Effects of zinc and other divalent metals on deoxyribonucleic acid binding and hormone-binding activity of human alpha 1 thyroid hormone receptor expressed in Escherichia coli

Full-length human thyroid hormone receptor alpha 1 (hTR alpha 1) was expressed in Escherichia coli using a T7 expression system. While present in large amounts, the receptor was highly enriched in the insoluble fraction after cell lysis. We describe here the successful solubilization and refolding of the expressed receptor in a functional form in the presence of Zn2+. Using a DNA-cellulose binding assay and gel shift assay, we found that treatment of expressed receptor with 1 mM EDTA in the denaturing agent (5 M guanidine-HCl) results in the formation of aporeceptor that does not specifically recognize target DNA, while it does retain T3-binding activity. This aporeceptor recovered DNA-binding activity by adding Zn2+ during refolding. Zinc-induced restoration of DNA-binding activity occurred in a dose- and time-dependent manner. Moreover, once recovered, this DNA-binding activity persisted without Zn2+, even in the presence of 1 mM EDTA. These data indicate that the hTR alpha 1 molecule has a high affinity for Zn2+, and this metal coordination is essential for proper folding of TR protein into its native active structure.     

Hemodynamic Parameters Influencing Clinical Performance of Novacor Left Ventricular Assist System

The interrelationships between hemodynamic variables including right ventricular (RV) performance with filling/ejection dynamics of the Novacor left ventricular assist system (LVAS) were determined in 10 of 11 patients who received LVAS as a bridge to heart transplant. Nine were successfully transplanted. Data were obtained intraoperatively, at periodic intervals up to 48 h postimplant and at explant. The hypotheses investigated included (a) RV performance influences LVAS filling characteristics and (b) LVAS pump output is influenced by systemic vascular resistance (SVR). During the period of LVAS support (2-126 days), pumping characteristics included a mean filling volume of 51 ml (range, 24-70), residual volume of 4.9 ml (range, 1-18), pump rate of 113/min (range, 63-175), and pump output of 5.81/min (range, 2.8-8.2). Multiple regression analysis identified pulmonary vascular resistance (PVR), RV stroke work index (RVSWI), and pulmonary capillary wedge pressure, but not RV ejection fraction, pulmonary artery pressure, or central venous pressure (CVP) as the most important correlates with LVAS filling volume (p less than 0.001, R2 = 0.6). In addition, LVAS pump output was influenced mainly by RVSWI, PVR, and SVR (p less than 0.001, R2 = 0.7). It was concluded that LVAS performance is highly dependent on RV function and systemic/pulmonary vascular resistances.     

Electrically evoked brainstem responses in cochlear implant recipients

Electrical stimulation of the auditory nerve in the profoundly deaf population through implanted cochlear prostheses has increased the need for reliable electrophysiologic assessment tools. We have recorded electrically evoked brainstem responses (EABRs) in 21 subjects who have received a 3M/House cochlear implant. Recordings have been made, both intraoperatively and postoperatively, in the laboratory setting. The recording technique, methods of stimulus artifact suppression, and results of our measurements are described. Clinical applications of this technology are suggested.     

Radiotherapeutic management of an orocutaneous defect with a balloon-retaining stent.

Maxillary resection defects have traditionally been technically difficult to treat with conventional radiation therapy. The irregular contours of the defect lead to an uneven distribution of the radiation doses. The potential to undertreat defects of microscopic involvement may leave residual disease. Tissue morbidity is high at isolated "hot spots" or at locations of excessive dose distribution. The use of a water-filled balloon bolus in conjunction with a balloon-retaining stent can significantly improve treatment efficacy and decrease tissue morbidity. This article describes a technique for the fabrication of a balloon-retaining stent for an orocutaneous defect.     

Sonographic diagnosis of breast carcinoma by a 7.5 MHz high-resolution electronic linear array transducer

The diagnostic quality with 7.5 MHz real-time, transducer was evaluated in 175 cases of breast lesions including 106 carcinoma. All cases were histologically confirmed by surgery or excisional biopsy. In the prospective study, accuracy in carcinoma was 84.6% with true positive ratio of 86.8%. There were 14 false negative cases with carcinoma and 13 false positive cases. Five "early" breast carcinoma within 1 cm in diameter were misdiagnosed as benign breast lesions. The false positive cases were composed of 7 degenerated fibroadenomas, 2 mastopathies and 4 inflammatory changes. In the retrospective study, accuracy in carcinoma was 83.4% with true positive ratio of 93.4%. Compared with the prospective study, false negative cases decreased and false positive cases increased in retrospect. The predictive value of positive results with carcinoma were about 80% in each malignant findings on the criterion of breast carcinoma. The 7.5 MHz electronic, linear-array transducer has several advantages over the 7.5 MHz polymer transducer in detection and also precise observation of the breast lesions.     

Reduced-intensity unrelated cord blood transplantation for patients with advanced malignant lymphoma.

We report the results of reduced-intensity unrelated cord blood transplantation (RI-UCBT) in patients with advanced malignant lymphoma. Twenty patients (median age, 46.5 years; range, 27-66 years) underwent RI-UCBT with a preparative regimen consisting of fludarabine 125 mg/m2 , melphalan 80 mg/m 2 , and 4 Gy of total body irradiation. The median infused total cell dose was 2.75 x 10(7)/kg (range, 2.3-3.4 x 10(7)/kg). Graft-versus-host disease (GVHD) prophylaxis was composed of cyclosporine or tacrolimus alone. Fifteen patients achieved primary neutrophil engraftment after a median of 20 days. Eight patients developed grade II to IV acute GVHD, and 2 developed chronic GVHD. Of the 16 patients with evaluable disease, 10 achieved a complete response. Primary disease recurred in 1 patient, and transplant-related mortality within 100 days occurred in 8 of 20 patients. The estimated 1-year probability of progression-free survival was 50%. These data suggest that RI-UCBT is a feasible option for patients with refractory lymphoma who lack an HLA-matched donor.     

Immunoelectron microscopic study of HTLV-II-producing cells with an anti-envelope gp46 monoclonal antibody

Summary We observed expression of envelope gp46 protein on an HTLV-II-producing T-cell line (Mo) cells by an immunoelectron microscopical method using a monoclonal antibody against HTLV-II gp46. gp46 reactivity was observed on virus-like particles, extracellular vesicles, cell membrane, and partially in nuclear membrane and endoplasmic reticulum.     

Changes in fibre-type composition and myosin heavy-chain IId isoform in rat soleus muscle during recovery period after hindlimb suspension

We examined the changes in myosin heavy-chain (HC) isoforms and fibre-type composition in rat soleus muscle using both myosin adenosine triphosphatase staining and sodium dodecyl sulphate/polyacrylamide gel electrophoresis (SDS-PAGE) analyses during the recovery period after 4 weeks of hindlimb suspension. Although there was no change in type IIc fibres after the suspension, an increase in this type of fibres was observed during the 1- to 4-week recovery period. The increase in type Ilc fibres was considered to be due to a shift from type Ila to IIc fibres. The SDS-PAGE analysis revealed the presence of the HC IId isoform, which was not observed in the control muscle, after a 4-week hindlimb suspension. The HC IId isoform gradually decreased over 3 weeks of recovery and disappeared in the 4th week of recovery after the suspension. These results suggest that the hypogravity conditions induced by hindlimb suspension stimulated the synthesis of the HC IId isoform, whereas an increase in mechanical load to the muscle accelerated the degradation of the HC IId isoform and the synthesis of type Ilc fibres during the recovery period after hindlimb suspension.