Development and Characterization of Plant-derived Aristatoside C and Davisianoside B Saponin-loaded Phytosomes with Suppressed Hemolytic Activity.

Saponins are glycosides widely distributed in the plant kingdom and have many pharmacological activities. However, their tendency to bind to cell membranes can cause cell rupture, limiting their clinical use. In the previous study, aristatoside C and davisianoside B were isolated from Cephalaria species. Cytotoxicity assays showed that they are more active on A-549 cell lines than doxorubicin but caused hemolysis. In the current research, aristatoside C and davisianoside B were loaded to phytosomes called ALPs and DLPs respectively, and characterized for particle size, zeta potential, encapsulation efficiency, release kinetic, hemolytic activity, and cytotoxicity on A-549 cell line. DLPs maintained the cytotoxic activity of the free saponins against A-549 cells with IC50 of 9,64±0,02 µg/ml but dramatically reduced their hemolytic activity. Furthermore, temperature and time-dependent stability studies based on the size and zeta potential of ALPs and DLPs revealed that the phytosomes have sustained release properties over 2 weeks. Overall, DLPs displayed cytotoxicity against A-549 cells with minimal hemolysis and sustained release, highlighting their potential as nanotherapeutics for clinical applications.

A novel Lactiplantibacillus plantarum strain: probiotic properties and optimization of the growth conditions by response surface methodology.

The objective of this study is to explore the probiotic properties and optimal growth conditions of Lactiplantibacillus plantarum BG24. L. plantarum BG24 exhibited a remarkable ability to utilize lactose, and to grow under acidic conditions and in the presence of high levels of bile salts. The strain showed the highest antibacterial activity against L. monocytogenes Scott A (zone of inhibition: 26 mm). L. plantarum BG24 was found to be resistant to 8 of the tested 19 antibiotics using the disc diffusion method.and its multiple antibiotic resistance (MAR) index was calculated as 0.421. The adhesion rate to human intestinal epithelial Caco-2 cells was determined as 37.51%. The enzyme profile of L. plantarum BG24 was investigated using API ZYM test kit and the highest enzymatic activities were found for Leucine arylamidase, ß-glucosidase, Valine arylamidase, ß-galactosidase and N-acetyl-ß-glucosaminidase. L. plantarum BG24 strain showed higher microbial growth under static conditions (6.60 OD600) compared to 100 rpm (5.73 OD600) and 200 rpm (5.02 OD600) shaking speed due to its facultative anaerobic characteristic. However, different inoculation rates and glucose addition did not make a statistically significant difference on biomass formation (p > 0.05). The specific growth rate of L. plantarum BG24 was 0.416 h-1, the doubling time was 1.67 h, and the biomass productivity value was 0.14 gL-1 h-1 in the original MRS broth (pH 5.7) while higher values were found as 0.483 h-1, 1.43 h and 0.17 gL-1 h-1, respectively, in MRS broth (pH 6.5) medium enriched with 5 g/L yeast extract. The stirred tank bioreactor was used to optimise the growth of BG24 strain. The process variables was optimized at 0.05 vvm of aeration rate, 479 rpm of agitation speed, 3% of inoculation rate and 18 h of incubation time. The maximum biomass (g/L) production was obtained as 3.84 g/L at the optimized conditions.

Synthesis and biological evaluation of novel salicylidene uracils: Cytotoxic activity on human cancer cell lines and inhibitory action on enzymatic activity.

