New benzimidazole-oxadiazole derivatives as potent VEGFR-2 inhibitors: Synthesis, anticancer evaluation, and docking study.

We report herein, the design and synthesis of benzimidazole-oxadiazole derivatives as new inhibitors for vascular endothelial growth factor receptor-2 (VEGFR-2). The designed members were assessed for their in vitro anticancer activity against three cancer cell lines and two normal cell lines; A549, MCF-7, PANC-1, hTERT-HPNE and CCD-19Lu. Compounds 4c and 4d were found to be the most effective compounds against three cancer cell lines. Compounds 4c and 4d were then tested for their in vitro VEGFR-2 inhibitory activity, safety profiles, and selectivity indices using the normal hTERT-HPNE and CCD-19Lu cell lines. It was determined that compound 4c was the most effective and safe member of the produced chemical family. Vascular endothelial growth factor A (VEGFA) immunolocalizations of compounds 4c and 4d were evaluated relative to control by VEGFA immunofluorescence staining. Compounds 4c and 4d inhibited VEGFR-2 enzyme with half-maximal inhibitory concentration values of 0.475 ± 0.021 and 0.618 ± 0.028 µM, respectively. Molecular docking of the target compounds was carried out in the active site of VEGFR-2 (Protein Data Bank: 4ASD).

The mechanism of anticancer effects of some pyrrolopyrimidine derivatives on HT-29 human colon cancer cells.

In the present work, the mechanism of anticancer activity of some pyrrolopyrimidine derivatives was evaluated. Compounds 5 and 8 exhibiting significant antiproliferative activity against HT-29 cells with IC50 values of 4.17 µM and 2.96, arrested the cells at the G2/M phase and significantly induced apoptosis. The apoptotic potential of the compounds has been verified via ELISA assay, which resulted in increased BAX, PUMA, BIM, and cleaved caspase 3 expression and decreased BCL-XL and MCL-1 protein levels in HT-29 cells. Moreover, the immunofluorescence technique showing that compounds 5 and 8-treatment reduced Ki67 immunolocalization and increased the caspase 3 and p53 immunolocalization confirmed the apoptotic activity. While treatment of HT-29 cells to compounds 5 and 8 inhibited Akt and ERK1/2, there are no alterations in JNK and p38 signaling pathways. According to molecular docking results, compounds 5 and 8 occupied the active site of Akt kinase and showed important hydrogen bonding interactions with key amino acids. Also, siRNA-mediated depletion of BIM, PUMA, and BAX/BAK expression decreased apoptotic response in HT-29 cells upon exposure to compound 5 and compound 8. Compounds 5 and 8 trigger the activation of mitochondrial apoptosis in HT-29 cells. Additionally, we found that proapoptotic BH3-only proteins BIM and PUMA are required for the full engagement of mitochondrial apoptosis signaling. However, p53 was dispensable for compound 5- or compound 8-induced apoptosis in HT-29 cells.

Does the use of low-molecular-weight heparin during pregnancy change the expression of PD-1 and PDL-1 in women with recurrent pregnancy loss?

Objective: The programed cell death gene-1 ligand (PDL-1) is expressed by villous syncytiotrophoblasts, cytotrophoblasts, and fetal cells in close contact with maternal tissue and blood. Programmed cell death gene-1 (PD-1) and the PDL-1 pathway cooperate with human leucocyte antigen-G (HLA-G), expressing intermediate trophoblastic cells and syncytiotrophoblasts to inhibit the function of activated T-cells. With this mechanism, the immunosuppressive microenvironment protects the placenta. This study investigated changes in PD-1 and PD-L1 gene expression in patients with a history of recurrent pregnancy loss (RPL). Materials and Methods: Sixty patients participated in the study and were divided into three groups. Group 1 (G1): healthy pregnancy, G2: RPL but not low-molecular-weight heparin (LMWH), and G3: RPL and LMWH. PD-1 gene expression in placental tissue samples was measured by reverse-transcriptase polymerase chain reaction and PD-L1 Elisa assay, and the study was supported by histopathology. Results: The PD-L1 value decreased significantly in G2. A significant difference was observed between the groups in PD-1 gene expression levels in G1 and G2. It was observed that vascularization increased and the villi structures intensified in G3. In G2, there was villus dysplasia in the placenta, enlargement in the intervillous region, and fibrin deposition. It was observed that the villi structures in G3 returned to a morphology similar to that of G1. Conclusion: T-cells are activated in patients using LMWH, and a new therapeutic strategy can be developed to prevent pregnancy loss by targeting the PD-1 pathway.

