Browsing by Author "Baloglu M."

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Ceftriaxone and phenylalanine combination as broad spectrum antimicrobials therapy
(2017-08-01) Sayiner H.; Ganim M.; Altunoglu Y.; Baloglu M.; Kandemirli F.
Background: Ceftriaxone belongs to the third-generation β-lactam antibiotics and it is useful for the treatment of a number of infectious diseases caused by both aerobic and anaerobic Gram-positive and Gram-negative bacteria. Phenylalanine is an essential aromatic amino acid of human being, from which dopamine and norepinephrine neurotransmitters are being synthesized. In the present study, we examined their combined efficacy against different types of pathogenic bacterial strains. Objective: The aim of the study was designed to investigate the effects of ceftriaxone combination with phenylalanine on antimicrobial activity Method: Gram positive-bacteria and Gram negative- including Klebsiella pneumoniae, - ATCC 25923, Proteus vulgaris, Escherichia coli, Serratia marcrescens, Staphylococcus epidermis, Alpha haemolyticus streptococcus, Enterococcus faecium, Pseudomonas aeruginosa, Listeria monocytogenes ATCC 7644, Enterococcus durans, Salmonella kentucky, Enterobacter aerogenes ATCC 13048 and Candida albicans ATCC 26555 were exposed to ceftriaxone and phenylalanine based on disk-diffusion method, and Minimum inhibition concentration (MIC) was determined with ceftriaxone. Results: 0.3 mol/L ceftriaxone and 0.3 mol/L phenylalanine solutions were mixed and observed greater zone of inhibition than ceftriaxone or phenylalanine alone against above mentioned bacterial strains. These results might open up a new avenue for using phenylalanine in combination with ceftriaxone to lower MIC level for better antibacterial effect, to reduce side effects of antibiotics, and to reduce emerging threats of antibiotic resistance bacteria. Conclusion: In this study, combined use of phenylalanine and ceftriaxone has revealed increased antimicrobial sensitivity against some selected both Gram positive and Gram negative bacteria in vitro.
Genome-wide analysis of the bZIP transcription factors in cucumber
(2014-04-23) Baloglu M.; Eldem V.; Hajyzadeh M.; Unver T.
bZIP proteins are one of the largest transcriptional regulators playing crucial roles in plant development, physiological processes, and biotic/abiotic stress responses. Despite the availability of recently published draft genome sequence of Cucumis sativus, no comprehensive investigation of these family members has been presented for cucumber. We have identified 64 bZIP transcription factor-encoding genes in the cucumber genome. Based on structural features of their encoded proteins, CsbZIP genes could be classified into 6 groups. Cucumber bZIP genes were expanded mainly by segmental duplication rather than tandem duplication. Although segmental duplication rate of the CsbZIP genes was lower than that of Arabidopsis, rice and sorghum, it was observed as a common expansion mechanism. Some orthologous relationships and chromosomal rearrangements were observed according to comparative mapping analysis with other species. Genome-wide expression analysis of bZIP genes indicated that 64 CsbZIP genes were differentially expressed in at least one of the ten sampled tissues. A total of 4 CsbZIP genes displayed higher expression values in leaf, flowers and root tissues. The in silico micro-RNA (miRNA) and target transcript analyses identified that a total of 21 CsbZIP genes were targeted by 38 plant miRNAs. CsbZIP20 and CsbZIP22 are the most targeted by miR165 and miR166 family members, respectively. We also analyzed the expression of ten CsbZIP genes in the root and leaf tissues of drought-stressed cucumber using quantitative RT-PCR. All of the selected CsbZIP genes were measured as increased in root tissue at 24th h upon PEG treatment. Contrarily, the down-regulation was observed in leaf tissues of all analyzed CsbZIP genes. CsbZIP12 and CsbZIP44 genes showed gradual induction of expression in root tissues during time points. This genome-wide identification and expression profiling provides new opportunities for cloning and functional analyses, which may be used in further studies for improving stress tolerance in plants. © 2014 Baloglu et al.
Genome-wide in silico identification and comparison of Growth Regulating Factor (GRF) genes in Cucurbitaceae family
(2014-01-01) Baloglu M.