A series of salicylidene uracil (1-18) derived from 5-aminouracil and substituted salicylaldehydes were analyzed for cytotoxic activity and enzyme inhibitory potency. Nine out of eighteen derivatives (6-8, 10, 12-15, 18) are novel molecules synthesized for the first time in this work, and other derivatives were previously synthesized by our group. The compounds were characterized by Proton nuclear magnetic resonance, carbon nuclear magnetic resonance, fourier transform infrared spectroscopy, and elemental analysis. All compounds were tested for their in vitro cytotoxicity against PC-3 (human prostate adenocarcinoma), A549 (human alveolar adenocarcinoma), and SHSY-5Y (human neuroblastoma) cancer cell lines and the nontumorigenic HEK293 (human embryonic kidney cells) cell line. The 3,5-di-tert-butylsalicylaldehyde derived compound (8) was toxic to PC-3 human prostate adenocarcinoma cells, showing a promising IC50 value at 7.05 ± 0.76 µM. The present study also aimed to evaluate the inhibitory effects of the compounds against several key enzymes, namely carbonic anhydrase I and II (CA I and CA II), acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and glutathione reductase (GR), which are implicated in various global disorders, such as Alzheimer's disease, epilepsy, cancer, malaria, diabetes, and glaucoma. The inhibitory profiles of the tested compounds were assessed by determining their Ki values, which ranged from 2.96 to 9.24 nM for AChE, 3.78 to 12.57 nM for BChE, 8.42 to 25.74 nM for CA I, 7.24 to 19.74 nM for CA II, and 0.541 to 1.124 µM for GR. Molecular docking studies were also performed for all compounds. Most derivatives exhibited much more effective inhibitory action compared with clinically used standards. Thus, our findings indicate that the salicylidene derivatives presented in this study are promising drug candidates that need further evaluation.

Chemical characteristics and biological activity screening of Pistacia lentiscus mastic gum and leaves from Türkiye.

BACKGROUND: Mastic gum is a resin that is produced by Pistacia lentiscus. It has many traditional uses, dating from ancient times, such as the treatment of gastrointestinal disorders and as a food additive. In this study, the leaves and mastic gum of trees of different ages from Karaburun and the Cesme peninsula in Türkiye were examined chemically and biologically. Flavonoids, and phenolic and fatty acid components were evaluated by a liquid chromatography system coupled with high resolution mass spectrometry (LC-HRMS) and gas chromatography with flame ionization detection (GC-FID). Cytotoxicity was screened against several cancer and healthy cell lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Inducible nitric oxide synthase (iNOS) inhibition was determined on lipopolysaccharide (LPS)-induced murine macrophage cell line (RAW 264.7) cells. Antiviral activity was measured against avian coronavirus using an in ovo virucidal antiviral activity assay. RESULTS: The main phenolic constituents of the gum were found to be salicylic, rosmarinic, and caffeic acids whereas the most abundant compounds detected were flavonoids in the leaf extracts. The most abundant fatty acids in hexane extracts were palmitic and oleic acids. All gum extracts except 3-year-old gum had significant cytotoxic activity on HeLa (IC50 1.74 ± 0.03-4.76 ± 0.95) and PC-3 (0.64 ± 0.25-6.22 ± 1.40) cells. Moreover, reducing virus activity by fivefold or sixfold logarithmically between the range of 5-10 µg g-1 of 30-year-old gum extracts underscored the biological activity. CONCLUSION: In ovo antiviral activity studies on the P. lentiscus were conducted for the first time. The mastic gum and leaves obtained from P. lentiscus may have strong potential in terms of their chemical content and antiviral and cytotoxic activity. As a consequence of these properties, it is a sustainable, renewable natural resource that can be used as an additive and flavoring in the food and pharmaceutical industries. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Preparation, characterization, and toxicity evaluation of microemulsion formulation containing prunetin for potential oral applications.

Phytochemicals as therapeutic alternatives can have a fundamental impact on the various stages of inflammation and its resolution. Prunetin is a naturally occurring isoflavone and has been claimed to have numerous therapeutic potentials. The objective of this study is preparation, characterization, and toxicity evaluation of microemulsion formulation containing prunetin (PMF) for potential oral applications. With this research, it was targeted to emphasize the way of improving the therapeutic efficacy of natural biomolecules with a nontoxic and effective formulation. In the study, the pseudo-ternary phase diagram was developed and PMF was characterized by conductivity, droplet size, viscosity and pH. Effects against to cytokines (IL-1ß and IL-6) and TNF-α levels of the PMF were determined by ELISA technique. Genotoxicity and acute oral toxicity tests were carried out according to OECD guidelines. The results showed that PMF is a colloid system that reduced proinflammatory cytokine levels in LPS-induced macrophage cells compared to the control group. PMF demonstrated no mutagenic activity against TA98, TA100, TA1535, and TA1537 Salmonella strains. The in vivo oral acute toxicity test results indicated that PMF did not show mortality or significant side effects even at 2000 mg/kg bw. This study represents PMF showed a good safety profile in animal study. It is thought that this formulation may have anti-inflammatory potential with further in vivo testing.

