HeLa Research Articles

Molecular Actions of Enicostemma hyssopifolium Whole Plant Extract on HPV18-Infected Human Cervical Cancer (HeLa) Cells.

Enicostemma hyssopifolium (E. hyssopifolium) contains several bioactive compounds with anti-cancer activities. This study was performed to investigate the molecular effects of E. hyssopifolium on HPV18-containing HeLa cells. The methanol extract of E. hyssopifolium whole plant was tested for cytotoxicity by MTT assay. A lower and higher dose (80 and 160 μg/mL) to IC50 were analyzed for colonization inhibition (Clonogenic assay), cell cycle arrest (FACS analysis), and induction of apoptosis (AO/EtBr staining fluorescent microscopy and FACS analysis) and DNA fragmentation (comet assay). The HPV 18 E6 gene expression in treated cells was analyzed using RT-PCR and qPCR. A significant dose-dependent anti-proliferative activity (IC50 - 108.25±2 μg/mL) and inhibition of colony formation cell line were observed using both treatments. Treatment with 80 μg/mL of extract was found to result in a higher percent of cell cycle arrest at G0/G1 and G2M phases with more early apoptosis, while 160 μg/mL resulted in more cell cycle arrest at SUBG0 and S phases with late apoptosis for control. The comet assay also demonstrated a highly significant increase in DNA fragmentation after treatment with 160 μg/mL of extract (tail moments-19.536 ± 17.8), while 80 μg/mL of extract treatment showed non-significant tail moment (8.152 ± 13.0) compared to control (8.038 ± 12.0). The RT-PCR and qPCR results showed a significant reduction in the expression of the HPV18 E6 gene in HeLa cells treated with 160 μg/mL of extract, while 80 μg/mL did not show a significant reduction. The 160 μg/mL methanol extract of E. hyssopifolium demonstrated highly significant anti-cancer molecular effects in HeLa cells.

Rational design, synthesis, and anticancer evaluation of pyridine and substituted aryl linked 1,3,4-oxadiazole derivatives

A new series of Pyridine and substituted aryl linked 1,3,4-oxadiazole derivatives (7a-j) were designed, synthesized and evaluated for their anticancer activities against HeLa, MDA-MB-231, MCF-7, and A549 cell lines by using standard MTT reduction assay protocol with doxorubicin as standard drug. It is very important to mention that all these synthesized derivatives do not show any cytotoxicity on HEK-293 kidney cell lines. Among them, the compound 7d showed better activity against HeLa and MCF-7 cell lines with IC50 values of 1.8 ± 0.3 µM and 1.3 ± 0.11 µM.

Evaluation of a Radioiodinated G‐quadruplex Binder in Cervical Cancer Models

We herein describe the radiosynthesis of a 125I‐labeled acridine orange derivative ([125I]‐C8), acting as a G‐quadruplex binder, and its biological evaluation in cervical cancer models, aiming to enlighten its potential as a radioligand for Auger Electron Radiopharmaceutical Therapy (AE‐RPT) of cancer. [125I]‐C8 was synthesized with a moderate radiochemical yield (ca. 60 %) by a [125I]iodo‐destannylation reaction. Its evaluation in cervical cancer HeLa cells demonstrated that the radiocompound has a significant cellular internalization with a notorious accumulation in the cell nucleus. In line with these results, [125I]‐C8 strongly compromised the viability of HeLa cells in a dose‐dependent manner, inducing non‐repairable DNA lesions that are most probably due to the AEs emitted by 125I in close proximity to the DNA. Biodistribution studies in a murine HeLa xenograft model showed that [125I]‐C8 has fast blood clearance and high in vivo stability but poor tumor uptake, after systemic administration. The respective supramolecular conjugate with the AS1411 aptamer ([125I]‐C8/AS1411) led to a slower blood clearance in the same animal tumor model, although without improving the tumor uptake. To take advantage of the radiotoxicity of [125I]‐C8 against cervical cancer cells other strategies need to be studied, based namely on alternative nanodelivery carriers and/or intratumoral injection approaches.

