Changes in prostacyclin and thromboxane biosynthesis and their catabolic enzyme activity in kidneys of aging rats

The effects of aging on the prostacyclin and thromboxane biosynthesis and prostaglandin catabolic enzyme activity in rat kidney were investigated. The prostacyclin biosynthesis, using arachidonic acid as substrate, was the greatest in young kidneys (2 months old) and then progressively decreased in mature (12 months old) and old (24 months old) kidneys, while thromboxane biosynthetic activity showed no significant change as a function of age. When prostaglandin H2 was used as substrate, the prostacyclin and thromboxane biosynthesis showed similar results as when arachidonic acid was used as substrate; the prostacyclin biosynthesis progressively decreased and thromboxane biosynthesis showed no significant change as a function of age. The fatty acid cyclooxygenase in kidney was measured by a specific radioimmunoassay. No significant change in renal fatty acid cyclooxygenase as a function of age was found. Thus, we concluded that the progressive decrease in renal prostacyclin biosynthesis as a function of age is due to a defect in prostacyclin synthetase in aged kidneys. The prostaglandin catabolic enzyme, NAD+-dependent 15-hydroxyprostaglandin dehydrogenase, in kidneys was also investigated. The enzyme activity progressively decreased as a function of age, which suggested a decrease in the metabolism of thromboxane A2 in aged kidneys. The present results, indicating a decrease in renal prostacyclin biosynthesis and renal 15-hydroxyprostaglandin dehydrogenase activity with aging, might contribute to a plausible explanation of the progressive decrease in renal functions in the elderly.     

Cytochemical study of the DNA and histone in the tissues of the lily anther in the course of microsporo- and gametogenesis]

The DNA and histone content in the microsporocytes, microspores, generative and vegetative cells of the pollen grain of Lilium candidum L., as well as in the anther wall tissues, was estimated by double wavelength cytophotometry. The lack of histone, as compared with DNA content, was demonstrated in the microsporocytes at the late premeiotic interphase and early meiotic prophase, as well as in the young microspores and anther wall tissues. The analysis of hydrolysis curves suggests the increase of non-condensed chromatin during endothelial cell differentiation.     

Histochemical investigations on the localization of the purple acid phosphatase in the bovine spleen

Summary The localization of the purple tartrate-resistant, iron-containing acid phosphatase in the bovine spleen was studied by enzyme histochemistry at the light and electron microscopic levels as well as by immunohistochemistry. The purple phosphatase was localized only in lysosome-like organelles of cells belonging to the reticulo-phagocytic system. The same cells were identified as containing large iron(III)-deposits as ferritin in homogeneously granular accumulations and freely in the cytoplasm, or as hemosiderin in siderosomes. The phagocytosing cells containing purple phosphatase and ferritin often had close contact with clusters of aged and deformed erythrocytes.A possible catabolic role of the purple enzyme as a phosphatase degrading phosphoproteins of the erythrocyte membrane and the cytoskeleton was assumed.     

"Aachen 3-D-finger". Development of a 3-D-digitizer for use in dental, oral and maxillary treatment

The "AACHEN 3D Finger" is a three-dimensional measuring system for use in all fields of dentistry. The system can equally as well be installed on a plane table, as fixed to the head of a patient. The measuring device is computer-assisted, and is able to localize, register and calculate any combination of points in the oral and maxillofacial area. The reference system can be changed at any time. The "AACHEN 3D Finger" can be used as a computer-a ded system in dentistry as well as in implantology or dental and maxillofacial surgery.     

Platelet and cardiovascular activity of the hydantoin BW245C, a potent prostaglandin analogue

