Search

Testicular torsion leads ischaemic injury and generates reactive oxygen species. Reactive oxygen species triggers lipid peroxidation, protein degradation and DNA damage. These biochemical processes trigger tissue damage. Heat shock proteins (HSPs) are important in spermatogenesis, and this work elucidates role of HSPs at the testicular torsion-detorsion process. A proton-pump inhibitor, omeprazole, tested to reveal the drug's curative effect since HSP functions through ATP hydrolysis. Thirty-two male Wistar Albino rats were divided into four groups: sham, control, omeprazole and serum physiologic groups. Right testis was torsed, while left ones remained untorsed. Protein peroxidation, DNA damage and lipid hydroperoxide levels as well as HSP expression were measured. Further, the effects were visualised with histopathologic imaging. HSP expression increases at the torsed right testis compared to the contralateral testis. Although HSP70 and HSP90 help antioxidant enzymes to keep their native structure, their anti-apoptotic properties accelerate the tissue damage. Omeprazole a proton-pump inhibitor employed to impair electron transfer chain and to inhibit HSP ATPase function. Omeprazole effectively inhibits HSPs and alleviates lipid peroxidation and DNA damage levels both at molecular and at tissue level, and the drug has profound curative effect on testicular torsion recovery.

Background Fibronectin (FN) is an indispensable part of the extracellular matrix. During regeneration or wound healing, the plasma form of FN is incorporated into the fibrin clots to form a temporary fibrin-FN matrix, and also locally synthesized cellular FN migrates to the clot to regenerate the injured tissue. We aimed to examine wound tissue FN EIIIB and plasma FN EIIIB expression levels in an experimental wound healing model in rabbits. Methods Plasma and tissue EIIIB splice variant expressions were measured serially with RT-qPCR in a cutaneous wound model of rabbits. Results Tissue FN expression increased as beginning on day 3 and continued to increase on days 6 and 9, reaching maximum expression at day 12 before starting to decrease. On the contrary to the tissue levels, plasma FN levels gradually decreased until day 15 when expression returned to the initial values. Conclusion The findings of the current study support that tissue EIIIB expression level increases during wound healing; and plasma EIIIB expression level decreases minimal changed. This is in contrast to reports where plasma FN provisionally helps ECM formation. Therefore, our data show an essential role of EIIIB at the tissue level in accelerating the wound healing process. The RT-qPCR method in our experimental setup can provide more accurate and precise results compared to the antibody-based methods.

Background: HSP70 is a survival factor for tumor cells in transformation and in tumor progression as well as in anti-apoptotic response. Objective: Several inhibitors targeting HSP70 ATPase function displayed off-target affect but PES which targets HSP70 substrate binding domain prevents tumor cell survival prominently. However, PES may not bind HSP70 in the absence of nucleotide. This research aimed to design a unique inhibitor molecule that work both in the presence and absence of nucleotides to amplify inhibition. Methods: A set of chimeric coumarine-pyrazole derivatives determined by in silico techniques and synthesized to elucidate their inhibitory effects. Cell viability experiments displayed KBR1307 as the most efficient inhibitor. A set of characterization experiments performed, and results compared to that of PES agent. Binding constant, ATP hydrolysis rate, and percent aggregation determined in the presence and absence of inhibitors. Results: In silico docking experiments showed that only KBR1307 binds the HSP70 substrate binding domain and interacts with cochaperone interface. Binding experiments indicated that KBR1307 binds HSP70 both in the presence and absence of nucleotides, but PES does not. Both inhibitors significantly lower HSP70 ATPase activity and substrate protein disaggregation activity. However, KBR1307 displays a lower IC50 value at the MCF-7 cell line compared to PES. Both inhibitors do not alter HSP70 secondary structure composition and overall stability. Conclusion: KBR1307 effectively inhibits HSP70 compared to PES and provides promising template for novel anticancer drug development.

