Back to Old Books Toward Affordable Research: Homemade Phenol-Based Reagent for Triphasic RNA Purification.

Covid-19 crisis did hit many socio-economic aspects in the whole world. In the scientific research, the problem is getting even worse, since most of materials and consumable are allocated to the health sector. Many research laboratories around the world have big delay in receiving their purchases to accomplish their research projects. In the developing countries, the situation is much more difficult, since most of the funding resources are directed to the Covid-19 crisis and there is a notable increase in reagents' prices. Therefore, the aim of the present study is to make a homemade reagents for RNA purification from eukaryotic cells/tissues. The homemade phenol-based RNA extraction reagents were prepared using saturated phenol pH 4.3 (adjusted by 0.5 M citrate buffer) and guanidine thiocyanate. To validate the phenol-based reagent, RNA was purified from different biological samples (cell line, tissues, and fungi) using homemade phenol-based versus a commercial one. Concentration of RNA samples extracted from the same number of cells were compared to assess the homemade preparation of phenol-based reagent. In conclusion, homemade phenol-based reagent is cost effective and comparable to the commercial one. Using homemade phenol-based, RNA extraction was successfully purified from different biological sources.

Secretome Analysis for a New Strain of the Blackleg Fungus Plenodomus lingam Reveals Candidate Proteins for Effectors and Virulence Factors.

The fungal secretome is the main interface for interactions between the pathogen and its host. It includes the most important virulence factors and effector proteins. We integrated different bioinformatic approaches and used the newly drafted genome data of P. lingam isolate CAN1 (blackleg of rapeseed fungus) to predict the secretion of 217 proteins, including many cell-wall-degrading enzymes. All secretory proteins were identified; 85 were classified as CAZyme families and 25 were classified as protease families. Moreover, 49 putative effectors were predicted and identified, where 39 of them possessed at least one conserved domain. Some pectin-degrading enzymes were noticeable as a clustering group according to STRING web analysis. The secretome of P. lingam CAN1 was compared to the other two blackleg fungal species (P. lingam JN3 and P. biglobosus CA1) secretomes and their CAZymes and effectors were identified. Orthologue analysis found that P. lingam CAN1 shared 14 CAZy effectors with other related species. The Pathogen-Host Interaction database (PHI base) classified the effector proteins in several categories where most proteins were assigned as reduced virulence and two of them termed as hypervirulence. Nowadays, in silico approaches can solve many ambiguous issues about the mechanism of pathogenicity between fungi and plant host with well-designed bioinformatics tools.

Synthesis, characterization and antimicrobial activity of a novel chitosan Schiff bases based on heterocyclic moieties.

Three heteroaryl pyrazole derivatives; namely 1-phenyl-3-(thiophene-2-yl)-1H-pyrazole-4-carbaldehyde, 1-phenyl-3-(furan-2-yl)-1H-pyrazole-4-carbaldehyde and 1-phenyl-3-(pyridine-3-yl)-1H-pyrazole-4-carbaldehyde were synthesized and reacted with chitosan to form Schiff bases of chitosan. All newly synthesized compounds have been characterized by solubility tests, elemental analysis, spectral (FTIR, 1H NMR) analyses, thermogravimetric analysis and X-ray diffraction (XRD). The Schiff bases were screened for their biological activity against gram-negative bacteria (Escherichia coli and Klebsiella pneumonia), gram-positive bacteria (Staphylococcus aureus and Streptococcus mutans) and fungi (Asperagillus fumigatus and Candida albican). The results indicated that the antimicrobial activity was dependent on the type of the Schiff base moiety. Cytotoxicity of the prepared chitosan derivatives was evaluated by MTT assay and the results indicated the absence of cytotoxic activity.

Mucormycosis in Cairo, Egypt: review of 10 reported cases.

We report on 10 cases of mucormycosis, as defined by The European Organization for Research and Treatment of Cancer and Mycoses Study Group (EORTC/MSG) standards of invasive fungal diseases, among patients with a recent history of neutropenia, prolonged use of corticosteroids and treatment with immunosuppressants. They were all observed at the Ain Shams University Specialized Hospital in Cairo, Egypt, during the year 2010. These cases were categorized as 50% proven and 50% probable, with none considered to be possible mucormycosis. The median age of the patients discussed in this report was 50 years (range 22-68 years), of which 80% were male and 20% were female. Uncontrolled diabetes with ketoacidosis was noted in 60% of cases, while 40% of the patients had undergone liver transplantations. Pulmonary mucormycosis was the predominant presentation as it was noted in 80% of cases, but there was only 20% sinus involvement. Members of the genus Lichtheimia were the most common etiologic agents (40% of all cases), whereas Rhizopus ssp. were recovered from 30% of cases, Syncephalastrum spp. in 20%, and 10% of patients were infected with Rhizomucor. Liposomal formulation of amphotericin B (LAMB) was successfully used to treat all the cases described in this report. We concluded that the incidence of mucormycosis was relatively high during the study period in this one-center study and that additional studies looking into the diagnosis and the control of mucormycosis in Egypt are required.