Current Pharmaceutical Biotechnology - Current Issue

Antibiofilm agents serve as an essential tool in the fight against antibiotic resistance, and natural products provide a promising source for potential drug leads.

This study investigates the activity of twenty Bangladeshi medicinal plants against Staphylococcus aureus and Pseudomonas aeruginosa biofilms and predicts the interactions of selected phytochemicals from five of the best performing plants with the active sites of transcriptional regulatory proteins SarA of S. aureus and LasR of P. aeruginosa.

The plant extracts were tested by microtiter plate-based assay against S. aureus and P. aeruginosa biofilms. Molecular docking and molecular dynamics simulation (MD) were conducted using PyRx and GROMACS, respectively.

The best activity was identified for Cassia fistula and Ananas comosus, showing ≥ 75% inhibition of biofilm formation. ent-Epicatechin-(4α→8)-epiafzelechin (EEE) of C. fistula, cyanidin-3,3',5-tri-O-β-D-glucopyranoside (CTG) of A. comosus, and 7-O-(4-hydroxy-E-cinnamoyl)-spinoside of A. spinosus showed the best predictive binding affinity (-7.6, -7.6 and -7.7 kcal/mol, respectively) for SarA. EEE was the only ligand to exhibit a stable ligand-protein complex with SarA in the MD simulation of 200 ns (binding energy of MMPBSA analysis -39.899 kJ/mol). Chrysophanol, epicatechin and physcion, of C. fistula (-10.5, -10.5, and -11.0 kcal/mol, respectively) and auraptene of F. limonia (-10.8 kcal/mol) showed the best predictive binding affinity for LasR. Epicatechin showed the most stable ligand-protein complex with LasR (binding energy of MMPBSA analysis -63.717 kJ/mol).

Epicatechin and its derivative EEE could be used as scaffolds for the development of new antibiofilm agents against P. aeruginosa and S. aureus, respectively.

To identify potential phytochemical-based drugs for both wild-type and Omicron variants of SARS-CoV-2 using virtual screening and molecular dynamic simulation.

Coronavirus disease 2019 (COVID-19) is an infectious viral disease caused by SARS-CoV-2. Since 2019, multiple variants have been reported from all over the world and the emergence of new variants of SARS-CoV-2 is a major concern.

To identify potential phytochemicals that can be used as drugs against different variants of SARS-CoV-2.

In our present study, we have selected 594 phytochemicals and performed virtual screening to identify potential drug candidates. The screening commenced with molecular docking techniques with both ACE2 (Human) and Spike protein (wild-type and Omicron variant), followed by prediction of pharmacokinetics parameters and toxicity. The Schrodinger tools, Swiss ADME, and ProTox-II accomplish the analysis. Further, molecular dynamics simulation, binding free energy calculation and meta-dynamics study was performed for best protein-ligand complexes of both proteins using GROMACS and gmx_MMPBSA to validate the stability of the docked complexes.

We have identified 6 and 4 drugs as spike protein inhibitors for wild-type and Omicron variants, respectively. 6 drugs were identified as ACE2 receptor inhibitors. We have identified silymarin as a common drug inhibitor for both pathogen (Wild-type, and Omicrons spikes) as well as host (human ACE2) protein that reflects its ability to inhibit the host-pathogen interaction and prevent infection.

We have found some potential Phytochemicals that can be used against different variants of SARS-CoV-2 such as silymarin.

The aim of this study was to develop a clinical application model for the rational use of caffeine.

Caffeine is related to the incidence of neuro immune gastrointestinal diseases. Coffee consumption needs to be optimized in order to reduce the incidence rate.

By using KEEG analysis to explore potential molecular signaling pathways involved in the progression of neurological immune gastrointestinal diseases, and analyzing the details of this signaling Pathway using molecular simulation results, which can support AI system for doctor.

The research team designed a controlled experiment to analyze the differences in reward and reinforcement of Brain pleasure/addiction and dopamine related signaling pathways function between multiple groups of people with different coffee drinking habits and a blank control group. The study team used molecular dynamics methods to investigate the signaling route that links coffee with the binding of dopamine receptor D3.AI is used to predict the prevalence of gastric reflux disease.

Human experiments have shown a correlation between caffeine intake and gastroesophageal reflux disease. AI algorithm results can provide clinical support, and molecular simulation results are consistent with human experimental results. Caffeine and DRD3 protein have a stable interaction system.

The research team elucidated the intermolecular interaction between caffeine and DRD3, and AI algorithms can predict the likelihood of disease occurrence, providing a new strategy for clinical practice. This study has passed ethical approval at Chifeng Cancer Hospital, and the ethical documents for this study have been submitted to the World Health Organization for filing.

Background: Melanoma is considered the deadliest form of skin cancer. While monoclonal-antibodies and molecular targets marked milestones in melanoma therapy, more research is needed to overcome the advanced stages of this disease.

