PBMC Cell Lines Research Articles

In-house glutamine based compound loaded poly-(D-L-lactide-coglycolide) nanoparticles as a possible candidate for the management of chronic myeloid leukemia

The development of targeted medicines such tyrosine kinase inhibitors (TKIs) have turned chronic myeloid leukemia (CML) from a potentially fatal illness into a manageable chronic condition for many individuals. But in case of TKIs, some serious drug induced adverse events as well as drug resistance were reported. In continuation of our earlier work, we have synthesized two p-nitrobenzenesulfonyl-D-glutamine derivatives which were screened against human CML cell line (K562). In comparison with earlier reported p-nitrobenzenesulfonyl-L-glutamine derivatives (Compound 1 and Compound 2), newly synthesized compounds showed more than 3 times better activity against K562 cell line. The best active compound (Compound 3) was encapsulated into Poly-(D-L-lactide-coglycolide) nanoparticles (NPs). The NPs were characterized and screened against K562 and human PBMC cell line. The IC50 value of NP of Compound 3 was found in the nanomolar range (522 nM) against K562 cell line which is more than 10 times better active than Compound 3. DNA deformation assay also indicates cell shrinkage and nuclear fragmentation which are associated with apoptosis-mediated cell death. The prepared NPs of Compound 3 (NP3) are very much less toxic against normal human PBMC (IC50 value > 500 µM). The current study implies that NP3 holds promise for managing chronic myeloid leukemia.

Statistical optimization and characterization of fucose-rich polysaccharides extracted from pumpkin (Cucurbita maxima) along with antioxidant and antiviral activities

Biologically active phytochemicals from pumpkin reveal versatile medical applications, though little is known about their antiviral activity. The fucose-rich polysaccharide extraction conditions were optimized through Box-Behnken design and purified by column chromatography. The purified fucose-rich polysaccharide was characterized through SEM, FT-IR, 1H NMR, XRD, TGA, and GS-MS. The analysis results revealed an irregular and porous surface of the purified polysaccharide with high fucose, rhamnose, galactose, and glucose contents. The tested fucose-rich polysaccharides revealed significant antioxidant and anti-inflammatory activity at very low concentrations. The purified fucose-rich polysaccharides exerted a broad-spectrum antiviral activity against both DNA and RNA viruses, accompanied by high safety toward normal cells, where the maximum safe doses (EC100) were estimated to be about 3–3.9 mg/mL for both Vero and PBMC cell lines. Treatment of HCV, ADV7, HSV1, and HIV viruses with the purified polysaccharides showed a potent dose-dependent inhibitory activity with IC50 values of 95.475, 20.96, 5.213, and 461.75 μg/mL, respectively. This activity was hypothesized to be through inhibiting the viral entry in HCV infection and inhibiting the reverse transcriptase activity in HIV. The current study firstly reported the antioxidant, anti-inflammatory, and antiviral activities of Cucurbita maxima fucose-rich polysaccharide against several viral infections.

MiR-182 antagonist alleviates glucocorticoid-induced secondary bone degeneration and osteoclast differentiation.

In this study, the expression of miR-182 in secondary bone degeneration was investigated, and the effect of its antagonist on glucocorticoid-induced osteoclast differentiation and its mechanism was studied. For this purpose, PBMC cell lines were selected for cultivation, and the changes were observed by hematoxylin-eosin (HE) staining. Real-time fluorescence quantitative (qRT-PCR) was used to detect mRNA expression. The protein expressions of RANKL, OPG and CXCL10 were detected by Western blot. CCK-8 and flow cytometry was used to detect cell proliferation and apoptosis. The results showed that protein expression levels of RANKL, OPG and CXCL10 in the miR-182 group were significantly higher than those in other groups (P>0.01). The miR-182 can promote RANK signal transduction in osteoclasts by regulating RANKL/NFκB signaling pathway, accelerating osteoclast proliferation and differentiation, and slowing down the process by miR-182 inhibitor. In general, miR-182 alleviates OP by inhibiting the activity of osteoclast via RANKL/NFκB signaling.

