Влияние Лазерного Облучения на Активность Бактерий Bacillus Subtilis и Pseudomonas Fluorescens

The paper discusses the problem of increasing the activity of phytopathogen-antagonistic bacteria under the effect of laser irradiation. It has been shown that short-term treatment of cells of Bacillus subtilis and Pseudomonas fluorescens with coherent light can increase bacterial growth rate and improve their fungicidal activity. It ensures high efficiency of environmentally safe measures for biological monitoring of plant diseases, which is in line with the principles of organic farming.El artículo analiza el problema de aumentar la actividad de las bacterias antagonistas de fitopatógenos bajo el efecto de la irradiación con láser. Se ha demostrado que el tratamiento a corto plazo de las células de Bacillus subtilis y Pseudomonas fluorescens con luz coherente puede aumentar la tasa de crecimiento bacteriano y mejorar su actividad fungicida. Asegura una alta eficiencia de las medidas ambientalmente seguras para el monitoreo biológico de las enfermedades de las plantas, lo cual está en línea con los principios de la agricultura orgánica.Обсуждается проблема повышения активности бактерий-антагонистов фитопатогенов под действием лазерного облучения. Показано, что кратковременная обработка когерентным светом клеток Bacillus subtilis и Pseudomonas fluorescenсs способна повысить скорость размножения бактерий и усилить их фунгицидную активность. Это обеспечивает высокую эффективность экологически безопасных мероприятий по биоконтролю болезней растений, что соответствует принципам органического земледелия

Raver1, a dual compartment protein, is a ligand for PTB/hnRNPI and microfilament attachment proteins

By screening a yeast two-hybrid library with COOH-terminal fragments of vinculin/metavinculin as the bait, we identified a new protein termed raver1. Raver1 is an 80-kD multidomain protein and widely expressed but to varying amounts in different cell lines. In situ and in vitro, raver1 forms complexes with the microfilament-associated proteins vinculin, metavinculin, and α-actinin and colocalizes with vinculin/metavinculin and α-actinin at microfilament attachment sites, such as cell–cell and cell matrix contacts of epithelial cells and fibroblasts, respectively, and in costameres of skeletal muscle. The NH2-terminal part of raver1 contains three RNA recognition motifs with homology to members of the heterogeneous nuclear RNP (hnRNP) family. Raver1 colocalizes with polypyrimidine tract binding protein (PTB)/hnRNPI, a protein involved in RNA splicing of microfilament proteins, in the perinucleolar compartment and forms complexes with PTB/hnRNPI. Hence, raver1 is a dual compartment protein, which is consistent with the presence of nuclear location signal and nuclear export sequence motifs in its sequence. During muscle differentiation, raver1 migrates from the nucleus to the costamere. We propose that raver1 may coordinate RNA processing and targeting as required for microfilament anchoring in specific adhesion sites

Создание новых форм томата с генами устойчивости к грибным болезням на основе маркерной селекции

