DTT protein equalization and Tryptophan protein quantification as a powerful tool in analytical proteomics.

Assessing total protein levels in biological samples is a common procedure in biochemistry and molecular biology. In this study, we compare tryptophan fluorescence (WF) with Bradford and BCA assays to determine total protein in serum samples. Our results indicate that tryptophan fluorescence spectrometry is an efficient, sensitive, and straightforward technique for quantifying proteins in serum. We observed minimal variation between the three methods: BCA de one with the lowers LOD and LOQ. The tryptophan method offers the possibility of reusing the intact sample that does not need colourimetric reagents for quantification. Consequently, free tryptophan serves as a reliable universal standard. This assay can be performed using a conventional fluorescence spectrometer with cuvettes or in a 96-well plate format with a plate reader. The method was successfully used as proof of concept, using serum from patients diagnosed with myeloma and serum from healthy donors

Navigating the molecular diversity of SARS-CoV-2: early pandemic insights from comparative phylogenetic analysis: SARS-CoV-2 Clades Phylogeny

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in December 2019 precipitated the onset of the COVID-19 pandemic, which swiftly spread across more than 214 countries and territories, posing a significant global health crisis. In response, laboratories worldwide have embarked on extensive efforts to characterize the genomic landscape of the virus, employing a myriad of sophisticated genomic analysis techniques. This study endeavors to undertake a comprehensive exploration into the genetic diversity, geographical distribution, and virulence determinants of SARS-CoV-2 clades across 11 diverse countries, employing advanced computational biology methodologies. Leveraging molecular data sourced from prominent international databases, the analysis aims to unravel the intricate phylogenetic relationships and mutational dynamics exhibited by various viral strains circulating worldwide. The findings of this investigation promise to yield invaluable insights into the evolutionary trajectory of SARS-CoV-2, shedding light on potential therapeutic targets and informing strategies for mitigating the impact of the ongoing pandemic on global public health. Results highlight significant genetic diversity among SARS-CoV-2 strains across different countries, with phylogenetic analysis revealing distinct subclass groupings within each country. A manual comparison of sequences identified numerous mutations, with certain mutations associated with increased virulence. Comparison of clade G and clade O sequences revealed differences in mutation profiles, suggesting potential links to virulence and transmissibility. These findings underscore the dynamic nature of SARS-CoV-2 evolution and the importance of monitoring genetic changes for public health interventions

Ultrasound-Assisted Extraction- and Liquid Chromatography-Based Method Development and Validation for Obtaining and Qualitative Determination of Apple Pomace Three Triterpene Acids using Analytical Quality by Design

Background: Apple pomace has garnered significant attention within the life sciences domain due to its underutilized status as a waste material from apple processing. It represents a cost-effective and abundant source of triterpene acids due to its multifunctional clinical, nutritional, and pharmaceutical benefits. Purpose: The present study aimed to develop and validate a new, selective, effective, robust and reproducible laboratory methodology based on extraction, purification and analytical procedures to obtain and determining three major triterpene acids – Ursolic acid (UA), Oleanolic acid (OA) and Betulinic acid (BA) into the dry extracted product from apple pomace. Method: A new, cost-efficient, rapid, selective and high-yield two-stage ultrasound-assisted extraction procedure was developed and the effect of critical parameters: ultrasonic power, extraction time, solvent volume, temperature, and the amount of raw material on the extraction process were investigated. The dry column vacuum chromatography technique was used for purification to remove unwanted non-polar and polar impurities from the target bioactive compounds; A new, effective, specific, sensitive, and rapid HPLC analytical procedure was developed using analytical quality by design (AQbD) approach and validated according to ICH guidelines. Conclusion: The method has a good accuracy (the mean recovery >95 %) and linearity (R2>0.999).  The limit of quantitation (LOQ) is 0.0001 mg/mL for UA, 0.00005 mg/mL for OA and 0.000025 mg/mL for BA.  The validation results confirm that the method is specific, precise and robust. The purity of the extracted and purified target product from apple pomace is not less than 93 %. The developed laboratory methodology is capable of being considered for industrial purposes and through the appropriate technology transfer process can be successfully transferred to the industrial scale

Computational Screening of Natural Compounds as Antiviral Candidates Targeting the SARS-CoV-2 Main Protease

The onset of the global pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in Wuhan in December 2019 has led to an urgent need for effective preventive and therapeutic solutions. Among the various approaches explored, natural products have shown potential in the fight against COVID-19. This study employed computational techniques to screen and evaluate six natural antiviral compounds for their effectiveness against the SARS-CoV-2 Main Protease (Mpro). By using molecular docking simulations, the interactions between these natural compounds and the target proteins were predicted and analyzed, focusing on factors such as binding affinity, interaction patterns, and structural compatibility within the active sites. The analysis indicated that Cladosin C and Rhodatin formed the most stable interactions with Mpro, engaging with several critical residues. Cannabidiol, Capsaicin, and Kappa-Carrageenan also demonstrated promising interactions, though with some variability. On the other hand, Astaxanthin exhibited the least stable binding, suggesting limited antiviral potential. This research provides insights into the possible roles of these natural compounds as antagonists of the SARS-CoV-2 Mpro enzyme. Further in vitro and in vivo studies are necessary to confirm the antiviral properties of these compounds, and future research should investigate their broader antiviral applications

Antibiotic resistance and the one health approach. The importance of teaching OMICS at Universities.

Letter to the edito

INSIGHTS INTO THE USE OF MICROSAMPLING FOR OMICS STUDIES

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Computational Nanomedicine: Helping Us Get Through COVID-19

This is a letter to the editor so there is no astract