HRS Facilitates Newcastle Disease Virus Replication in Tumor Cells by Promoting Viral Budding

Newcastle disease virus (NDV) is a highly pathogenic avian infectious disease agent and also a promising oncolytic virus with broad application prospects. The Endosomal Sorting Complex Required for Transport (ESCRT) machinery has been increasingly recognized for its crucial role in the life cycles of enveloped viruses, influencing processes such as viral entry, replication, and budding. In this study, we employed an RNA interference screening approach to identify key ESCRT components that regulate NDV replication in tumor cells. qPCR, immunofluorescence, and Western blot assays demonstrated that knockdown of HRS, CHMP4A, CHMP4B, and CHMP4C significantly impaired NDV replication in HeLa cells, with HRS exhibiting the most pronounced inhibitory effect. Additionally, HRS knockout significantly inhibited viral budding and suppressed NDV-induced cell death in HeLa cells. Notably, NDV infection was shown to significantly upregulate HRS gene and protein expression in a time-dependent manner. In conclusion, this study systematically identifies critical ESCRT components involved in NDV replication within tumor cells, with a particular focus on the role of HRS in promoting NDV’s replication by promoting viral budding, offering new insights for the development of NDV-based oncolytic therapies.

Relative biological effectiveness of clinically relevant photon energies for the survival of human colorectal, cervical, and prostate cancer cell lines

Abstract Objective: Relative biological effectiveness (RBE) differs between radiation qualities. However, an RBE of 1.0 has been established for photons regardless of the wide range of photon energies used clinically, the lack of reproducibility in radiobiological studies, and outdated reference energies used in the experimental literature. Moreover, due to intrinsic radiosensitivity, different cancer types have different responses to radiation. This study aimed to characterize the RBE of clinically relevant high and low photon energies in vitro for three human cancer cell lines: HCT116 (colon), HeLa (cervix), and PC3 (prostate). Approach: Experiments were conducted following dosimetry protocols provided by the American Association of Physicists in Medicine. Cells were irradiated with 6 MV x-rays, an 192Ir brachytherapy source, 225 kVp and 50 kVp x-rays. Cell survival post-irradiation was assessed using the clonogenic assay. Survival fractions were fitted using the linear quadratic model, and the survival curves were generated for RBE calculations. Main results: The cell killing was more efficient with decreasing photon energy: the HCT116 RBESF0.1 for 6 MV x-rays, 192Ir, and 50 kVp x-rays using 225 kVp x-rays as the reference were 0.89 ± 0.03, 0.95 ± 0.03, and 1.24 ± 0.04; the HeLa RBESF0.1 were 0.95 ± 0.04, 0.97 ± 0.05, and 1.09 ± 0.03, and the PC3 RBESF0.1 were 0.84 ± 0.01, 0.84 ± 0.01, and 1.13 ± 0.02, respectively. HeLa and PC3 cells had varying radiosensitivity when irradiated with 225 and 50 kVp x-rays. Significance: This difference supports the notion that RBE may not be 1.0 for all photons through experimental investigations that employed precise dosimetry. It highlights that different cancer types may not have identical responses to the same irradiation quality. Additionally, the RBE of clinically relevant photons was updated to the reference energy of 225 kVp x-rays.

Pyrrolidine, Piperazine, and Diazinane Alkaloids from the Marine Bacterium Strain Vibrio ruber ZXR-93

Four new alkaloids, vibripyrrolidine A (1), vibripiperazine A (2), and vibridiazinane A, B (3, 4), comprising one pyrrolidine, one piperazine, and two diazinane alkaloids, along with two known analogs (5, 6), were isolated from the marine bacterium Vibrio ruber ZXR-93 cultured in ISP2 medium. Their chemical structures were elucidated by analysis of their 1D and 2D NMR, mass spectra, and electronic circular dichroism (ECD) calculations. Compounds 1 and 3–6 showed vigorous antibacterial activity against Staphylococcus aureus, with MIC values ranging from 0.96 to 7.81 μg/mL. Moreover, compound 1 exhibited robust anti-inflammatory activity in vitro using the LPS-induced RAW264.7 macrophage model. All compounds also showed moderate antineoplastic activity against cervical cancer cells (HeLa) and gastric cancer cells (SGC-7901).