WOS: 000425757900001, PubMed ID: 29466082, Objective: The aim of this study was to determine the optimum operating parameters (pulse duration, energy levels, and application time) to promote induction of osteoblast and fibroblast cell proliferation and to maintain cell viability treated with low-intensity pulsed ultrasound (LIPUS) and low-level laser therapy (LLLT). Background data: The positive effects of LIPUS and LLLT on cellular activity have been reported in recent years. Comparisons between experimental parameters of previous studies are difficult because scientific studies reported frequencies and the duty cycles of LIPUS and wavelengths and doses of LLLT in a wide range of parameters. However, optimum amount of energy and optimum time exposure must be determined to induce bone and tissue cell proliferation for effective healing process and to avoid cell damage. Material and methods: Fibroblast and osteoblast cell cultures were irradiated with LIPUS (10-50% pulse and continuous mode at 1 and 3MHz for 1, 3, and 5min) and LLLT (4, 8, and 16J at 50, 100, 200, 300, 400, and 500mW). Cell cultures were analyzed using XTT assay. Results: For both cell types, LIPUS treatment with 10% pulse (1:9 duty cycle), 3MHz, and for 1min and LLLT treatment over 100mV for 4, 8, and 16J modalities contributed to the growth, and may help bone repair and tissue healing process optimally. Conclusions: Bio-stimulating effects of LLLT irradiation promote proliferation and maintain cell viability better than LIPUS treatment without causing thermal response for both cell types, and the therapeutic modality above 200mV has maximum effectiveness., Scientific Research Projects Unit of Cumhuriyet University, Sivas, Turkey [DIS-130], This study was funded by Scientific Research Projects Unit of Cumhuriyet University, Sivas, Turkey (Project No. DIS-130).

WOS: 000386021200007, Carbon monoxide (CO) is an important signaling molecule which plays significant roles in the pathogenesis of cancer. CO is produced by enzymatic degradation of heme in mammals. Heme oxygenase 1 (HO-1) catalyzes the breakdown of heme into CO, ferrous iron, and biliverdin. CO induces HO-1 and inhibits cell proliferation. Cancer cells exposed to several stress factors (hypoxia, reactive oxygen species, cis-platin, and oxidative stress), and HO-1 displays cytoprotective role against oxidative stress and inhibits apoptosis, metastases, angiogenesis, and cell proliferation processes. Therefore, metal containing CO-releasing molecules (CORMs) have been designed as an effective cancer treatment strategy. CORMs are responsible for releasing controlled amounts of CO to cells and tissues. Thus, we synthesized [Mn(CO)(3)(bpy)L]X manganese containing CORMs [bpy=2,2-bipyridine, X=hexafluorophosphate (PF6), trifluoromethanesulfonate (OTf), L=imidazole, methylimidazole, benzimidazole, N-benzylbenzimidazole, N-(4-chlorobenzyl)benzimidazole] to release CO in human invasive ductal breast (MCF-7) cell line. In vitro experiments indicated that the compounds inhibited cell proliferation and exhibited cytotoxic effect on breast cancer cells. Moreover, side groups of the compounds enhanced the anticancer effects in MCF-7 cell line. These manganese containing CORMs gave promising results and may be used as a drug template for effective treatment of invasive ductal breast carcinoma. [GRAPHICS] ., Scientific and Technological Research Council of Turkey (TUBITAK) [112T320], This work was supported by the Scientific and Technological Research Council of Turkey (TUBITAK) [grant number 112T320].

Hindawi Limited, Introduction. Strontium salts are anti-irritants for chemically induced sensory irritation. Interstitial cystitis is a painful disease without definitive therapy. The aim of the study was to determine the effect of strontium in bladder with experimental interstitial cystitis model. Material and Methods. Rats' bladders in control group were instilled with NaCl. Second group was instilled with E. coli LPS. Third group was instilled with strontium. Fourth group was initially instilled with strontiumand then LPS. Fifth group was instilled with LPS initially and then strontium. Urine of rats was collected at the beginning and end of the study. Results. Histamine and TNF-P changes were statistically significant in the second group but were not significant in the third group.When we compared the histamine levels of second via fourth and fifth groups the changes were statistically not significant.When we compared the TNF- P levels of second via fourth and fifth groups the changes were statistically significant. Conclusions. In our model, strontiumdid not make any significant changes in histopathology or histamine levels; however, it significantly reduced the levels of TNF-P. Given the role of TNF-P in the physiopathology of interstitial cystitis, these results suggested that further studies are required to evaluate the potential use of strontium in the management of interstitial cystitis., Korgali, E.; Department of Urology, Urology Clinic, Cumhuriyet UniversityTurkey; email: estkorgali@hotmail.com