Growth-regulating factor (GRF) genes play a regulatory role for plant growth and development. The recently available cucumber, melon and watermelon genomes provide an opportunity to conduct a comprehensive overview of the GRF gene family. In the present study, identification and analysis of the GRF gene family was conducted using bioinformatics methods. Totally, 24 potential GRF genes were identified in cucumber, melon and watermelon. Cucumber and watermelon GRF gene members were physically mapped to their corresponding chromosomes. All GRF genes contain an intron whose number ranging from 2 to 3. Phylogenetic analysis categorized the cucurbit GRF proteins into 2 distinct classes. GRF proteins of cucurbits and Arabidopsis were clustered together in a joined tree and grouped into the same cluster with high bootstrap values. WRC and QLQ motifs, specific for GRF proteins, were found in all predicted GRF proteins. Gene Ontology analysis showed that majority of the GRFs was predicted to function in response to biological regulation and binding activity. In addition, predicted GRF proteins were localized in the nucleus. These results provide information about the relationship between evolution and functional divergence in the GRF family. We assume that systematic characterization of these GRF genes will enable researcher to open new insights for further exploration into the functions of this significant gene family in Cucurbitaceae family members.
Investigation of chemical profile, biological properties of Lotus corniculatus L. extracts and their apoptotic-autophagic effects on breast cancer cells
(2019-09-10) Yerlikaya S.; Baloglu M.; Diuzheva A.; Jekő J.; Cziáky Z.; Zengin G.
This study aimed to reveal chemical profiles and biological activities of ethyl acetate (EA), methanol (MeOH), and water extracts of Lotus corniculatus. Ethnobotanical reports have indicated the importance of phytochemical properties of the genus Lotus. In this study, the effects of medicinal plant extracts on antioxidant (DPPH, ABTS, CUPRAC, FRAP, phosphomolybdenum, and metal chelating assays), enzyme inhibitory (on cholinesterase, tyrosinase, a-amylase and a-glucosidase), DNA protection and anticancer properties (including anti-proliferative, cell death and telomerase activity marker gene analysis, apoptotic DNA fragmentation analysis, cell migration test) were evaluated. According to chemical analysis, quercetin derivatives geraldol, isorhamnetin and kaempferol-O-coumaroylhexoside-O-deoxyhexoside isomers were dominant in the extracts. MeOH extracts showed the highest total flavonoids capacity with 21.13 mg RE/g. EA extract showed the strongest anti-amylase activity among the tested extracts. Water extract had the most protective activity against plasmid DNA. To indicate cell survival, MTT test was performed against human MCF-7 and MDA-MB-231 breast cancer cells. Half-maximal inhibitory concentration for cells were calculated and used for detection of mechanisms behind the cancer cell death. EA extract showed up-regulation of Bax proapoptotic gene and apoptotic DNA fragmentation activity on highly invasive MDA-MB-231 cells. Beclin-1 and LC3-II autophagy genes were higly expressed after treatment of MCF-7 cells with EA extracts. EA and MeOH extracts inhibited cell migration ability of both cancer cells. Linoleamide, was dominant component in EA extract and caused apoptosis on MDA-MB-231 breast cancer cells via increasing intranuclear Ca²+. The detailed mechanism behind the anticancer properties should be further investigated.
Investigations into the therapeutic potential of Asphodeline liburnica roots: In vitro and in silico biochemical and toxicological perspectives
(2018-10-01) Locatelli M.; Yerlikaya S.; Baloglu M.; Zengin G.; Altunoglu Y.; Cacciagrano F.; Campestre C.; Mahomoodally M.; Mollica A.
This study aims to establish the biological and chemical profile of Asphodeline liburnica (Scop.) Rchb. root. The antioxidant, antimicrobial, enzyme inhibitory, DNA protection, apoptotic DNA ladder fragmentation analysis, and anti-proliferative of A. liburnica were established using standard assays. In silico study was also performed to understand interactions between quantified anthraquinones and key enzymes of clinical relevance. Total phenolic and flavonoid contents were found to be 9.67 mgGAE/g and 1.48 mgRE/g extract, respectively. Chrysophanol was detected as a major anthraquinone. The extract exhibited radical scavenging ability against DPPH and ABTS with values of 13.23 and 66.99 mgTE/g extract, respectively. Good inhibitory activity against tyrosinase was recorded. In silico experiments showed that the anthraquinones were able to establish coordinative bonds with the copper atoms present in the enzymatic cavity of tyrosinase. MTT cell viability test on MDA-MB-231 cells showed that at 0.1 and 1 μg of extracts induced anti-proliferative effect. Apoptotic DNA fragmentation analysis indicated nuclear condensation resulting in DNA fragmentation, which exhibited apoptotic cell death in the presence of A. liburnica. This study has provided insights on the potential usage of A. liburnica which could open new avenues for research and stimulate future interest for the development of safe novel biopharmaceuticals.