Preventive effects of Capparis Spinose extract on experimental periodontitis in rats: a histopathological and biochemical study.

This study aimed to assess the effectiveness of Capparis Spinose (CS) in preventing the initiation and progression of experimental periodontitis and to evaluate the effect of its on systemic oxidative stress in rats by experimental periodontitis model. Twenty-four male rats were equally divided into; Ligatured (L), non-ligatured (NL), and Ligatured with CS (11 days/day per 20 mg/kg) (LC) groups. Experimental periodontitis was induced with the silk suture technic. Alveolar bone loss was examined, and total antioxidant capacity(TAOC), total oxidant status(TOS), and oxidative stress index(OSI) were analyzed in rat serum. Although; alveolar bone loss showed statistically significant lower values in the LC group compared to L (p < 0.05), not NL. In the LC group, osteoclast and osteoblast numbers were statistically significant compared to L, but there were no statistical differences between LC and NL. Serum TAOC levels were significantly lower in group L compared to others and also LC group showed significant differences from NL. TOS and OSI levels were significantly higher in group L than in other groups. Within the limitation of the present study, it can be said that the destruction via local inflammation that may occur after the experimental periodontitis can be prevented by using CS.

Novel Bioactive Glass/Graphene Oxide-Coated Surgical Sutures for Soft Tissue Regeneration.

The combination of a commercially available PGLA (poly[glycolide-co-l-lactide]), 90:10% suture material with bioactive bioglass nanopowders (BGNs) and graphene oxide (GO)-doped BGNs offers new opportunities for the clinical application of biomaterials in soft tissue engineering. In the present experimental work, we demonstrate that GO-doped melt-derived BGNs were synthesized via the sol-gel process. After that, novel GO-doped and undoped BGNs were used to coat resorbable PGLA surgical sutures, thereby imparting bioactivity, biocompatibility, and accelerated wound healing properties to the sutures. Stable and homogeneous coatings on the surface of the sutures were achieved using an optimized vacuum sol deposition method. The phase composition, morphology, elemental characteristics, and chemical structure of uncoated and BGNs- and BGNs/GO-coated suture samples were characterized using Fourier transform infrared spectroscopy, field emission scanning electron microscopy, associated with elemental analysis, and knot performance test. In addition, in vitro bioactivity tests, biochemical tests, and in vivo tests were performed to examine the role of BGNs and GO on the biological and histopathological properties of the coated suture samples. The results indicated that the formation of BGNs and GO was enhanced significantly on the suture surface, which allowed for enhanced fibroblast attachment, migration, and proliferation and promoted the secretion of the angiogenic growth factor to speed up wound healing. These results confirmed the biocompatibility of BGNs- and BGNs/GO-coated suture samples and the positive effect of BGNs on the behavior of L929 fibroblast cells and also showed for the first time the possibility that cells can adhere and proliferate on the BGNs/GO-coated suture samples, especially in an in vivo environment. Resorbable surgical sutures with bioactive coatings, such as those prepared herein, can be an attractive biomaterial not only for hard tissue engineering but also for clinical applications in soft tissue engineering.

Death receptor-dependent apoptosis and cell cycle delay induced by bioymifi in human cervical cancer cells.

In chemotherapy applied against cervical cancer, non-specific cytotoxicity and drug resistance that develops over time are trying to be overcome. Therefore, the development of effective and innovative chemotherapeutic drugs for the treatment is among the priority issues in the medical field. The anticancer activity of the Bioymifi, which can activate apoptosis by inducing DR-5 clustering and aggregation against the human cervical cancer cell line, was investigated in the current study. The cytotoxic activity of Bioymifi on the HeLa cell line was identified using XTT assay. The pathway of the cell death mechanism was analyzed through the cell cycle and Annexin V assays by the flow cytometry. DAPI staining assay was applied under fluorescence microscopy to examine the nuclear morphology. Bioymifi appeared to have a remarkable IC50 value (11.75µM) against HeLa cells. The cell cycle analysis demonstrated the increase of Bioymifi cured HeLa cells in the S phase. And also, 11.75µM of Bioymifi caused a significantly higher apoptotic effect compared to control. In addition, in vitro immunofluorescence experiments of this study represented that Bioymifi reduced Ki-67 localization in HeLa cells. Bioymifi has significantly anticancer actions in Human cervix cancer in vitro and can be combined with standard treatment.