WOS: 000326985700002, PubMed ID: 24261974, Toxoplasma gondii is ubiquitous obligate intracellular parasite and is one of the most important pathogen for humans and animals. In humans, T. gondii has two life forms: tachyzoites and bradyzoites. Tachyzoites form of T. gondii can cause acute infection, and it is called toxoplasmosis. The development of latent bradyzoites from rapidly growing tachyzoites has been linked to cellular and environmental stresses which are associated with heat shock proteins (Hsps). Hsps play a protective role against stressors. Hsp40 is an important member of Hsp family and T. gondii has 36 predicted Hsp40 family members. Therefore, we studied the cloning and biochemical characterization of the T. gondii RH strain Hsp40 protein-Gok1. Hsp40 prevents protein aggregation and induce refolding. Consequently, Hsp40s may play essential roles in the mechanisms of bradyzoite development and survival in the host organism. Hsp40-Gok1 functional and structural properties may facilitate drug design and protein targeting against toxoplasmosis., Scientific and Technological Research Council of Turkey, TUBITAK [110T928]; TUBA-GEBIP [2008-29], The authors gratefully acknowledge the financial support received from the Scientific and Technological Research Council of Turkey, TUBITAK (Grant number 110T928) and partial support from TUBA-GEBIP for YT (2008-29).

WOS: 000320347100024, PubMed ID: 23664495, In this work novel organic based compounds, acyl thiourea derivatives were synthesized and their anticancer activities were investigated. A new series of acyl thiourea derivatives containing pyrazole ring were prepared in good yield through one pot reaction of 4-benzoyl-1, 5-diphenyl-1H-pyrazole-3-carbonyl chloride with ammonium thiocyanate and various amines. The structures of the newly synthesized compounds were confirmed by IR, H-1 NMR, C-13 NMR and elemental analysis. Anticancer activities of synthesized compounds were evaluated on human colon, liver and leukemia cancer cell lines. Cell culture studies have demonstrated significant toxicity of the compounds on the cell lines, and the levels of toxicity have altered in the presence of various side groups. These results confirm that novel pyrazolyl acyl thioureas derived compounds may be utilized for cancer treatment. Furthermore, these compounds have a great potential and significance for further investigations. (C)2013 Elsevier Ltd. All rights reserved., Turkish National Academy of Sciences [TUBA GEBIP 2008-29], We deeply appreciate expert Mehmet Gilmus for helping on the analysis of IR spectra. This work was funded through a seed grant from the Turkish National Academy of Sciences (TUBA GEBIP 2008-29 for Y.T.).

WOS: 000399456900007, Carbon monoxide is an important signaling molecule which is produced by heme oxygenase-1. CO shows antiproliferative activity against cancer cells; hence, activation of HO-1 is a significant inhibition strategy against tumor formation and survival of cancer cells. In this work, manganese-based CO-releasing molecules (CORMs) were designed and synthesized to inhibit breast cancer cell proliferation. Human invasive ductal breast cancer cells (MCF-7) were treated with the synthesized CORMs to investigate the effect of the complexes on breast cancer survival under UV light. In vitro experiments indicated that the complexes inhibited breast cancer cell proliferation, and further, the antiproliferative effects were increased under UV light. Thus, these novel CORMs may provide a drug template for the treatment of invasive ductal breast cancer., Scientific and Technological Research Council of Turkey (TUBITAK) [112T320], This work by Scientific and Technological Research Council of Turkey (TUBITAK, Project No: 112T320) is gratefully acknowledged.

