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BMC Microbiology Apr 2024Leishmaniasis as a neglected tropical disease (NTD) is caused by the inoculation of Leishmania parasites via the bite of phlebotomine sand flies. After an infected bite,...
BACKGROUND
Leishmaniasis as a neglected tropical disease (NTD) is caused by the inoculation of Leishmania parasites via the bite of phlebotomine sand flies. After an infected bite, a series of innate and adaptive immune responses occurs, among which neutrophils can be mentioned as the initiators. Among the multiple functions of these fighting cells, neutrophil extracellular traps (NETs) were studied in the presence of Leishmania major promastigotes and salivary gland homogenates (SGH) of Phlebotomus papatasi alone, and in combination to mimic natural conditions of transmission.
MATERIAL & METHODS
The effect of L. major and SGH on NETs formation was studied in three different groups: neutrophils + SGH (NS), neutrophils + L. major (NL), neutrophils + L. major + SGH (NLS) along with negative and positive controls in 2, 4 and 6 h post-incubation. Different microscopic methods were used to visualize NETs comprising: fluorescence microscopy by Acridine Orange/ Ethidium Bromide staining, optical microscopy by Giemsa staining and scanning electron microscopy. In addition, the expression level of three different genes NE, MPO and MMP9 was evaluated by Real-Time PCR.
RESULTS
All three microscopical methods revealed similar results, as in NS group, chromatin extrusion as a sign of NETosis, was not very evident in each three time points; but, in NL and especially NLS group, more NETosis was observed and the interaction between neutrophils and promastigotes in NL and also with saliva in NLS group, gradually increased over times. Real-time reveals that, the expression of MPO, NE and MMP9 genes increased during 2 and 4 h after exposure, and then decreased at 6 h in most groups.
CONCLUSION
Hence, it was determined that the simultaneous presence of parasite and saliva in NLS group has a greater impact on the formation of NETs compared to NL and NS groups.
Topics: Animals; Humans; Phlebotomus; Leishmania major; Matrix Metalloproteinase 9; Extracellular Traps; Neutrophils; Salivary Glands
PubMed: 38575882
DOI: 10.1186/s12866-024-03270-z -
Journal of Cancer Research and... Oct 2023Cisplatin and platinum-based compounds have been used successfully to treat various cancers. However, their use is often restricted due to the acquired resistance by...
BACKGROUND
Cisplatin and platinum-based compounds have been used successfully to treat various cancers. However, their use is often restricted due to the acquired resistance by cancer cells. Over-expression of p53 and inhibition of NF-kB sensitize several cancer cells towards cisplatin-induced apoptosis. Quinacrine, a cytotoxic drug with predictable safety revealed to concurrently suppress NF-kB and activate p53, which may be an attractive adjuvant in cisplatin chemotherapy. Therefore, the objective of the present study was to establish the role of quinacrine as an adjuvant in lowering the dose of cisplatin during cancer therapy to circumvent its toxic effects.
MATERIALS AND METHODS
The colon cancer (HCT-8) cells were cultured and cell survival assays were performed using standard procedures. Cell cycle arrest and the extent of apoptosis were determined using a muse cell analyzer. Cancer survival proteins were analyzed using western blotting techniques.
RESULTS AND CONCLUSION
We demonstrated that concomitant use of quinacrine with cisplatin increased cell apoptosis, suppressed cell proliferation and inhibited colony formation in a colorectal cancer cell line. Moreover, cell cycle arrest in the G0/G1 and G2/M phases and upregulation of p53 expression were observed. There was also downregulation of NF-kB and Bcl-xL protein expressions, both of which are associated with enhanced cell apoptosis and an increase in the sensitivity of cancer cells to cisplatin, overcoming its chemoresistance. Overall, the results of the present study and available literature clearly indicate that the use of quinacrine as an adjuvant with cisplatin may enhance its anti-cancer activity and reduce chemoresistance.
Topics: Humans; Cisplatin; Quinacrine; NF-kappa B; Tumor Suppressor Protein p53; Radiation-Sensitizing Agents; Antineoplastic Agents, Alkylating; Apoptosis; Cell Line; Colonic Neoplasms
PubMed: 38376308
DOI: 10.4103/jcrt.jcrt_902_22 -
The Journal of Maternal-fetal &... Dec 2023To explore the suitable regimens of induced termination of second-trimester pregnancy in women with prior cesareans.
