Studies on some specific Ap4A-degrading enzymes with the use of various methylene analogues of P1P4-bis-(5'',5''''''-adenosyl) tetraphosphate.

Six new methylenephosphonate analogues of P1P4-bis-(5',5'-adenosyl) tetraphosphate, Ap4A, having P2-P3 carbon bridges CF2, CCl2 and CH2CH2 or P1-P2 and P3-P4 carbon bridges CF2, CCl2 and CH2CH2 in the tetraphosphate chain, were examined as substrates or inhibitors for two specific Ap4A-degrading enzymes: (asymmetrical) Ap4A hydrolase (EC 3.6.1.17) from yellow-lupin seeds and (symmetrical) Ap4A hydrolase (EC 3.6.1.41) from Escherichia coli. All analogues in which the central oxygen atom was replaced by a stable carbon bridge were hydrolysed by the asymmetrical hydrolase (CF2 greater than CCl2 greater than O greater than CHBr greater than CH2 greater than CH2CH2). As expected, these analogues were not hydrolysed by the symmetrical hydrolase, which was also unable to act on analogues having P1-P2 and P3-P4 carbon bridges.     

Dynamics of changes in the cardiovascular response to submaximal exercise during low-intensity endurance training with particular reference to the systolic time intervals

Eighteen male volunteers (aged 20-23 years), not involved in any sporting activities, were submitted to 13 weeks of training consisting of 30 min exercise [at 50%-75% maximal oxygen intake (VO2max)] on a cycle ergometer, performed 3 times a week. Every 4 weeks cardiac function was evaluated by measuring the systolic time intervals at rest and during submaximal cycle exercise. Stroke volume (SV), heart rate (HR) and blood pressure (BP) responses to submaximal exercise, VO2max and anaerobic threshold (AT) were also determined. Significant increases in VO2max, increases in AT and SV at the submaximal exercise intensities, as well as decreases in HR and BP were found after 4 weeks of training. Resting systolic time intervals were not affected by training, but during the submaximal cycle exercise the values of the pre-ejection period (PEP) and isovolumic contraction time (ICT) corresponding to HR of 100 beats.min-1 were significantly lowered after 13 weeks of training, whereas PEP, ICT and total electromechanical systole corresponding to HR of 130 beats.min-1 were significantly shortened by the 4th week. The ratios of PEP:LVET (left ventricular ejection time) and ICT:LVET during submaximal exercise were significantly lowered by training starting from the 8th week. These changes might be interpreted as evidence of the training-induced enhancement of the "contractility reserve", i.e. the ability to increase heart muscle contractility with increasing exercise intensity.     

Hydrolysis of sunflower oil by means of hydrophobic membrane with lipolytic activity

Polytetrafluoroethylene (PTFE) membranes of different porosity were used for lipase from Rhizopus immobilization. The immobilized enzyme applied to sunflower oil hydrolysis achieved the activity of 1228 U/m² of the membrane area and the half-life time was calculated to be 7 days.     

Immunohistology of gastrointestinal carcinoid tumors

DAKO quick staining kits were used to immunostainings for Epithelial Membrane Antigen (EMA) Neuron Specific Enolase (NSE) and S-100 protein (S-100) in 18 carcinoid tumors of the gastrointestinal tract classified according to Soga and Tazawa. EMA was always absent in carcinoid tumors, being at the same time present within glandular epithelium. 88% of cases showed positive immunostaining for NSE. S-100 immunostaining showed immunopositive stellate cells present within the tumor especially within type A carcinoids. In addition in one carcinoid tumor thick, strongly S-100 positive bundles were noticed at the periphery of nests of tumor cells. Combined immunohistochemical and ultrastructural studies are needed to elucidate interrelation of neoplastic and neural elements within carcinoid tumorsa.     