Relevance. The presented studies are aimed at obtaining new forms of tomato with a complex of genes for resistance to fungal diseases in combination with a standard type of bush and dark coloring of fruits based on marker-mediated selection.Methodology. The biological objects of the study are varieties and hybrid forms of tomato from the collection of the Michurinsky SAU. Molecular genetic analysis was performed using the following methods. DNA extraction was carried out from young leaves using a kit for isolation of NC Sample NC manufactured by Agrodiagnostika LLC according to the manufacturer's protocol. Fermentas production kits were used for PCR. Identification of the cladosporosis resistance gene was Cf-19 performed using the DNA marker R7. The presence of a fusarious wilting resistance gene was determined by a I-2/5 marker. The amplification results were visualized by agarose gel electrophoresis.Results. During the research, a collection of varieties and hybrid forms of tomato of the Michurinsky GAU was analyzed in order to identify genes for resistance to cladosporiosis Cf-19 and fusarium wilt I-2. A total of 52 genotypes were analyzed. It was found that most samples (41 samples) are characterized by a heterozygous state of the Cf-19 gene. All indeterminant and semi-determinant forms had both alleles. Of the 23 determinant forms presented in the collection, 10 had only one allele corresponding to recessive homozygote. Among all analyzed tomato genotypes, no dominant homozygous forms were noted. The study of the collection revealed several alleles of the I-2 gene. In total, four fragments corresponding to various alleles were amplified. A total of 50 resistant genotypes have been identified in the collection. Two alleys of the I-2 gene (633/693 bp) were identified in 42 tomato samples. Four varieties are homozygous in one allele (633 bp), which determines resistance. Three varieties have a second resistance allele (566 bp). One genotype has only an allele defining susceptibility (693 bp). On the basis of molecular analysis, as well as an assessment of the type of bush and fetal color, initial forms were selected with subsequent hybridization. 67 hybrid tomato plants were obtained. Evaluation of the presence of resistance genes showed that most of the resulting hybrids are resistant to cladosporiosis and fuzariosis. This is due to the presence of dominant alleles of Cf-19 and I-2 genes in a heterozygous state. Among the resulting hybrids, plants with a bark type of bush were identified. A total of 13 such plants were obtained.Conclusion. Thus, the work carried out allowed to obtain hybrid forms of tomato combine the signs of resistance to two pathogens of fungal diseases and the stem type of the bush. These forms are planned to be used in further selection work.Цель. Исследования направлены на получение новых форм томата с комплексом генов устойчивости к грибным болезням в сочетании со штамбовым типом куста и темной окраской плодов на основе маркер-опосредованной селекции.Методы. Объект исследований – сорта и гибридные формы томата из коллекции Мичуринского ГАУ. Молекулярно-генетический анализ проводили с использованием следующих методов. Экстрагирование ДНК осуществляли из молодых листьев с применением набора для выделения НК «Проба НК» производства ООО «Агродиагностика» согласно протоколу производителя. Для проведения ПЦР использованы наборы производства компании Fermentas. Идентификацию гена устойчивости к кладоспоризу Cf-19 проводили с использованием ДНК-маркера Р7. Наличие гена устойчивости к фузариозному увяданию определяли с помощью маркера I-2/5. Визуализацию результатов амплификации осуществляли с помощью электрофореза в агарозном геле.Результаты. При проведении исследований была проанализирована коллекция сортов и гибридных форм томата Мичуринского ГАУ с целью идентификации генов устойчивости к кладоспориозу Cf-19 и фузариозному увяданию I-2. Всего проанализировано 52 генотипа. Установлено, что для большинства образцов (41 образец) характерно гетерозиготное состояние гена Cf-19. Все индетерминантные и полудетерминантные формы имели оба аллеля. Из 23 представленных в коллекции детерминантных форм у 10 отмечен только один аллель, соответствующий рецессивной гомозиготе. Среди всех анализируемых генотипов томата не отмечено доминантных гомозиготных форм. Изучение коллекции позволило выявить нескольких аллелей гена I-2. Всего амплифицировано четыре фрагмента, соответствующих различным аллелям. Всего устойчивых генотипов в коллекции выделено 50. У 42 образцов томата идентифицированы два аллея гена I-2 (633/693 п.н). Четыре сорта гомозиготны по одному аллелю (633 п.н.), обуславливающему устойчивость. Три сорта имеют второй аллель (566 п.н.) устойчивости. Один генотип имеет только аллель определяющий восприимчивость (693 п.н.).На основании молекулярного анализа, а также оценки типа куста и окраски плода был проведен отбор исходных форм с последующей гибридизацией. Получено 67 гибридных растений томата. Оценка наличия генов устойчивости показала, что большинство полученных гибридов являются устойчивыми к кладоспориозу и фузариозу. Это обусловлено наличием доминантных аллелей генов Cf-19 и I-2 в гетерозиготном состоянии. Среди полученных гибридов выделены растения со штамбовым типом куста. Всего таких растений получено 13. Проведенная работа позволила получить гибридные формы томата, сочетающие признаки устойчивости к двум возбудителям грибных болезней и штамбовый тип куста. Эти формы планируется использовать в дальнейшей селекционной работе

α-Spectrin and integrins act together to regulate actomyosin and columnarization, and to maintain a monolayered follicular epithelium.

The spectrin cytoskeleton crosslinks actin to the membrane, and although it has been greatly studied in erythrocytes, much is unknown about its function in epithelia. We have studied the role of spectrins during epithelia morphogenesis using the Drosophila follicular epithelium (FE). As previously described, we show that α-Spectrin and β-Spectrin are essential to maintain a monolayered FE, but, contrary to previous work, spectrins are not required to control proliferation. Furthermore, spectrin mutant cells show differentiation and polarity defects only in the ectopic layers of stratified epithelia, similar to integrin mutants. Our results identify α-Spectrin and integrins as novel regulators of apical constriction-independent cell elongation, as α-Spectrin and integrin mutant cells fail to columnarize. Finally, we show that increasing and reducing the activity of the Rho1-Myosin II pathway enhances and decreases multilayering of α-Spectrin cells, respectively. Similarly, higher Myosin II activity enhances the integrin multilayering phenotype. This work identifies a primary role for α-Spectrin in controlling cell shape, perhaps by modulating actomyosin. In summary, we suggest that a functional spectrin-integrin complex is essential to balance adequate forces, in order to maintain a monolayered epithelium.BFN was supported by Singapore Ministry of Education; GKS by the BBSRC; IAG by the Wellcome Trust, the BBSRC and the Isaac Newton Trust (Cambridge, UK); CSCM and MDMB by the “Ministerio de Economía y Competiti vidad”,the FEDER programme (BFU2013-48988-C2-1-P), and Junta de Andalucía (Proyecto de Excelencia P09-CVI-5058); IG by a JAE-DOC (CSIC); and IMP by the Wellcome Trust, the BBSRC, the Department of Zoology (Cambridge) and the University of Cambridge.This is the final version of the article. It first appeared from the Company of Biologists via https://doi.org/10.1242/dev.13007