The platelet anti-aggregating, cardiovascular and gastro-intestinal actions of a hydantoin prostaglandin analogue, BW245C have been compared with prostacyclin and PGD2 in several species. In human plasma in vitro, BW245C was 0.2 times as active as prostacyclin and 8 times as active as PGD2 in inhibiting platelet aggregation. In rat and rabbit plasma, BW245C was only weakly active but was more potent in sheep and horse plasma. Since the activity of PGD2 varied in a parallel fashion, BW245C may interact with PGD2 binding sites on platelets. The potency of BW245C as a vasodepressor also varied between species, being 0.5, 0.1, 0.06 and less than 0.02 times as active as prostacyclin in the anaesthetised dog, monkey, rat and rabbit respectively. The relative activity of BW245C as an inhibitor of platelet aggregation ex vivo was more comparable, being 0.08, 0.04 and 0.05 times as active as prostacyclin following intravenous infusion in the rabbit dog and monkey respectively. In the rabbit, BW245C was a highly selective platelet inhibitor with minimal cardiovascular actions, whereas in the dog and monkey, BW245C lowered BP in anti-aggregating doses. The potent platelet anti-aggregating actions of BW245C following parenteral or oral administration makes this hydantoin a potentially-useful anti-thrombotic prostaglandin analogue.     

Histochemical investigations on the localization of the purple acid phosphatase in the bovine spleen

The localization of the purple tartrate-resistant, iron-containing acid phosphatase in the bovine spleen was studied by enzyme histochemistry at the light and electron microscopic levels as well as by immunohistochemistry. The purple phosphatase was localized only in lysosome-like-organelles of cells belonging to the reticulo-phagocytic system. The same cells were identified as containing large iron(III)-deposits as ferritin in homogeneously granular accumulations and freely in the cytoplasm, or as hemosiderin in siderosomes. The phagocytosing cells containing purple phosphatase and ferritin often had close contact with clusters of aged and deformed erythrocytes. A possible catabolic role of the purple enzyme as a phosphatase degrading phosphoproteins of the erythrocyte membrane and the cytoskeleton was assumed.     

Genetic toxicology: Findings and challenges

The review highlights the history of genetic toxicology as a distinct research area, as well as the issues of genetic toxicology and development of its methodology. The strategies and testing patterns of genotoxic compounds are discussed with the purpose of identifying potential human carcinogens, as well as compounds capable of inducing heritable mutations in humans. The main achievements of genetic toxicology in the 20th century are summarized and the challenges of the 21st century are discussed.     

Hereditary defects in both germ cells and the blood-testis barrier system in as-mutant rats: evidence from spermatogonial transplantation and tracer-permeability analysis

The rat mutant allele as is located on chromosome 12. Homozygous (as/as) males show arrested spermatogenesis, mainly at the pachytene spermatocyte stage. It is not clear whether this defective spermatogenesis is caused by a failure in a somatic cell component that supports spermatogenesis or in the germ cell itself. Spermatogonial transplantation was performed to identify the genetically defective site in the as/as testis. In experiment 1, germ cells collected from as/as testes were transplanted into the testes of immunodeficient mice and normal rats. In experiment 2, normal rat germ cells were transplanted into as/as testes. The results of experiment 1 showed arrest of spermatogenesis at the pachytene spermatocyte stage, accompanied by a characteristic morphological feature, i.e., the formation of inclusion-like bodies in the cytoplasm, in both rat and mouse recipients. These results revealed the intrinsic effect of the mutant gene(s) on germ cells. In experiment 2, no restoration of spermatogenesis was detected in the recipient testes despite thorough histological examination. These results suggest that defects in a somatic cell component in as/as testes prevent the donor germ cells from colonizing and regaining their spermatogenetic ability. When the seminiferous epithelium of the as/as testis was examined by electron microscopy, no morphological abnormalities, including the formation of ectoplasmic specializations between adjacent Sertoli cells, were observed in the somatic cell components. However, when cytochrome c was applied as a tracer material, it penetrated the tight junctions between the Sertoli cells, indicating dysfunction of the blood-testis barrier in the as/as testis. The lack of restoration of spermatogenesis in the as/as testis after transplantation of normal germ cells may have been caused by the unfavorable environment in the seminiferous epithelium resulting from the incomplete barrier system between adjoining Sertoli cells. The gene(s) at the as locus may have a role in both germ cell differentiation and the establishment of the blood-testis barrier.     