Coexpression of vimentin and desmin in gastric leiomyomas. An immunohistochemical study in paraffin sections

The aim of the study was to evaluate the incidence of the reactions for vimentin and desmin in gastric leiomyomas routinely processed in formalin and embedded i paraffin. The material studied included four benign leiomyomas, seven malignant leiomyomas and three malignant epithelioid leiomyomas. A positive reaction to vimentin was found in 13 out 14 leiomyomas under study. The number of neoplastic cells showing vimentin expression was larger in malignant, especially epithelioid leiomyomas than in non-malignant leiomyomas. A positive reaction for desmin in neoplastic cells was found in 9 leiomyomas 64%. One non-malignant leiomyoma showed a moderate reaction. In the remaining eight cases the reaction was weak and occurred in single neoplastic cells. Coexpression of vimentin and desmin in neoplastic cells occurred in 8 out of 9 leiomyomas with a positive reaction for desmin. Coexpression of vimentin and desmin occurred also in the smooth muscle cells of blood vessels in all 14 cases. A weak reaction for desmin or its lack in the tumour cells of leiomyomas with its marked expression in the smooth muscle cells of the blood vessels and gastric wall outside the tumor points rather to a small number of desmin filaments in the neoplastic cells than to their destruction by fixation in formalin. The occurrence of the reaction to desmin only in a limited number of neoplastic cells questions the reliability of its use in the oligopiopsy material.     

Expression of nonspecific cross-reacting antigen species in myeloid leukemic patients and healthy subjects

The reactivity of two monoclonal antibodies recognizing NCA-95 and NCA-55 (MAb 47 and MAb 192, respectively) with a polyclonal anti-NCA serum in myeloid leukemic cells isolated by density gradient centrifugation was compared using an immunofluorescence test (IF). It was observed that the blood myeloid cells in 78.8% of the patients with different types of myelocytic leukemias and all granulocytes of 15 normal donors showed similar expression of the NCA species studied. The leukocytes of the remaining patients did not synthesize the NCA-95 species regardless of the maturation stage of the cells studied. In two patients, synthesis of this NCA form was limited to the fractions containing myelocytes and metamyelocytes. We have found that all anti-NCA antibodies studied recognized different antigenic epitopes in a myeloid cell series. A relationship between the patient's survival and the proportion of NCA-containing cells was also observed.     

Formation and function of accessory nuclei in the oocytes of the bird louse,Eomenacanthus stramineus (Insecta,Mallophaga)

In the oocytes of Eomenacanthus stramineus accessory nuclei arise by budding from the nuclear envelope. It is suggested that microtubules and the thick layer of the nuclear lamina are involved in this process. Newly formed accessory nuclei contain aggregations of fibrillogranular material. These aggregations are slightly Feulgen positive, RNA negative and stain positively with the AgNOR method. During later developmental stages one dense, RNA-positive inclusion appears in each accessory nucleus. These inclusions consist of on Ag-NOR-positive cortical layer and an Ag-NOR-negative core. The function of accessory nuclei in the species investigated is discussed in the light of these results.     

Linkage disequilibrium between mutation and RFLP haplotype at the phenylalanine hydroxylase locus in the German population

Summary Restriction fragment length polymorphism (RFLP) haplotypes at the phenylalanine hydroxylase (PAH) locus have been determined in 60 German families with PAH deficiency. Similar to the Danish population, about 90% of the mutant alleles are confined to four distinct haplotypes. There are however, differences in the frequency distributiion of these haplotypes among the mutant alleles between the two populations. Using an oligonucleotide probe for the splicing mutation associated with mutant haplotype 3 in the Danish population, a tight association between the mutation and the RFLP haplotype has also been observed in Germany. The results provide strong evidence that the splicing mutation occurred on a haplotype 3 chromosome and that the mutant allele has spread into different populations smong Caucasians.     

Intergeneric relationships ofLolium,Festuca, andVulpia (Poaceae) and their phylogeny

Morphological and seed protein analyses of 26 species of the generaLolium, Festuca andVulpia confirmed their close systematic affinities. Six inflorescence characters readily differentiatedFestuca fromLolium. Protein similarities betweenFestuca of sect.Bovinae and cross-pollinated species ofLolium, coupled with cytogenetic and crossability data, substantiate that they should be united into one genus.Vulpia had phenetic similarities with sect.Scariosae, Montanae andOvinae ofFestuca. Lolium, Festuca, andVulpia are most likely derived from a common ancestral form which was close toFestuca pratensis andLolium perenne.