Breast cancer is the leading cause of death among women. Natural compounds play an important role in the prevention of breast cancer. In this study, the anticancer properties of Lobaria pulmonariawere investigated. Cell viability and proliferation were assessed using MTT test, apoptotic cell percentages were determined by AnnexinV/PI assay, protein levels of BCL2, CASP3 and P53 were measured by western blot. Additionally, F-actin levels were analyzed using Immunofluorescence staining. It was determined that L. pulmonariareduced the viability of MCF7 cells. While the percentage of apoptotic cells did not change, the percentage of non-apoptotic cell death increased. Moreover, there were no significant changes in BCL2, CASP3, and P53 protein levels, nor in the amount of F-actin. In conclusion, L. pulmonariadid not induce apoptosis in MCF7 breast cancer cells but appeared to trigger a non-apoptotic cell death pathway, such as necrosis.

Background and Aims: Plant extracts are an important source of cytotoxic compounds and have consistently been an interesting field of research. The aim of this study is to investigate the cytotoxic and apoptotic effects of Thymus vulgaris (T. vulgaris) extracts on human breast cancer cell lines. Methods: This study was carried out using human breast cancer cell lines (MCF-7 and MDA-MB-231) as experimental groups and the healthy human fibroblast cell line (PCS-201-012) as the control group. Petroleum ether and ethanol extracts were obtained from T. vulgaris. The extracts were applied to MCF-7 and MDA-MB-231 human breast cancer cell lines and human breast cancer stem cells. Cytotoxicity studies were performed using the RTCA iCELLigence system (Agilent Technologies), and apoptosis studies were performed using terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) and 4',6-diamidino-2 phenylindole (DAPI) methods. Results: The T. vulgaris extracts were found to have concentration-dependent cytotoxic effects on human breast cancer cells. The growth of breast cancer stem cells was also determined to be inhibited when an effective concentration (45 mu g/ mL) of the extracts was applied. Lastly, specific morphological changes related to apoptosis were detected in the cells that had been treated with the effective concentration. Conclusion: The T. vulgaris extracts were found to inhibit the proliferation of human breast cancer cells and human breast cancer stem cells selectively and concentration-dependently via an apoptosis-dependent pathway. The results suggest that the extracts may make promising sources for developing drugs for breast cancer therapy.

p-Chloroamphetamine (PCA), a serotonin depletor, was given to rats at least 24 h prior to testing in an open field or a shock avoidance Y-maze task. In the open field PCA groups showed hypoactivity and increased defecation up to 30 days after drug administration. These same animals plus independent groups of PCA animals, showed facilitated avoidance acquisition in the Y-maze up to 15 days after PCA administration. At the beginning of behavioral testing serotonin levels in PCA animals were reduced 70 percent and were still reduced 41 percent after 38 days in whole brain. These results suggest a separation between shock and non-shock effects of brain serotonin depletion. The facilitated avoidance also provides support for the role of serotonin as an inhibitory neurotransmitter mediating a behavioral suppression system.

The pathogenic mechanisms of these diseases must be well understood for the treatment of neurological disorders such as Huntington's disease. Huntington's Disease (HD), a dominant and neurodegenerative disease, is characterized by the CAG re-expansion that occurs in the gene encoding the polyglutamine-expanded mutant Huntingtin (mHTT) protein. Genome editing approaches include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and Clustered Regularly Interspaced Short Palindromic Repeats/Caspase 9 (CRISPR/Cas9) systems. CRISPR/Cas9 technology allows effective gene editing in different cell types and organisms. Through these systems are created isogenic control of human origin induced pluripotent stem cells (iPSCs). In human and mouse models, HD-iPSC lines can be continuously corrected using these systems. HD-iPSCs can be corrected through the CRISPR/Cas9 system and the cut-and-paste mechanism using isogenic control iPSCs. This mechanism is a piggyBac transposon-based selection system that can effectively switch between vectors and chromosomes. In studies conducted, it has been determined that in neural cells derived from HD-iPSC, there are isogenic controls as corrected lines recovered from phenotypic abnormalities and gene expression changes. It has been determined that trinucleotide repeat disorders occurring in HD can be cured by single-guide RNA (sgRNA) and normal exogenous DNA restoration, known as the single guideline RNA specific to Cas9. The purpose of this review in addition to give general information about HD, a neurodegenerative disorder is to explained the role of CRISPR/Cas9 system with iPSCs in HD treatment.

