Flowering Biology of Rhododendron pulchrum

To study the flowering biology of Rhododendron pulchrum, we used scanning electron microscopy (SEM) and paraffin sectioning to observe the microstructures of its floral organs, a methyl thiazolyl tetrazolium (MTT) colorimetric assay to detect pollen viability in different periods, continuous observations to study flowering phenology, and artificial pollination and a benzidine-hydrogen peroxide method to determine stigma receptivity. R. pulchrum exhibited a centralized flowering phenology. The protogynous stigmas of R. pulchrum were able to receive pollen before flowering. The pollen grains of R. pulchrum fused into tetrads, the average ratio of the polar axis length to the equatorial axis length (P/E) was 1.05, and the pollen viability was highest in the initial flowering period, reaching 88.98%. The pollen/ovule (P/O) ratio was 266–328, and the outcrossing index (OCI) was 4; the vitality of R. pulchrum pollen remained high in the initial flowering and blooming periods. Compared with the lifespan of a single flower, pollen vitality remained high for most of the experimental period, thereby improving male fitness. The P/O ratio suggests that R. pulchrum may have a facultative outcrossing breeding system. The OCI estimation suggests that R. pulchrum is partially self-compatible, most likely requiring pollinators to complete pollination.

Pyrrole Plasma Polymer-Coated Electrospun Scaffolds for Neural Tissue Engineering

Promising strategies for neural tissue engineering are based on the use of three-dimensional substrates for cell anchorage and tissue development. In this work, fibrillar scaffolds composed of electrospun randomly- and aligned-oriented fibers coated with plasma synthesized pyrrole polymer, doped and undoped with iodine, were fabricated and characterized. Infrared spectroscopy, thermogravimetric analysis, and X-ray diffraction analysis revealed the functional groups and molecular integration of each scaffold, as well as the effect of plasma polymer synthesis on crystallinity. Scanning microscopy imaging demonstrated the porous fibrillar micrometric structure of the scaffolds, which afforded adhesion, infiltration, and survival for the neural cells. Orientation analysis of electron microscope images confirmed the elongation of neurite-like cell structures elicited by undoped plasma pyrrole polymer-coated aligned scaffolds, without any biochemical stimuli. The MTT colorimetric assay validated the biocompatibility of the fabricated composite materials, and further evidenced plasma pyrrole polymer-coated aligned scaffolds as permissive substrates for the support of neural cells. These results suggest plasma synthesized pyrrole polymer-coated aligned scaffolds are promising materials for tissue engineering applications.

Photodynamic Therapy Using Toluidine Blue O (TBO) Dye as a Photosensitizer against Leishmania major

Background: Photodynamic therapy (PDT) is alternative treatment of cutaneous leishmaniasis (CL), and phenolthiazine dyes such as Toluidine Blue O (TBO) have the potential role in PDT and notably affect parasites inactivation. This study aimed to evaluate the effectiveness of PDT by using TBO and a light-emitting diode (LED) in the treatment of zoonotic CL (ZCL). Methods: The study was conducted in Iran University of Medical Sciences, Tehran, Iran in 2018-2020. Different concentration (7.8 µg/ mL up to 500 µg/ mL) of TBO as a photosensitizer and a 630 nm LED light as a source of light were used for antileishmanial activity against both forms of Leishmania major promastigotes and intracellular amastigotes. Effective concentration (EC50) and cell cytotoxicity (CC50) were calculated in both infected and non-infected J774.A1 macrophages, respectively. As well as inhibitory concentration (IC50) was quantified in L. major promastigotes for 2 h, 24 h, and 48 h after incubation using a MTT colorimetric assay. Results: TBO dye in combination with the PDT significantly decreases the L. major promastigotes and intracellular amastigotes viability when compared with TBO alone. Both TBO dye in combination with the PDT and TBO alone had no toxic effects on the mice macrophages; however, it significantly killed the entered parasites inside the cells. Our results in the current study established satisfactory findings in clearing intracellular L. major parasites in in-vitro conditions. Conclusion: TBO dye in combination with the PDT can be considered as a harmless, effective and importantly perfect treatment against L. major, causative agent of ZCL, in an in-vitro situation without any negative toxicity to the mice macrophages.