Identification of novel regulatory pathways across normal human bronchial epithelial cell lines (NHBEs) and peripheral blood mononuclear cell lines (PBMCs) in COVID-19 patients using transcriptome analysis

The SARS-CoV-2 is one of the most infectious and deadly coronaviruses, which has gripped the world, causing the COVID-19 pandemic. Despite the numerous studies being conducted on this virus, many uncertainties are with the disease. This is exacerbated by the speedy mutations acquired by the viral strain, which enables the disease to present itself differently in different people, introducing new factors of uncertainty. This study aims at the identification of regulatory pathways across two cell lines, namely, the peripheral blood mononuclear cell line (PBMC) and the normal human bronchial epithelial (NHBE) cell line. Both the above-mentioned cell lines were considered because they support viral replication. Furthermore, the NHBE cell line captures vital changes in the lungs, which are the main organs affected by the COVID-19 patients, and the PBMC cell line is closely linked to the body's immune system. RNA-Seq analysis, differential gene expression and gene set enrichment analysis for pathway identification were followed. Pathway analysis throws light upon the various systems affected in the body due to the COVID-19. Gene regulatory networks associated with the significant pathways were also designed. These networks aid in identifying various gene targets, along with their interactions. Studying the functionality of the pathways and the gene interactions associated with them, aided by long COVID studies, will provide immense clarity about the current COVID-19 scenario. In the long term, this will help in the design of therapeutic approaches against the SARS-CoV-2 and can also contribute to drug repurposing studies. Ultimately, this study identifies and analyses the relationship of various undiscovered or lesser explored pathways in the human body to the SARS-CoV-2 and establish a clearer picture of the association to help streamline further studies and approaches.

To evaluate the Effects of Lactic Acid Bacteria Fermented Lemon Juice from Limon and Eureka Varieties of Taiwan on Anti-pathogenic Bacteria and Anti-allergy

Lemon juice contains many bioactive compounds that have potential health benefits. In this study, two different lactic acid bacteria (LAB) strains were used as starter cultures, change in pH, viable cell count during fermentation and antimicrobial, immune boosting, anti-allergy properties were monitored. LAB strains were able to grow in the juice and their viable cell reached to 8.0 log CFU/mL after 72 h at 30°C. Fermentation of lemon juice with LAB had a 2+ inhibitory ability against Vibrio parahaemolyticus and 3+ inhibitory ability against uropathogenic Escherichia coli. The inhibitory action of lactic acid fermentation against the pathogens may be due to the accumulation of main primary metabolites and pH changed during the fermentation. In the adhesion assay, PL LP10069 and TL PM229 exhibited stronger adherence to the Caco-2 cell line. With the anti-allergy test, the expression of anti-allergy cytokine of lemon juice fermentation in PBMC cell lines. The result indicated the significant enhanced to IL-12 by 1% NE PM229. The conclusion showed that fermentation of Lemon juice by probiotic bacteria not only increase antibacterial and anti-allergy effects but also enhance health benefit.

Parasporin A13-2 of Bacillus thuringiensis Isolates from the Papaloapan Region (Mexico) Induce a Cytotoxic Effect by Late Apoptosis against Breast Cancer Cells

The protein A13-2 was obtained from Bacillus thuringiensis strains isolated from the Papaloapan watershed region (Oaxaca, Mexico). The cytotoxic activity of parasporal inclusions was studied against breast cancer cell line (MCF-7) and normal cell (human peripheral blood mononuclear cells). The MTT, the formation of reactive species, nitric oxide, free cell DNA, and the type of death cellular were assessed. The protein A13-2 shows the highest cytotoxic activity against MCF-7 (13% cell viability at 6 µg/mL), the extracellular DNA increases, and it shows no stress for reactive species or nitric oxide. Besides, the A13-2 parasporin shows no toxicity to peripheral blood mononuclear cells, and it does not generate changes in nitric oxide levels or free cell DNA. Due to that, the cytotoxic effect of A13-2 was specific for MCF-7, and it does not affect normal cells. According to microscopy and flow cytometry, A13-2 parasporin leads to the death of MCF-7 cells by late apoptosis together with necrosis and without allowing the triggering of the survival mechanisms. When analyzed together, our results show for the first time that the A13-2 protein isolated from Mexican strains of B. thuringiensis preferentially kills MCF- 7 (cancer cells) over HEK 293 and PBMC cell lines (normal cells), thus representing a promising alternative for the treatment of cancer breast.