The use of phthalates as plasticizers has been omnipresent, especially in cosmetics and food packaging, despite the proven effects on some organs of humans and animals. Therefore, alterations in living organisms due to phthalate exposure attract the attention of many scientists. Here, we demonstrate a mechanical and chemical investigation of the mentioned effects of di(2-ethylhexyl)phthalate (DEHP) and dibutyl phthalate (DBP) on rat liver by utilizing scanning acoustic microscopy (SAM), Raman spectroscopy (RS) and inductively coupled plasma optical emission spectrometry (ICP-OES) for the first time in the literature, as far as we know. The combined analysis gives insights into the degree of modification in the tissue components and which chemicals lead to these modifications. Our study shows that the acoustic impedance values of tissues of DEHP and DBP delivered mother rats are higher than those of tissues of the control mother rat, while the acoustic impedance values of tissues of offspring rats of DEHP and DBP delivered mother rats do not differ significantly from those of tissues of the control offspring rats of the control mother rat. Besides, RS analysis shows how the incorporation of DEHP into liver tissues changes the configuration and conformation of lipids and fatty acids. ICP-OES results show increased element levels within the tissues of DEHP and DBP delivered rats. Therefore, we can say that phthalates cause modifications within the liver. This study is a preliminary effort to investigate tissues with a mechano-chemical probe.

Objective: Phthalates, despite their endocrine disrupting effects, are widely used as plastifiants. Environmental exposure of phthalates was demonstrated to cause fetal death and reproductive toxicity in human beings, as well as in laboratory animals. However, underlying mechanisms are not clear. Material and Methods: Here, we examine the impact of di(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP)on rat lungs, brain and heart by scanning acoustic microscopy (SAM) and inductively coupled plasma optical emission spectroscopy (ICP-OES). First, we evaluate tissues of mother rats and we show that the acoustic impedance values oftissues of DEHP and DBP delivered rats differ from those of tissues of the control rat. Then, element level analyses withinthese tissues are done and element levels within tissues of DEHP and DBP delivered pregnant rats are found to be higherthan those within tissues of the control pregnant rat. We then evaluate the tissues of offspring female rats. Results: It is shown that acoustic impedance values of tissues of offspring rats of DEHP and DBP delivered mother rats are higher than those of tissues of the control offspring rats of the control mother rat. Besides, element analysis revealshigher element levels in the tissues of offspring rats of DEHP and DBP delivered mother rats. Conclusion: Therefore, we can conclude that phthalates cause structural and functional changes within rat internal organs such as lungs, brain and heart. In summary, both modalities are confirmatory in a way that tissues of DEHP and DBP exposed pregnant rats and their offspring rats are differentiated by different acoustic impedance values obtained by SAM and higher element levels specified by ICP-OES.

Hereditary spherocytosis (HS) is characterized by the morphological transformation of erythrocytes into a spherical shape due to a hereditary defect in cell membrane proteins (ghosts) associated with disruption of erythrocyte skeletal structures. Contrary to the literature, pores were detected in the erythrocytes of a patient with HS. The aim of the present study was to determine the affected proteins and genes that were responsible for the pores. Ghost isolation was performed to determine the proteins responsible for the pores observed on the erythrocytes of the patient. Erythrocyte membrane proteins were visualized using SDS‑PAGE. Exome and matrix‑assisted laser desorption/ionization time‑of‑flight mass spectrometry (MALDI TOF MS) analyses were used to identify the genes and proteins responsible for the observed defect. Quantitative protein assessments were performed using MALDI TOF MS. A difference was detected in the components of the erythrocyte membrane proteins. Band 3 and protein 4.2, which serve a particular role in membrane structure, decreased 4.573 and 4.106 fold, respectively. Through proteomic analyses, a non‑synonymous exonic mutation region was identified in the Golgi membrane protein 1 (GOLM1) gene (Chr9 rs142242230). Sorting Intolerant From Tolerant and Polymorphism Phenotyping Scores, Likelihood Ratio Tests and MutationTaster revealed that the mutation was deleterious. The pores observed in the morphology of the erythrocytes may have developed due to the decrease in these proteins, which reside in the erythrocyte membrane structure. Furthermore, genetic profiling of the patient with HS and her family was conducted in the present study. Next‑generation sequencing was used, and the genetic source of HS was identified as a GOLM1 gene mutation. The assessment of specific molecular defects is often not performed as the majority of mutations are unique to a family. However, molecular analyses should be performed in severe cases where prenatal diagnosis is required, or for unique HS phenotypes to aid scientific investigation.