Conformations of oligonucleotides in solution as determined by sequence-specific antibodies

The conformations of some related oligoribonucleotides in solution were investigated immunochemically with antisera specific for two synthetic oligonucleotide sequences, A-A-U and A-A-U-U. Radioimmunoassay showed differences of as much as three or more orders of magnitude in binding among oligonucleotides with commonly shared sequences. These large differences, which reflect the loss of many points of contact with antibody because of changes in overall conformation, allow the following conclusions: (1) A-A-U and A-A-U-U have conformations distinct from any present in the Un family of oligomers. (2) Conformations of A-A-U and A-A-U-U differ markedly from those of oligomers of A. The dinucleotide A-A, in particular, bears little resemblance in conformation to the A-A sequence in A-A-U and A-A-U-U. (3) The recognizable conformational unit appears to be the triplet A-A-U, which binds as well as A-A-U-U and far better than its component dimers. Interactions between non-adjacent bases may be a factor here, as well as in codon recognition. The immunological data support the conclusion that, in oligonucleotides, as in polypeptides, primary sequence can determine conformation in solution.     

Hedgehog signalling: how to get from Smo to Ci and Gli

The secreted morphogens of the Hedgehog family have important roles in normal development as well as in associated pathologies, including cancer. The Hedgehog signalling pathway has been studied in Drosophila and is thought to be conserved in vertebrates. Hedgehog elicits a signalling response that activates Smoothened (Smo). There is evidence of differences between Drosophila and vertebrates concerning signalling downstream of Smo, as well as in Smo itself. Here, we discuss this evidence and its importance for investigations of the pathway and related biology, as well as for the development of drugs targeting components of the pathway for treatment of associated pathologies.     

Norway spruce galactoglucomannans exhibiting immunomodulating and radical-scavenging activities

Wood-derived naturally acetylated galactoglucomannans (AcGGM) can be recovered even in ton-scale at mechanical pulp mills using spruce as raw material. These cell wall polysaccharides have a great potential as hydrocolloids and bioactive polymers in food and pharmaceutical applications, or as starting material for production of functional polymers. The immunostimulatory activity of both AcGGM and its deacetylated form (GGM) was now in vitro tested. The biological response of both AcGGM and GGM in the lymphocyte transformation test was dose-dependent. The direct mitogenic as well as comitogenic activities of the AcGGM were comparable to those of the immunogenic corn cob xylan used as control, and GGM showed significantly higher biological responses also at lower doses. In contrast to GGM, AcGGM possessed also DPPH radical-scavenging activity. The results suggested that the spruce AcGGM and GGM are potentially important as additives with immuno-potentiating and antioxidant properties in food products and pharmaceutical formulations.     

Genetic networks regulated by ASYMMETRIC LEAVES1 (AS1) and AS2 in leaf development in Arabidopsis thaliana: KNOX genes control five morphological events

The asymmetric leaves 1 ( as1 ) and as2 mutants of Arabidopsis thaliana exhibit pleiotropic phenotypes. Expression of a number of genes, including three class-1 KNOTTED -like homeobox ( KNOX ) genes ( BP , KNAT2 and KNAT6 ) and ETTIN / ARF3 , is enhanced in these mutants. In the present study, we attempted to identify the phenotypic features of as1 and as2 mutants that were generated by ectopic expression of KNOX genes, using multiple loss-of-function mutations of KNOX genes as well as as1 and as2 . Our results revealed that the ectopic expression of class-1 KNOX genes resulted in reductions in the sizes of leaves, reductions in the size of sepals and petals, the formation of a less prominent midvein, the repression of adventitious root formation and late flowering. Our results also revealed that the reduction in leaf size and late flowering were caused by the repression, by KNOX genes, of a gibberellin (GA) pathway in as1 and as2 plants. The formation of a less prominent midvein and the repression of adventitious root formation were not, however, related to the GA pathway. The asymmetric formation of leaf lobes, the lower complexity of higher-ordered veins, and the elevated frequency of adventitious shoot formation on leaves of as1 and as2 plants were not rescued by multiple mutations in KNOX genes. These features must, therefore, be controlled by other genes in which expression is enhanced in the as1 and as2 mutants.     

The action of beta-galactosidase (Escherichia coli) on allolactose.

