The involvement of RIPK4 in TNF-α-stimulated IL-6 and IL-8 production by melanoma cells.

PURPOSE: The receptor-interacting protein kinase (RIPK4) has an oncogenic function in melanoma, regulates NF-κB and Wnt/ß-catenin pathways, and is sensitive to the BRAF inhibitors: vemurafenib and dabrafenib which lead to its decreased level. As its role in melanoma remains not fully understood, we examined the effects of its downregulation on the transcriptomic profile of melanoma. METHODS: Applying RNA-seq, we revealed global alterations in the transcriptome of WM266.4 cells with RIPK4 silencing. Functional partners of RIPK4 were evaluated using STRING and GeneMANIA databases. Cells with transient knockdown (via siRNA) and stable knockout (via CRISPR/Cas9) of RIPK4 were stimulated with TNF-α. The expression levels of selected proteins were assessed using Western blot, ELISA, and qPCR. RESULTS: Global analysis of gene expression changes indicates a complex role for RIPK4 in regulating adhesion, migration, proliferation, and inflammatory processes in melanoma cells. Our study highlights potential functional partners of RIPK4 such as BIRC3, TNF-α receptors, and MAP2K6. Data from RIPK4 knockout cells suggest a putative role for RIPK4 in modulating TNF-α-induced production of IL-8 and IL-6 through two distinct signaling pathways-BIRC3/NF-κB and p38/MAPK. Furthermore, increased serum TNF-α levels and the correlation of RIPK4 with NF-κB were revealed in melanoma patients. CONCLUSION: These data reveal a complex role for RIPK4 in regulating the immune signaling network in melanoma cells and suggest that this kinase may represent an alternative target for melanoma-targeted adjuvant therapy.

The Combined Effects of Azoxystrobin and the Biosurfactant-Producing Bacillus sp. Kol B3 against the Phytopathogenic Fungus Fusarium sambucinum IM 6525.

This study aimed to evaluate how the combined presence of the synthetic fungicide azoxystrobin (AZ) and the biosurfactant-producing Bacillus sp. Kol B3 influences the growth of the phytopathogenic fungus Fusarium sambucinum IM 6525. The results showed a noticeable increase in antifungal effectiveness when biotic and abiotic agents were combined. This effect manifested across diverse parameters, including fungal growth inhibition, changes in hyphae morphology, fungal membrane permeability and levels of intracellular reactive oxygen species (ROS). In response to the presence of Fusarium and AZ in the culture, the bacteria changed the proportions of biosurfactants (surfactin and iturin) produced. The presence of both AZ and/or Fusarium resulted in an increase in iturin biosynthesis. Only in 72 h old bacterial-fungal co-culture a 20% removal of AZ was noted. In the fungal cultures (with and without the addition of the bacteria), the presence of an AZ metabolite named azoxystrobin free acid was detected in the 48th and 72nd hours of the process. The possible involvement of increased iturin and ROS content in antifungal activity of Bacillus sp. and AZ when used together are also discussed. Biosurfactants were analyzed by liquid chromatography with tandem mass spectrometry (LC-MS/MS). Microscopy techniques and biochemical assays were also used.

Identification of the selected soil bacteria genera based on their geometric and dispersion features.

The visual analysis of microscopic images is often used for soil bacteria recognition in microbiology. Such task can be automated with the aid of machine learning and digital image processing techniques. The best results for soil microorganism identification usually rely on extracting features based on color. However, accommodating in the latter an extra impact of lighting conditions or sample's preparation on classification accuracy is often omitted. In contrast, this research examines features which are insensitive to the above two factors by focusing rather on bacteria shape and their specific group dispersion. In doing so, the calculation of layout features resorts to k-means and mean shift methods. Additionally, the dependencies between specific distances determined from bacteria cells and the curvature of interpolated bacteria boundary are computed to extract vital geometric shape information. The proposed bacteria recognition tool involves testing four different classification methods for which the parameters are iteratively adjusted. The results obtained here for five selected soil bacteria genera: Enterobacter, Rhizobium, Pantoea, Bradyrhizobium and Pseudomonas reach 85.14% classification accuracy upon combining both geometric and dispersion features. The latter forms a promising result as a substitutive tool for color-based feature classification.

Arbuscular Mycorrhizal Fungi as an Important Factor Enabling the Adaptation of Anthyllis vulneraria L. to Zn-Pb-Polluted Tailings.