Use of agricultural waste (coconut shell) for the synthesis of silver nanoparticles and evaluation of their antibacterial activity against selected human pathogens

Green synthesis of silver nanoparticles (AgNPs) is environmentally satisfactory because of their low cost and safe to nature. In the present study, extract of an agricultural waste, coconut (Cocos nucifera) shell is used to synthesize AgNPs and their antibacterial effect was investigated against selected human pathogens Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Salmonella typhimurium. The AgNPs synthesized using coconut shell extract (CSE-AgNPs) were characterized using UV–Visible spectroscopy (absorption peak at 432 nm), Transmission Electron Microscopy (spherical shaped particles size of 14.2–22.96 nm), Fourier-Transform Infrared Spectroscopy indicating the CSE capping around the AgNPs (Peaks 1384, 1609 and 3418 corresponds to organic molecules) and X-Ray Diffraction (Peak at 32.078 and 2-Theta). CSE-AgNPs exhibited zone of inhibition against S. aureus (15 mm), E. coli (13 mm), S. typhimurium (13 mm) and L. monocytogenes (10 mm) and minimum inhibitory concentration (MIC) of 26, 53, 106 and 212 μg/ml, respectively. Growth curve assay showed the effectiveness of CSE-AgNPs to inhibit the selected pathogens when compared to amphicillin control and extract. Scanning electron microscopy results indicated that the cell wall degradation might be the possible mechanism of antibacterial action of CSE-AgNPs. Different concentrations of AgNPs (0.078–2.5 mg/ml) showed no toxicity against human PBMC cell line. Hence, such highly effective CSE-AgNPs could be explored as antibacterial agent.

The role of biopolymer matrix films derived from carboxymethyl cellulose, sodium alginate and polyvinyl alcohol on the sustained transdermal release of diltiazem

Due to changing lifestyles of modern world, cardiac failures are increasing day by day. Drug delivery systems that can overcome the drawbacks of conventional drug administration are highly desired. Diltiazem hydrochloride (DTZ) is a common and effective drug used for cardiac failures. However, its efficient loading, high bio availability and sustained transdermal release from polymer matrix are of high demand. Herein, the main objective was to fabricate a transdermal drug delivery system (TDDS) capable of efficient DTZ loading with sustained release. Owing to the high hydrophilicity of DTZ, a hydrophilic matrix comprising of poly ethylene glycol coated vinyl trimethoxy silane-g-chitosan (PEG@VTMS-g-CS) was developed. DTZ encapsulated copolymer was dispersed in matrices like sodium alginate (ALG), carboxy methyl cellulose (CMC) and poly vinyl alcohol (PVA). Economic viability and cosmetic attractiveness of the films were evaluated and optimum results were obtained for PVA matrix. The in vitro skin penetration study of DTZ on rat skin further demonstrated the efficacy of PVA based film which yielded more than 40.0% cell viability on HaCaT and PBMC cell line with no histological changes on the skin which further confirmed the practical utility of the prepared film.

Pyranochromones from Dictyoloma vandellianum A. Juss and Their Cytotoxic Evaluation.

One new chromone 3,3-dimethylallylspatheliachromene methyl ether (1), as well as five known chromones, 6-(3-methylbut-2-enyl) allopteroxylin methyl ether (2), 6-(3-methylbut-2-enyl) allopteroxylin (3), 3,3-dimethylallylspatheliachromene (4), 5-O-methylcneorumchromone K (5) and spatheliabischromene (6), two alkaloids, 8-methoxy-N-methylflindersine (7) and 8-methoxyflindersine (8), and two limonoids, limonin diosphenol (9) and rutaevin (10), were isolated from Dictyoloma vandellianum A. Juss (Rutaceae). Cytotoxic activities towards tumor cell lines B16-F10, HepG2, K562 and HL60 and non-tumor cells PBMC were evaluated for compounds 1-6. Compound 1 was the most active showing IC50 values ranging from 6.26 to 14.82μg/ml in B16-F10 and K562 cell lines, respectively, and presented IC50 value of 11.65μg/ml in PBMC cell line.

