Analysis of Physarum proteins throughout the cell cycle by two-dimensional PAGE

The accumulation of several hundred proteins during the nuclear division cycle of Physarum polycephalum was measured by digital image processing of silver-stained two-dimensional (2D) polyacrylamide gels. In contrast to previous studies, we have used an organism with a naturally synchronous cell cycle, so there are no uncertainties concerning synchronization artifacts or cell-sorting artifacts, and we have measured the specific amounts of each protein rather than the rate of synthesis. Since one-dimensional SDS-PAGE shows no significant fluctuations in the most abundant plasmodial proteins, we have loaded 2D gels so that proteins of low-to-moderate abundance appear in the linear range of the silver stain standard curve. Only five proteins showed reproducible, measureable fluctuations during the cell cycle. One of these proteins was tubulin. Full quantitative information was obtained by analysing the digital images of silver-stained gels by a general image processing system.     

Final height after growth hormone therapy in peripubertal boys with a subnormal integrated concentration of growth hormone.

The aim of this study was to test the effect of growth hormone (GH) therapy on final height in peripubertal boys with idiopathic short stature in whom a subnormal integrated concentration of GH (< 3.2 micrograms/l) was found. Twenty-eight peripubertal children were studied. Height was below 2 SD for age, growth velocity was < 4.5 cm/year, bone age was more than 2 SD below mean for age and GH response to provocative tests was more than 10 micrograms/l. Eleven subjects (group B) were treated with recombinant GH 0.75 unit/kg/week, divided into 3 weekly doses for 2 years, and then the same weekly dose divided into daily injections was administered until final height was attained. Seventeen untreated children (group A) who were followed until cessation of growth served as controls. The GH-treated patients reached their target heights (-2.1 +/- 0.5, mean +/- SD in SDS) and predicted heights (-1.8 +/- 0.8) determined by the Bayley and Pinneau method, while the final heights of the untreated patients were significantly lower than their target heights and their predicted final heights (-2.7 +/- 0.7, -1.8 +/- 1.0 and -2.7 +/- 0.7, respectively). The main effect of GH was observed during the 1st year of treatment when height velocity was significantly higher in the GH-treated group than in the untreated one (9.3 +/- 2.1 vs. 5.3 +/- 1.1, respectively, p < 0.001). The high cost of the treatment in this specific age group should be weighed against the results.     

Regression analysis of the spontaneous spike activity of neurons in snail ganglia

Regression analysis of the spontaneous spike activity of neurons in Helix pomatia was carried out with the aim to establish the statistical parameters of this activity under constant experimental conditions and during longer time intervals. The activity of 38 randomly chosen neurons in visceral and parietal ganglia, penetrated by microelectrodes and activated either endogenously by pacemaker potentials or by synaptic inputs, was recorded during time intervals lasting from 20 min to 3 h. The main results of the statistical analyses are presented in the table where the parameters of both cell types are listed. The validity of the regression analysis applied here is discussed from the point of the possibility it offers for carrying on the data processing quickly and without applying complex calculating means. The results are also considered regarding the current interest of our research group.     

Bacterial bioluminescence as an early indication of marine fish spoilage

Summary The relationships between different microbiological and biochemical parameters and the development of bacterial luminescence associated with the spoilage of marine fish from the Mediterranean-Sea was studied during storage at different temperatures. The bioluminescence level of the bacterial suspensions that were taken from the fish skin increased during the storage; at 20°–25°C the growth and luminescence of the luminuous bacteria correlated well with the total bacterial count while at 5°C the bacterial proliferation was not accompanied by a parallel increase in luminescence.The shift in storage temperature from 25°C to 5°C stabilized the level of the luminescence of bacterial suspension taken from the winter fish which were comprised mainly by Photobacterium phosphoreum, and caused a drop in the luminescence of bacterial suspension taken from the fish caught in the summer which were comprised mainly by Beneckea barveyi. The increase in the bioluminescence level appeared earlier than the increase in trimethylamine level and occured approximately at the same time as the increase in the hypoxanthine concentration. The potential value of the use of bacterial bioluminescence as an early indication for marine fish spoilage is discussed.     

DNA interpolyelectrolyte complexes as a tool for efficient cell transformation.

A tool was developed for enhancement of plasmid penetration into an intact cell, based on increasing DNA hydrophobicity via inclusion into a soluble interpolyelectrolyte complex (IPC) with polycations. The characteristics of formation of DNA IPC with synthetic polycations [poly(N-ethyl-4-vinylpyridinium)bromide (PVP) and PVP modified with 3% of N-cetyl-4-vinylpyridinium units (PVP-C)] were studied using ultracentrifugation and polyacrylamide gel electrophoresis methods. The conditions were established under which the mixing of DNA and polycation aqueous solutions results in the self-assembly of soluble IPC species. Incorporation of DNA into IPC results in the enhancement of DNA binding with isolated Bacillus subtilis membranes. A considerable increase in the efficiency of transformation of B. subtilis cells with pBC16 plasmid resulted from incorporation of the plasmid into the IPC with PVP and CVP.     

Orcein staining and the study of the effect of chemicals on chromosomes

Summary Plant materials of both dicotyledonous and monocotyledonous groups were subjected to treatment with alloxan followed by coumarin, phenols, oxyquinoline etc. for 3 to 4 hours.Root tips were then heated in an orcein-acid mixture for 10 to 40 seconds and studied after mounting in 1% orcein. Clear metaphase plates were obtained after 10 seconds of heating, whereas after 20 and 30 seconds considerable erosion and fragmentation respectively were recorded. Complete loss of stainability resulted after heating for 40 seconds.Fragmentation is caused only by orcein under heat conditions. This is demonstrated by control series checking all variables.Pretreatment with the chemicals stated is of absolute necessity for fragmentation. It is suggested that this pretreatment causes depolymerisation of DNA and lability of nucleoprotein linkage at certain segments from which the DNA becomes detached during subsequent heating.Of all the chemicals tested for pretreatment effects, marked positive results were obtained with alloxan (1 hour) followed by coumarin (2 hours at 16–20 ° C.). Phenols also gave positive results. The other chemicals tried need slightly longer time of treatment for fragment induction.     

A Modest Protective Effect of Thyrotropin against Bone Loss Is Associated with Plasma Triiodothyronine Levels

Background

The independent skeletal effect of thyrotropin (thyroid stimulating hormone, TSH) has been suggested in animal studies. However, clinical data on the association between bone loss and variations in TSH levels is inconsistent. This study aimed to investigate the relationship between TSH levels and bone mineral density (BMD).

Methods

We conducted a cross-sectional study with 37,431 subjects (33,052 cases with euthyroidism and 4,379 cases with subclinical thyroid dysfunction) aged over 35 years. We performed thyroid function tests and measured BMD at the lumbar spine, femur neck, and total hip.

Results

Levels of TSH and T3 were positively correlated in women (r = 0.076, P = 0.001) and uncorrelated in men. In both men and women, TSH levels correlated positively and T3 levels correlated negatively with BMD at all skeletal sites in age and body mass index adjusted analyses. BMD increased steadily with TSH levels from the subclinical hyperthyroid to subclinical hypothyroid range in subjects with T3 levels in the highest tertile (119.5–200.0 ng/dL), but was no longer significant in subjects with lower plasma T3 levels.

Conclusions

The variations in TSH levels within the euthyroid and subclinical range were positively correlated with BMD in healthy men and women. The negative effect of T3 on BMD appears to be compensated for by increased TSH in subjects with plasma T3 levels in the upper normal range.