OBJECTIVE
To explore the suitable regimens of induced termination of second-trimester pregnancy in women with prior cesareans.
METHODS
A total of 204 s-trimester pregnant women with prior cesareans at the Third Affiliated Hospital of Zhengzhou University from January 2019 to December 2020 were included in this retrospective study. Group A included pregnant women who were administered mifepristone with misoprostol, Group B included those administering mifepristone with misoprostol as well as a transcervical Cook double-balloon catheter, Group C included those receiving mifepristone with an intra-amniotic injection of ethacridine lactate, and Group D included those receiving mifepristone, transcervical Cook double-balloon catheter, and intra-amniotic injection of ethacridine lactate. Their characteristics, clinical outcomes, and complications among the four groups were compared.
RESULTS
All women had similar profiles in maternal age, gravidity, and previous cesarean delivery ( > .05). There was no significant difference in successful abortion among the four groups ( > .05). Group C had a significantly shorter induction-to-abortion interval than Group D ( < .01). The blood loss after abortion at 2 h in Group B was much less than Group A ( < .05). It made a significant difference between Group B and Group D regarding the blood loss after abortion at 2 h ( < .01). With regard to total incidences of adverse reactions, there were much fewer in the group B than the group A ( < .05).
CONCLUSION
The four regimens are all effective for the termination of second-trimester pregnancy in women with prior cesareans. The use of transcervical Cook double-balloon could reduce the risks caused by misoprostol, and the combination of these is feasible to induce second-trimester pregnancy termination in women with prior cesareans.
Topics: Pregnancy; Female; Humans; Pregnancy Trimester, Second; Cesarean Section; Ethacridine; Mifepristone; Misoprostol; Retrospective Studies
PubMed: 37654101
DOI: 10.1080/14767058.2023.2249187 -
BMC Complementary Medicine and Therapies Nov 2023Screening of herbal plants for various therapeutic properties is the hour as it shows promising activity. Scientific evidence of the pharmacological activity of the...
BACKGROUND
Screening of herbal plants for various therapeutic properties is the hour as it shows promising activity. Scientific evidence of the pharmacological activity of the plant strengthens the traditional application of plants.
METHODS
Rose flowers (Rosa chinensis) were procured and grounded into a coarse powder. The DNA was isolated from rose flower and molecular identification was performed by rbcL-BF and rbcL-724R primers. Antibacterial activity was evaluated by using disc and agar diffusion methods and the anti-cancer effect of the rose flower extract (RE) was examined using MTT assay in lung cancer cell line. The mechanism of cell death induced by RE was qualitatively measured using Acridine orange/Ethidium bromide staining and Hoechst staining. GC-MS analysis was performed using GC-MS-5975C.
RESULT
The RE showed potent antimicrobial activity against various ATCC cultures. The rose extract strongly inhibits the growth of ESBL resistant organism along with inhibition of biofilm formation in the ESBL resistant organism. The extract caused apoptotic and necrotic cell death in lung cancer cells. GC-MS analysis demonstrated the presence of several biologically active compounds such as Clindamycin, Phytol, Octanoic acid, and Stigmasterol which might be the reason for the therapeutic properties of the plant.
CONCLUSION
This study shows the antimicrobial and biofilm inhibition activity against the clinical isolates of Klebsiella pneumonia. The study shows the cytotoxic and apoptotic activity in A549 cancer cell line. Thus, the plant may act as a potent antimicrobial drug against resistant strains.
Topics: Humans; Rosa; Lung Neoplasms; Plant Extracts; Acetone; Anti-Infective Agents; A549 Cells
PubMed: 37950173
DOI: 10.1186/s12906-023-04222-2 -
Cureus Aug 2023To investigate the cytocompatibility effect and wound healing activity of chitosan thiocolchicoside lauric acid (CTL) nanogel using human gingival fibroblast (hGF) cells.
AIM
To investigate the cytocompatibility effect and wound healing activity of chitosan thiocolchicoside lauric acid (CTL) nanogel using human gingival fibroblast (hGF) cells.
