The complete amino acid sequence of bovine skeletal muscle acylphosphatase

The primary structure of bovine skeletal muscle acylphosphatase was determined by performing the sequence analyses of the complete series of tryptic peptides. The amino acid composition of the entire series of peptic peptides was used to reconstruct the sequence by the overlapping method. The proposed structure is further confirmed by analogy with known amino acid sequences of acylphosphatase from skeletal muscle of other vertebrate species. The length of the polypeptide chain is 98 residues, identical to the length of the enzymes from other known mammalian species, but different from that found in turkey. The enzyme is NH2-acetylated and a comparison with the analogous molecular forms from other vertebrate species indicates that there are several long polypeptide stretches strictly conserved (93-97% identical position among mammals, and about 80% between calf and turkey enzymes).     

Dynamics of the accumulation of the basic immunoglobulin classes in the intestinal contents in food toxinfections]

The work presents the data on the dynamics of accululation of the main classes of immunoglobulins (A, G and M) in coprofiltrates obtained from patients with alimentary toxicoinfections (bacteriologically confirmed salmonellosis and diseases of unknown etiology). The levels of immunoglobulins of all classes (mainly IgA and IgG) were shown to be elevated in the process of the disease. The dynamics of the increase in the level of IgA (both general and secretory) was supposed to indicate the formation of local immunity in the intestinal wall. The presence of serum IgA and the characteristic IgG dynamics seemed to be indicative of destructive processes occurring in the intestinal wall. Thus, the dynamics of accumulation of immunoglobulins in coprofiltrates obtained from patients with alimentary toxicoinfections reflects the main local pathological and immunological processes.     

Immunological reactions in the secretions of volunteers vaccinated against influenza by aerosol and intranasally]

Aerosol and intranasal methods of influenza vaccination were studied on volunteers. Aerosol vaccination induced intensive S-IgA-producing reaction and increase of the specific antibody titres in the saliva and nasal secretions. Intranasal vaccination led to increase of influenza antibodies in the washings from the nasopharynx. The presence of a wide spectrum of antiviral and antibacterial normal secretory antibodies was revealed in the fluids under study. Along with stimulation of specific secretory antibodies, the methods of influenza vaccination under study led to increase of the titre of antibodies nonspecific of the vaccine used.     

A phylogeny reconstruction of the Dendrophylliidae (Cnidaria,Scleractinia) based on molecular and micromorphological criteria,and its ecological implications

Recent molecular phylogenetic studies have shown that most traditional families of zooxanthellate shallow‐water scleractinians are polyphyletic, whereas most families mainly composed of deep‐sea and azooxanthellate species are monophyletic. In this context, the family Dendrophylliidae (Cnidaria, Scleractinia) has unique features. It shows a remarkable variation of morphological and ecological traits by including species that are either colonial or solitary, zooxanthellate or azooxanthellate, and inhabiting shallow or deep water. Despite this morphological heterogeneity, recent molecular works have confirmed that this family is monophyletic. Nevertheless, what so far is known about the evolutionary relationships within this family, is predominantly based on skeleton macromorphology, while most of its species have remained unstudied from a molecular point of view. Therefore, we analysed 11 dendrophylliid genera, four of which were investigated for the first time, and 30 species at molecular, micromorphological and microstructural levels. We present a robust molecular phylogeny reconstruction based on two mitochondrial markers (COI and the intergenic spacer between COI and 16S) and one nuclear (rDNA), which is used as basis to compare micromorphogical and microstructural character states within the family. The monophyly of the Dendrophylliidae is well supported by molecular data and also by the presence of rapid accretion deposits, which are ca. 5 μm in diameter and arranged in irregular clusters, and fibres that thicken the skeleton organized in small patches of a few micrometres in diameter. However, all genera represented by at least two species are not monophyletic, Tubastraea excluded. They were defined by traditional macromorphological characters that appear affected by convergence, homoplasy and intraspecific variation. Micromorphogical and microstructural analyses do not support the distinction of clades, with the exception of the organization of thickening deposits for the Tubastraea clade.     

Errors in determining the flow rate of Hirst-type pollen traps

Standardisation of methods of pollen monitoring networks is vital for data quality. In pollen monitoring networks in Europe, the Hirst-type trap is standard. Hirst traps are calibrated with handheld rotameters. We detected a systematic error in the flow rate calibrated by these standard handheld rotameters. We measured the flow rate of 19 Hirst traps from three commercial brands during calibration but also during monitoring. We used three different rotameters supplied by the manufacturers of the traps, respectively. The actual air flow rate was measured using an electronic heat anemometer with negligible air flow resistance. After calibration to 10 l/min, the rotameter was removed, which led to a significant increase in the flow rate in the range of 10.5–17.2 l/min, a systematic error between 5 and 72%. No significant difference was found between the different commercial trap brands. The analysis revealed that the error depended on the type of the rotameter and the individual trap. The error may be explained by the additional air flow resistance of each rotameter. The total resistance of the system—trap plus rotameter—is higher during calibration when the rotameter is held on the inlet compared to the routine monitoring without the rotameter. Depending on the characteristic curve of the suction pump in the trap (fan), the air flow rate increases to values considerably higher than 10 l/min. Thus, monitoring is done under a higher flow rate than that was calibrated. In order to obtain comparable data within a monitoring network, a solution for correction of this systematic error seems advisable, preferably in cooperation with the manufacturers.     

Generation of monoclonal antibodies to human acylphosphatase (muscular isoenzyme) and application in solid-phase immunoassay

Monoclonal antibodies to human acylphosphatase (muscle isoenzyme) were generated by an improved hybridoma technique. Immunization consisted of four antigen administrations in an overall period of 15 weeks. After cell fusion and repeated subcloning of positive lines, seven monoclonal antibodies with good affinity and specificity were selected. These antibodies were characterized for their affinity constant and immunoreactivity. The latter was determined using peptides generated by CNBr cleavage of the antigen. One of the selected antibodies had an affinity constant such that it could be used to develop a competitive enzyme-linked immunosorbent assay. In our test, the antigen that was coated on the matrix, and the free one, competed for the antibody-horseradish peroxidase conjugate. No cross-reactivity with the erythrocyte iso-enzyme was found, and the test showed a limit in sensitivity of 0.32 ng/ml of antigen. We expect that the enzyme immunoassay could be useful for clinical application.