Unimpaired formation of hormone-stimulated inositol trisphosphate in human mesangial cells under hyperglycemic conditions.

The relationship between bulk cellular myo-inositol content and phosphatidylinositol metabolism was evaluated in a human mesangial cell line under euglycemic and hyperglycemic conditions. Mesangial cells maintained in high glucose medium displayed a concentration-dependent fall in myo-inositol as measured by gas-liquid chromatography. Measurements of phosphatidylinositol, phosphatidylinositol 4-monophosphate and phosphatidylinositol 4,5-bisphosphate mass revealed slight but statistically insignificant increases in cells exposed to high glucose containing medium. CDP-diacylglycerol: myo-inositol 3-phosphatidylinositol transferase activity, measured in plasma membranes from mesangial cells grown under control and hyperglycemic conditions, was kinetically similar with Michaelis constants (Km values) for myo-inositol of 2.9 and 2.1 mM, respectively. Finally, hormone-stimulated intracellular calcium mobilization and myo-inositol 1,4,5-trisphosphate mass was measured from mesangial cells grown under normal and hyperglycemic conditions. Both intracellular calcium and inositol trisphosphate formation were unchanged in cells previously exposed to high glucose conditions (400 mg/dl) compared to cells grown under normal glucose concentration (100 mg/dl). These data indicate that bulk changes in myo-inositol induced by hyperglycemia are neither associated with alterations in basal levels of inositol containing glycerolipids nor with changes in hormone-stimulated calcium mobilization and inositol trisphosphate formation under conditions of short term changes in extracellular glucose.     

Scrapie I. Transmission and Pathogenesis

Homogenized brain, extracts, and residues of brain from normal and scrapie sheep were inoculated into 116 sheep. Of 72 sheep inoculated with scrapie material 27 developed the disease, whereas four of 44 inoculated with ;normal' brain material showed symptoms similar to those of scrapie. The scrapie agent survived extraction for 18 hours with diethyl ether followed by water for 24 hours. Similarly the agent survived extraction in a Soxhlet apparatus with 95 per cent ethanol containing 2.0 per cent 4 M HC1 for 24 hours. There was no evidence of demyelination in sections of brain and spinal cord supporting the view that allergic encephalomyelitis is not responsible for the scrapie symptoms. Liver function, as measured by the bromsulfalein test, remained normal until just before death. These results preclude that liver dysfunction contributes to the disease but do not exclude the possibilities of other metabolic derangements.     

Effect of exogenous gibberellin A4/7 on tracheid production, longitudinal growth and the levels of indole‐3‐acetic acid and gibberellins A4, A7 and A9 in the terminal shoot of Pinus sylvestris seedlings

Synchronously dividing cultures of the unicellular green alga Scenedesmus obtusiusculus were cultivated for 24 or 70 h in medium high (1000 μM) or low (60 μM) in phosphorus. Aliquots of AlCl₃ (0, 37, 74, 111, 148, 185, or 222 μmol) were added daily to 1 l cell suspension at the end of the cell division phase. Algae were also grown in media with different pH, adjusted with HCl, in the absence of AlCl₃.
Effects of Al on cell metabolism vary with the intracellular Al concentration and with the concentration of Al available per cell. When the concentration of phosphorus is low, internal concentrations of Al are high and the chlorophyll content and the net dry matter production per cell increase, whereas the photosynthesis and the cell division are increased. Presence of Al in a low P medium decreases the pH of the medium down to 4.5. There are only small effects of Al in the presence of P, due to precipitation of most of the Al with P in the medium.
Despite the Al-induced decrease of the pH of the culture medium, effects caused by Al cannot be explained as a pH effect. Instead, the Al effect may, at least to some extent, be related to a decrease in availability of P in the metabolism, due to formation of aluminium phosphate inside the cell.     

Detection of Salmonella typhi by polymerase chain reaction

A rapid and sensitive method for detection of Salmonella typhi would help in preventing the spread of outbreaks and in clinical diagnosis. In order to develop unique PCR primers to detect Salm. typhi , ribosomal RNA genes from Salm. typhi (Rawlings) were cloned in pUC18. The resulting clone was confirmed by sequencing. The cloned DNA fragment contained the 5S, part of the 23S rRNA genes and the 5S-23S spacer region (EMBL/GenBank accession No. U04734).
It was expected that the 5S-23S spacer region is divergent unlike the highly conserved 23S+5S genes. This was confirmed by comparison with the rRNA gene sequences in the EMBL/GenBank database. A pair of PCR primers specific for Salm. typhi was obtained, based on this spacer region sequence. The specificity of this pair of primers was tested with 54 Salm. typhi strains (of 27 different phage types). All these Salm. typhi strains showed the positive 300 bp PCR product with this pair of primers. Six other Salmonella species as well as six other non- Salmonella bacteria were tested and none showed the 300 bp PCR product. The sensitivity of the detection level was 0·1 pg of pure Salm. typhi genomic DNA, or approximately 40 Salm. typhi cells in a spiked food sample. This pair of primers therefore has the potential for development into a diagnostic tool for the rapid diagnosis of typhoid fever.