Structural and in vitro anticancer properties of the kaempferol–lactoferrin complex

AbstractLactoferrin and polyphenols are common natural functional compounds. Their interactions and the consequential alterations in functional activity have received widespread attention. The work aimed to investigate the interaction between lactoferrin and kaempferol, as well as evaluate the in vitro anticancer properties of the lactoferrin–kaempferol complex. The results of the spectra experiments revealed that lactoferrin and kaempferol are capable of forming complexes to quench the endogenous fluorescence of lactoferrin. Further insight into the binding mechanism was gained through molecular docking and molecular dynamics simulations. These analyses suggest that both hydrophobic interactions and hydrogen bonding are essential factors in the interaction between lactoferrin and kaempferol. Furthermore, the MTT assay and apoptosis by flow cytometry were conducted, revealing a synergistic effect of kaempferol and lactoferrin on the inhibition of HeLa cell proliferation. The findings from this investigation could improve our understanding of lactoferrin's interaction with polyphenols and the role of the lactoferrin–kaempferol complex in inhibiting cancer cell proliferation.

Exploring Tryptophan-based Short Peptides: Promising Candidate for Anticancer and Antimicrobial Therapies

Background: Ultra-short peptides are essential therapeutic agents due to their heightened selectivity and reduced toxicity. Scientific literature documents the utilization of dipeptides, tripeptides, and tetrapeptides as promising agents for combating cancer. We have created a range of tryptophan-based peptides derived from literature sources in order to assess their potential as anticancer drugs. Methods: We present the results of our study on the antibacterial and anticancer effectiveness of 10 ultra-short peptides that were produced utilizing microwave-assisted solid phase peptide synthesis. The synthesized peptides underwent screening for in vitro antibacterial activity using the agar dilution method. Results: HPLC, LC-MS, 1H NMR, and 13C NMR spectroscopy were used to analyze the synthesized peptides. In tests using the HeLa and MCF-7 cell lines, the synthesized peptides' anticancer efficacy was assessed. The study found that two peptides showed potential median inhibitory concentration (IC50) values of 3.9±0.13 μM and 1.8±0.09 μM, respectively, and showed more activity than the reference medication doxorubicin. Conclusion: The antibacterial activity of synthesized peptides 3b and 4b was found to be better than the other synthetic peptides. MIC value of roughly 5–50 μg/mL for peptides 3a, 4c, and 4d showed strong antifungal activity against Candida albicans. The synthesized peptides were also evaluated for their anticancer activity against HeLa and MCF-7 cell lines, and found that peptides 3e and 4e were more potent than other peptides against doxorubicin.

Dual folate/biotin-decorated liposomes mediated delivery of methylnaphthazarin for anti-cancer activity

Chemotherapy is an effective strategy for mitigating the global challenge of cancer treatment, which often encounters drug resistance and negative side effects. Methylnaphthazarin (MNZ), a natural compound with promising anti-cancer properties, has been underexplored due to its poor aqueous solubility and low selectivity. This study introduces a novel approach to overcome these limitations by developing MNZ-encapsulating liposomes decorated with folate and biotin (F/B-LP-MNZ). This dual-targeting strategy aims to enhance the anti-cancer efficacy and specificity of MNZ delivery. Our innovative F/B-LP-MNZ formulation demonstrated excellent physicochemical properties, stability, and controlled drug release profiles. In vitro studies revealed that MNZ-loaded liposomes attenuate the toxicity associated with free MNZ while F/B-LP-MNZ significantly increased cytotoxicity against HeLa cells, which express high levels of folate and biotin receptors, compared to non-targeted liposomes. Enhanced cellular uptake and improved dynamic flow attachment further confirmed the superior specificity of F/B-LP in targeting cancer cells. Additionally, our results revealed that F/B-LP-MNZ effectively inhibits HeLa cell migration and adhesion through EMT suppression and apoptotic induction, indicating its potential to prevent cancer metastasis. These findings highlight the potential of dual folate and biotin receptors-targeting liposomes as an effective delivery system for MNZ, offering a promising new avenue for targeted cancer therapy.