A digital image analysis study on the disintegration kinetics of reticular fibers in the ethylene glycol-induced rat liver tissue.

Ethylene glycol (EG), the raw material of polyethylenterephthalate, which is the most consumed plastic in the world, has low toxicity, but its metabolites are toxic. EG metabolites can cause acidosis, fibrosis, and eventually cirrhosis in the liver. This study aimed to investigate the effect of EG on rat liver and to determine the quantitative values of the disintegration of reticular fibers (RF) in the liver with the dose duration and to investigate the changes by digital image analysis (DIA). For this purpose, Wistar albino rats were divided into control, and five different daily experimental groups. The control group received saline, and the experimental groups received EG. At the end of experiments, liver tissues of all euthanized rats were removed, and sections were taken, and RF was shown by silver staining. It was observed that the RF fragments in the experimental groups were less than the control group. DIA of RF fragments was then performed with Olympus cellSensDimension 1.15 software and number, area, and ROI% values of the fragments were determined. Statistical analysis revealed that there was a significant difference between control and all experimental groups. RF fragments showed first-order disintegration kinetics, mean disintegration rate constant, and half-time values were 0.1 day-1 and 7 days, respectively. Consequently, the digital image analysis approach can be a useful tool for the biologist, pathologist, fibrosis-cirrhosis specialist, and computer scientist to understand the effects of toxic chemicals in the liver and analyze reticular fiber disintegration.

A study of digital image analysis on the acrylamide derivative monomers induced apoptosis in rat cerebrum.

Nowadays, apoptosis is mostly evaluated visually in histological studies. By using the quantitative digital image analysis, this study aimed to investigate the effect of acrylamide-based monomers (acrylamide [AAm], methacrylamide [MAAm], N-isopropylacrylamide [NIPAm]) on the cerebrum tissues in rats, which are the most common water-soluble monomers in the production of polymeric hydrogels used as biomaterials. The Wistar albino rats weighing ~220-240 g were divided into control and three test groups. The control group received 1 mL of saline, and the test groups received 1 mL of aqueous 50 mg/kg/day intramuscular injection of AAm, MAAm, and NIPAm, respectively. At the end of the experiments, brain tissues of all rats euthanized by intramuscular injection of sodium pentobarbital were removed. Terminal deoxynucleotide transferase dUTP nick and labeling (TUNEL) method was applied to brain tissue sections. The monomers have been shown to cause apoptosis due to oxidative stress in cerebrum tissue. Based on apoptosis by tunneling method, quantitative digital image analysis of cell fragments was performed with Olympus cellSens Dimension 1.15 software, and the number, total count area, selected area, average area, and ROI% values of the fragments were found. In addition, the total area and ROI% values of the fragments increased linearly with increasing the molar mass of monomers from the digital image analysis data. Quantitative digital image analysis can facilitate the monitoring of apoptosis caused by the oxidative stress of monomers used in the production of the biomaterials.

Role of pentoxifylline and iloprost in the prevention of ischemia-reperfusion injury in an experimental model of intestine ischemia-reperfusion in rats.

