Molecular Detection and Characterization of Ehrlichia ruminantium

Ehrlichia ruminantium is an obligate intracellular bacterium that can cause a disease in ruminants known as heartwater. The vertebrate host becomes infected when infected Amblyomma ticks (nymphs or adults) feed on it. When the host survives the infection it becomes a carrier. Ticks become infected with E. ruminantium when they feed on an infected host. The infection is transmitted from one developmental stage to the other except from adult to eggs for as far as is known now. Detection of E. ruminantium is difficult, especially in carrier animals, and is based on either detection of antibodies (MAP1-B ELISA) or detection of bacterial DNA using PCR techniques. The purpose of the research was to molecularly characterize E. ruminantium and to develop new or improved diagnostic tests for the detection of E. ruminantium. For the detection the MAP1-B ELISA was further validated using the TG-ROC method to determine cutoff values with equal sensitivity and specificity, and a new assay based on the reverse line blot (RLB) technique was developed. The developed RLB assays can simultaneously detect and identify 8 Anaplasma and Ehrlichia species. For the characterization several 16S rRNA sequences were determined, which were also used for a reorganization of the taxonomic classification of several rickettsial species. Finally, the transcription of the map1 multigene family was studied in different environments and it was found that map1-1 was transcribed in vitro and in vivo in the vector tick whereas it was not transcribed at a detectable level in vitro in bovine endothelial cells

Принципи управління персоналом сільськогосподарських підприємств (на прикладі Луганської області)

У статті проаналізовано сучасний стан сільськогосподарського виробництва в Луганській області та здійснена оцінка перспектив реформування управління персоналом на підприємствах АПК. Регресійним аналізом оцінено ступінь впливу деяких факторів на рентабельність персоналу. Рекомендується використання SWOT-аналізу для дослідження й формування раціонального управління персоналом підприємств АПК.Performed analysis of the current state of agriculture in the Luhansk region, evaluated the prospects for personnel resources reforming for the agricultural enterprises. The degree of factors influencing on profitability of personnel is appraised by the regressive analysis. SWOT-analysis is recommended for research and forming of agrarian enterprises rational management of a personnel

A murine and a porcine coronavirus are released from opposite surfaces of the same epithelial cells

Epithelial cells are important target cells for coronavirus infection. Earlier we have shown that transmissible gastroenteritis coronavirus (TGEV) and mouse hepatitis coronavirus (MHV) are released from different sides of porcine and murine epithelial cells, respectively. To study the release of these viruses from the same cells, we constructed a porcine LLC-PK1 cell line stably expressing the recombinant MHV receptor cDNA (LMR cells). The MHV and TGEV receptor glycoproteins were shown by immunofluorescence to appear at the surface of the cells and to be functional so that the cells were susceptible to both MHV and TGEV infection. Both coronaviruses entered polarized LMR cells only through the apical surface. Remarkably, while the cells remained susceptible to TGEV for long periods, infectability by MHV decreased with time after plating of the cells onto filters. This was not due to a lack of expression of the MHV receptor, since this glycoprotein was still abundant on the apical surface of these cells. TGEV and MHV appeared to exit LMR cells from opposite sides. Whereas TGEV was released preferentially at the apical membrane, MHV was released preferentially at the basolateral surface. These results show that vesicles containing the two coronaviruses are targeted differently in LMR cells. We propose that the viruses are sorted at the Golgi complex into different transport vesicles that carry information directing them to one of the two surface domains. The apical release of TGEV and the basolateral release of MHV might be factors contributing to the difference in virus spread found between TGEV and MHV in their respective natural hosts, the former causing mainly a localized enteric infection, the latter spreading through the body to other organs

Coronaviruses in polarized epithelial cells

Coronaviruses have a marked tropism for epithelial cells. In this paper the interactions of the porcine transmissible gastroenteritis virus (TGEV) and mouse hepatitis virus (MHV-A59) with epithelial cells are compared. Porcine (LLC-PK1) and murine (mTAL) epithelial cells were grown on permeable supports. By inoculation from the apical or basolateral side both TGEV and MHV-A59 were found to enter the polarized cells only through the apical membrane. The release of newly synthesized TGEV from LLC-PK1 cells occurred preferentially from the apical plasma membrane domain, as evidenced by the accumulation of viral proteins and infectivity in the apical culture fluid. In contrast, MHV was released preferentially from the basolateral membrane of mTAL cells. The apical release of TGEV and the basolateral release of MHV may explain the in vivo establishment of a local and systemic infection, respectivel

A pan-inflammatory microRNA-cluster is associated with orbital non-Hodgkin lymphoma and idiopathic orbital inflammation

Ophthalmic researc

A pan‐inflammatory microRNA‐cluster is associated with orbital non‐Hodgkin lymphoma and idiopathic orbital inflammation

Ophthalmic researc