Effects of Rhizopus Nigricans Exopolysaccharide on Proliferation, Apoptosis, and Migration of Breast Cancer MCF-7 Cells and Akt Signaling Pathway

Objective. To study the effect of Rhizopus nigricans exopolysaccharide EPS1-1 on the proliferation, apoptosis, and migration of breast cancer MCF-7 cells. Methods. Human breast cancer MCF-7 cells were cultured in vitro and treated with different concentrations of EPS1-1. The effect of EPS1-1 on cell proliferation was tested by the CCK-8 experiment, and the effect of EPS1-1 on cell apoptosis was determined by flow cytometry. And the scratch test was used to detect the impact of EPS1-1 on cell migration. Western blot then was used to measure the expression changes of related proteins in the Akt signaling pathway. Results. Compared with the control group, treatment with EPS1-1 significantly reduced the proliferation, migration, and invasion ability of MCF-7 cells and promoted the apoptosis of MCF-7 cells in a dose-dependent manner. In terms of the underlying mechanism, EPS1-1 can significantly inhibit the phosphorylation of Akt at threonine 308 and serine 473 and cause the expression changes of downstream proliferation-related genes CCND1 and p21, apoptosis-related genes Bcl-2 and Bax, and migration-related genes Vimentin and E-cadherin in terms of their protein levels. Conclusion. EPS1-1 can inhibit the proliferation, migration, and invasion of breast cancer MCF-7 cells and promote the apoptosis of MCF-7 cells by inhibiting the activation of the Akt signaling pathway. Therefore, EPS1-1 can be used as a potential new drug or adjuvant drug for the treatment of breast cancer.

Rubesanolides F and G: Two Novel Lactone-Type Norditerpenoids from Isodon rubescens

A phytochemical investigation of the leaves of the medicinal plant Isodon rubescens led to the isolation of the two new degraded abietane lactone diterpenoids rubesanolides F (1) and G (2). Their structures were elucidated based on the analyses of the HRESIMS and 1D/2D NMR spectral data, and their absolute configurations were determined by ECD spectrum calculations and X-ray single crystal diffraction methods. Compounds 1 and 2, with a unique γ-lactone subgroup between C-8 and C-20, were found to form a carbonyl carbon at C-13 by removal of the isopropyl group in an abietane diterpene skeleton. Rubesanolide G (2) is a rare case of abietane that possesses a cis-fused configuration between rings B and C. The two isolates were evaluated for their biological activities against two cancer cell lines (A549 and HL60), three fungal strains (Candida alba, Aspergillus niger and Rhizopus nigricans) and three bacterial strains (Escherichia coli, Staphylococcus aureus and Bacillus subtilis).

Microbiota Fúngica en Quesos Artesanales en Saltillo, Coahuila, México

El queso es un alimento importante en la cocina mexicana, el consumo per cápita en México es de 6 kg; sin embargo, puede transportar numerosos hongos dañinos para las personas, como micotoxigénicos. El objetivo del trabajo fue evaluar la microbiota fúngica presente en los quesos artesanales del mercado abierto en Saltillo, Coahuila, México, se obtuvieron cinco tipos de quesos por semana de diferentes orígenes; uno de Saltillo, dos del general Cepeda (A y B), uno de Mazapil, Zacatecas y uno de Ciudad Fernández, San Luis Potosí. Cada queso se cortó en tres niveles (réplicas), y se tomó una muestra de 1 cm en el norte, sur, este, oeste y centro del queso, con cuatro réplicas, que se colocaron en placas de Petri con medio de cultivo PDA mantenido a 25 °C ± 2 °C durante 120 h. Los hongos se identificaron utilizando la técnica de Medina y colaboradores. Geotrichum candidum, Fusarium verticillioides, Rhizopus nigricans, Cladosporium sp. y Penicillium sp. fueron identificados morfológicamente, con una incidencia entre 0 y 100%. Fusarium verticillioides y Penicillium sp. son hongos toxigénicos, y Geotrichum candidum causa infección en los bronquios, pulmones y membranas mucosas, llamada geotricosis, por lo que es importante implementar medidas de calidad para garantizar la producción segura de quesos para el consumo humano.