In vitro growth inhibitory efficacy of some target specific novel drug molecules against Theileria equi

The in vitro growth inhibitory efficacies of five drug molecules against Theileria equi were evaluated in in vitro cultured parasites. A continuous microaerophilic stationary-phase culture (MASP) system was established for propagation of T. equi parasites. This in vitro culture system was used to assess the growth inhibitory effect of harmaline hydrochloride dihydrate (HHD), hexadecyltrimethylammonium bromide (HDTAB), hesparidin methyl chalcone (HMC), andrographolide and imidocarb dipropionate against T. equi. The 50% inhibitory concentration value of HHD, HDTAB, HMC, and imidocarb dipropionate for T. equi growth were 17.42μM, 14.00μM, 246.34μM and 0.279μM (equivalent to 0.139μg/ml), respectively (P<0.05). The andrographolide was not effective in inhibiting in vitro growth of T. equi in the present study. Furthermore, the in vitro cytotoxicity of these five drugs was evaluated on horse PBMC. At 2000μM concentration of HHD, HDTAB, HMC, andrographolide and imidocarb dipropionate were 8.34, 46.44, 58.53, 31.06, 15.14% cytotoxic on PBMC, respectively. Out of our four tested drug molecules, HHD was having low IC50 value along with least cytotoxicity, as compared to reference drug imidocarb dipropionate. The difference in IC50 value of HDTAB and HHD was significant, but HDTAB was moderately more cytotoxic on PBMC cell lines. HHD and HDTAB are selective inhibitor for heat shock protein 90 (Hsp90) and choline kinase pathway. It can be concluded that HHD and HDTAB are potential drug molecules against T. equi parasite by acting on Hsp90 and choline kinase pathway.

Ent-Kaurane diterpenes from the stem bark of Annona vepretorum (Annonaceae) and cytotoxic evaluation

This work describes a novel ent-kaurane diterpene, ent-3β-hydroxy-kaur-16-en-19-al along with five known ent-kaurane diterpenes, ent-3β,19-dihydroxy-kaur-16-eno, ent-3β-hydroxy-kaur-16-eno, ent-3β-acetoxy-kaur-16-eno, ent-3β-hydroxy-kaurenoic acid and kaurenoic acid, as well as caryophyllene oxide, humulene epoxide II, β-sitosterol, stigmasterol and campesterol from the stem bark of Annona vepretorum Mart. (Annonaceae). Cytotoxic activities towards tumor B16-F10, HepG2, K562 and HL60 and non-tumor PBMC cell lines were evaluated for ent-kaurane diterpenes. Among them, ent-3β-hydroxy-kaur-16-en-19-al was the most active compound with higher cytotoxic effect over K562 cell line (IC50 of 2.49μg/mL) and lower over B16-F10 cell line (IC50 of 21.02μg/mL).

Microwave‐Assisted Synthesis of 3,5‐Dibenzyl‐4‐amino‐1,2,4‐triazole and its Diazo Ligand, Metal Complexes Along with Anticancer Activity

Synthesis of 3,5‐dibenzyl‐4‐amino‐1,2,4‐triazole was accomplished via a conventional method as well as microwave irradiation method, followed by diazotization and coupling with 2,4‐pentanedione. The dinucleating ligand was isolated and complexed with Ni(II), Cu(II) and Ru(III) chlorides. These complexes were screened on Jurkat, Raji &amp; PBMC cell lines for anticancer activity. Ruthenium complexes showed potential anticancer activities.

Actinomycetes from Moroccan habitats: isolation and screening for cytotoxic activities

In our screening for actinomycetes showing cytotoxic activities, 8 samples were collected from various Moroccan habitats, 136 isolates were tested for their capacity to produce antibacterial compounds against gram positive bacteria. Thirty-seven strains of these isolates were active against Gram-positive bacteria. Using the following steps of primary screening: antibacterial activity, confrontation between the isolates and toxicity to Artemia salina; fifteen different isolates were used for further investigation. The aqueous extracts of Streptomyces sp. T5 and Streptomyces sp. AS8 were selected for their cytotoxic activity against Hep2, BSR and P815 cell lines, and two active compounds were observed on HPLC. The two isolates exhibited high activity against human cancer cell lines and were inactive on PBMC cell lines. Furthermore, the Streptomyces sp. T5 extract showed a proliferative activity.