally effective as second-line therapy: 8/25 (32%), suggesting that intensive regimens provide an effective rescue therapy, as well in previous responders as in nonresponders. Initial intensive regimens have not substantially altered long-term survival in the whole group of treated patients, Arm II (CFPMV----CA----CFP) 17.6 months; Arm III (CA----CFP----CFPMV) 12.3 months when compared with initial moderate regimens Arm I (CFP----CFPMV----CA) 16.6 months (P = 0.24). The same lack of difference in survival was noticed in responder patients in each arm: Arm II 19.0 months; Arm III 16.0 months versus Arm I 22.0 months (P = 0.13). Our data suggest that a moderate regimen is as effective as more intense regimens for induction therapy in metastatic breast cancer, with less toxicity, preserving the opportunity for an effective rescue therapy with intensive regimens in second or third-line chemotherapy.

Adult↗

Effect of axillary nodal status on the long-term survival following mastectomy for breast carcinoma: nodal metastases may not always suggest systemic disease.

Records of 215 patients receiving radical mastectomies from 1958 to 1968 at Roswell Park Memorial Institute were reviewed for the significance of axillary nodal status on long-term survival and recurrence. Ten-year disease-free interval rates were 83%, 47%, and 17%, respectively, for patients with negative nodes, 1-3 positive nodes, and greater than or equal to 4 positive nodes. Fifteen-year rates were 80%, 37%, and 8%, respectively. Survival and disease-free interval curves for the 3 nodal status groups were significantly different from each other (P less than 0.0001). Curves for three subclasses of the greater than or equal to 4 positive node group (4-6, 7-12, and greater than or equal to 13) were similar in overall survival, but were significantly (P = 0.04) different for disease-free interval, due to a rapid rate of recurrence in the greater than or equal to 13 positive node group. Hazard rates of treatment failure during each successive 3-year period for 9 years following mastectomy for those with 1-3 positive nodes or greater than or equal to 4 positive nodes decreased with time, whereas the rates for those with negative nodes were low and relatively constant throughout follow-up. These findings support the thesis that a significant proportion of negative nodes patients are cured by mastectomy and that a smaller group of node-positive patients have regional disease with a chance of cure by mastectomy.

Axilla↗

Purification and characterization of a nonhormone-binding component of the nontransformed glucocorticoid receptor from rat liver.

The nontransformed glucocorticoid receptor (GR) and an 88-kDa protein in rat liver cytosol were selectively adsorbed on protamine- and arginine-Sepharose. The 88-kDa protein was purified from rat liver cytosol to homogeneity by precipitation with protamine sulfate, followed by DEAE-ion exchange chromatography, gel chromatography, and DEAE ion-exchange high-performance liquid chromatography. The 88-kDa protein appeared to be present as a dimer at both low and high salt concentrations. The physicochemical properties and the amino acid composition of the 88-kDa protein were almost the same as those of heat-shock protein 90 of HeLa cells and yeast. Preincubation of the GR with the polyclonal antibody raised against the 88-kDa protein increased the sedimentation coefficient of the nontransformed GR, but did not change that of the transformed GR. These results indicate that heat-shock protein 90 is associated with rat liver nontransformed GR and is responsible for the interaction of GR with protamine.

Adrenalectomy↗