Adjuvant effects of novel water/oil emulsion formulations on immune responses against infectious bronchitis (IB) vaccine in mice.

Vaccines have long made use of adjuvants to boost the immune response of the body and reduce the amount of vaccine needed as well as the expense of producing the vaccine. Many vaccine adjuvants are in development, but their application in veterinary vaccinations is restricted due to their lack of efficacy or undesirable side effects. For this reason, it is essential to develop novel adjuvants. To address the issue that the currently available infectious bronchitis (IB) vaccine often fails to produce sufficient immune responses, Coral Biotechnology tested two of their newly developed water-in-oil (W/O) type emulsion adjuvants (Coralvac RZ 528 and Coralvac RZ 506) in the IB vaccine. These adjuvants were tested in a mouse model to determine whether it worked with an inactive IBV H120 vaccine. Vaccine formulations were prepared by combining a virus concentration of 1 × 106 EID50/0.1 ml with an emulsion of the W/O type in a specific ratio. Once the formulations were ready, it was injected intramuscularly as a single dosage, and the mice were monitored for 21 days afterwards. The results showed that anti-IB antibody titer (IgG and IgG1), CD3+ CD8+ T cell responses as well as IFN- γ cytokine production, and splenocyte proliferation were all considerably higher in the IBV H120 with Coralvac RZ 528 and IBV H120 with Coralvac RZ 506 formulation groups than in the viral control group. According to our findings, the humoral and cellular immune responses of mice were significantly enhanced by these novel vaccine adjuvants. Thus, our results provide evidence that the W/O type emulsion adjuvants developed by Coral Biotechnology may be a useful adjuvant in IBV vaccines.

Anthemis tricolor Containing Unusual Totarol with Cytotoxic and Acetylcholinesterase-Inhibitory Activity.

Anthemis tricolor is an endemic species of Cyprus, and there is very limited information on its chemistry and pharmacological activities. The study aims to identify the in-vitro cytotoxic and acetylcholinesterase activities of Anthemis tricolor. The compounds responsible for the activities were also identified. Potent extracts of A. tricolor were subjected to preparative isolation and spectral structure determination studies. The chloroform extract contained many components, and due to the small quantity of extract available, enough pure compound(s) cannot be obtained for structure determination studies, though the n-hexane extract afforded two known compounds, totarol (1) and taraxasterol (2). The structures of the compounds (1 and 2) were determined by 1 H and 13 C NMR experiments. The pure compounds were also tested for their acetylcholinesterase inhibitory properties. For compound 1, the IC50 value was found to be 87.88 µg/mL. However, no inhibition was seen for 2. Anthemis tricolor was established to be a valuable source of pharmacologically active compounds and requires further studies.

Single and repeat-dose toxicity and local tolerance assessment of newly developed oil emulsion adjuvant formulations for veterinary purposes.

Adjuvants are components of vaccines that boost the intensity, duration, and breadth of the immune response. Insight into the mechanisms responsible for the immunotoxicity of both local and systemic adverse reactions following the use of adjuvants has been gained through research over the past twenty years. In the present study, single and repeated-dose toxicity and local tolerance of newly developed Water-in-Oil (W/O) and Water-in-Oil-in-Water (W/O/W) Emulsion adjuvants (Coralvac RZ 528, Coralvac RZ 506, Coralvac AT 318, Coralvac AT 318 SIS and Coralvac 252) by Coral Biotechnology Industry and Trade Incorporated Company were demonstrated after intramuscular injection in mice. In both toxicity studies, no adverse reactions such as death, general appearance, behavior, or weight loss were observed in the mice in the experimental groups. The results indicate that clinical chemistry parameters demonstrated normal function of the major organs and no irreversible damage to the mice in all adjuvant groups compared to the control group. In histopathologic investigation of single dose toxicity study, inflammation, edema, and large amounts of lipid droplets were observed on the 7th day in all experimental groups. On the 14th day, when the control group and the experimental groups were compared, it was seen that inflammation and edema had decreased considerably. Similarly, repeated dose toxicity study showed mild inflammation and edema in the control group, while quite widespread and severe inflammation, edema, and diffuse lipid droplets of varying sizes were observed in all adjuvant groups compared to the control group. These observations would be useful for the future development of oil-based adjuvants and their use in veterinary inactive vaccines.

Four new triterpene saponins from Cephalaria speciosa and their potent cytotoxic and immunomodulatory activities.

