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.     

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.     

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.     

Development of ovarian pathology after hysterectomy without oophorectomy

This study was done to determine the occurrence of disease in retained ovaries after hysterectomy. A retrospective analysis of patient charts was performed, comparing the patient reports of women who had secondary ovarian lesions with those whose ovaries showed no pathologic findings during the ten year period of observation (1980 to 1990). The study included 1,265 women with at least one ovary saved after hysterectomy for benign indications. Main outcome measures were ovarian pathologic findings after hysterectomy requiring repeat operation. The overall incidence of lesions in retained ovaries was 3.95 percent. There was a 3 percent risk of having secondary ovarian pathologic findings within three years after hysterectomy, with a decreased risk for the following seven years (mean follow-up time of 60 months, range of three to 120 months). Histologic findings at reoperation included common benign conditions of the ovary. No instance of carcinoma of the ovary was found. The risk of having pathology in the retained ovaries after hysterectomy was significantly higher in women who had only one ovary saved, compared with those who had both ovaries saved (7.63 versus 3.47 percent; p < 0.05). The mean age at hysterectomy was significantly lower in women who had ovarian disorders subsequent to hysterectomy than in those who did not (39.3 versus 43.9 years; p < 0.001). In the group of women with secondary ovarian lesions, mean parity was significantly lower than in those without reoperation (1.22 versus 1.94; p < 0.0001). Women with unilateral oophorectomy at the time of hysterectomy had twice the risk of secondary ovarian lesions, compared with those without oophorectomy at hysterectomy. Determinants, such as age, parity and gravidity must be considered when deciding whether or not to perform oophorectomy at hysterectomy.     

Antitubercular activity of lomefloxacin in an experiment

The effect of lomefloxacin was studied on mice with experimental infection due to Mycobacterium tuberculosis. The antibiotic was administered in doses of 100 and 200 mg/kg. It was shown that the use of lomefloxacin for a month provided a lower death rate of the animals with progressing acute generalized tuberculosis, a lower level of the lesions in the internal organs and a lower number of the Mycobacterium isolates from them. The efficacy of the treatment depended on the drug dose. When lomefloxacin was used in a dose of 200 mg/kg, the survival rate was much higher and the number of the epithelial unicellular granulomas in the tissue of the lung and spleen was markedly decreased while with the lower dose the indices did not differ from those in the control.     

Task-relevant late positive component in rats: is it related to hippocampal theta rhythm?

In the last 2 years (1994-95), two symposium volumes and three reviews have been published that were fully devoted to peptide antibiotics (antibacterial peptides or antimicrobial peptides). Since the field has been growing rapidly, this review is largely a follow-up of new results published in the last 2 years. Sequencing of the 16S RNA of the small ribosomal subunit indicate that the microbial world is much larger than generally appreciated. The importance of the natural flora is stressed and its effect on the evolution of peptide antibiotics and immunity in general is discussed.     

Decision by the Principle Board of the Polish Allergy Society

A review of isolates of Neisseria meningitidis and other laboratory evidence of meningococcal infection was carried out for the period 1990-1995. Thirty-three patients with evidence of meningococcal disease were identified over the six year period. Group B organisms were the most commonly identified, but an increase in Group C has occurred and is almost equal in number. Data from clinical, laboratory and public health sources should be centralized in a National Surveillance Centre.     

Experimental methods in hepatology. Guidelines of the German Association for the Study of the Liver (GASL). Liver perfusion--technique and applications

Purified cell-envelope polyphosphatase as well as polyphoshatase activities of cytosol and isolated vacuoles, of nuclei and mitochondria of the yeast Saccharomyces cerevisiae were compared. The polyphosphatases of cell envelope and cytosol are similar, the polyphosphatases of nuclei, vacuoles and mitochondria differ in their kinetic properties, substrate specificity, requirements in divalent cations and in some effector actions both from these and from each other.     

Responses in growth, food intake and food conversion efficiency to different dietary protein concentrations in meat-type lines of Japanese quail

1. A total of 360 Japanese quail of 4 commercial meat-type lines and two diet treatments (260 and 216 g/kg or 238 and 195 g/kg of crude protein (CP) in the starter and grower diet, respectively) were used. 2. The positive effect of a high CP diet on body weight was significant only for the first 4 weeks after hatching. 3. The mean age at inflection point of the growth curve (t+) across lines and sexes was 1.4 d (0.6 to 2.8 d within line/sex groups) earlier for quail fed on a high CP diet than in quail receiving a low CP diet. The inflection (y+) and asymptotic (A) weights were similar under both dietary protein concentrations. Nevertheless, the shape of the growth curve, characterised by the ratio y+/A and parameter of the maturing rate k, was significantly influenced by diet. 4. A higher food intake and less efficient food conversion were found for quail fed on a high CP diet in the period from 15 to 28 d of age. 5. The fattening traits such as body weight, cumulative food intake and food conversion, were not affected by dietary CP content at the age of 5 weeks. 6. The effects of line on body weight, food intake and food conversion are discussed.     

Glial development in the Drosophila CNS requires concomitant activation of glial and repression of neuronal differentiation genes

Two classes of glial cells are found in the embryonic Drosophila CNS, midline glial cells and lateral glial cells. Midline glial development is triggered by EGF-receptor signalling, whereas lateral glial development is controlled by the gcm gene. Subsequent glial cell differentiation depends partly on the pointed gene. Here we describe a novel component required for all CNS glia development. The tramtrack gene encodes two zinc-finger proteins, one of which, ttkp69, is expressed in all non-neuronal CNS cells. We show that ttkp69 is downstream of gcm and can repress neuronal differentiation. Double mutant analysis and coexpression experiments indicate that glial cell differentiation may depend on a dual process, requiring the activation of glial differentiation by pointed and the concomitant repression of neuronal development by tramtrack.