Purification of the cardiac sarcoplasmic reticulum membrane protein phospholamban from recombinant Escherichia coli

Phospholamban (PLN) was expressed in Escherichia coli as a protein fusion with glutathione S-transferase (GST). GST-PLN was mostly present in the insoluble protein fraction and accounted for approximately 50% of total insoluble protein. Attempts to suppress inclusion body formation or to use GST as an affinity-purification tag failed. A successful purification method is based on preparative SDS/PAGE and electrodialysis. From 1 g cells we typically purified 13.5 mg fusion protein with a PLN content of 2.8 mg. We genetically inserted an enterokinase (EK) protease site just in front of the PLN sequence and demonstrated the proteolytical liberation of PLN from the carrier protein. The approach described represents a substantial advancement in PLN expression and purification.     

High performance of silicided silicon-sidewall source and drain (S4D) structure

A silicided silicon-sidewall source and drain (S⁴D) structure is proposed for sub-0.1-μm devices. The merit of the S⁴D structure is that the series resistance of the source and drain is significantly reduced since the silicide layer is attached very close to the gate electrode and the silicon sidewall can be doped very highly. Thus, very high drain current drive can be expected, Another advantage of this structure is that the source and drain extensions are produced by the solid-phase diffusion of boron from the highly doped silicon-sidewall. Thus, shallow extensions with very high doping can be realized. A 75-nm gate length pMOSFET fabricated with this structure is shown to exhibit excellent electrical characteristics     

46,XX/69,XXX diploid-triploid mixoploidy with hypothyroidism and precocious puberty

We report a 20 month old female patient with diploid-triploid mixoploidy (46,XX/69,XXX) syndrome with hypothyroidism and precocious puberty. The triploid cell line was only expressed in the fibroblast culture and comprised the majority (95%) of the cells. Chromosome analysis of the fetal blood sample and peripheral blood sample were normal. The patient shows typical features of full triploidy (growth and severe mental retardation, cranial and facial dysmorphism, complete syndactyly of fingers 3/4, partial syndactyly of toes 2/3) and facial but no body asymmetry. At the age of 5 months central hypothyroidism and precocious puberty were diagnosed. Thin pigmented streaks were visible on the wrists and legs of the patient at the age of 16 months. This is the first patient reported so far with 46,XX/69,XXX mixoploidy suffering from hypothyroidism and precocious puberty.     

Transgenesis by adenovirus-mediated gene transfer into mouse zona-free eggs

Zona-free mouse eggs at the pronucleus stage were infected with a replication-defective adenovirus vector containing a nuclear-targeted lacZ gene. Exogenous beta-galactosidase activity was detected in almost all eggs at the two-cell stage. Of 27 mice that developed from infected eggs, three carried the integrated exogenous gene mediated by the adenovirus. Two of the three expressed the lacZ gene, and all three mice transmitted the adenovirus-mediated transgene to F1 progeny Southern blot analysis was consistent with single copy integration. This finding should accelerate the development of new strategies for transgenesis and assist studies on the function of cloned genes in vivo.     

Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in pig ovaries

The porcine antral follicles, 3-6 mm in diameter, were dissected from the ovaries of mature pigs, and then granulosa and cumulus cells were isolated from each follicle. In atretic follicles, high activity of neutral Ca2+/Mg2+-dependent endonuclease and DNA ladder formation, estimated by electrophoresis, were noted in granulosa cells but not in cumulus cells. Extremely low activity of the endonuclease and no DNA ladder formation were observed in both types of cells obtained from healthy follicles. Moreover, apoptotic cells were observed histochemically among granulosa cells only. A good correlation (r = 0.987) between the endonuclease activity of granulosa cells and the progesterone/estradiol ratio of follicular fluid in each follicle was found. These results suggest that apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in pigs.     

Hemodynamic effects of spinal anesthesia in the elderly: single dose versus titration through a catheter

Protective immunity against infection with Mycobacterium tuberculosis is imparted by T cells rather than antibodies, but B cells can play a role as antigen-presenting cells and in granuloma formation. We re-evaluated the role of B cells in the course of tuberculous infection in mu-chain knock-out (Ig-) mice. Surprisingly, the organs of M. tuberculosis-infected Ig- mice were found to have three- to eight-fold elevated counts of viable bacilli compared with normal littermates at 3-6 weeks post-infection. Splenic interferon-gamma responses to whole antigen were unimpaired, whilst proliferation to certain mycobacterial peptides was found to be diminished. However, bacille Calmette-Guérin (BCG) vaccination significantly reduced the infection in Ig- mice. The mechanisms by which B cells can influence primary tuberculous infection need further study.     

On the determination of optimal transition profile of a stepped bar in tension or in torsion by BEM

The objective is to have uniformly distributed tangential stresses on the transition profile of a stepped bar subjected to tensile and torsional loading using an axisymmetric boundary element formulation. The transition curve is represented by the Langragian interpolation polynomial with progressive degrees to avoid shape distortion during the optimization procedure. The calculated result is compared with that obtained by fluid dynamics given in the literature. It is seen that the transition curve obtained in this paper is reasonable.