Novel phosphatidylinositol flippases contribute to phosphoinositide homeostasis in the plasma membrane.

Phosphatidylinositol is a precursor of various phosphoinositides, which play crucial roles in intracellular signaling and membrane dynamics and have impact on diverse aspects of cell physiology. Phosphoinositide synthesis and turnover occur in the cytoplasmic leaflet of the organellar and plasma membranes. P4-ATPases (lipid flippases) are responsible for translocating membrane lipids from the exoplasmic (luminal) to the cytoplasmic leaflet, thereby regulating membrane asymmetry. However, the mechanism underlying phosphatidylinositol translocation across cellular membranes remains elusive. Here, we discovered that the phosphatidylcholine flippases ATP8B1, ATP8B2, and ATP10A can also translocate phosphatidylinositol at the plasma membrane. To explore the function of these phosphatidylinositol flippases, we used cells depleted of CDC50A, a protein necessary for P4-ATPase function and ATP8B1 and ATP8B2, which express in HeLa cells. Upon activation of the Gq-coupled receptor, depletion of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] was accelerated in CDC50A knockout (KO) and ATP8B1/8B2 double KO cells compared with control cells, suggesting a decrease in PtdIns(4,5)P2 levels within the plasma membrane of the KO cells upon stimulation. These findings highlight the important role of P4-ATPases in maintaining phosphoinositide homeostasis and suggest a mechanism for asymmetry of phosphatidylinositol in the cytoplasmic leaflet of the plasma membrane.

Tuning The Intracellular Distribution of [3+2+1] Iridium(III) Complexes In Bacterial And Mammalian Cells By iClick Reaction With Biomolecular Carriers Functionalized With Alkynone Groups.

Three iridium(III) triazolato complexes of the general formula [Ir(triazolatoR,R')(ppy)(terpy)]PF6 with ppy=2-phenylpyridine and terpy=2,2':6',2''-terpyridine were efficiently prepared by iClick reaction of [Ir(N3)(ppy)(terpy)]PF6, with alkynes and alkynones, which allowed facile introduction of biological carriers such as biotin and cholic acid. In contrast to the precursor azido complex, which decomposed upon photoexcitation on a very short time scale, the triazolato complexes were stable in solution for up to 48 h. They emit in the spectral region around 540 nm with a quantum yield of 15-35 % in aerated acetonitrile solution and exhibit low cytotoxicity with IC50 values >50 μM for most complexes in L929 and HeLa cells, demonstrating their high suitability as luminescent probes. Cell uptake studies with confocal luminescence microscopy in prokaryotic Gram-positive S. aureus and Gram-negative E. coli bacteria as well as eukaryotic mammalian L929 and HeLa cells showed significant uptake in particular of the cholic acid conjugates iridium(III) moiety and distinct intracellular distribution modulated by the nature of the peripheral functional groups that can easily be modified by the iClick reaction.

The Effects of Sulphanomide-E Derivative on Carnitine Metabolism of Cervix Cancer Cells