WOS: 000383003900024, PubMed ID: 27376491, Invasive ductal carcinoma is the most common breast malignancies tumors and has tendency to bone metastases. Many oncogenic client proteins involved in formation of metastatic pathways. Stabilization, regulation, and maintenance of these oncogenic client proteins are provided with Heat Shock Protein 90 (Hsp90). Hsp90 perform these processes through its ATP binding and subsequent hydrolysis energy. Therefore, designing Hsp90 inhibitors is a novel cancer treatment method. However, many Hsp90 inhibitors have solubility problems and showed adverse effects in clinical trials. Thus, we designed and synthesized novel pyrimidinyl acyl thiourea derivatives to selectively inhibit Hsp90 alpha in human invasive ductal breast (MCF-7) and human bone osteosarcoma (Saos-2) cell lines. In vitro experiments showed that the compounds inhibited cell proliferation, ATP hydrolysis, and exhibited cytotoxic effect on these cancer cell lines. Further, gene expression was analyzed by microarray studies on MCF-7 cell lines. Several genes that play vital roles in breast cancer pathogenesis displayed altered gene expression in the presence of a selected pyrimidinyl acyl thiourea compound. Molecular docking studies were also performed to determine interaction between Hsp90 ATPase domain and pyrimidinyl acyl thiourea derivatives. The results indicated that the compounds are able to interact with Hsp90 ATP binding pocket and inhibit ATPase function. The designed compounds powerfully inhibit Hsp90 by an average of 1 mu M inhibition constant. And further, the compounds perturb Hsp90 N terminal domain proper orientation and ATP may not provide required conformational change for Hsp90 function as evidenced by in silico experiments. Therefore, the designed compounds effectively inhibited both invasive ductal breast carcinoma and bone metastasis. Pyrimidinyl acyl thiourea derivatives may provide a drug template for effective treatment of invasive ductal breast carcinoma and its bone metastasis as well as new therapeutic perspective for drug design. (C) 2016 Elsevier Masson SAS. All rights reserved., Bozok University [FBE/T127]; Scientific and Technological Research Council of Turkey [TUBITAK 114Z365], This work was funded through a seed grand from Bozok University (Project No:FBE/T127) and through The Scientific and Technological Research Council of Turkey (TUBITAK 114Z365).

Heat shock protein 90 (Hsp90) is an ATP dependent highly conserved protein that provides client proteins to reach the correct conformation. Hsp90 uses ATPase activity to perform function such as folding for substrate proteins, misfolded protein degradation, cell cycle, and signal transduction.

Stress induces radicals and this process triggers several biochemical pathways but most essential problem is the damage caused by the radicals on lipids, proteins, and nucleic acids. To perform biochemical functions, these macromolecules must be working properly. Proteins must be correctly folded so that they can regulate pathways and transmit signals. Under stress, alteration in cellular environment cause proteins to lose their structure. Then, the proteins either lose or alter their function. To scavenge radicals and to chaperone substrate proteins, a set of mechanic proteins are employed.

WOS: 000354756000013, PubMed ID: 25728232, Toxoplasma gondii is an intracellular parasitic protozoon which infects human and most warm-blooded animals. Almost one-third of the world's population is affected by life-threatening infection of T. gondii tachyzoites form. Slow growing, transmissible and encysted bradyzoites forms are composed after tachyzoites stage. Cellular and environmental stresses induce conversion of tachyzoites from bradyzoites and this condition is associated with Heat Shock Protein (Hsps) family. Hsp100 is a member of this protein family, and coordinates to disassemble protein aggregates with Hsp70 and Hsp40 in an ATP dependent manner. Several proteins are involved during this stage differentiation and Hsp100 may help them to be in their native soluble form to perform their function as observed in other organisms. For this purpose, Hsp100-Batul was isolated from T gondii RH strain to characterize its biochemical properties in this current study. Hsp100 proteins play a role in survival and virulence of pathogens as shown in the literature. Therefore, manipulation of protein protein interaction may perturb T. gondii infection and impair conversion to tachyzoites by inhibiting Hsp100 function. Therefore, results of this work present a potential route for vaccination or immunotherapy. (C) 2015 Elsevier Inc. All rights reserved., Scientific and Technological Research Council of Turkey, TUBITAK [110T928]; TUBA-GEBIP (Turkish National Academy of Science Young Scientist Grant) [2008-29], The authors gratefully acknowledge the financial support received from the Scientific and Technological Research Council of Turkey, TUBITAK (Grant # 110T928) and partial support from TUBA-GEBIP (Turkish National Academy of Science Young Scientist Grant # 2008-29) for YT.