The old Zn-Pb-contaminated (calamine) tailings in southern Poland are spontaneously colonized by metal-tolerant Anthyllis vulneraria L. (Fabaceae), which can form simultaneously symbiotic association with nitrogen-fixing rhizobia and phosphorus-acquiring arbuscular mycorrhizal fungi (AMF). So far, fungal colonization and the AMF diversity of calamine-inhabiting legumes have been poorly studied. Thus, we determined AMF spore density in the substratum and the mycorrhizal status of nodulated A. vulneraria plants occurring on calamine tailings (M) and on a reference non-metallicolous (NM) site. The results indicate the presence of the Arum-type of arbuscular mycorrhiza in the roots of both Anthyllis ecotypes. Despite the presence of AM fungi in M plant roots, the dark septate endophyte (DSE) fungi (hyphae and microsclerotia) were occasionally also detected. Metal ions were accumulated mainly in the nodules and intraradical fungal structures rather than thick plant cell walls. Mycorrhization parameters (frequency of mycorrhization and intensity of root cortex colonization) for M plants were markedly higher and differed in a statistically significant manner from the parameters for NM plants. Heavy metal excess had no negative effect on the number of AMF spores, the amounts of glomalin-related soil proteins and AMF species composition. Molecular identification of AMF using PCR-DGGE analysis based on the 18S rDNA ribosomal gene by nested-PCR with primers AM1/NS31 and NS31-GC/Glo1 revealed similar genera/species of AMF in the roots of both Anthyllis ecotypes: Rhizophagus sp., R. fasciculatus, and R. iranicus. The results of this work indicate the presence of unique fungal symbionts, which may enhance A. vulneraria tolerance to heavy metal stress and plant adaptation to extreme conditions on calamine tailings.

Correlation among some phenological and biochemical traits in date palm (Phoenix dactylifera L.) germplasm.

Date palm is an economically important fruit crop in the arid and semi-arid regions of the world. The knowledge of genetic variation, genotype-by-traits comparison, as well as the relationship among several phenological traits is effective for developing breeding populations by choosing the best allelic combinations and employing selection approaches. Information regarding the genetic variability and inter-relationships among fruit characteristics is facilitating the plant breeders to efficiently exploit the date palm germplasm for the introduction of elite genotypes. The present study was conducted to examine genetic variation among different date palm cultivars -collected from two different research stations in Pakistan as well as the relationship among their phenological and biochemical characteristics. Biplot analysis revealed that the cultivars 'Dhakki', 'Chohara', and 'Kupra' possessed the highest fruit and pulp weights. Moreover, the fruits of the cultivars 'Champa Kali' and 'Shakri' had the maximum TSS (total soluble solids) and total sugar content. Similar variations were observed in the mean values of the studied phenological and biochemical characteristics as in the biplot analysis. Cluster analysis on the basis of phenological and biochemical characteristics divided 50 cultivars into seven clusters, showing differences in the studied characters. A correlation matrix revealed that pulp weight had a strong relationship with fruit weight, length, and diameter. A strong correlation also existed among antioxidant capacity, antioxidant activity, and total phenolic content. These results demonstrated a huge diversity in phenological and biochemical characteristics of date palm cultivars as well as a correlation among several characteristics of the studied germplasm, which can be exploited in future breeding programs.

The role of RIPK4 in epidermis physiology / Rola bialka RIPK4 w fizjologii naskórka.

Proteins which regulate morphogenesis of the epidermis ensure its proper construction and function and mutations or abnormal expression of those proteins impact epidermal function. One recently described protein is Receptor Interacting Serine/Threonine Kinase 4 (RIPK4). Mutations in RIPK4 cause the autosomal-recessive form of Bartsocas-Papas syndrome and Popliteal Pterygium Syndrome the Aslan type. In mice, deletion of Ripk4 (Ripk4-/-) leads to premature death of neonates caused by numerous skin adhesions. RIPK4 regulates development and maintenance of differentiation and proliferation homeostasis of keratinocytes, as well as inflammation. RIPK4 also appears to act as a tumor suppressor in skin, since expression is decreased in squamous skin carcinoma and inducible deletion in mice facilitates induction and growth of papillomas and squamous skin carcinomas in chemical carcinogenesis. The review describes the RIP family, the engagement of RIPK4 in differentiation of the epidermis and consequences resulting from its improper expression.

Identification and assessment of genetic similarity of soil bacterial isolates of Pseudomonas spp. using molecular techniques.

Bacteria of the genus Pseudomonas are often components of bioproducts designed to enhance the condition of the soil and plants. The use of Pseudomonas bacteria in bioproducts must be preceded by the acquisition, characterization and selection of beneficial strains living in the soil. A prerequisite for the selection of bacterial strains for use in bioproducts is to be able to identify the isolates rapidly and accurately. To identify and differentiate 15 bacterial isolates obtained from the soil surrounding the roots of sour cherry trees and to assess their genetic similarity, the rep-PCR technique and restriction analysis of the 16S rRNA gene and the 16S-ITS-23S rRNA operon were used. In addition, a sequence analysis of the 16S rRNA gene was performed. The analyses made it possible to divide the isolates into four clusters and to confirm their affiliation with the Pseudomonas species. RFLP analysis of the 16S-ITS-23S rRNA operon enabled greater differentiation of the isolates than RFLP of the 16S rRNA gene. The greatest differentiation of isolates within the clusters was obtained after using the rep-PCR technique. However, none of the techniques was able to discriminate all the isolates, which indicates very high genetic similarity of the Pseudomonas isolates found in the same sample of soil from around the roots of sour cherry trees. The tests performed will find application for distinguishing and identifying Pseudomonas strains collected from the soil in order to select the most valuable bacterial strains that produce beneficial effects on plants.