Introduction: Cervical cancer is one of the most common types of cancer in women worldwide. Recent studies have shown that metabolic programming can support cervical cancer treatment by increasing sensitivity to chemotherapy and radiotherapy. Therefore, the investigation of new treatment agents targeting the metabolism of cervical cancer is of great importance in terms of improving treatment outcomes and developing new strategies. In our previous studies, we determined that Sulfanomide-E derivative has an apoptotic effect on HELA cells. In this study, we aimed to investigate the effects of Sulfanomide-E on carnitine metabolism in HELA cells. Method: HELA cells were used in our study and the cytotoxic and apoptotic effects of Sulfanomide-E were investigated. A dose of 25 μg/ml Sulfanomide-E was applied to HELA cells and the cells were incubated for 24 hours. In order to determine the changes in carnitine metabolism, a total of 27 carnitine and acylcarnitine derivatives were analyzed using LC-MS/MS. The obtained data were analyzed with SPSS 25.0 program. Findings: As a result of the analyses, a significant increase was observed in all carnitine and acylcarnitine derivatives in the Sulfanomide-E applied groups compared to the control group. These findings indicate that Sulfanomide-E has an effect on carnitine metabolism and these metabolic changes may be important in terms of response to treatment. Conclusion: This study reveals that Sulfanomide-E affects carnitine metabolism in HELA cells and exhibits apoptotic effects. The obtained data suggest that these changes in carnitine metabolism may provide a new understanding of the potential benefits of metabolic programming in the treatment of cervical cancer. Therefore, the role of agents such as Sulfanomide-E in the treatment of cervical cancer should be supported by more detailed studies.

Amine-Rich Carbon Dots Synthesized from Kappa-Carrageenan and l-Lysine as a Dual Probe for Detection of Folic Acid and Tumor-Targeted Delivery of Therapeutics.

Strategically designed, heteroatom-rich surface functionalized blue fluorescent carbon dots (CDs) were synthesized for high-throughput detection of folic acid (vitamin B9). The highly stable CDs could particularly detect vitamin B9 in the presence of 35 analytes, even up to 40 nM of the vitamin. The versatile CDs were found to have a high affinity for folic acid in wastewater, folic acid tablets, and food samples enriched with folic acid. The hemocompatibility of the CDs was also studied by using a hemolysis assay, confirming the CDs to be nontoxic to human blood samples up to 400 μg/mL. The CDs were then covalently conjugated to biotin, which possesses receptors that are overexpressed in tumor cells. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide dye) assay and confocal bioimaging studies proved the biotin-modified CDs (CDBT) were remarkably nontoxic in healthy cell lines (HEK-293) and highly target-specific toward tumor cells (HeLa), including triple-negative breast cancer cells (MDA-MB-231). The cytotoxicity assay of 5-fluorouracil encapsulated CDs (CDBTFu) showed the IC50 value to be 81 μM in HeLa cells and 185 μM in MDA-MB-231 cells, respectively, and significantly higher in HEK-293 cells (over 300 μM), owing to high specificity toward tumor cells.

Biosynthesis of Silver Nanoparticles Using Barleria albostellata C.B. Clarke Leaves and Stems: Antibacterial and Cytotoxic Activity