Objective: Breast cancer is the type of cancer that starts in breast cells and has the highest incidence in the world after lung cancer. Doxorubicin is widely used in the treatment of some leukemia and Hodgkin’s lymphoma, as well as in the treatment of bladder, breast, stomach, lung, ovary, thyroid, soft tissue sarcoma, multiple myeloma and other cancers. We want to determine the binding interaction of Doxorubicin in the active site of the Galectin-3 with ASN-48, ARG-32, ASN-62 and GLU-72 residues, conformation and docking score energies. Material and Method: We used docking methods to detect the efficiency of Doxorubicin at breast cancer cells, clarify the role of Galectin-3 and elucidate the interaction between Galectin-3 and Doxorubicin. Results: The docking score obtained for Doxorubicin in the active side of the Galectin-3 protein was -5.32 kcal/mol. In to the active site of protein, Doxorubicin was bound with strong hydrogen bond by the residue ASN-48, ARG-32, ASN-62 and GLU-72, and a salt bridge with the same amino acid residue were established and stability was achieved. Conclusion: The development of specific therapies targeting cancer stem cells may provide hope for prolonging the life span and improving quality.

BACKGROUND:X-rays are potential mutagenic agents that can cause both the gene mutations and chromosomal aberrations. AIMS:In this study, the micronucleus (MN) test and the comet assay methods are implemented in order to observe the damage that can occur in the cell nucleus and in the structure of DNA of the patients who underwent a panoramic examination. METHODS AND MATERIALS:In our study, buccal mucosa swabs were obtained just before the radiography and 2 weeks after the radiography from 30 volunteer patients who had to take radiographs due to dental diagnosis. Changes in the nuclei of 1,000 cells of each swab sample had been counted under a light microscope and recorded. Besides, 100 cells of each other swab samples were analyzed by the comet assay. Comet assay parameters namely tail length and percentage of DNA in tail, which indicate the level of DNA damage were analyzed and compared in both groups. Statistical analysis was performed by using the Statistical Package for the Social Sciences (Version 21). RESULTS:In our study, the results of percentage of DNA in tail and tail length before and after X-ray exposure were statistically significant (P < 0.001). Likewise, increase in the MN frequency observed in buccal mucosa cells after X-ray exposure was found significant (P < 0.001). CONCLUSIONS:As a result, panoramic radiographs taken during dental diagnosis and treatment cause cytotoxicity and DNA damage in oral mucosal cells. Panoramic radiographs should be applied only when necessary, using an accurate radiographic technique and radioprotection criteria.

Microcalcification is the precursor of vulnerability of plaques in humans. Visualization of such small structures in vivo with high spatial resolution is an unsolved issue. The goal of this study is to evaluate the potential of scanning acoustic microscopy (SAM) in the determination of atherosclerotic plaques with calcifications by validating this technique with micro-computer tomography (micro-CT), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS). The fibrocalcific plaques were obtained from 12 different patients and initially examined with micro-CT. The images exhibited calcifications within these plaques. For imaging with SAM, approximately 5 μ m thick slices were prepared. Sound speed values within calcified regions were measured to be greater than the ones in collagen-rich regions. These fibrocalcific plaques were also examined with SEM and EDS revealing collagen and calcium deposition within these samples. The consistency of the results obtained by all of the modalities involved in our study is an indication of the potential of SAM as a clinical tool for the diagnosis of vulnerable plaques.