Cytotoxic Impacts of N-Oleoylethanolamine in Bone Cancer Cells

Background: Cancer is a complex disease that is derived from the uncontrolled proliferation of cells. Bone cancer is a type of prevalent cancer that occurs both in youngsters and adults. Bone cancer is mostly common in the long bones of the pelvis, arms, and legs. Statistically, more than 200 cases of osteosarcoma have been reported annually in our country. Classical treatment with chemotherapeutics remains ineffective for the cure of this cancer. Recent studies have shown that ceramide induces apoptosis due to its increased levels in the cells. Thus, many studies have been conducted for the accumulation of ceramide molecules in the cell by different ways to induce apoptosis. NOE (N-oleoylethanolamine) is a specific inhibitor of ceramidase enzymes that hydrolyse intracellular ceramides and prevent apoptosis. Objective: This study investigates the cytotoxic and apoptosis-inducing activities of NOE on human osteosarcoma Saos-2 cells. Methods: Cytotoxic effects were investigated by MTT colorimetric assay. For the detection of morphological and ultrastructural indicators of apoptosis, confocal and TEM techniques were used, respectively. Results: Our finding indicated that NOE is effective in the inhibition of the growth of Saos-2 cells. Confocal and TEM findings showed morphological and ultrastructural changes as chromatin condensation, fragmentations of nuclei and mitochondria, as well as damaged cytoskeleton and cell shrinkage. Conclusion: The results revealed that NOE exhibits its cytotoxicity on Saos-2 cells by changing the ultrastructure and morphology of cells with clear apoptotic sparks.

F127/Cisplatin Microemulsions: In Vitro, In Vivo and Computational Studies

The development of effective strategies for local administration of chemotherapeutic drugs, thus minimizing the adverse side effects to patients, is one of the key challenges in biomedicine and cancer research. This work reports the formulation and characterization of PluronicF127 microemulsions to enhance the bioavailability of Cisplatin (Cis). The size of Cis microemulsion was about 12.0 nm, as assessed by dynamic light scattering analysis. In vitro cytotoxic activity of free Cis and F127/Cis microemulsions were studied on malignant (C152 and MCF7) and normal (HUVEC) cells via tetrazolium (MTT) colorimetric assay. Cell morphology was also monitored. In vitro assessments revealed thatF127/Cis microemulsions induced cytotoxicity/morphological changes to a lesser extent than free Cis. Regarding in vivo experiments, F127/Cis microemulsions were injected intraperitoneally at 7 and 14 mg/kg doses into adult male Wistar rats to assess histologic and biochemical changes. In this case, the bulk Cis group caused severe histopathological changes and significant increases in serum liver enzymes and serum kidney function markers. The group treated with the 14 mg/kg dose of F127/Cis microemulsions also showed severe fatty changes and significant increases in serum liver enzymes, blood urea nitrogen, and creatinine levels. The group treated with the low dose of nano-Cis showed a significant increase in serum liver enzymes levels accompanied by mild fatty changes of the liver. Theoretical surveys were performed to get an understanding of the interplay between F127 and Cis. Results reveal that hydrogen bonding (HB) interactions with F127have an influence on the molecular properties of Cis and may playa role in the lower toxicity of F127/Cis in comparison to free Cis.

Preparation, Spectral Characterization and Anticancer Potential of Cinnamic Esters

Cinnamic acid and its derivatives show a remarkable variety of biological activities and are often studied in search of the development of new and highly effective drugs. This work aims to synthesize, characterize and evaluate the cytotoxic activity of esters derived from cinnamic acid. Eighteen esters were synthesized through Steglich’s esterification, of which eleven were not reported in the literature. All compounds were fully characterized by Fourier transform infrared epectroscopy (FTIR), nuclear magnetic resonance (1H and 13C NMR) and high-resolution mass spectrometry (HRMS) data. The cytotoxic activity of esters obtained was evaluated using four human tumor cell lines: SNB-19 (astrocytoma), HCT-116 (colon carcinoma, human), PC3 (prostate) and HL60 (promyelocytic leukemia) through the 3-(4,5-dimethyl-2-thiazolyl)- 2,5‑diphenyl-2H‑tetrazolium (MTT) colorimetric assay. These studies showed that the compound 3-methoxybenzyl (E)‑3‑(4‑methoxyphenyl)acrylate (12) is the most potent against HCT-116, PC3 and SBN-19 cells, with the lowest half maximal inhibitory concentration (IC50) value of 16.2 μM in the HCT-116 strain. The derivatives were obtained in good yields (76.6-95%), except for compounds 5-isopropyl-2-methylphenyl (E)-3-(3-hydroxy-4-methoxyphenyl)acrylate (17) (18.6%) and 2-isopropyl-5-methylphenyl (E)-3-(3-hydroxy-4-methoxyphenyl)acrylate (18) (15.5%).