The Molecular Evolution of the p120-Catenin Subfamily and Its Functional Associations

p120-catenin (p120) is the prototypical member of a subclass of armadillo-related proteins that includes δ-catenin/NPRAP, ARVCF, p0071, and the more distantly related plakophilins 1–3. In vertebrates, p120 is essential in regulating surface expression and stability of all classical cadherins, and directly interacts with Kaiso, a BTB/ZF family transcription factor.To clarify functional relationships between these proteins and how they relate to the classical cadherins, we have examined the proteomes of 14 diverse vertebrate and metazoan species. The data reveal a single ancient δ-catenin-like p120 family member present in the earliest metazoans and conserved throughout metazoan evolution. This single p120 family protein is present in all protostomes, and in certain early-branching chordate lineages. Phylogenetic analyses suggest that gene duplication and functional diversification into “p120-like” and “δ-catenin-like” proteins occurred in the urochordate-vertebrate ancestor. Additional gene duplications during early vertebrate evolution gave rise to the seven vertebrate p120 family members. Kaiso family members (i.e., Kaiso, ZBTB38 and ZBTB4) are found only in vertebrates, their origin following that of the p120-like gene lineage and coinciding with the evolution of vertebrate-specific mechanisms of epigenetic gene regulation by CpG island methylation.The p120 protein family evolved from a common δ-catenin-like ancestor present in all metazoans. Through several rounds of gene duplication and diversification, however, p120 evolved in vertebrates into an essential, ubiquitously expressed protein, whereas loss of the more selectively expressed δ-catenin, p0071 and ARVCF are tolerated in most species. Together with phylogenetic studies of the vertebrate cadherins, our data suggest that the p120-like and δ-catenin-like genes co-evolved separately with non-neural (E- and P-cadherin) and neural (N- and R-cadherin) cadherin lineages, respectively. The expansion of p120 relative to δ-catenin during vertebrate evolution may reflect the pivotal and largely disproportionate role of the non-neural cadherins with respect to evolution of the wide range of somatic morphology present in vertebrates today

Desmoglein 3, via an Interaction with E-cadherin, Is Associated with Activation of Src

Desmoglein 3 (Dsg3), a desmosomal adhesion protein, is expressed in basal and immediate suprabasal layers of skin and across the entire stratified squamous epithelium of oral mucosa. However, increasing evidence suggests that the role of Dsg3 may involve more than just cell-cell adhesion.To determine possible additional roles of Dsg3 during epithelial cell adhesion we used overexpression of full-length human Dsg3 cDNA, and RNAi-mediated knockdown of this molecule in various epithelial cell types. Overexpression of Dsg3 resulted in a reduced level of E-cadherin but a colocalisation with the E-cadherin-catenin complex of the adherens junctions. Concomitantly these transfected cells exhibited marked migratory capacity and the formation of filopodial protrusions. These latter events are consistent with Src activation and, indeed, Src-specific inhibition reversed these phenotypes. Moreover Dsg3 knockdown, which also reversed the decreased level of E-cadherin, partially blocked Src phosphorylation.Our data are consistent with the possibility that Dsg3, as an up-stream regulator of Src activity, helps regulate adherens junction formation

Plakophilin-3 Is Required for Late Embryonic Amphibian Development, Exhibiting Roles in Ectodermal and Neural Tissues

The p120-catenin family has undergone a significant expansion during the evolution of vertebrates, resulting in varied functions that have yet to be discerned or fully characterized. Likewise, members of the plakophilins, a related catenin subfamily, are found throughout the cell with little known about their functions outside the desmosomal plaque. While the plakophilin-3 (Pkp3) knockout mouse resulted in skin defects, we find larger, including lethal effects following its depletion in Xenopus. Pkp3, unlike some other characterized catenins in amphibians, does not have significant maternal deposits of mRNA. However, during embryogenesis, two Pkp3 protein products whose temporal expression is partially complimentary become expressed. Only the smaller of these products is found in adult Xenopus tissues, with an expression pattern exhibiting distinctions as well as overlaps with those observed in mammalian studies. We determined that Xenopus Pkp3 depletion causes a skin fragility phenotype in keeping with the mouse knockout, but more novel, Xenopus tailbud embryos are hyposensitive to touch even in embryos lacking outward discernable phenotypes, and we additionally resolved disruptions in certain peripheral neural structures, altered establishment and migration of neural crest, and defects in ectodermal multiciliated cells. The use of two distinct morpholinos, as well as rescue approaches, indicated the specificity of these effects. Our results point to the requirement of Pkp3 in amphibian embryogenesis, with functional roles in a number of tissue types