Verbascum cinsi halk tıbbında geleneksel olarak kullanılan ve anti-enflamatuvar, antioksidan ve antibakteriyal aktiviteleri bilinen bir cinstir. Verbascum inulifolium türü Türkiye’de yetişen endemik bir türdür. Bu çalışmanın amacı V. inulifolium türünün toprak üstü kısımlarından elde edilen metanol ekstresinin insan meme kanseri hücre hattı (MCF7) üzerine antiproliferatif ve apoptotik etkilerinin değerlendirilmesidir. Ekstrenin farklı konsantrasyonlarının (1, 5, 25 ve 125µg/ml) hücre canlılığı ve proliferasyonu üzerine etkisi MTT ve Xcellange yöntemi ile test edilmiştir. Daha sonra hücreler Annexine V/propodium iodür (PI) ile boyanmış ve apoptozun safhaları FACS cell counter metodu ile belirlenmiştir. Ayrıca apoptotik ve anti-apoptotik protein ekspresyon düzeyleri western blot metodu ile belirlenmiştir. Son olarak phallodine boyama metodu ile hücrelerde aktin flamentleri boyanmış ve floresan mikroskobunda incelenmiştir. MCF-7 hücre canlılığı ve proliferasyonun doz artışına bağlı olarak azaldığı belirlenmiştir. Ancak apoptozun erken veya geç dönemlerinde MCF-7 hücre sayılarında bir değişim gözlenmemiştir. P53, Bcl-2 ve kaspaz-3 protein ekspresyon düzeylerinde anlamlı bir değişim gözlenmemiştir. Ayrıca aktin flamentlerinin yoğunluğu sadece 125 mg/ml dozda hafif azalmıştır. Sonuç olarak V. inulifolium türünün methanol ekstresi MCF-7 hücreleri üzerine anti-proliferatif etki göstermiştir, fakat sonuçlar bu etkinin apoptotik yolakla ilişkili olmadığı göstermiştir.

The harmful effects of tobacco use and nicotine are well known. There is a strong epidemiological evidence for smoking as a risk factor for lung cancer and oral cavity cancer. The evidence for second-hand smoke as a risk factor for breast cancer is rapidly accumulating. The single-cell gel electrophoresis (Comet) assay has been widely used to measure DNA damage in human breast cancer cells in a variety of physiological and pathological conditions. These damages occured in DNA are some structural damages such as single- and double- strand breaks, oxidative base damage, and DNA-DNA, DNA-protein, DNA- Drug crosslinking and mutations in DNA. Single cell gel electrophoresis (SCGE) or so-called “Comet assay” is a rapid, simple and very sensitive fluorescent microscopic method to examine DNA damage at individual cell level in living populations. We investigated the effects of nicotine on breast cancer cells (MCF-7) in vitro and examined breast cancer cells line by using the Comet assay. This was seen most clearly when changes in the Comet tail length were measured when there were discrete undamaged and damaged populations. After increasing doses of nicotine, an increasing proportion of cells were found in the damaged population. These observations led to the hypothesis that nicotine might be playing a direct role in the promotion and progression of human breast cancers. Here, we study on the role and the effects of nicotine on breast cancer cell growth and physiology. However, the mechanisms by which nicotine promotes cancer development and metastasis are not fully understood. We determined the interaction between nicotine and the receptor is a protein (galectin-3) by molecular docking technique. It has recently been determined that development of breast cancer is originated from cancer stem cells, which are a minor population of breast cancer. In the present study; it is thought that the DNA damage is related to cancer stem cell number increase. Keywords : Nicotine, breast cancer, docking, comet assay, DNA damage