To explore the possible use of the yeast cytosine deaminase::uracil phosphoribosyltransferase fusion enzyme/5-fluorocytosine (CD::UPRT/5FC) suicide gene (SG) system for human melanoma.

In eight metastatic human melanoma cell lines, we determined: cytotoxicity, lipofection efficiencies, colony forming capacity and bystander effects due to soluble and/or particulate factors secreted to the conditioned media after treatments.

CD::UPRT induced cell death in a prodrug (5FC) concentration-dependent manner and was able to eliminate the sub-population of surviving cells with clonogenic capacity. Compared with human interferon-β gene transfer or the herpes simplex virus thymidine kinase/ganciclovir system, at 100 µM 5FC, CD::UPRT was more efficient in inducing cell death. The strong cytotoxic response contrasted with the low lipofection efficiencies (<5%), indicating a potent bystander effect. We analyzed the contribution of soluble and particulate factors released by SG lipofected cells to the conditioned media (CM) finding that they were able to deliver CD::UPRT genetic information and/or recombinant enzyme to recipient cells. When exposed to 5FC, the cells that received either supernatant or 12000×g pellet fractions of CM, efficiently activated the prodrug because of the acquired CD::UPRT activity and caused cell death.

This suicide gene therapy approach, amplified by the release of free 5-fluorouracil and soluble and particulate factors containing CD::UPRT genetic information and/or enzyme, could have a great clinical potential for malignant melanoma.

Prostate cancer is an androgen-dependent malignancy, and the use of androgen deprivation therapies frequently results in treatment resistance, relapse, and the development of aggressive castration-resistant tumors. Patchouli alcohol, a tricyclic sesquiterpene derived from Pogostemon cablin of the Labiatae family, has demonstrated potential in modulating inflammatory responses and tumor progression. This study aimed to investigate the mechanisms through which patchouli alcohol influences inflammatory pathways associated with prostate cancer using bioinformatics and experimental validation.

Differentially Expressed Genes (DEGs) were identified from the GSE46602 dataset, containing 36 prostate cancer and 14 normal prostate biopsy samples, using the GEO2R tool (adjusted P < 0.05). Functional annotation was performed using GO and KEGG databases, while PPI networks were constructed via STRING and Cytoscape. Key hub genes were identified. To validate the bioinformatics findings, qPCR and Western blotting were employed to confirm the differential expression of selected hub genes in DU145 prostate cancer cells treated with patchouli oil.

Bioinformatic analysis revealed 71 DEGs, including 35 upregulated and 36 downregulated genes. Thirteen hub genes were identified (DCK, APRT, ADK, KCNK9, ADSL, PKM, KCNK3, S100A10, ENTPD2, PKLR, ARHGEF38, TPK1, and AK5), which were enriched in pathways, such as MAPK, PI3K-Akt, Ras, and Rap1. Experimental validation confirmed the upregulation of DCK, APRT, KCNK9, ADSL, PKM, S100A10, ENTPD2, PKLR, ARHGEF38, and AK5, and the downregulation of ADK, KCNK3, and TPK1 at both the mRNA and protein levels.

Patchouli alcohol appears to influence multiple hub genes associated with prostate cancer progression through its modulation of key cellular signaling and metabolic pathways. These findings support its potential role as a therapeutic agent for prostate cancer.

Nanoparticles used in enzyme immobilization offer a high surface area-to-volume ratio, high chemical and thermal stability, and resistance to microbial attack.

The present investigation demonstrates the immobilization of Aspergillus oryzae β-galactosidase on silica nanoparticles via covalent binding. A greater yield of enzyme immobilization (89%) was attained on the developed nanobiocatalyst.

It was observed that the immobilized and soluble enzymes had optimal pH and temperature values of 50°C and 4.5, respectively. It was monitored that at pH 4.0, soluble β-galactosidase (SβG) exhibited 59% activity. However, the immobilized enzyme showed 92% activity under identical conditions. Similarly, 41% enzyme activity was retained at 70 °C by the free enzyme. Conversely, immobilized β-galactosidase (IβG) retained 70% activity under similar experimental conditions. Additionally, it was observed that at 5% galactose concentration, IβG showed 55% activity under one hour of incubation. However, under comparable experimental conditions, SβG showed 24% activity.

It was observed that the immobilized enzyme was reusable, maintaining 90% of its activity even after five uses. The soluble enzyme demonstrated 62% and 70% lactose hydrolysis under the same conditions after 8 hours, while IβG demonstrated 74% and 85% lactose hydrolysis at 40°C and 50°C, respectively, in a controlled batch reactor experiment that was run for 10 hours.

Hence, owing to the greater reusability (90% after 5^th repeated use) and excellent conversion of lactose at higher temperatures, the developed nanosupport may be used to produce lactose-free dairy products in continuous reactors on a large scale in biotechnology industries.