Cytotoxic Effects of Valsartan Organotin(IV) Complexes on Human Lung Cancer Cells

The organotin(IV) compounds used in chemotherapy due to its lipophilicity, affected by the number of carbon atoms and the cytotoxicity. These are affected by the obtainability of Sn coordination bond and bond stabilization between ligand and tin. Two novel organotin(IV) complexes were synthesized, characterized, and tested against human lung cancer cells (A549). The cytotoxic effect of the prepared organotin(IV) complexes against human lung cancer cells (A549) was investigated using the MTT colorimetric assay. Apoptosis was investigated by flow cytometry. The cytotoxicity assay reveals that the Bu2SnL2 complex is more active to inhibit the growth of A549 cells compared to the Ph2SnL2, and doxorubicin, nevertheless at high concentration (50 and 100) µg/mL the doxorubicin was more affective to inhibit the viability of A549 cells.

Immunological Activities of Isoprinosine Inhibition on Viral Infections Inhuman

Isoprinosine is a combination of inosine used as antiviral drug without effect on viral particle itself, but instead only and acts as on immunostimulant and also acts indirectly by activation of immune cells. Aim of this study was to determine level of interferon-alpha (INF-α) with parainfluenza viruses HPIV-2, and adenoviruses HAdV-2 replication. In the present study, cytotoxic effect of isoprinosine was assessed using A549 cell line exposed to different concentrations of compound (isoprinosine: 50-800μg/mL) for 48 hours. Cytotoxic effect was examined visually using light, inverted microscopy Olympus CK2 under 400x magnification and by the MTT colorimetric assay. The yield re­duction assay (YRA), which evaluates the ability of the isoprinosine (50-800 μg/mL) to inhibit virus multiplication in cell cultures, was applied. The cytopathic effect of the virus was evaluated 48 h after infection of A549 cell cultures with viruses by means of light, inverted microsco­py. The YRA method was used to determine the 50% end point (IC50) in the presence of Isoprinosine with the controlled one. MTT cytotoxicity assay confirmed microscopic observations. There were no morphological changes, as assessed visually, in cell cultures treated with isoprinosine. After conducting the experi­ments and analyzing the results we noticed that higher concentrations of isoprinosine strongly inhibited multiplication of all viruses. HPIV-2 and HAdV-2 showed the highest sensitivity to the antiviral activity of isoprinosine as compared with the control, however, increasing concentrations of isoprinosine up to 800 μg /ml slightly enhanced the antiviral activity of 400 μg/ml isoprinosine. Our study was conducted that HAdV-2 and HPIV-2 have the highest sensitivity to the antiviral activity of isoprinosine from all tested viral strains.

Prostaglandin E2 Induces Expression of Mineralization Genes by Undifferentiated Dental Pulp Cells

Prostaglandin E2 (PGE2) is a lipid mediator usually released during inflammation. This study aimed to investigate the potential of soluble or microsphere-loaded PGE2 on inducing differentiation of dental pulp stem cells. PGE2-loaded microspheres (MS) were prepared using an oil-in-water emulsion solvent extraction-evaporation process and were characterized. Mouse dental pulp stem cells (OD-21) were stimulated with soluble or PGE2-loaded MS (0.01 and 0.1 µM). Cell viability was determined by MTT colorimetric assay. Ibsp, Bmp2 and Runx2 expression was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) after 3, 6, and 24 h. The results showed that the soluble PGE2 reduced dental pulp stem cells viability after 24 h of stimulation whereas PGE2-loaded MS did not. Soluble PGE2 up-regulated Ibsp and Bmp2 at 3 h, differently from PGE2-loaded MS. On the other hand, PGE2-MS induced Bmp2 and Runx2 at 6 h and Ibsp at 24 h. In conclusion, our in vitro results show that PGE2, soluble or loaded in MS are not cytotoxic and modulateIbsp,Bmp2, andRunx2gene expression in cultured OD-21 cells.