P-cadherin expression in breast cancer: a review

P-cadherin is frequently over-expressed in high-grade invasive breast carcinomas and has been reported to be an enhancer of migration and invasion of breast cancer cells, being correlated with tumour aggressiveness. In addition, expression of P-cadherin is well established as an indicator of poor prognosis in human breast cancer, which has stimulated our interest in studying its role in this setting. This review describes the most important findings on P-cadherin expression and function in normal mammary tissue and breast cancer cells, emphasizing that further research is required to elucidate the role played by this protein in human mammary tumours

Immunoglobulin, glucocorticoid, or combination therapy for multisystem inflammatory syndrome in children: a propensity-weighted cohort study

Background: Multisystem inflammatory syndrome in children (MIS-C), a hyperinflammatory condition associated with SARS-CoV-2 infection, has emerged as a serious illness in children worldwide. Immunoglobulin or glucocorticoids, or both, are currently recommended treatments. Methods: The Best Available Treatment Study evaluated immunomodulatory treatments for MIS-C in an international observational cohort. Analysis of the first 614 patients was previously reported. In this propensity-weighted cohort study, clinical and outcome data from children with suspected or proven MIS-C were collected onto a web-based Research Electronic Data Capture database. After excluding neonates and incomplete or duplicate records, inverse probability weighting was used to compare primary treatments with intravenous immunoglobulin, intravenous immunoglobulin plus glucocorticoids, or glucocorticoids alone, using intravenous immunoglobulin as the reference treatment. Primary outcomes were a composite of inotropic or ventilator support from the second day after treatment initiation, or death, and time to improvement on an ordinal clinical severity scale. Secondary outcomes included treatment escalation, clinical deterioration, fever, and coronary artery aneurysm occurrence and resolution. This study is registered with the ISRCTN registry, ISRCTN69546370. Findings: We enrolled 2101 children (aged 0 months to 19 years) with clinically diagnosed MIS-C from 39 countries between June 14, 2020, and April 25, 2022, and, following exclusions, 2009 patients were included for analysis (median age 8·0 years [IQR 4·2–11·4], 1191 [59·3%] male and 818 [40·7%] female, and 825 [41·1%] White). 680 (33·8%) patients received primary treatment with intravenous immunoglobulin, 698 (34·7%) with intravenous immunoglobulin plus glucocorticoids, 487 (24·2%) with glucocorticoids alone; 59 (2·9%) patients received other combinations, including biologicals, and 85 (4·2%) patients received no immunomodulators. There were no significant differences between treatments for primary outcomes for the 1586 patients with complete baseline and outcome data that were considered for primary analysis. Adjusted odds ratios for ventilation, inotropic support, or death were 1·09 (95% CI 0·75–1·58; corrected p value=1·00) for intravenous immunoglobulin plus glucocorticoids and 0·93 (0·58–1·47; corrected p value=1·00) for glucocorticoids alone, versus intravenous immunoglobulin alone. Adjusted average hazard ratios for time to improvement were 1·04 (95% CI 0·91–1·20; corrected p value=1·00) for intravenous immunoglobulin plus glucocorticoids, and 0·84 (0·70–1·00; corrected p value=0·22) for glucocorticoids alone, versus intravenous immunoglobulin alone. Treatment escalation was less frequent for intravenous immunoglobulin plus glucocorticoids (OR 0·15 [95% CI 0·11–0·20]; p<0·0001) and glucocorticoids alone (0·68 [0·50–0·93]; p=0·014) versus intravenous immunoglobulin alone. Persistent fever (from day 2 onward) was less common with intravenous immunoglobulin plus glucocorticoids compared with either intravenous immunoglobulin alone (OR 0·50 [95% CI 0·38–0·67]; p<0·0001) or glucocorticoids alone (0·63 [0·45–0·88]; p=0·0058). Coronary artery aneurysm occurrence and resolution did not differ significantly between treatment groups. Interpretation: Recovery rates, including occurrence and resolution of coronary artery aneurysms, were similar for primary treatment with intravenous immunoglobulin when compared to glucocorticoids or intravenous immunoglobulin plus glucocorticoids. Initial treatment with glucocorticoids appears to be a safe alternative to immunoglobulin or combined therapy, and might be advantageous in view of the cost and limited availability of intravenous immunoglobulin in many countries. Funding: Imperial College London, the European Union's Horizon 2020, Wellcome Trust, the Medical Research Foundation, UK National Institute for Health and Care Research, and National Institutes of Health