Four new triterpene saponins, namely speciosides A-D (1-4) along with six known saponins were isolated from the n-butanol extract of Cephalaria speciosa. In addition to these, three new prosapogenins (2a-4a) were obtained after alkaline hydrolysis. Elucidation of the structures of the isolated compounds was carried out by 1D, 2D NMR, HR-ESI/MS and GC-MS analyses. Cytotoxic activity was investigated on A549, CCD34-Lu, MDA-MB-231, PC-3, U-87MG, HeLa, HepG-2 cells by MTT method. Additionally, the immunomodulatory effect of compounds was evaluated for macrophage polarization with/without inactivated IBV D274 antigen treatment on THP-1 cells originated macrophage cells in terms of M1 or M2. According to the cytotoxicity results, compound 1 and prosapogenin 2a exhibit significant cytotoxicity than doxorubicin by comparison. The results demonstrated that saponin molecules treated THP-1 originated macrophages were induced M1 and/or M2 polarization. Additionally, macrophage cells treated with/without IBV D274 antigen contained saponin compounds were triggered significantly M2 polarization relative to M1. Notably, monodesmosidic saponins (1 and 2a-4a) in comparison with bisdesmosidic ones (2-4) demonstrated the most effect on M2 polarization. In conclusion, the results showed that all the isolated new saponins and their prosapogenins have immunomodulatory potential on macrophage cells increasing immune response without significant cytotoxic effect on THP-1 originated macrophages.

The solubility, pH value, chemical structure, radiopacity, and cytotoxicity of four different root canal sealers: an in vitro study.

OBJECTIVE: The aim of this study was to investigate solubility, pH value, chemical structure, radiopacity, and cytotoxicity of AH Plus BC, TotalFill BC, AH Plus, and AH Plus Jet sealers. MATERIALS AND METHODS: Cytotoxicity analysis with direct and extraction tests at 3 different concentrations (1:1, 1:2, 1:4 v/v%) and time (24 h, 48 h, and 72 h) on Saos-2, PdLF, and THP-1 cell lines, chemical structure with scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) analysis, solubility, pH, and radiopacity values of AH Plus BC, TotalFill BC, AH Plus, and AH Plus Jet were evaluated. For statistical analyses of the groups, repeated measures, factorial, and one-way ANOVA tests were used. The statistical significance level was set at p < .05. RESULTS: Resin-based sealers showed higher cytotoxicity values than the bioceramic-based sealers (p < 0.05). Time and concentrations were effective on the cell viabilities for cell lines. Higher peaks of calcium were detected bioceramic-based sealers and higher amount of zirconium was detected in AH Plus BC (p < 0.05). AH Plus BC showed similar radiopacity value with AH Plus, AH Plus Jet, whereas TotalFill BC showed the lowest radiopacity (p < 0.05). Bioceramic-based sealers had higher pH values in all experiment periods, and the difference between resin- and bioceramic-based sealer groups was significant (p < 0.05). However, the solubility values of the tested root canal sealers revealed no differences (p > 0.05). CONCLUSIONS: The newly produced AH Plus BC Sealer showed similar properties with TotalFill BC, and their biological properties were better than AH Plus and AH Plus Jet. CLINICAL RELEVANCE: AH Plus BC could be a possible alternative to other bioceramic- or resin-based sealers.

In Vivo Investigation of Supportive Immunotherapeutic Combination of Bifidobacterium infantis 35624 and Doxorubicin in Murine Breast Cancer.

The aim of the study is to investigate the anti-tumor effect of Bifidobacterium infantis 35624 in a xenograft model in BALB/c mice injected with 4T1 cells as a support for chemotherapeutic treatments of doxorubicin in vivo. The MTT assay was used to determine the cytotoxicity of doxorubicin against cancer cells, and apoptosis was analyzed by using flow cytometry. 4T1 cells (2 × 104 cells/mouse) were injected to BALB/c mice, and mice were fed with/without gavage B. infantis milk (108 CFU/mL) for 14 days and treated with doxorubicin on 5th and 10th days. The weights of the mice were recorded during the study, and the tumor sizes were measured by caliper at the 14th day. CD8 + T cell response was analyzed by using flow cytometer, and the results were compared to control and tumor control groups. The IC50 value for doxorubicin on 4T1 cell lines was determined as 0.053 ± 0.012 µg/mL. The apoptotic effect of doxorubicin at IC50 concentration was determined as 82.3% of cells to late apoptosis, 3.6% of cells to pro-apoptosis, and 6.2% of cells to necrosis. The treatment of doxorubicin, B. infantis milk, and the combination of them inhibited the tumor volumes by 55.50%, 40.69%, and 75.95%, respectively. B. infantis administration significantly enhanced the PHA-induced splenocyte proliferation (P < 0.05). It was shown that IFN-γ was effective in tumor growth and regression of metastasis. Consequently, the combination of B. infantis milk and doxorubicin showed the best anti-tumor effect.