Silver nanoparticles (AgNPs) have increasingly gained attention owing to their distinctive physicochemical and biological properties. The objective of the investigation was to biologically synthesize AgNPs using plant extracts from Barleria albostellata. The synthesized AgNPs, obtained from B. albostellata (leaves and stems), were characterized through various techniques including UV-visible spectroscopy, scanning electron microscopy (SEM), high-resolution transmission electron microscopy (HRTEM), energy-dispersive X-ray analysis, Fourier transform infrared (FTIR) spectral analysis, and nanoparticle tracking analysis (NTA). The antibacterial efficacy of the synthesized AgNPs was evaluated utilizing the disk diffusion method. The cytotoxicity effects of the synthesized AgNPs were determined using the MTT assay (3-[(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide]) in cervical cancer (HeLa), embryonic kidney (HEK293), and breast adenocarcinoma (MCF-7) cell lines. The results indicate that B. albostellata extracts could serve as eco-friendly biofactories for the synthesis of AgNPs. UV-vis spectroscopy of the leaf and stem extracts revealed absorption peaks within the range of 400–450 nm, thereby confirming the synthesis of AgNPs. Elemental Ag was highest in the methanol leaf extracts (16.87 ± 0.89%) and lowest in the powdered stem extracts (7.13 ± 1.44%). Synthesized AgNPs were predominantly spherical in shape. HRTEM revealed that synthesized AgNPs from the methanolic stem extracts (34.32 ± 16.99 nm) were larger in size, while those from the powdered stem extracts were smaller (16.57 ± 5.55 nm). AgNPs synthesized from both the leaf and stem extracts exhibited zeta potential values between −8.8 and −32.1 mV, with hydrodynamics diameters ranging from 34.3 to 111.3 nm. FTIR spectroscopy confirmed the presence of various functional groups on the AgNPs. AgNPs synthesized from the leaf and stem extracts displayed significant antibacterial activity and were sensitive to Gram-negative and Gram-positive bacteria. AgNPs exhibited significant antibacterial activity (diameter of the zone of inhibition) against Pseudomonas aeruginosa (21.67 ± 2.87 mm) in the leaf methanolic extract. Synthesized AgNPs exhibited selective in vitro cytotoxicity against HEK293, HeLa, and MCF-7 cell lines. The IC50 values of the AgNPs synthesized from the various extracts were all above 9 µg/mL. Significant cytotoxic levels (IC50 16.11 and 27.23 µg/mL) were observed for the MCF-7 cell line upon exposure to the methanolic leaf and stem AgNPs. This study recommends the use of medicinal plant extracts in producing economically effective AgNPs, due to their efficient capping. Overall, AgNPs synthesized from B. albostellata extracts comprised novel antibacterial and anticancer agents, and warrant further investigation. Bio-synthesized AgNPs show great potential in the area of nanotechnology and may be used as an affordable, eco-friendly alternative for the delivery of conventional therapeutics.

Organic acid-fractionated lignin silver nanoparticles: Antimicrobial, anticancer, and antioxidant characteristics

Bioactive substances are utilized to treat a variety of diseases. Green lignin-mediated silver nanoparticles (L-Ag-NPs) have significant promise as a building block in the production of bio-renovation materials. The work optimized organic acid extraction to remove lignin from residual fermented hybrid Napier grass byproducts. We subsequently produced L-Ag-NPs. FTIR, XRD, DLS, and STEM characterized the sample. L-Ag-NPs were tested for antioxidant activity with the DPPH, DMPD, FRAP, and ABTS assays, as well as antibacterial activities. Antimicrobial activity was evaluated using four pathogenic bacteria (Klebsiella pneumonia, Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli). In contrast, cytotoxicity and ROS production assays were carried out using the HeLa cell line. The findings showed that L-Ag-NPs had high antioxidant efficiency. For each bacteria isolate, the antimicrobial activity showed favorable growth inhibition, with significant variations in L-Ag-NPs. L-Ag-NPs were reported to have an IC50 of 43.61 g/mL in the cytotoxicity test, and a significant increase in ROS generation was seen. In conclusion, L-Ag NPs have an excellent prospect in the pharmaceutical and biomedical industries and can be a dependable and environmentally safe material for their potential use.

An Enzyme-Encapsulated MOF@MOF Nanocomposite for Detecting H2O2 Derived From Superoxide Anion Released by Mitochondria of HeLa Cells.

In this work, a horseradish peroxidase (HRP)-encapsulated metal organic framework (MOF)@MOF nanocomposite is developed for detecting H2O2 converted by dismutation of superoxide anions released from live HeLa mitochondria. Initially, an HRP-polyacrylic acid cluster is incorporated on a mesoporous, peroxidase-like Cu/Co-1,4-benzenedicarboxylate (BDC) MOF platform to avoid structural change and deactivation of HRP through its interactions with MOF metal ions. Additionally, a Cu/Co-BDC(HRP)@1,3,5-benzenetricarboxyate (BTC) core-shell MOF/MOF structure, also with peroxide-like properties, serves as a protective matrix for HRP. Then, ultrathin porous carbon shells (UPCS) are adopted to improve the electrical conductivity of the MOF@MOF. The Cu/Co-BDC(HRP)@BTC|UPCS sensing platform exhibits two linear ranges of 0.05-1 µM and 1-1000 µM with a sensitivity of 172mA mM-1 cm-2 and 1.63mA mM-1 cm-2, respectively. A limit of detection of 0.057 µM, good selectivity and stability over 35 days for H2O2 detection are also achieved. After treating the mitochondrial complex with specific inhibitors, amperometric results at the sensing platform confirmed complex I and III within mitochondria as the main electron leakage sites in the electron transfer chains. Therefore, this sensing platform provides a tool that may aid in predicting and even developing treatments for some oxidative stress diseases caused by mitochondrial abnormalities.