The parameters involved in the action of beta-galactosidase (EC 3.2.1.23) (Escherichia coli) on allolactose, the natural inducer of lac operon in E. coli, were studied. At low allolactose concentrations only galactose and glucose were formed, while at high allolactose concentrations transgalactolytic oligosaccharides were also produced. Detectable amounts of lactose were not formed. The V and Km values (49.6 U/mg and 0.00120 M, respectively) indicated that allolactose is as good if not a better substrate of beta-galactosidase as lactose. The pH optimum with allolactose (7.8-7.9) as well as its activation by K+ (as compared to activation by Na+) were similar to the case with lactose as substrate. The alpha-anomer of allolactose was hydrolyzed about two times as rapidly as was the beta-anomer.     

THE MEMBRANE SYSTEM OF STREPTOMYCES CINNAMONENSIS

The plasma membrane bounding the cytoplasm immediately inside the hyphal wall of Streptomyces cinnamonensis may not retract from the hyphal wall. When it does retract from the wall, it appears as a single dark line in some sections and as 2 dark lines separated by a light zone in others. The membrane system consists of mesosomes and endomembrane structures. The mesosomes are those membrane structures whose derivatives appear to be the plasma membrane. The endomembrane structures, in the present report, are those that appear to have been derived from either the cytoplasm or the limiting membranes of the pre-existing membrane structures. All membranes seem capable of proliferation, a mechanism obviously responsible for the growth of the individual membrane structures and for the origin of many new ones. The mesosomes, according to their limits, are of 2 distinct types, the open mesosomes and the closed mesosomes. The open mesosomes are partially enclosed by limiting membranes, leaving the unenclosed sides limited by the wall. These mesosomes, when old, usually in aerial hyphae, may become attached to the wall and somewhat deleted from their limiting membranes. The individual membranes in their interior may appear disfigured. The closed mesosomes are completely surrounded by the limiting membranes. These mesosomes, as well as endomembrane structures, retain their original positions in the cytoplasm even in the older aerial hyphae, and the membranes in their interior usually remain practically as distinct in the aerial hyphae as they are in the substratal hyphae. New mesosomes and endomembrane structures are being formed continuously as the mycelium develops. The mesosomes, as a rule, occupy more or less the peripheral regions of the cytoplasm, while the endomembrane structures distribute themselves widely in the cytoplasm and also in the nucleoids. The appearance of the unit membranes, being double-layered (2 dark lines separated by a light line), is not consistent. The membranes as a whole are more resistant to degeneration than the cytoplasm and the nucleoids.     

Crystal structure of cathepsin X: a flip-flop of the ring of His23 allows carboxy-monopeptidase and carboxy-dipeptidase activity of the protease

BACKGROUND: Cathepsin X is a widespread, abundantly expressed papain-like mammalian lysosomal cysteine protease. It exhibits carboxy-monopeptidase as well as carboxy-dipeptidase activity and shares a similar activity profile with cathepsin B. The latter has been implicated in normal physiological events as well as in various pathological states such as rheumatoid arthritis, Alzheimer's disease and cancer progression. Thus the question is raised as to which of the two enzyme activities has actually been monitored. RESULTS: The crystal structure of human cathepsin X has been determined at 2.67 A resolution. The structure shares the common features of a papain-like enzyme fold, but with a unique active site. The most pronounced feature of the cathepsin X structure is the mini-loop that includes a short three-residue insertion protruding into the active site of the protease. The residue Tyr27 on one side of the loop forms the surface of the S1 substrate-binding site, and His23 on the other side modulates both carboxy-monopeptidase as well as carboxy-dipeptidase activity of the enzyme by binding the C-terminal carboxyl group of a substrate in two different sidechain conformations. CONCLUSIONS: The structure of cathepsin X exhibits a binding surface that will assist in the design of specific inhibitors of cathepsin X as well as of cathepsin B and thereby help to clarify the physiological roles of both proteases.     

Relationship Between Physiological Status and Formation of Extracellular Polysaccharide Glycocalyx in Pseudomonas atlantica

Marine pseudomonads, such as Pseudomonas atlantica, are readily isolated from sediments. These organisms form extracellular polysaccharide polymers (glycocalyx). The factors affecting the composition and amount of glycocalyx in batch culture of these organisms were examined. The formation of glycocalyx was stimulated by the inclusion of galactose as the carbon source and by increased surface area resulting from addition of sand to the medium. The composition of the glycocalyx changed during the growth cycle, with a marked increase in the proportions and absolute amounts of uronic acids as the rate of synthesis increased. In estuarine sediments, the glycocalyx contained a carbon content at least as great as in the microbes themselves. The greatest accumulation of these polymers occurred late in the stationary phase when the physiological status of the cells, as measured by the adenylate energy charge, showed maximal stress. Maximal formation of glycocalyx possibly could be used as an estimate of the nutritional status of these microbes.     