MATERIALS AND METHODS
hGF cells were established from gingival tissue as per the standard cell isolation protocol. The cytocompatibility effect was assessed using an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide) assay. A scratch wound healing assay was performed to assess the wound-healing potential of CTL nanogel. For the nuclear morphological changes analysis, acridine orange staining was used in gingival fibroblast cells. The stained nuclei were viewed under a fluorescent microscope. ANOVA with posthoc analysis was performed using GraphPad Prism 5 software (Dotmatics, Boston, Massachusetts). The significance level (p-value) was expressed as <0.05. Results: CTL nanogel did not show any significant cytotoxicity at concentrations 10-80 µl/ml (p<0.05). CTL nanogel at a concentration of 40µl/ml has a cytocompatibility effect on hGF cells and increases cell viability. In vitro scratch wound healing assay resulted in faster wound healing and cell migration with CTL nanogel when compared to the control group.
CONCLUSION
CTL nanogel has a significant effect on cell proliferation at various concentrations, which suggests its use as a safe and effective drug delivery system.
PubMed: 37727182
DOI: 10.7759/cureus.43727 -
Chemico-biological Interactions Jun 2024Ionic liquids (ILs) are a class of low melting point salts with physicochemical properties suitable for a range of industrial applications such as chemical processing...
Ionic liquids (ILs) are a class of low melting point salts with physicochemical properties suitable for a range of industrial applications such as chemical processing and battery design. Major challenges to the wide-scale adoption of ILs in industry include their eco- and cytotoxic effects, however, this opens up the possibility of the use of ILs use as novel anticancer agents. Understanding the structural features that promote IL cytotoxicity is therefore important. Key structural features that can impact IL cytotoxicity include size and lipophilicity of the cationic head group. In this study, the cytotoxic effects of acridinium-based ILs containing relatively large tri- and tetracyclic cations were evaluated. It was found that 9-phenylacridinium-based ILs are potent cytotoxic agents that reduce the viability of human MDA-MB-231 breast cancer cells with IC concentrations in the nanomolar range. In mechanistic studies, it was found that unlike the pyridinium-based analogue, [CPy][I], acridinium-based ILs did not inhibit oxidative phosphorylation or induce reactive oxygen species formation, and may instead target other mitochondrial processes or components such as mitochondrial DNA.
Topics: Humans; Ionic Liquids; Acridines; Structure-Activity Relationship; Cell Line, Tumor; Reactive Oxygen Species; Cell Survival; Antineoplastic Agents; Oxidative Phosphorylation
PubMed: 38735455
DOI: 10.1016/j.cbi.2024.111042 -
International Journal of Molecular... Aug 2023Metallic nanoparticles (mNPs) are widely used as food additives and can interact with gliadin triggering an immune response, but evaluation of the effects on crypts,...
Metallic nanoparticles (mNPs) are widely used as food additives and can interact with gliadin triggering an immune response, but evaluation of the effects on crypts, hypertrophic in celiac subjects, is still lacking. This study evaluated the effects of gold and silver mNPs in combination with gliadin on crypt-like cells (HIEC-6). Transmission electron microscopy (TEM) was used to evaluate gliadin-mNP aggregates in cells. Western blot and immunofluorescence analysis assessed autophagy-related molecule levels (p62, LC3, beclin-1, EGFR). Lysosome functionality was tested with acridine orange (AO) and Magic Red assays. TEM identified an increase in autophagic vacuoles after exposure to gliadin + mNPs, as also detected by significant increments in LC3-II and p62 expression. Immunofluorescence confirmed the presence of mature autophagosomes, showing LC3 and p62 colocalization, indicating an altered autophagic flux, further assessed with EGFR degradation, AO and Magic Red assays. The results showed a significant reduction in lysosomal enzyme activity and a modest reduction in acidity. Thus, gliadin + mNPs can block the autophagic flux inducing a lysosomal defect. The alteration of this pathway, essential for cell function, can lead to cell damage and death. The potential effects of this copresence in food should be further characterized to avoid a negative impact on celiac disease subjects.
Topics: Humans; Gold; Glutens; Silver; Gliadin; Autophagy; Acridine Orange; Nanoparticles; ErbB Receptors
PubMed: 37685847
DOI: 10.3390/ijms241713040 -
Marine Drugs Sep 2023With the emergence of drug resistance and the consequential high morbidity and mortality rates, there is an urgent need to screen and identify new agents for the...
