Glucose-6-phosphate-dependent pyruvate kinase in Streptococcus mutans.

Pyruvate kinase of Streptococcus mutans JC 2 had an absolute and specific requirement for glucose-6-phosphate. Inorganic phosphate was a strong inhibitor. The enzyme required K+ or NH4+ and Mg2+ or Mn2+. S. mutans FIL and E 49, Streptococcus bovis ATCC 9809, and Streptococcus salivarius ATCC 13419 had also glucose-6-phosphate-dependent pyruvate kinases, whereas Streptococcus sanguis NCTC 10904 had an enzyme activated by fructose-1,6-diphosphate.     

The temperature sensitive mutant 72c

Summary The spontaneous temperature sensitive mutant 72c is shown to be more tolerant to fusidic acid, but less tolerant to trimethoprim on plates at permissive temperature, than is the parental strain. The poor growth of the mutant on amino acids supplemented plates, as well as its inability to grow on broth plates at 40°, can be compensated by sublethal amounts of chloroamphenicol. Also some mutations to Rif-R or Str-R improve growth of the mutant under certain conditions.Reversion and other genetic analysis strongly suggest, that the pleiotropic behaviour of the mutant is due to a single mutation in a gene, which is designated fusB and is closely cotransducible with lip at min 14 of the E. coli chromosome. The gene order is lip-fusB-supE.     

Screening for carcinoma of the prostate by digital rectal examination in a randomly selected population.

OBJECTIVE--To study the acceptability, costs, psychosocial consequences, and organisation of screening for carcinoma of the prostate. DESIGN--A randomly selected population was personally invited for digital rectal examination by a urologist and a general practitioner. Further examinations were performed if induration was felt. Each man completed a questionnaire on his response to the examination. SETTING--General practices in the area of Norrköping. PATIENTS--1494 Men aged 50-69 randomly selected from a population of 9026. MAIN OUTCOME MEASURE--Prostates having a firm nodular consistency. RESULTS--Carcinoma of the prostate was suspected in 45 of 1163 patients examined; in 10 by the general practitioners, in 10 by the urologists, and in 25 by both. Forty four men had a fine needle aspiration biopsy, and carcinomas were found in 13 cases. Of these, one had been suspected by the general practitioner, four by urologists, and eight by both. The cost for each man was 11.60 pounds, and the cost for each case of carcinoma detected and treated by potentially curative methods was 2477 pounds. Of the 13 men with carcinoma, 10 underwent radical prostatectomy and one radiotherapy. One man had advanced disease and was given endocrine treatment, another was not treated. Only 193 men felt distress during the initial examination. Of the 44 men who had an aspiration biopsy, 25 experienced anxiety. CONCLUSIONS--Screening for carcinoma of the prostate by a urologist or a general practitioner using digital rectal examination is a cost effective method of early diagnosis. Whether such screening leads to prolonged survival, however, remains doubtful.     

Organization and expression of mouse Hox3 cluster genes

Summary The three yolk protein genes (yp) of Drosophila melanogaster are transcribed in a sex- and tissue-limited fashion. We have searched for cis-regulatory sequences in regions flanking yp1 and yp2 to identify the elements that confer female-specific expression in the fat body. One such 127 by element has previously been identified in this region. We show here the existence of two additional regions which confer female fat body-specific expression on an Adh reporter gene and on the native yp2 gene, respectively. This suggests some redundancy in the regulation of expression of the yp genes. Computer searches for putative binding sites for the DSX protein, which regulates sex-specific expression of the yp genes, revealed several such sites in our constructs. However, the significance of these is unclear since many such sites also occur in genes which one would not expect to be regulated in a sex-specific manner (e.g. Adh, Actin 5C). We suggest that DSX acts in concert with other proteins to mediate sex- and tissue-specific expression of the yp genes.     

Low Density Lipoproteins Promote Unstable Calcium Handling Accompanied by Reduced SERCA2 and Connexin-40 Expression in Cardiomyocytes