BACKGROUND: Intestinal ischemia-reperfusion (I/R) injury can lead to multiple organ failure and death. The aim of this study was to investigate the effects of pentoxifylline and iloprost administered before reperfusion in intestinal ischemia. METHODS: In total, 25 male Wistar Albino rats weighing 250-300 g were divided into five groups each comprising five subjects: control group (n=5), sham group (n=5, no I/R), I/R group (n=5, 45 min ischemia, and 120 min reperfusion), I/R + pentoxifylline group (n=5, 45 min ischemia following intraperitoneal 50 mg/kg pentoxifylline and 120 min reperfusion), and I/R + iloprast group (n=5, 45 min ischemia followed by intraperitoneal 2 mcg /kg iloprost and 120 min reperfusion). At the end of the experiment, ileum specimens were stained using hematoxylin-eosin and histopathologically evaluated using the Chiu score. Isometric contraction-relaxation responses were recorded using organ baths for contraction-relaxation responses. RESULTS: Pentoxifylline provided a significant improvement in response to histopathological and contraction-relaxation responses. Although iloprost provided recovery in reperfusion injury, it was not statistically significant. CONCLUSION: Our findings demonstrate that pentoxifylline may be promising in preventing small bowel ischemia-reperfusion injury. We concluded that further clinical and experimental studies for iloprost are needed.

The effects of monomers used in polymeric biomaterials on renal tissue / Efectos de los monómeros de los biomateriales poliméricos utilizados en tejido renal

SUMMARY: Biomaterials are mostly polymers and are used in artificial organ production in contemporary medicine. They are prepared by the polymerization reaction of many monomers. There are many monomers used in biomaterial production. In this study, we investigated whether acrylamide (AAm), methacrylamide (MAAm), N-isopropylacrylamide (NIPAm) and acrylic acid (AAc) used in polymeric biomaterial production had histopathological effects on renal tissue. In the present study, Wistar albino rats weighing ~ 250-300 g were used. Following the intramuscular injections of 1 mL aqueous monomer solutions at 50 mg/kg concentrations, acrylamide group animals were sacrificed at 1st, 2nd and 3rd weeks, the other monomer group animals were sacrificed at 1st, 2nd, 4th and 6th weeks. One mL serum physiologic were injected intramuscularly to the control group animals at the same time intervals with the experimental group animals. After histological follow-up, serial sections were prepared for evaluation under light microscope. In addition, the diameters of glomeruli and glomeruli space (Bowman's space) are measured, and the changes of the values of all groups with the exposure time were investigated. Acrylamide and its derivatives cause glomerular, arteriolar and tubule interstitial damage in the renal tissue. The narrowing glomeruli space, increasing diffuse mesangial matrix and tubular dilation was observed in some groups. In addition, dilatation, dissociation of tubular epithelium, thickening basement membranes and glycogenic vacuolization was also noted. These adverse results may be due to residual monomer. There should be no monomer residue in the polymer used as biomaterials.

RESUMEN: Los biomateriales en su mayoría son polímeros utilizados en la producción de órganos artificiales en la medicina contemporánea. Éstos son preparados mediante la reacción de polimerización de varios monómeros. Existe una gran cantidad de monómeros usados en la producción de biomateriales. En este estudio se investigó si la acrilamida (AAm), la metacrilamida (MAAm), la N-isopropilacrilamida (NIPAm) y el ácido acrílico (AAc) utilizados en la producción de biomateriales poliméricos tuvieron efectos histopatológicos en el tejido renal. En el presente estudio, se utilizaron ratas Wistar albinas que pesaban 250-300 g. Después de las inyecciones intramusculares (1 ml) de soluciones acuosas de monómero a concentraciones de 50 mg / kg, los animales del grupo de la acrilamida se sacrificaron a la 1ª, 2ª y 3ª semanas, los otros animales del grupo monómero se sacrificaron a las 1ª, 2ª, 4ª y 6ª semanas. Se inyectaron intramuscularmente 1 ml de suero fisiológico a los animales del grupo control en los mismos intervalos de tiempo que los animales del grupo experimental. Después del análisis histológico, se prepararon secciones en serie para su evaluación bajo microscopio óptico. Además, se midieron los diámetros de los glomérulos y el espacio glomerular, y se investigaron los cambios de los valores de todos los grupos con el tiempo de exposición. La acrilamida y sus derivados causaron daño intersticial glomerular, arteriolar y tubular en el tejido renal. El estrechamiento del espacio de los glomérulos, el aumento de la matriz mesangial difusa y la dilatación tubular se observó en algunos grupos. Además, también se observó dilatación, disociación del epitelio tubular, membranas basales espesantes y vacuolización glicogénica. Estos resultados adversos pueden deberse al monómero residual. No debe haber residuo de monómero en el polímero utilizado como biomateriales.