Molecular characterization, 3D model analysis, and expression pattern of the CmUBC gene encoding the melon ubiquitin-conjugating enzyme under drought and salt stress conditions
(2014-02-01) Baloglu M.; Patir M.
Ubiquitin-conjugating (UBC) enzyme is a key enzyme in ubiquitination. Here, we describe the cloning, characterization, and expression pattern of a novel gene, CmUBC, from a melon. Comparison of the deduced amino acid sequences allowed the identification of highly conserved motifs. Synteny analysis between Cucumis sativus L. and Arabidopsis demonstrated that homologs of several Cucumis UBC genes were found in corresponding syntenic blocks of Arabidopsis. The homology structure model of the CmUBC protein was constructed. UBCs from melon, yeast, and Arabidopsis were highly conserved in their three-dimensional folding. CmUBC was ubiquitously expressed in all melon tissues. Increased transcript levels of CmUBC were observed during drought and salinity stresses, which suggested that the expression of the CmUBC gene in melon plants is responsive to physiological water stress. These results suggested that the CmUBC gene might play an important role in the modulation of the ubiquitination pathway. © 2013 Springer Science+Business Media New York.
Multi-targeted potential of Pittosporum senacia Putt.: HPLC-ESI-MSⁿ analysis, in silico docking, DNA protection, antimicrobial, enzyme inhibition, anti-cancer and apoptotic activity
(2019-12-01) Mahomoodally M.; Picot-Allain C.; Hosenally M.; Ugurlu A.; Mollica A.; Stefanucci A.; Llorent-Martínez E.; Baloglu M.; Zengin G.
Pittosporum senacia (PS) Putt. (Pittosporaceae), indigenous to the Mascarene Islands, is a common ingredient in traditional medicines. However, there is currently a dearth of studies to validate some of these traditional claims. Given the broad traditional uses of PS against several diseases, we aimed to provide a comprehensive insight into the biological and chemical profile of P. senacia. The antioxidant, enzyme inhibitory activity, anticancer, and phytochemical composition of the methanolic extract of P. senacia leaf extracts were studied. The possible interaction and binding mode of the most abundant phytochemicals were studied via in silico docking experiments on tyrosinase and α-glucosidase. The mechanism behind the cytotoxic property of P. senacia extract for MDA-MB-231 was also examined using different methods including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability test checking apoptosis-associated genes, and wound healing assays. Twenty-six compounds were identified, of which caffeoylquinic acid derivatives, ferulic acid derivative, cinnamoylquinic acid derivative and two other polyphenols (oleuropeine and isoramnetin glucoside) being abundant, have been tested using in silico studies, against α-glucosidase and tyrosinase. The extract (IC50 = 118.8 μg/ml) exhibited time and dose dependent anti-proliferative effect on human breast cancer cell line, MDA-MB-231. According to the expression profile of apoptosis inhibitors and apoptosis promoters genes, expression of Bax and Bak genes were significantly increased compared to Bcl-2 and Birc5 genes. Based on wound healing analysis, cell migration was inhibited after the application of the plant extract. The present findings suggested that PS might be a good candidate as sources of bioactive compounds for designing functional applications.
Synergistic and Antagonistic Effects of Phenylalanine and Various Antibiotics on the Growth of Pathogenic Bacteria
(2019-06-15) Sen F.; Ganim M.; Baloglu M.; Aygun A.; Sayiner H.; Altunoglu Y.; Kandemirli F.; Demirkan B.; Kuyuldar E.; Bulut E.