Exopolysaccharides from the Green Microalga Strain Coelastrella sp. BGV—Isolation, Characterization, and Assessment of Anticancer Potential

Algal metabolites have been extensively studied as potential anticancer therapeutics. Among them, polysaccharides have attracted much attention because of their beneficial biological effects and safety. In the present research, the chemical characteristics, antitumor, and proapoptotic activities of extracellular polysaccharides (EPS) isolated from a new Bulgarian strain of the green microalga Coelastrella sp. BGV were investigated. A fast and convenient method of precipitation with cold ethanol was used to isolate EPS from the culture medium. The chemical characteristics of the isolated EPS were examined by colorimetric and spectrophotometric analyses, HPSEC-RID and HPLC-UV chromatography, and FT-IR spectroscopy. The results showed that the isolated EPS sample consists of three carbohydrate fractions with different molecular weights (11.5 × 104 Da, 30.7 × 104 Da, and 72.4 × 104 Da, respectively) and contains 7.14 (w/w%) protein. HPLC-UV analysis revealed the presence of galactose and fucose. The total uronic acid content in the sample was 4.5 (w/w%). The IR-FT spectrum of EPS revealed the presence of various functional groups typical of a polysaccharide (or proteoglycan) composed primarily of neutral sugars. The anticancer potential of the obtained EPS was assessed using cell lines with cancerous and non-cancerous origins as in vitro experimental models. The results of the performed MTT assay showed that EPS reduced the viability of the cervical and mammary carcinoma cell lines HeLa and MCF-7, while the control non-cancer cell lines BALB/3T3 and HaCaT were less affected. The HeLa cell line showed the highest sensitivity to the effects of EPS and was therefore used for further studies of its anticancer potential. The ability of EPS to inhibit cancer cell migration was demonstrated by wound-healing (scratch) assay. The cell cycle FACS analysis indicated that the EPS treatment induced significant increases in the sub G1 cell population and decreases of the percentages of cells in the G1, S, and G2-M phases, compared to the control. The fluorescent microscopy studies performed using three different staining methods in combination with Annexin V-FITC flow cytometric analysis clearly demonstrate the ability of EPS to induce cancer cell death via the apoptosis pathway. Moreover, an altered pattern and intensity of the immunocytochemical staining for the apoptosis- and proliferation-related proteins p53, bcl2, and Ki67 was detected in EPS-treated HeLa cancer cells as compared to the untreated controls. The obtained results characterize the new local strain of green microalgae Coelastrella sp. BGV as a producer of EPS with selective antitumor activity and provide an opportunity for further studies of its pharmacological and biotechnological potential.

Radical-Scavenging and Anti-inflammatory Activities of Fermented Eucheuma cottonii from Lombok

Eucheuma cottonii, a widely cultivated seaweed from the Island of Lombok, Indonesia, contains various bioactive compounds. Its utilization, however, is mainly limited based on its high carbohydrate content. Nevertheless, plant fermentation using lactic acid bacteria is renowned for increasing bioactive compounds and enhancing bioactivities. This study unveiled the potential of Lactobacillus plantarum-fermented E. cottonii as a functional food. E. cottonii was fermented using L. plantarum for 24 h. The folin-Ciocalteau method was used to determine the total phenolic content. The antioxidant capacity was measured using a 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay. Anti-inflammatory activity was carried out using cyclooxygenase (COX) inhibition assay against cervical cancer (HeLa) and colon cancer (WiDr) cells. Overall, fermentation successfully enhanced the bioactivities of E. cottonii. Fermented products exhibited higher antioxidant capacity than unfermented ones. Interestingly, the bioactivities only showed a moderate correlation with total phenolic content. Regarding anti-inflammatory activity, fermented extracts exhibited higher cyclooxygenase inhibition against HeLa cells, whereas no significant differences were observed between the fermented and unfermented products in WiDr cells. These findings indicate that L. plantarum-fermented E. cottonii holds promise to be a profitable functional food and has the potential to be utilized as an additional food therapy for cancer treatment. Keywords: Eucheuma cottonii; fermentation; Lactobacillus plantarum; antioxidant, anti-inflammation.

