Cortisol production rate measurement by stable isotope dilution using gas chromatography-negative ion chemical ionization mass spectrometry.

Presented here is a stable isotope dilution technique for determining cortisol production rate (CPR). The method involves extraction and derivatization of cortisol isoforms from serum (0.5 ml), separation of derivatives by gas chromatography, and detection by using negative ion chemical ionization mass spectrometry. This method provides 50-100-fold greater sensitivity than positive ion mass spectrometry and allows for estimations of cortisol production rate with the use of small amounts of pooled serum, even in the presence of high concentrations of lipophilic contaminants. The area under the curve for the total selected ion chromatogram of fluoroacyl derivatives of cortisol (d0, m/z 782) and deuterated cortisol (d3, m/z 785) were used to determine the isotopic dilution ratio in three types of samples: 1) standards: d0/d3 ratios ranging from 1 to 8%; 2) controls: d3-cortisol added to serum with known cortisol concentration; 3) subjects: 24-h pooled serum samples (q 30 min over 24 h) from healthy children (male 10-13 years; female 7-11 years) receiving continuous infusions of d3-cortisol at 2-4% of their estimated CPR. Recovery after the solid phase extraction and derivatization process was >90%, as determined by thin-layer chromatography. Expected versus measured ratios for d3/d0 in standards and serum controls were highly correlated (r2(standard) = 0.99; r2(control) = 0.99) over a wide range of d3-cortisol enrichment (1.0-10.0%). Mean 24-h CPRs were 4.8 +/- 0.6 mg/m2/24 h (mean +/- SEM, n = 7) in male children and 4.4 +/- 0.5 mg/m2/24 h in female children (n = 4). These CPR values are lower than those derived by radio tracer methods, but are in agreement with previous isotopic dilution studies. This technique is an important tool for assessing CPRs in a wide range of disease states affecting cortisol production.     

Shrimps and eutrophication in the waterbodies of a coastal grazing marsh

The influence of shrimps on turbidity was examined in laboratory and field experiments. Palaemonetes varians in the laboratory stimulated populations of both rotifers and Anabaena, which was the main cause of increased turbidity. In field experiments, P. varians increased and Neomysis integer decreased turbidity. In feeding trials P. varians took cladocerans and culicids in preference to copepods. It is considered that shrimps are likely to be important predators of cladocerans in shallow, coastal waterbodies and result in phytoplankton blooms, with reduced water transparency. This revised version was published online in July 2006 with corrections to the Cover Date.     

Content of significant amounts of a cytotoxic end-product of lipid peroxidation in human semen

(E)-4-Hydroxy-2-nonenal (HNE), a cytotoxic end-product of lipid peroxidation, is present in significant amounts in human semen (0.902 +/- 0.190 microM; mean +/- s.e.; n = 18). The addition of the divalent cation ionophore A23187 to suspensions of human spermatozoa resulted in increased production of HNE. Exogenous HNE was powerfully spermicidal and as little as 50 microM caused an irreversible loss of motility of human spermatozoa within minutes. The addition of human seminal plasma protected spermatozoa from the toxic effects of HNE.     

Occurrence of thermophilic actinomycetes in natural substrates in Nigeria

Thermophilic actinomycetes were isolated from 163 (48.95%) of 333 samples of vegetable substrates and soil from different sites in Anambra and Enugu States in Nigeria.Thermoactinomyces (Tha.) vulgaris was the most common, occurring in 32.4% of samples whileTha. thalpophilus was isolated from 20.1%.Tha. sacchari, Saccharomonospora (Sam.) viridis andSaccharopolyspora (Sap.) rectivirgula were isolated from 3–10.5% of the samples examined.Streptomyces (Stm.) thermovulgaris occurred in 2.7% andThermomonospora spp. in 0.6% of samples. Except forStm. thermovulgaris and, perhaps,Thermomonospora spp., all the species isolated are known to be allergenic in man. The five known allergenic species were abundant in hay and maize silage.T. vulgaris andT. thalpophilus were also frequently isolated from rice straw.Tha. vulgaris, Tha. thalpophilus andSap. rectivirgula were all most numerous in hay whileTha. sacchari was most numerous in maize silage. The widespread occurrence of thermophilic actinomycetes in Nigeria emphasizes the need for investigation of their role in allergic respiratory disorders.     

A model for the coevolution of the genetic code and the process of protein synthesis: Review and assessment

The contemporary genetic code and the process of protein biosynthesis most assuredly evolved from a simpler code and process. We believe that there was obligatory coevolution of the two and that the earlier code and process must have involved a more direct linkage between the amino acids and the informational macromolecule. We propose that an early form of translating existed in which amino acids were attached directly to the ‘messenger’ RNA along the backbone as 2'OH aminoacyl esters. These esters then condensed with each other on the RNA backbone yielding a peptide covalently attached to the RNA, without the use of tRNAs and ribosomes. This presentation is concerned with experimental data which indicate that such a simple translation system is possible and must have involved the following steps: (1) formation of the aminoacyl adenylate anhydride, (2) transfer of the amino acid from the adenylate to imidazole, (3) transfer of the amino acid from imidazole to 2'OH groups along the backbone of RNAs (4) condensation of the amino acids to yield peptides. Steps (1)–(3) have been confirmed in chemical systems. Our preliminary evidence indicates step (4) is also possible. The aminoacylation of polyribonucleotides and the subsequent formation of peptides is a dynamic and experimentally accessible system for studying genetic coding specifities and our present studies are now concentrated on step (4), looking for such specifities.     

Was there a universal tRNA before specialized tRNAs came into existence?

It is generally true that evolving systems begin simply and become more complex in the evolutionary process. For those who try to understand the origin of a biochemical system, what is required is the development of an idea as to what simpler system preceeded the present one. Here we present an hypotheis that a universal tRNA molecule, capable of reading many codons may have preceeded the appearance of individual tRNAs. Evidence seems to suggest that this molecule may have been derived from a common ancestor of the contemporary 5S rRNAs and tRNAs.     

Effects of Microwave Cookery on the Bacterial Counts of Food

A method is described by which foods drawn from the normal hospital kitchen can be reheated with infrared and microwave irradiation before it is served to patients in an "ultraclean"unit. A considerable reduction in bacterial content has been achieved in all the samples tested and sterility was obtained in some. There was no obvious effect on the palatability of the foodstuffs tested.