Broad-spectrum antibiotics have been widely used in the treatment of many systemic and local infections in humans and animals. Herein, we aimed to determine the synergistic and antagonistic effects of phenylalanine with antibiotics cefoxitin, amoxicillin, vancomycin, lincomycin, and bacitracin against 14 pathogenic bacteria. The effect of antibiotics, either alone or in combination with this biomolecular liquid, was tested using the disk diffusion method against different bacteria. The addition of phenylalanine to antibiotic disks directly affected their antimicrobial activity. All the antibiotics used did not show any antimicrobial activity against Staphylococcus haemolyticus when used alone. However, in combination with phenylalanine, each antibiotic inhibited the growth of S. haemolyticus. The use of this biomolecular liquid together with amoxicillin and vancomycin also increased the antimicrobial activity against Enterococcus durans. The use of phenylalanine in combination with antibiotics also resulted in antagonistic effects on some pathogens. Further, the effects of antibiotics in combination with phenylalanine on different bacterial pathogens were investigated in vitro. Results provide valuable information to further our understanding of the molecular mechanism of action of antibiotics and to improve their efficacy against bacterial pathogens.
Syzgium coriaceum Bosser & J. Guého—An endemic plant potentiates conventional antibiotics, inhibits clinical enzymes and induces apoptosis in breast cancer cells
(2020-01-01) Mahomoodally M.; Ugurlu A.; Llorent-Martínez E.; Nagamootoo M.; Picot-Allain M.; Baloglu M.; Altunoglu Y.; Hosenally M.; Zengin G.
Syzygium species are renowned for being important reservoirs of phytochemicals with pharmaceutical and biomedical potential. However, no attempt has been made to delineate the pharmacological potential and phytochemical profile of Syzgium coriaceum Bosser & J. Guého, an endemic plant to Mauritius. The present study aimed to determine the antibacterial, antioxidant, cytotoxicity, enzyme inhibitory and phytochemical profile of the ethyl acetate and methanol extracts of S. coriaceum. Preliminary qualitative phytochemical study of the extracts showed the presence of phenol, tannins, and alkaloids. Chemical characterisation showed the presence of derivatives of tannins, gallic acids, quercetin, and kaempferol. Potentiating activity between S. coriaceum extracts and antibiotics (ampicillin and streptomycin) using the checkerboard method showed additive interaction. The extracts showed potent 2,2-diphenyl-1-picrylhydrazyl (DPPH) (2.95 and 2.93 mmol trolox equivalent (TE)/g sample for ethyl acetate and methanol extracts, respectively) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) (4.09 and 3.83 mmol TE/g sample for ethyl acetate and methanol extracts, respectively) scavenging abilities. Syzygium coriaceum extracts were active inhibitors of α-glucosidase (about 47 mmol acarbose equivalent/g sample for ethyl acetate and methanol extract). S. coriaceum methanol extract caused maximum inhibition against human breast adenocarcinoma (MDA-MB-231) cancer cells after 48 h treatment with the IC50 value of 53.41 μg/mL. Expression of anti-apoptotic Bcl2 and BIRC5 genes were down-regulated. It can be concluded that S. coriaceum extracts lead to MDA-MB-231 cells apoptosis. This investigation has provided a comprehensive report of the biological and chemical profile of S. coriaceum. Collected scientific evidences can open new avenues for research and contributes towards establishing primary data on Syzygium species endemic to Mauritius for bioprospection of novel phytopharmaceuticals.
Vector construction strategies for transformation of wheat plant
(2013-07-08) Baloglu M.; Battal A.; Aydin G.; Eroglu A.; Oz M.; Kavas M.; Oktem H.; Yucel M.
Gene cloning and vector construction for plant genetic transformation are routine procedures in plant functional genomic studies. The availability of effective transformation vector is one of the pre-requisites for plant transformation studies. Here, we describe the construction of a series of transformation vectors through different genetic engineering techniques for wheat transformation. For this purpose, NAC-type transcription factor genes TaNAC69-1andTtNAM-B2 were isolated from T. aestivum and T. turgidum, respectively. Then they were cloned into different types of cloning and expression vectors. Besides traditional restriction enzyme digestion and ligation method, Gateway cloning technology which is a fast and reliable alternative cloning method were used for construction of wheat transformation vectors. The transformation vectors constructed in this study are suitable for use in both particle bombardment (biolistic) and Agrobacterium based transformation protocols for wheat plant.