Comparative Study of Antiviral, Cytotoxic, Antioxidant Activities, Total Phenolic Profile and Chemical Content of Propolis Samples in Different Colors from Turkiye.

Propolis is a valuable natural substance obtained by honey bees after being collected from the bark, resin of trees, plant leaves and mixed with their saliva, and has been widely used for various biological activities. The properties of propolis can vary widely by botanical origin, location of the hives and colony population. It is thought that the color of propolis is one of the main factors determining its acceptability and originates from the flower markers, pollen and nectar of some plants and is directly related to its chemical content. It is important to compare and standardize the colors, chemical content and biological activities of propolis in our country, which has a rich endemic plant diversity. Thus, in this study, the color indexes of 39 propolis samples from different locations in Turkiye were determined by Lovibond Tintometer, for the first time. The color index, total phenolic content, cytotoxic and antioxidant activities relationship of propolis and two commercial propolis samples were also investigated by HCA and PCA. Turkish propolis, which is defined by its color indices, chemical contents and many different activity potentials, such as antioxidant, antiviral and cytotoxic activity, will find use in many fields from medicine to cosmetics with this study.

IL-1ß, IL-6 and TNF-α expression levels of macrophage cells induced by benzydamine hydrochloride, benzydamine hydrochloride with chitosan, calcium hydroxide and chlorhexidine medicaments: An ELISA study.

The aim of the present study was to evaluate IL-1ß, IL-6 and TNF-α expression levels of macrophage cells induced by benzydamine hydrochloride (BNZ), BNZ with chitosan, calcium hydroxide (CH) and chlorhexidine (CHX) medicaments. Half maximal inhibitory concentrations (IC50) were assessed on THP-1, Saos-2, and CRL-2014 cells using MTT assay. THP-1 cells were differentiated into macrophages with phorbol12-myristate13-acetate and activated with lipopolysaccharide. IL-1ß, IL-6 and TNF-α levels in supernatants were determined using enzyme-linked immunosorbent assay (ELISA). The data were examined with one-way ANOVA and Tukey's multiple comparison test (p=0.05). At the selected concentrations, the cell viability was higher than 50% for chitosan and CH, whereas CHX presented lower IC50 values than BNZ and BNZ+chitosan. According to ELISA results, the lowest IL-1ß, IL-6 and TNF-α values were observed with BNZ+Chitosan 50 µg/mL and BNZ 50 µg/mL. BNZ+chitosan 50 µg/mL combination has revealed promising anti-inflammatory effects. Nevertheless, these findings need to be examined in clinical conditions.

Modern venomics-Current insights, novel methods, and future perspectives in biological and applied animal venom research.

Venoms have evolved >100 times in all major animal groups, and their components, known as toxins, have been fine-tuned over millions of years into highly effective biochemical weapons. There are many outstanding questions on the evolution of toxin arsenals, such as how venom genes originate, how venom contributes to the fitness of venomous species, and which modifications at the genomic, transcriptomic, and protein level drive their evolution. These questions have received particularly little attention outside of snakes, cone snails, spiders, and scorpions. Venom compounds have further become a source of inspiration for translational research using their diverse bioactivities for various applications. We highlight here recent advances and new strategies in modern venomics and discuss how recent technological innovations and multi-omic methods dramatically improve research on venomous animals. The study of genomes and their modifications through CRISPR and knockdown technologies will increase our understanding of how toxins evolve and which functions they have in the different ontogenetic stages during the development of venomous animals. Mass spectrometry imaging combined with spatial transcriptomics, in situ hybridization techniques, and modern computer tomography gives us further insights into the spatial distribution of toxins in the venom system and the function of the venom apparatus. All these evolutionary and biological insights contribute to more efficiently identify venom compounds, which can then be synthesized or produced in adapted expression systems to test their bioactivity. Finally, we critically discuss recent agrochemical, pharmaceutical, therapeutic, and diagnostic (so-called translational) aspects of venoms from which humans benefit.