With the emergence of drug resistance and the consequential high morbidity and mortality rates, there is an urgent need to screen and identify new agents for the effective treatment of cancer. Terphenyls-a group of aromatic hydrocarbons consisting of a linear 1,4-diaryl-substituted benzene core-has exhibited a wide range of biological activities. In this study, we discovered a terphenyllin derivative-CHNQD-00824-derived from the marine compound library as a potential anticancer agent. The cytotoxic activities of the CHNQD-00824 compound were evaluated against 13 different cell lines with IC values from 0.16 to 7.64 μM. Further study showed that CHNQD-00824 inhibited the proliferation and migration of cancer cells, possibly by inducing DNA damage. Acridine orange staining demonstrated that CHNQD-00824 promoted apoptosis in zebrafish embryos. Notably, the anti-cancer effectiveness was verified in a doxycin hydrochloride (DOX)-induced liver-specific enlargement model in zebrafish. With Solafinib as a positive control, CHNQD-00824 markedly suppressed tumor growth at concentrations of 2.5 and 5 μM, further highlighting its potential as an effective anticancer agent.
Topics: Animals; Cell Line, Tumor; Zebrafish; Cell Proliferation; Drug Screening Assays, Antitumor; Antineoplastic Agents; Apoptosis; DNA Damage; Structure-Activity Relationship; Molecular Structure
PubMed: 37888447
DOI: 10.3390/md21100512 -
Journal of Oral and Maxillofacial... 2023Polymorphonuclear neutrophils are the most abundant leukocytes in humans and are key host cells in defence against invading microorganisms. The oral neutrophil count may...
BACKGROUND AND OBJECTIVES
Polymorphonuclear neutrophils are the most abundant leukocytes in humans and are key host cells in defence against invading microorganisms. The oral neutrophil count may be an indicator of the periodontal health status, which correlates with the severity of periodontal disease. This study attempts to quantify orogranulocytes utilising an oral rinse and to assess the usefulness of this method in evaluating the oral inflammatory load much the same way the circulating neutrophils are used to screen for patients with infection in extra-oral sites.
METHODS
A total of 125 participants were divided into five groups with 25 subjects in each group. The groups consisted of healthy, gingivitis, mild periodontitis, moderate periodontitis, and severe periodontitis. Participants were asked to rinse with 10 mL of 0.9% saline for 30 s and to expectorate. Samples were centrifuged at 2000 RPM for 10 min. The supernatant removed was suspended in 5 mL of Hanks's balanced salt solution. One millilitre of this suspension was mixed with 4 μL of acridine orange. A 10 μL aliquot of this suspension was then assessed on a haemocytometer, and the oral PMNs were counted using fluorescence microscopy.
RESULTS
The mean number of oral neutrophils (100,000 cells/mL) was the lowest in the healthy group and increased in ascending order across the different groups with the highest for severe periodontitis group.
CONCLUSION
The oral neutrophil counts increased with the severity of periodontal inflammation. This is an easy, safe, reliable, and non-invasive method of quantification of oral neutrophils.
PubMed: 38304508
DOI: 10.4103/jomfp.jomfp_418_21 -
Genes and Environment : the Official... Apr 2024An in vitro micronucleus assay is a standard genotoxicity test. Although the technique and interpretation of the results are simple, manual counting of the total and...
BACKGROUND
An in vitro micronucleus assay is a standard genotoxicity test. Although the technique and interpretation of the results are simple, manual counting of the total and micronucleus-containing cells in a microscopic field is tedious. To address this issue, several systems have been developed for quick and efficient micronucleus counting, including flow cytometry and automated detection based on specialized software and detection systems that analyze images.
RESULTS
Here, we present a simple and effective method for automated micronucleus counting using image recognition technology. Our process involves separating the RGB channels in a color micrograph of cells stained with acridine orange. The cell nuclei and micronuclei were detected by scaling the G image, whereas the cytoplasm was recognized from a composite image of the R and G images. Finally, we identified cells with overlapping cytoplasm and micronuclei as micronucleated cells, and the application displayed the number of micronucleated cells and the total number of cells. Our method yielded results that were comparable to manually measured values.
CONCLUSIONS
Our micronucleus detection (MN/cell detection software) system can accurately detect the total number of cells and micronucleus-forming cells in microscopic images with the same level of precision as achieved through manual counting. The accuracy of micronucleus numbers depends on the cell staining conditions; however, the software has options by which users can easily manually optimize parameters such as threshold, denoise, and binary to achieve the best results. The optimization process is easy to handle and requires less effort, making it an efficient way to obtain accurate results.
PubMed: 38659010
DOI: 10.1186/s41021-024-00305-9