Screening of Microflora Associated with Brinjal Seeds (Solanum melongena L.) in Western Part of India

Total 110 seed samples of brinjal (Solanum melongena L.) were collected from 12 major growing districts of Rajasthan and subjected to dry seed examination (DSE). DSE revealed asymptomatic (07.75-97.5%), moderately discolored (04.50-67.50%) and shriveled discolored (03.25-38.75%) seeds. Symptomatic seeds in DSE showed various types of discolorations, deformation like white crust on seed surface. The seeds with water soaked symptoms and spots on seed surface were also observed such seeds on incubation yielded bacterial species. The incubation of symptomatic seeds about 21 fungal species of 14 genera and 3 bacterial species were observed viz. Alternaria alternata, A. solani, Aspergillus flavus, A. niger, Cladosporium oxysporium, Curvularia lunata, Fusarium oxysporium, F. solani, Rhizoctonia bataticola and Rhizopus nigricans etc and bacterial species like Xanthomonas axonopodis var. vesicatoria, Ralstonia solanacearum and Pseudomonas aeruginosa. The microflora severely affects seed germination (failure or delayed germination), wilting and rotting of seedlings, bacterial oozing, collapse of hypocotyls and cotyledonary leaves which resulting seedling mortality. High yield of microflora was obtained on standard blotter method (SBM) and agar plate method (APM).

Mycological profiling of raspberry cultivars in storage

Economic losses incurred during raspberry storage are primarily a result of fungal spoilage. This study aimed to characterise the micromycete phytopathogenic complex in stored raspberries. The fungal microbiome was assessed in six Russian raspberry cultivars (Cleopatra, Novost’ Kuzmina, Sulamiph, Fregat, Shakhrazada, Yarkaya) as well as three foreign varieties (Glen Lyon, Cascade Delight, Limonnaya). The economic and biological traits of the cultivars were studied within the “Programme and Protocol for Fruit, Berry and Nut Crops Varietal Testing”. Internal and surfacelevel contamination in infected fruit tissues was detected via the stimulation of microbial growth in a wet chamber. The micromycete species composition was assessed via commonly used methods. The following raspberry spoilage organisms were identified as being predominant. Rots: black (Alternaria alternata (Fr) Keissler), dry (Fusarium Link.), leather (Phytophthora cactorum (Leb. et Cohn) Schroet), grey (Botrytis cinerea Pers.); moulds: black (Cladosporium Link.), green-blue (Penicillium Link.), green-yellow (Aspergillus P. Micheliex Haller), grey capitate (Rhizopus nigricans Ehrenb.), capitate (Mucor spp.); drupe rot (Phragmidium rubi Wint.); bacterioses. Microbiotic profiles varied between cultivars in storage. The fungal genera Fusarium and Alternaria prevailed, with the former dominating in almost all samples. The phytopathogenic complex primarily consisted of the fungal genera Fusarium, Alternaria and Penicillium. Raspberry cultivars which had a weak (Sulamiph, Shakhrazada, Novost’ Kuzmina) or strong (Cleopatra) resistance to micromycetes in storage were identified.

Non-allergic severe asthma: is it really always non-allergic? The IDENTIFY project

Background This differential diagnosis of allergic vs non-allergic asthma is typically made on the basis of sensitization to allergens, such that absence of sensitization could result in a patient being managed as having non-allergic asthma. In Germany, the number of specific allergen tests is limited and non-standardized (across clinicians and laboratories), with the potential for false negative diagnoses. IDENTIFY aimed to gain data on sensitizations toward aeroallergens in patients with severe asthma who had tested negative to perennial aeroallergens in previous tests. Methods This was a single visit, non-randomized, non-interventional study conducted in 87 centers across Germany. The only inclusion criteria were that patients had to be adults (at least 18 years of age) with a diagnosis of severe asthma (receiving at least Global Initiative for Asthma Step IV therapy), and who had previously tested negative to perennial aeroallergens. Patients were then tested for sensitization to a panel of 35 perennial aeroallergens, with positive sensitization indicated by CAP ≥ 0.35 kU/L. Results Of 588 patients recruited, 454 had complete and valid data, and had previously tested negative to perennial aeroallergens. Overall, 43.6% of the analyzed patients tested positive for at least one of the included aeroallergens, with 18.7% testing positive for three or more, and 4.2% positive for more than ten. The five most common sensitizations were to Staphylococcus aureus enterotoxin B, Aspergillus fumigatus, Candida albicans, Dermatophagoides farinae, and Rhizopus nigricans, each of which tested positive in at least 9.7% of the population. Conclusions In this group of patients being managed as having non-allergic asthma (and who had all previously tested negative to perennial aeroallergens), a high proportion tested positive to a broad panel of aeroallergens. A diagnosis of allergic asthma therefore cannot be excluded purely on the basis of standard aeroallergen panels.

