Hemoglobin Dallas (alpha 97(G4)Asn-->Lys): functional characterization of a high oxygen affinity natural mutant.

Hemoglobin Dallas, an alpha-chain variant with a substitution of lysine for asparagine at position 97(G4), was found to have increased oxygen affinity (p1/2 = 1 mmHg at pH 7.3 and 20 degrees C), diminished cooperativity (n, the Hill coefficient = 1.7) and reduced Bohr effect (about 50%). Addition of allosteric effectors (such as 2,3-diphosphoglycerate, inositol hexakisphosphate and bezafibrate) led to a decrease in oxygen affinity and increase in cooperative energy. Kinetic studies at pH 7.0 and 20 degrees C revealed that (i), the overall rate of oxygen dissociation is 1.4-fold slower than that for HbA and (ii), the carbon monoxide dissociation rate is unaffected. The abnormal properties of this hemoglobin variant can be attributed to a more 'relaxed' T-state.     

Circulating immune complexes in the pathogenesis of recurrent erysipelas

The dynamics of circulating immune complexes (CIC) in comparison with the level of SH-groups of serum deproteinate and other characteristics of cell-mediated and humoral immunity (the reaction of the inhibition of antibodies, the levels of T-cells and their main subpopulations) was studied in 103 erysipelas patients and in 46 persons having had the disease at the acute period of this infection and at the periods between relapses. The elevated levels of CIC and SH-groups of serum deproteinate were found to be directly correlated with the inhibition index. The study showed that, as a rule, in patients with the elevated level of CIC the frequently relapsing form of erysipelas, accompanied by the formation of relative hypersuppressor-type secondary immunodeficiency and by a decrease in the functional activity of dermal macrophages, was observed.     

Hormonal regulation of anabolic processes and of protein utilization efficiency in the early postnatal period

The levels of thyroid, pituitary and steroid hormones-thyroxine, triiodothyronine and 11-hydroxycorticosteroids in the blood serum, somatotropin in the pituitary, and processes of protein assimilation were studied in rats in the early postnatal period. The highest endogenous production of thyroxine, triiodothyronine and somatotropin was detected in 15-day-old rats. The highest level of protein utilization was detected in 7 to 15-day-old rats, followed by the lowering of the utilization on changing over to definitive nutrition. Endogenous production of the anabolic hormones thyroxine, triiodothyronine and somatotropin was found to correlate with a high level of protein utilization in rats within the first days of life after birth.     

Comparison of cobalamin-independent and cobalamin-dependent methionine synthases from Escherichia coli: two solutions to the same chemical problem.

In Escherichia coli, two enzymes catalyze the synthesis of methionine from homocysteine using methyltetrahydrofolate as the donor of the required methyl group: cobalamin-dependent and cobalamin-independent methionine synthases. Comparison of the mechanisms of these two enzymes offers the opportunity to examine two different solutions to the same chemical problem. We initiated the research described here to determine whether the two enzymes were evolutionarily related by comparing the deduced amino acid sequences of the two proteins. We have determined the nucleotide sequence for the metE gene, encoding the cobalamin-independent methionine synthase. Our results reveal an absence of similarity between the deduced amino acid sequences of the cobalamin-dependent and cobalamin-independent proteins and suggest that the two have arisen by convergent evolution. We have developed a rapid one-step purification of the recombinant cobalamin-independent methionine synthase (MetE) that yields homogeneous protein in high yield for mechanistic and structural studies. In the course of these studies, we identified a highly reactive thiol in MetE that is alkylated by chloromethyl ketones and by iodoacetamide. We demonstrated that alkylation of this residue, shown to be cysteine 726, results in complete loss of activity. While we are unable to deduce the role of cysteine 726 in catalysis at this time, the identification of this reactive residue suggests the possibility that this thiol functions as an intermediate methyl acceptor in catalysis, analogous to the role of cobalamin in the reaction catalyzed by the cobalamin-dependent enzyme.     

Seasonal distribution of enteroviruses and adenoviruses in domestic sewage

A seasonal distribution of enteroviruses and adenoviruses in raw sewage effluents of Athens, Greece, was observed over a 15-month surveillance period. All 36 samples tested were positive for both virus groups. Adenovirus concentration levels ranged from 70 to 3200 cytopathic units per litre of sample, whereas the corresponding values for enteroviruses were 90-900 cytopathic units per litre. Peak values for adenoviruses were recorded during the months of April and June 1983, whereas for enteroviruses the peak was recorded in September 1983. All three types of poliovirus were present. Coxsackievirus types B-1, B-2, B-4, and B-5 and echovirus type 7 were also isolated. Adenovirus types 1, 2, 3, 5, 7, and 15 were detected as well.     

The Plastids and Pigments of Fresh and Dried Chinese Gooseberries (Actinidia chinensis)

The pericarp of Chinese gooseberries is green due to the presenceof low concentrations of chlorophylls. On a f. wt basis thereis about 1.5 times more tetrapyrollic pigments (chlorophyllsand related compounds) in the outer pericarp than in the innerpericarp, whereas the carotenoid pigment values only showed1.25 times more in the outer than in the inner part of the fruit.The drying of Chinese gooseberries at 40 °C for 40 h resultedin the loss of at least half the tetrapyrollic pigments andof carotenoids. Chlorophylls a and b were converted to chlorophyllides,pheophytins and pheophorbides. Chloroplasts in the outer pericarp are clustered closely aroundthe nucleus and have a well-defined grana and inter-granal membranesystem. In the inner pericarp the chloroplasts are again clusteredaround the nucleus and there is a proliferation of inter-granalmembranes. In dried tissue the limiting membrane of chloroplastswas completely dispersed whereas some of the internal membranesremained intact. Actinidia chinensis Planch., Chinese gooseberry, Kiwi fruit, pigments, chloroplast structure     

Human dynein and sperm pathology

Human spermatozoa with normal structure and with different axonemal deficiencies (absence of axoneme, of arms, or of central structures) were studied by electron microscopy, SDS-polyacrylamide gel electrophoresis, and ATPase activity measurements. Normal human sperm possess a complement of high molecular weight polypeptides with an electrophoretic migration similar to that of sea urchin and other mammalian sperm dyneins. Human high molecular weight bands are numbered one to four in order of increasing of electrophoretic mobility; all of them are absent in spermatozoa that lack axoneme. The absence of doublet arms, coincides with the absence of bands 2, 3, and 4; the absence of central structures coincides with a reduction in intensity of band 2. In the latter two abnormal conditions, band 1 has an increased intensity. The data are tentatively interpreted by attributing the polypeptides forming bands 3 and 4 to the arm structure, whereas band 2 is supposed to contain a mixture of polypeptides localized in the arms and in the central structures; these abnormal sperm contain modified polypeptides which gather in band 1. Histochemical ATPase stainings indicate that this enzyme is localized mainly in the doublet arms and, to a minor extent, in the central structures.