Tuning the surface charge and colloidal stability of hybrid gold-chitosan derivative nanoparticles for siRNA delivery

In recent years, the therapeutic potential of small interfering RNA (siRNA) has been confirmed by the emergence of commercially available siRNA-based drugs and by advances in the clinical trials phase. However, the many biological barriers faced by siRNA up to its delivery to the intended target make the advances in siRNA therapies highly dependent on the use of efficient gene vectors. In this study, the design of a new nanoparticle structured with a gold core and a diisopropylethylamine-chitosan shell was devised for application as a siRNA carrier. Thiol functionalized diisopropylethylamine-chitosan polymers were grafted onto gold nanoparticles (AuNPs) to obtain pH-responsive hybrid siRNA carriers (AuNP@polymers). The precise control of the diisopropylethylamine (DIPEA) graft and the tuning of the amount of polymer linked to the AuNPs enabled the assembly of hybrid carriers with good colloidal stability in physiological ionic strength (150 mmol L−1), sizes between 50 and 100 nm and positive zeta potentials (up to +17 mV), over a wide range of pH (5.5–7.4). The coated AuNPs were able to bind siRNA at N/P ratios in the range of 5–20, providing protection for the siRNA against RNAse degradation and making the hybrid vectors structurally and colloidally stable even in a protein-supplemented medium. The AuNP@polymers nanocarriers displayed non-cytotoxic effects in both 3T3/NIH fibroblast and HeLa-GFP cells up to an N/P ratio of 20 and the uptake of AuNP@polymers by RAW 264.7 macrophages was similar to Lipofectamine™ RNAiMAX. The zeta potential of the hybrid vectors was accurately adjusted by controlling the DIPEA graft and the pH of the formulation medium, which in turn drove the silencing efficiency of the siRNA nanocarrier as observed by the green fluorescent protein (GFP) knockdowns in HeLa cells. GFP knockdown levels close to that of Lipofectamine (up to 70 %) were achieved for the vectors formulated at pH 5.5. Overall, the results showed that the DIPEA-chitosan/AuNP association is a promising strategy to formulate hybrid nanocarriers for siRNA delivery with potential for in vivo studies.

Predicting of novel homoserine dehydrogenase inhibitors against Paracoccidioides brasiliensis: integrating in silico and in vitro approaches.

Aim: To search for potential inhibitors to homoserine dehydrogenase (HSD) in Paracoccidioides brasiliensis the causative agent of paracoccidioidomycosis, an infection with a high mortality rate in Brazil.Materials & methods: The enzyme was modeled and used in the virtual screening of the compounds. The library was first screened by the Autodock, in which 66 molecules were better ranked than substrate, and then, also evaluated by the Molegro and Gold programs.Results: The HS23 and HS87 molecules were selected in common by the three programs, and ADME/Tox evaluation indicates they are not toxic. The molecular dynamics of PbHSD bonded to ligands showed stable complexes until 50ns. To validate the results, compounds were purchased for assays of minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), synergic profile with Amphotericin B (AmB) and cytotoxicity. The two molecules presented MIC of 32μg/ml and MFC of 64μg/ml against the P. brasiliensis (strain Pb18). They also showed synergistic activity with AmB and a lack of toxicity against Hela and Vero cell lines.Conclusion: These results suggest that the HS23 and HS87 are promising candidates as PbHSD inhibitors and may be used as hits for the development of new drugs against paracoccidioidomycosis.