Monitoring of Soils Mycobiota of the Yaroslavl Region Agrocoenosis with Different Methods of Сultivation and Application of Fertilizers

Приведены результаты мониторинга состояния микобиоты почвы, начатого в 2005 г. с целью прогноза последствий сельскохозяйственного использования земель. В условиях многофакторного стационарного опыта по изучению поверхностно-отвальной системы обработки, заложенного методом расщеплённых делянок с рендомизированным размещением вариантов в повторениях, проведены наблюдения за динамикой видового состава и численности микромицетов в зависимости от используемых агротехнологических приёмов, включающих системы основной обработки почвы и удобрений. Всего за период 2011–2015 гг. выявлено 22 рода и 101 вид микроскопических грибов. Анализ показал, что в агроценозах комплекс грибов несколько отличается от типичного для дерново-подзолистых почв естественных биоценозов. Состав доминирующей группы в целом за время исследований почти не изменялся и был представлен видами р. Penicillium. В число содоминантов входили зигомицетовые сапротрофы (Rhizopus nigricans Ehrenb., Mucor hiemalis Wehmer и виды р. Mortierella), смена которых в большей степени зависела от внешних условий. Часто обнаруживались грибы рр. Alternaria, Aspergillus, Cladosporium, Fusarium, Verticillium, содержащие фитопатогенные виды. Однако их встречаемость и численность не проявляли чёткой зависимости от таких факторов, как обработка почвы и внесение удобрений. Расчёт индексов биоразнообразия Шеннона и Пиилу показал, что при поверхностно-отвальной системе обработки в год вспашки разнообразие грибных комплексов снижается, происходит резкая их перестройка за счёт перемещения видов, обитающих в разных по глубине слоях почвы. Подобные изменения касаются и численности микромицетов. Отмечено снижение КОЕ грибов, особенно на неудобренном фоне, после проведения вспашки. В целом, как при отвальной, так и поверхностно-отвальной системах обработки показано возрастание численности микромицетов при внесении соломы 3 т/га + NPK. Запаханная на глубину пахотного горизонта солома с минеральными удобрениями, являясь хорошим питательным субстратом для грибов, на большинстве вариантов приводила к увеличению их количества. The results of monitoring the state of soil mycobiota started in 2005 with the aim of predicting the consequences of agricultural land use are presented. Under the conditions of a complex stationary experiment on the study of a surface-moldboard tillage system, laid down by the method of split plots with a randomized block design of variants in repetitions, observations were made of the dynamics of the species composition and number of micromycetes, depending on the agricultural technological methods used, including the systems of basic tillage and fertilizers. Total for the period 2011–2015 22 genera and 101 species of microfungus have been identified. The analysis showed that in agrocoenosis the complex of fungi is slightly different from the natural biocoenosis typical for soddy podzolic soils. The composition of the dominant group as a whole remained almost unchanged during the studies and was represented by the species of the Penicillium. The codominants included zygomycete saprotrophs (Rhizopus nigricans Ehrenb., Mucor hiemalis Wehmer and species of Mortierella), the change of which was more dependent on external conditions. Fungi Alternaria, Aspergillus, Cladosporium, Fusarium, Verticillium containing phytopathogenic species were often found. However, their occurrence and number did not show a clear dependence on such factors as tillage and fertilization. The calculation of the Shannon and Pielou biodiversity indices showed that with a surface-moldboard system of tillage in a year of plowing, the variety of fungal complexes decreases, and their sharp restructuring occurs due to the movement of species living in layers of soil different in depth. Such changes concern the number of micromycetes. A decrease in CFU of fungi was noted, especially on an unfertilized background after plowing. In general, both moldboard and surface-moldboard tillage systems show an increase in the number of micromycetes with the introduction of straw 3 t/ha + NPK. Straw plowed under the depth of the plough-layer with mineral fertilizers, being a good nutrient substrate for mushrooms, in most cases led to an increase in their amount.

Objective: Antifungal activity of total ethanolic extract and fractions of Usnea varieties were proved against Aspergillus niger, Penicillium digitatum, and Rhizopus nigricans fungi. Methods: To perform the study of relative antifungal activity (AFR), flu

Objective: Antifungal activity of total ethanolic extract and fractions of Usnea varieties were proved against Aspergillus niger, Penicillium digitatum, and Rhizopus nigricans fungi. Methods: To perform the study of relative antifungal activity (AFR), fluconazole and ketoconazole were used as standards. The inhibition coefficient 50 (IC50) for each extract and the fractions was determined. Results: It was established that the highest activity was presented by the acetone fraction with a value of 58.53. In the study of antifungal activity, the fraction that presented the best activity against P. digitatum was ethanolic with a value of 44.33, while for R. nigricans was petrol extract, with a value of 75.35 and finally for A. niger was the total extract with a value of 35.48. Conclusions: Comparing the values obtained from the extract and the different fractions resulted in the dichloromethane fraction showing the best values.