A high-throughput method for quantifying gene expression data from early <Emphasis Type="Italic">Drosophila</Emphasis> embryos

We describe an automated high-throughput method to measure protein levels in single nuclei in blastoderm embryos of Drosophila melanogaster by means of immunofluorescence. The method consists of a chain of specific algorithms assembled into an image processing pipeline. This pipeline transforms a confocal scan of an embryo stained with fluorescently tagged antibodies into a text file. This text file contains a numerical identifier for each nucleus, the coordinates of its centroid, and the average concentrations of three proteins in that nucleus. The central algorithmic component of the method is the automatic identification of nuclei by edge detection with the use of watersheds as an error-correction step. This method provides high-throughput quantification at cellular resolution.Electronic Supplementary Material Supplementary material is available for this article at .H. Janssens and D. Kosman contributed equally to this paper.     

Temporal classification of Drosophila segmentation gene expression patterns by the multi-valued neural recognition method

In order to reconstruct the establishment of the body pattern over time in Drosophila embryos, we have developed automated methods for detecting the age of an embryo on the basis of knowledge about its gene expression patterns. In this paper we perform temporal classification of confocal images of expression patterns of genes controlling segmentation by means of a neural network based on multi-valued neurons (MVN). MVN are artificial neural processing elements with complex-valued weights and high functionality, which proved to be efficient for solving the image recognition problems. The results obtained by this method confirm its efficiency for image recognition and indicate that the method can detect characteristic features of expression patterns which mark their development over time.     

A new methodological approach for leptospira persistence studies in case of mixed leptospirosis

A new methodical approach for Leptospira persistence studies in case of mixed leptospirosis, based on the use of PCR test systems with different taxonomic specificity for the indication and identification of leptospires, was developed. Two PCR test systems (G and B) were used in experiments on BALB/c white mice to study patterns of the development of mixed infection caused by leptospires of serovar poi (genomospecies L. borgpeterseni) and grippotyphosa (genomospecies L. kirschneri). The conclusion was made of good prospects of this method application in studies on symbiotic relationships of leptospires both in vivo and in vitro.     

Membrane technology as a basis for creation of treatment-and-prophylactic equipment for inhalation therapy and normobaric hypoxia]

The principles of membrane technology of gas distribution have been considered, that has a wider adoption in different branches of economy and in medicine. One of the promising trends of membrane technology employment is production of respiratory gas mixtures for hypoxytherapy, non-medicamental method to increase the human organism resistance to numerous pathogenetic environmental factors. Gas distributors and devices with different surface mass exchange and wide range of operating stress using hollow selectively permeable fibers of poly-4-methylpentene-1 are designed and produced under conditions of scientific productions. The technology of hollow fibers formation has been tried out on production spinning machinery of enterprises of chemical fibers in Ukraine. Design of gas distributors permits supplying them as a part of different devices of normobaric hypoxia from individual ones to devices of joint use allowing for customer's demands.     

Effect of diazepam on the Na, K-ATPase state in a penicillin--induced hyperactivity focus in the cerebral cortex]

Intramuscular injection of diazepam to rats at doses of 0.01 and 2 mg/kg 25-30 min after penicillin application to the rat brain cortex leads to alteration of periodic appearance of epileptic seizures (ES), to changes in the seizure pattern, and to emergence of periodic acceleration of epileptiform discharges (ED). Injection of diazepam at a dose of 2 mg/kg 20 min before penicillin application results in the reduction of ED latency in the epileptogenic focus and in a decrease in their frequency before seizures as compared to the control animals without diazepam injection. ES appear irregularly, their quantity is markedly reduced while duration is increased. Diazepam injection leads to disappearance of the rat moving reaction during ER and ES. In vivo experiments diazepam (2 mg/kg) does not influence brain cortex Na, K-ATPase of crude synaptosomes. However, diazepam leads to an increase in Na, K-ATPase activity both in the primary and dependent secondary epileptogenic foci. It is suggested that the anticonvulsant action of diazepam may be underlain by its activating effect on Na, K-ATPase of neuronal membranes in the epileptogenic focus.     

Enzymatic micromethod for determining Na, K-ATPase activity in rat cerebral cortex membranes]

A method for the assay of Na,K-ATPase activity of unpurified synaptosomal fraction obtained from the microquantities (2--3 mg of fresh tissue) of the rat cerebral cortex is described. This method is based on the fluorimetric determination of ADP formed in the course of ATPase reaction. The method is highly sensitive and may be used to determine the membrane preparations Na,K-ATPase activity with the protein content of 0.05--10.0 microgram per sample.     

Importance of immunoelectrophoresis in agar and disc electrophoresis in polyacrylamide gel for characterizing different strains of Bordetella pertussis

The antigenic composition of typical and atypical B. pertussis strains obtained in the foci of pertussis infection, as well as experimentally obtained antibiotic-resistant B. pertussis strains, has been studied by the methods of immunoelectrophoresis in agar and electrophoresis in polyacrylamide gel (PAAG). Immunoelectrophoresis in agar has been found capable of differentiating B. pertussis culture from a group of unidentified morphologically similar Gram-negative bacilli by their antigenic composition and thus suitable for use as an additional criterion in the identification of atypical B. pertussis strains. PAAG electrophoresis has permitted finding differences in the set of protein antigens in the control strain and in its clones obtained by multiple subculturing in media with antibiotics added.