Design and evaluation of erucic acid-phytosphingosine structured cationic nanoemulsions as a plasmid DNA delivery system against breast cancer cells.

This study is focused on the preparation and characterization of erucic acid (EA) and phytosphingosine (PS) containing cationic nanoemulsions (NEs) for plasmid DNA (pDNA) delivery. Repurposing of cationic agents guided us to PS, previously used for enhanced interaction with negatively charged surfaces. It was reported that EA might act anti-tumoral on C6 glioma, melanoma, neuroblastoma, and glioblastoma. However, there is only one study about mixed oleic acid-EA liposomes. This gap attracted our interest in the possible synergistic effects of PS and EA on MDA-MB-231 and MCF-7 breast cancer cells. Three cationic NEs (NE 1, NE 2, and NE 3) were prepared and characterized in terms of droplet size (DS), polydispersity index (PDI), and zeta potential (ZP) before and after complexation with pDNA, long-term stability, SDS release, cytotoxicity, and transfection studies. The cationic NEs had DSs of <200 nm, PDIs <0.3, and ZPs > +30 mV. Long-term stability studies revealed that NE 2 and NE 3 were stable. NE 1-pDNA had appropriate particle properties. NE 2 reduced the viability of MDA-MB-231 cells to 11% and of MCF-7 cells to 13% and resulted in the highest number of transfected cells. To sum up, NE 2 containing PS and EA is appropriate for delivering pDNA.

Effects of 15-lipoxygenase overexpressing adipose tissue mesenchymal stem cells on the Th17 / Treg plasticity.

15-lipoxygenase (15-LOX) is a critical enzyme that allows the direction of arachidonic acid metabolism to change from inflammation into the resolution. This study aims to reveal how the immunomodulation properties of mesenchymal stem cells (MSC) alter by the 15-LOX overexpression. For this purpose, peripheral blood mononuclear cells (PBMCs) isolated from seven healthy volunteers, and both MSCs and 15-LOX overexpressing MSCs (15-LOXMSCs) were co-cultured at different cell ratios (1/1, 1/5 and 1/10). Alterations of CD4+Tbet+, CD4+Gata3+, CD4+RoRC2+, and CD4+FoxP3+ lymphocyte frequencies were detected by flow cytometry, and IFN-γ, IL-4, IL-6, IL-10, IL-17a, TGF-ß and LXA4 levels of medium supernatants were measured by ELISA method. According to our findings, MSC and 15-LOXMSCs have a suppressive effect on PHA activated PBMCs. However, as the ratio of PBMCs increased, the effects of 15-LOXMSCs increased significantly, while the effects of MSCs decreased. The most notable effect of the 15-LOX modification was the significant reduction in IL-6, IL-10 and IL-17a expression and the accompanying increase in TGF-ß and LXA4 levels. We also observed a similar situation between CD4+RoRC2+ and CD4+FoxP3+ cell frequencies. These data suggested that the effects of MSCs on the balance of Th17 / Treg could change by the 15-LOX overexpression, and this might be in favor of the Treg cells.

Assessment of in vitro Cytotoxic, iNOS, Antioxidant and Photodynamic Antimicrobial Activities of Water-soluble Sulfonated Phthalocyanines.