THE PHAGOCYTOSIS OF SOLID PARTICLES : III. CARBON AND QUARTZ.

1. The rates of ingestion of quartz and carbon particles by leucocytes, when both are in suspension in serum, was compared with the availability of the two particles as predicted from the calculated chances of collision with the leucocytes, and it was shown that carbon is ingested about 4 times as readily as quartz. 2. The greater ease of ingestion of carbon was verified by a new method of measuring phagocytosis, described as the film method in which the cells ingest particles as they creep about on a slide. 3. The relative rates of ingestion of carbon and quartz depend upon the condition of the cells, the difference increasing as the phagocytic activity of the cells decreases. 4. Sponge cells also ingest carbon about 3 times as readily as quartz. 5. The hypothesis is suggested that the cause of the more rapid ingestion of carbon may be identical with the cause of the greater instability of the carbon suspensions. 6. An inorganic analogy to this selective phagocytic action is offered. 7. The application to opsonins and agglutinins is discussed.     

Comparison between peroxidase-conjugated antigen or antibody and peroxidase-anti-peroxidase complex in a postembedding procedure.

The staining efficiency of peroxidase labeled immunoglobulin conjugate, used either as antigen or as antibody, has been compared with that of peroxidase-anti-peroxidase complex (PAP) on ultrathin sections of araldite embedded material. The conjugate gave positive results in a two layer method as well as in a three layer method when used as antibody. No staining was observed when it was used as antigen. The conjugation seemed to impair the antigenic reactivity of immunoglobulin. The conjugate when used as antibody in the three layer method gave approximately the same staining efficiency as PAP.     

Growth and enzyme synthesis during continuous culture of Trichosporon cutaneum on phenol

The soil yeast Trichosporon cutaneum was grown in continuous culture with phenol as the sole carbon source. The cultures were operated as carbon-limited chemostats or as steady-state continuous cultures without carbon limitation. Selected comparative runs were also conducted on glucose or acetate as carbon source. In addition to growth parameters, the activities of several intracellular enzymes were determined, comprising those directly involved in the degradation of phenol as well as auxiliary enzymes required for the generation of reducing power. All enzymes were assayed in detergent-permea-bilized cells. Phenol was found to serve as an excellent carbon source, comparable to glucose or acetate. The utilization of phenol in T. cutaneum is very efficient as indicated by a low maintenance requirement (0.01 g phenol/g cells.h). The cell yields obtained were on the order of 0.8 g cells/g phenol. Although the phenol-limited chemostats were run with fully phenol-induced cells, a further increase in the activities of isocitrate DH(NADP(+)), maleate DH and the phenol-degrading enzymes occurred after transition to nonlimiting condition. Enzyme activities increased in parallel with increasing phenol levels in the effluent, as well as with increasing toxicity. The significance of this phenomenon is discussed. The significance of this phenomenon is discussed. This elevation in enzyme activities in not related to an increase in specific growth rate.     

Development of biochemistry in the Biology Department of Kharkhov State University. Formation of a new direction--physiology and biochemistry of aging

The article presents some data on teaching Biochemistry at the Nature Department of Kharkiv Imperial University Physico-Mathematical Faculty, as well as restoration of biochemical education and scientific researches as a result of the University reorganization in the Soviet period and foundation of the Biochemistry Chair at the Biological Faculty in the renovated Kharkiv State University. The analysis of scientific biochemical investigations conducted in the Kharkiv University till now is revealed. The special attention is paid to development of such a scientific trend as age physiology and biochemistry. The article deals with the comprehensive information on scientific and pedagogical activity of the outstanding scientists such as O. V. Nagorny and I. M. Bulankin as founders of the Kharkiv school of age physiology and biochemistry. The work has utilized some archive data.