The damaging effects of high plasma levels of cholesterol in the cardiovascular system are widely known, but little attention has been paid to direct effects on cardiomyocyte function. We therefore aimed at testing the hypothesis that Low Density Lipoprotein (LDL) cholesterol affects calcium dynamics and signal propagation in cultured atrial myocytes. For this purpose, mRNA and protein expression levels were determined by real time PCR and western blot analysis, respectively, and intracellular calcium was visualized in fluo-4 loaded atrial HL-1 myocyte cultures subjected to field stimulation. At low stimulation frequencies all cultures had uniform calcium transients at all tested LDL concentrations. However, 500 µg LDL/mL maximally reduced the calcium transient amplitude by 43% from 0.30±0.04 to 0.17±0.02 (p<0.05). Moreover, LDL-cholesterol dose-dependently increased the fraction of alternating and irregular beat-to-beat responses observed when the stimulation interval was shortened. This effect was linked to a concurrent reduction in SERCA2, RyR2, IP3RI and IP3RII mRNA levels. SERCA2 protein levels were also reduced by 43% at 200 µg LDL/mL (p<0.05) and SR calcium loading was reduced by 38±6% (p<0.001). By contrast, HDL-cholesterol had no significant effect on SERCA expression or SR calcium loading. LDL-cholesterol also slowed the conduction velocity of the calcium signal from 3.2+0.2 mm/s without LDL to 1.7±0.1 mm/s with 500 µg LDL/mL (p<0.05). This coincided with a reduction in Cx40 expression (by 44±3%; p<0.05 for mRNA and by 79±2%; p<0.05 for Cx40 protein at 200 µg/ml LDL) whereas the Cx-43 expression did not significantly change. In conclusion, LDL-cholesterol destabilizes calcium handling in cultured atrial myocytes subjected to rapid pacing by reducing SERCA2 and Cx40 expression and by slowing the conduction velocity of the calcium signal.     

The effect of family structure on the likelihood for kin-biased distribution: an empirical study of brown trout populations

This study is aimed at evaluating the likelihood that kin-biased distribution will be expressed or detected in a range of brown trout ( Salmo trutta ) populations as a function of family size. Microsatellite analysis indicated that fewer full- and half-siblings were found in populations with larger effective population sizes, while more full- and half-siblings were found in populations with lower effective population size. It is suggested that kin-biased distribution and hence kin-biased behaviours are not likely to be expressed equally frequently in all populations since the number of close relatives will vary among populations and, hence, the opportunity for relatives to interact differs among populations. These findings can, at least partly, explain the discrepancy among previous studies of kin-biased distribution in wild salmonids under natural conditions. Effective population size could, hence, be used to predict the salmonid populations in which kin-biased distribution are more likely to occur and be detected.     

Movement patterns of temperate wrasses (Labridae) within a small marine protected area

The movement patterns of three commercially important wrasse (Labridae) species inside a small marine protected area (~ 0.15 km²) on the west coast of Norway were analysed over a period of 21 months. The mean distance between capture and recapture locations varied between 10 and 187 m, and was species and season specific. The extent of movement was not related to body size or sex. These results imply that a network of small strategically located marine protected areas can be used as management tools to protect wrasses from size- and sex-selective fishing mortality.     

Evaluation of activity of selected antioxidants on proteins in solution and in emulsions

Protection against protein oxidation by lipophilic and hydrophilic antioxidants in model systems using bovine serum albumin (BSA) in solution alone, or in an emulsion with linolenic acid methyl ester (LnMe) was found to be strongly dependent on the oxidation initiator. Tocopherol, Trolox, or the carotenoids astaxanthin and canthaxanthin were incubated with BSA or BSA/LnMe and oxidation was initiated either with the water-soluble azo-initiator 2,2′ azo-bis-(2-amidinopropane) hydrochloride (AAPH), or FeCl₃ and ascorbate, or the Fenton system using FeCl₂/EDTA/H₂O₂, or with the singlet oxygen generating species anthracene-9,10-dipropionic acid disodium 1,4 endoperoxide (NDPO₂).

The results show that all the antioxidants tested were inefficient in the system with FeCl₃/ascorbate. However, with the other initiating agents, the hydrophilic antioxidant, Trolox, was the most effective in preventing both protein and lipid oxidation. In contrast the lipophilic antioxidants were ineffective in preventing oxidation of BSA in aqueous solution, but did show some moderate antioxidative activity on protein and lipid in the BSA/LnMe system. Using the singlet oxygen generating system it was also demonstrated that Trolox always provided better protection of the protein than tocopherol and the carotenoids in both the BSA and the BSA/LnMe systems. In conclusion, prevention of protein oxidation using a water-soluble antioxidant has a protective effect on the lipid fraction and this approach deserves further attention in complex biological systems.     

Xenon for NMR biosensing – Inert but alert

NMR studies with hyperpolarized xenon as functionalized sensor or contrast agent recently made notable progress in developing a new approach for detecting molecular markers and parameters of biomedical interest. Combining spin polarization enhancement with novel indirect detection schemes easily enables a 10⁷-fold signal gain, thus having promising potential to solve the NMR sensitivity problem in many applications. Though an inert element, ¹²⁹Xe has exquisite NMR properties to sense molecular environments. This review summarizes recent developments in the production of hyperpolarized xenon and the design and detection schemes of xenon biosensors.     

Test System for Measurement of Translational Activity in Vivo

Abstract

A new plasmid which makes it possible to measure formation of a mature protein, as well as prematurely terminated gene products, has been constructed. It can be used for the analysis of translational efficiency of specified codon sequences in vivo.