Niğde Ömer Halisdemir Üniversitesi, Tıp Fakültesi, Biyofizik AD, 51240 Niğde, Türkiye Niğde Ömer Halisdemir Üniversitesi, Tıp Fakültesi Fizyoloji AD, 51240 Niğde, Türkiye Çukurova Üniversitesi, Tıp Fakültesi Biyofizik AD, 01330 Adana, Türkiye Adıyaman Üniversitesi, Tıp Fakültesi, Tıbbi Biyoloji ABD, 02200 Adıyaman, Türkiye İstanbul Üniversitesi, İstanbul Tıp Fakültesi, Biyofizik AD, 34093 İstanbul, Türkiye Haliç Üniversitesi, Tıp Fakültesi, Biyofizik AD, 34445 İstanbul, Türkiye Hacettepe Üniversitesi, Fen Fakültesi, Biyoloji Bölümü, Zooloji AD, 06800 Ankara, Türkiye Adıyaman Üniversitesi, Tıp Fakültesi, Genetik Anabilim Dalı, 02200 Adıyaman, Türkiye Gaziantep Üniversitesi, Nurdağı MYO. Tıbbi ve Aromatik Bitkiler ABD, 27840 Gaziantep, Türkiye Adıyaman Üniversitesi, Tıp Fakültesi, Biyoistatistik ve Tıp Bilişimi ABD, 02200 Adıyaman, Türkiye

Breast cancer is the most common type of cancer in females and the second most common cause of cancer mortality after lung cancer. Cancer stem cells represent a novel approach to target cancer and reduce cancer recurrence and metastasis. Many patients with breast cancer continue to smoke after receiving their diagnosis. Nicotine is a key factor in tobacco addiction and also changes some cellular functions, such as activation of mitogenic pathways, angiogenesis and cell proliferation. In the present study, the impact of nicotine was assessed in a population of MCF-7 human breast cancer cells. Cluster of differentiation (CD)44+CD24- cancer stem cell population of MCF-7 cells were evaluated using flow cytometry and scanning electron microscopy. Chemoresistance effects of nicotine were demonstrated in these cells. These findings demonstrated harmful effects of nicotine following metastasis of cancer, owing to the chemoresistance produced through uninterrupted smoking, which may impact the effectiveness of treatment.

We investigated the cytotoxic and apoptotic effects of a methanol extract of Centaurea nerimaniae, a plant endemic in Turkey, on HeLa and MDA-MB-231 cells. Eight concentrations of C. nerimaniae extract were applied to cells, and cytotoxic effects were measured using the xCELLigence system. The TUNEL assay was used to assess apoptotic cell death and immunohistochemistry was used to determine active caspase-3 using the effective cytotoxic doses of the extract. Doses of 1.42 mg/ml C. nerimaniae inhibited the growth of HeLa cells and 3.67 mg/ml C. nerimaniae inhibited the growth of MDA-MB-231 cells in a dose- and time-dependent manner. The apoptotic indexes for HeLa and MDA-MB-231 cells were increased significantly compared to control groups. Immunohistochemistry showed that the number of caspase-3 immunostained cells increased in the extract treatment groups for both HeLa and MDA-MB-231 cells. In the MDA-MB-231 cell line, caspase-3 immunostaining was observed in nuclei and/or cytoplasm in the extract treated group. Caspase-3 activation was greater in HeLa cells than in MDA-MB-231 cells. We found that the extract of C. nerimaniae had a strong antiproliferative effect and induced apoptosis via caspase-3; MDA-MB-231 cancer cells were more resistant than HeLa cells.

In the present study, we aimed to evaluate the anticancer activities of Cetraria islandica (C.islandica) extracts on MCF-7 breast cancer cell lines. Cell viability, protein levels, apoptotic cells number, F-actin distribution were measured. Cell viability of MCF-7 breast cancer cells was found to be reduced in a dose-dependent manner.EC50 values of C.islandica on MCF-7 cells were found to be 9.2047 E-5 g/ml (cell amount) by using intelligence system. Expressions of p53, caspase 3 and Bcl-2, were shown to be elevated after low doses of extract and diminished after high dose treatments. PPAR- protein level was decreased, although AMP-activated kinases-α1 (AMPK-α1) protein level was increasedin its extract groups. ERK1/2 protein level was also elevated in its extract groups. 125 mg/ml of extract treated cells show a low decrease in actin filament density. MCF-7 cells with C.islandica treatment for 24 h increased the apoptotic cell percentage, though the cells-treated with C.islandica for 48 was high necrotic cells percentage. Consequently, the C.islandica extract treatment causes to elevate ERK1/2 and AMPK-α1 protein levels, resulting in PPAR- and then triggers the apoptosis by modulation caspase-3 and P53 protein levels. Therefore, C.islandica might be a good candidate for anticancer tissue, especially soft tissue tumours.