Search

In Arabidopsis, the small signaling peptide (peptide hormone) RALF34 is involved in the gene regulatory network of lateral root initiation. In this study, we aimed to understand the nature of the signals induced by RALF34 in the non-model plant cucumber ( Cucumis sativus ), where lateral root primordia are induced in the apical meristem of the parental root. The RALF family members of cucumber were identified using phylogenetic analysis. The sequence of events involved in the initiation and development of lateral root primordia in cucumber was examined in detail. To elucidate the role of the small signaling peptide Cs RALF34 and its receptor Cs THESEUS1 in the initial stages of lateral root formation in the parental root meristem in cucumber, we studied the expression patterns of both genes, as well as the localization and transport of the Cs RALF34 peptide. CsRALF34 is expressed in all plant organs. CsRALF34 seems to differ from AtRALF34 in that its expression is not regulated by auxin. The expression of AtRALF34, as well as CsRALF34, is regulated in part by ethylene. CsTHESEUS1 is expressed constitutively in cucumber root tissues. Our data suggest that Cs RALF34 acts in a non-cell-autonomous manner and is not involved in lateral root initiation in cucumber.

The main role of RALF small signaling peptides was reported to be the alkalization control of the apoplast for improvement of nutrient absorption; however, the exact function of individual RALF peptides such as RALF34 remains unknown. The Arabidopsis RALF34 ( At RALF34) peptide was proposed to be part of the gene regulatory network of lateral root initiation. Cucumber is an excellent model for studying a special form of lateral root initiation taking place in the meristem of the parental root. We attempted to elucidate the role of the regulatory pathway in which RALF34 is a participant using cucumber transgenic hairy roots overexpressing CsRALF34 for comprehensive, integrated metabolomics and proteomics studies, focusing on the analysis of stress response markers. Cs RALF34 overexpression resulted in the inhibition of root growth and regulation of cell proliferation, specifically in blocking the G2/M transition in cucumber roots. Based on these results, we propose that CsRALF34 is not part of the gene regulatory networks involved in the early steps of lateral root initiation. Instead, we suggest that Cs RALF34 modulates ROS homeostasis and triggers the controlled production of hydroxyl radicals in root cells, possibly associated with intracellular signal transduction. Altogether, our results support the role of RALF peptides as ROS regulators.

CRISPR/Cas-mediated genome editing is a powerful tool of plant functional genomics. Hairy root transformation is a rapid and convenient approach for obtaining transgenic roots. When combined, these techniques represent a fast and effective means of studying gene function. In this review, we outline the current state of the art reached by the combination of these approaches over seven years. Additionally, we discuss the origins of different Agrobacterium rhizogenes strains that are widely used for hairy root transformation; the components of CRISPR/Cas vectors, such as the promoters that drive Cas or gRNA expression, the types of Cas nuclease, and selectable and screenable markers; and the application of CRISPR/Cas genome editing in hairy roots. The modification of the already known vector pKSE401 with the addition of the rice translational enhancer OsMac3 and the gene encoding the fluorescent protein DsRed1 is also described.

While in most higher plants, including the model system Arabidopsis thaliana , the formation of lateral root primordia is induced in the elongation zone of the parental root, in seven plant families, including Cucurbitaceae, an alternative root branching mechanism is established such that lateral roots are initiated directly in the apical meristem of the parental root. In Arabidopsis, the transcription factor GATA23 and MEMBRANE-ASSOCIATED KINASE REGULATOR4 (MAKR4) are involved in the gene regulatory network of lateral root initiation. Among all marker genes examined, these are the earliest known marker genes up-regulated by auxin during lateral root initiation. In this study, putative functional orthologs of Arabidopsis GATA23 and MAKR4 were identified in cucumber ( Cucumis sativus ) and squash ( Cucurbita pepo ). Both cucurbits contained 26 genes encoding GATA family transcription factors and only one MAKR4 gene. Phylogenetic and transcriptional analysis of up-regulation by auxin led to the identification of GATA23 putative functional orthologs in Cucurbitaceae - CpGATA24 and CsGATA24 . In squash, CpMAKR4 was up-regulated by naphthylacetic acid (NAA) and, similar to MAKR4 in Arabidopsis, indole-3-butyric acid (IBA). A detailed analysis of the expression pattern of CpGATA24 and CpMAKR4 in squash roots from founder cell specification until emergence of lateral root primordia was carried out using promoter-fluorescent reporter gene fusions and confocal microscopy. Their expression was induced in the protoxylem, and then expanded to founder cells in the pericycle. Thus, while the overall expression pattern of these genes was significantly different from that in Arabidopsis, in founder cells their expression was induced in the same order as in Arabidopsis. Altogether, these findings suggest that in Cucurbitaceae the putative functional orthologs of GATA23 and MAKR4 might play a role in founder cell specification and primordium positioning during lateral root initiation. The role of the protoxylem in auxin transport as a trigger of founder cells specification and lateral root initiation is discussed.

The objective of the present study was to estimate the informative value of the cytological investigation of nasal secretion in the children presenting with infectious and allergic rhinitis and sinusitis. This publication presents the results of comparative analysis of rhinocytograms obtained from the patients with the above diseases. It was shown that the rhinocytological technique can be applied for differential diagnostics of rhinitis and sinusitis in the children, the assessment of the severity of allergic diseases of the upper respiratory tract, and monitoring the efficacy of the treatment of rhinitis.

The organized benchmark analysis result treatments 152 sick with taken in perforated gastroduodenal ulcers. In the main group (87 sick) in complex treatment are used methods of ozonotherapy. In group of the comparison (65 sick) used standard scheme of the treatment to peptic ulcer. Methods of ozonotherapy promote more early and more full recovering the structure gastroduodenal mucous and provide efficient eradication of Helicobacter pylori, reduces the time of the healing the ulcers and reduces the risk of the relapse ulcers.

Strangulation colorectal obstruction was modeled in 60 Wistar rats. Necrotic segment of the intestine was resected under conditions of peritonitis and end-to-end intestinal anastomosis was performed on a PCV catheter conducted through the anus. Sodium hypochlorite and ozone solution were used for sanitation of the abdominal cavity and intestinal lavage, and the intestinal anastomosis was coated with Ozonide (ozonized oil). The use of physicochemical methods notably reduced the incidence of postoperative pyoinflammatory complications, incompetence of intestinal anastomosis sutures, and animal mortality.

To study collagen metabolism, 113 children aged 3 to 14 years afflicted with rheumatoid arthritis and 43 conventionally normal children were examined. The patients were examined for free and protein-bound hydroxyproline in the blood serum and synovial fluid and for total hydroxyproline in diurnal urine. The blood serum and synovial fluid showed a decline of free and a rise of protein-bound hydroxyproline, more marked in severe rheumatoid arthritis, its higher activity and damage to numerous joints. Excretion of free and total hydroxyproline with urine was significantly lower than in the normal children whatever the form of rheumatoid arthritis, its activity or number of the joints involved into the process. In view of the lack of hydroxyprolinuria in the sick children, the authors discuss a hypothesis of a potential initial decrease of collagen formation in children afflicted with rheumatoid arthritis, suggesting its relationship to the "ligand pathology".