In recent years, much effort has been devoted to the development of effective anticancer agents. In this manner, the utilization of water-soluble sulfonated phthalocyanines is crucial for many cancer cell lines. In this study, phthalonitrile and metallophthalocyanine compounds linked by benzenesulfonic acid groups have been prepared. Antimicrobial behaviors of those compounds were investigated by performing disk diffusion and photodynamic assays on gram-positive and negative bacteria. Indium phthalocyanine (InClPc) (3) showed inhibition activity against B. cereus, B. subtilis and S. aureus with disk diffusion assay. Also, gallium and indium phthalocyanines (2 and 3) exhibited inhibitory activity on both gram-positive and -negative microorganisms after light activation. Increasing the inhibitor concentration and light exposure time increased the inhibition activity for both molecules. GaClPc (2) demonstrated the maximum reducing power capacity among studied compounds, and CoPc (4) showed even better DPPH radical scavenging ability than the standard molecule Trolox at 2000 µg mL-1 concentration. The dose-dependent effect of compounds on cytotoxicity was studied against cancer cells PANC-1, MDA-MB-231, HepG2, A549, HeLa, CaCo-2 and non-tumorigenic cells HEK-293. All compounds showed no significant cytotoxic effect on any cell line up to the highest treated concentration at 50 µg mL-1 . However, all phthalocyanines had significant nitric oxide inhibition activity, and only in copper phthalocyanine (CuPc) (5), the MTT IC50 value was reached on LPS-activated RAW 264.7 macrophage cells. The lowest inducible nitric oxide synthase (iNOS) IC50 values were defined as 6 ± 1 µg mL-1 and 7 ± 0.5 µg mL-1 for CuPc (5) and InClPc (3), respectively.

Investigation of the supportive therapy potential of propolis extract and Lactobacillus acidophilus LA-5 milk combination against breast cancer in mice.

Immunotherapy has been applied in cancer treatments for many years as an alternative treatment method to radiotherapy, chemotherapy. It is well known that immunotherapy could suppress tumor formation by modulating the immune system of the host. The aim of the study is to investigate supportive therapy potential of acidophilus milk (AS) and propolis extract (PE) in the mouse xenograft breast cancer model. For this purpose, firstly cytotoxic effect of PE was determined by MTT assay against 4 T1 mouse breast cancer cells. Apoptotic effect of PE analyzed by flow cytometry. The antibacterial activity of PE was determined by the 96-well microplate broth-dilution method on Lactobacillus acidophilus LA-5. Then, Balb/c mice were injected subcutaneously with 4 T1 cells (2x105 cells/mouse) and also mice were given daily oral gavage with PE (66 mg/kg/day) and/or acidophilus milk (108 CFU/mL/mouse/day) for 14 days. The Balb/c mice were weighed throughout the study, and the tumor sizes were measured by caliper at the 14th day. The proliferation of splenocytes which collected spleen from mice was measured by MTT. CD8 + T cell response was analyzed by flow cytometry and results were evaluated in comparison with control and tumor control groups. The IC50 value for PE on 4 T1 cells was determined as 129.25 ± 1.90 µg/mL. The apoptotic effect of PE at IC50 concentration was determined as 3.3% of cells to late-apoptosis, 4.3% of cells to pro-apoptosis and 2.5% of cells to necrosis. The MIC and MBC values for PE on L. acidophilus LA-5 were 5000 ppm. The treatment of PE, AS and the combination of PE and AS were inhibited the tumor volumes by 59.16%, 28.29% and 63.39%, respectively. Acidophilus milk and PE combination significantly enhanced the ConA-, LPS- and PHA-induced splenocyte proliferation (P < 0.05). The acidophilus milk and PE combination were also found to stimulate IFN- γ production. In conclusion, the best anti-tumor effect was obtained by the combination of acidophilus milk and propolis.

A hederagenin-type triterpene saponin, sumbulianoside a from Cephalaria sumbuliana and its potent immunomodulatory activity against seasonal flu virus H3N2.

A new hederagenin-type triterpene saponin; hederagenin 28-O-ß-D-galactopyranosyl-(1→6)-ß-D-glucopyranosyl ester named sumbulianoside A (1), together with twelve known saponins were isolated from the n-butanol extract of Cephalaria sumbuliana (Caprifoliaceae) from which, one known saponin, dipsacus saponin A (2) was isolated, for the first time from Cephalaria species. The structures of the isolated compounds were elucidated by 1 D and 2 D NMR and HRESIMS analyses. Cytotoxic activities were investigated on A549, Hela, PANC1, SHSY5Y cells and non-cancerous cell HEK293 by MTT method and immunomodulatory activities were evaluated against activated H3N2 seasonal virus in whole blood by measuring IL-4, IFN-γ, IL-1ß cytokine level with ELISA. According to the cytotoxicity results, compounds 1 and 2 did not possess significant cytotoxicity, while only compound 2 induced significant IL-4 production (** p<.001) against H3N2 showing a possible Th2 response and antibody production. All compounds had no effect on cytokine release (p > 0.5).