WOS: 000319219800011, PubMed ID: 23848155, Two fish species, Cyprinion macrostomus macrostomus and Garra rufa obtuse, tolerate adverse conditions in the Kangal hot springs and cope with multiple stressors such as food deprivation, extreme temperature, toxins, protein degradation, hypoxia, and microbial damage. These fish have evolved strategies to counteract the stressors including the induction of heat shock proteins (Hsps). Hsps play an essential role in maintaining cellular homeostasis, and one of the key proteins in the mechanism is Hsp70. Hsp70 itself is exposed to the same stressors as all other proteins, and, hence, the stability of Hsp70 was investigated. For this purpose, Hsp70 ATPase activity was determined at different urea concentrations. It was found that the protein maintains considerable ATP hydrolysis activity at higher denaturant conditions. Temperature effects on the substrate peptide binding showed that Hsp70s bind prominently at elevated temperatures. Furthermore, temperature effects on Hsp70 aggregation indicated that the presence of nucleotides decreases the aggregation process. The present work has determined the stability and activity of cmHsp70 and grHsp70 themselves under extreme conditions. The stability of the Hsp70 proteins maintains substrate proteins in the native state, which may aid in the adaptation of the fish species to the hot spring environment., Turkish National Academy of Sciences [TUBA GEBIP 2008-29], This work was funded partly by the CUBAP (F 253) project and through a seed grant from the Turkish National Academy of Sciences (TUBA GEBIP 2008-29).

WOS: 000322647500001, PubMed ID: 23971043, The present work focuses on a local survey of free-living amoebae (FLA) that cause opportunistic and nonopportunistic infections in humans. Determining the prevalence of FLA in water sources can shine a light on the need to prevent FLA related illnesses. A total of 150 samples of tap water were collected from six districts of Sivas province. The samples were filtered and seeded on nonnutrient agar containing Escherichia coli spread. Thirty-three (22%) out of 150 samples were found to be positive for FLA. The FLA were identified by morphology and by PCR using 18S rDNA gene. The morphological analysis and partial sequencing of the 18S rDNA gene revealed the presence of three different species, Acanthamoeba castellanii, Acanthamoeba polyphaga, and Hartmannella vermiformis. Naegleria fowleri, Balamuthia mandrillaris, or Sappinia sp. was not isolated during the study. All A. castellanii and A. polyphaga sequence types were found to be genotype T4 that contains most of the pathogenic Acanthamoeba strains. The results indicated the occurrence and distribution of FLA species in tap water in these localities of Sivas, Turkey. Furthermore, the presence of temperature tolerant Acanthamoeba genotype T4 in tap water in the region must be taken into account for health risks., Turkish National Academy of Sciences (TUBA-GEBIP), This work was funded by a seed grant from the Turkish National Academy of Sciences (TUBA-GEBIP).

Amaç: Ülkemizde günümüze kadar su kaynaklarında serbest yaşayan amip (SYA) yaygınlığına yönelik kapsamlı bir çalışma yapılmamış, genellikle olgu sunumlarında SYA varlığı bildirilmiştir. Bu çalışmanın amacı Sivas ilinde musluk sularında ve çevresel su kaynaklarında SYA yaygınlığının belirlenmesidir. Yöntemler: Çalışma kapsamında şehir merkezinden, ilçelerden ve köylerden toplam 250 örnek toplanmıştır. Her bir kaynaktan iki litre su alınarak vakumlu filtrelerden süzülmüştür. Filtre kağıtları steril “Page's Amoeba Saline (PAS)” solüsyonunda bir gece bekletilmiştir. İnkübasyon sonrası filtreler çıkarılıp tüp santrifüj edildikten sonra dip kısımdan alınan bir iki damla örnek, Besleyici-Değeri Olmayan Agar (BDOA) plaklarına inoküle edilmiştir. İnkübasyonun üçüncü gününden itibaren besiyerlerinde üreme kontrolleri yapılmıştır. Naegleria spp. belirlenmesi için kamçı deneyi yapılmış, patojenik Acanthamoeba türleri için de 37°C, 42°C ve 52°C de ısı tolerans testi uygulanmıştır. Amiplerin kist ve trofozoit morfolojileri ışık mikroskobu altında incelenmiş ve morfolojik anahtarlar kullanılarak cins ayrımları yapılmıştır. Bulgular: İncelenen toplam 250 örneğin 75'inde (%30.0) SYA tespit edilmiştir. Bu türlerin mikroskobik olarak temel morfotiplendirme anahtarlarına göre incelemelerinde 11'inin (%4.4) Acanthamoeba spp., 25'inin (%10.0) Naegleria spp. ve 39'unun (% 15.6) Hartmannella spp., olduğu belirlenmiştir. Sonuç: Su örneklerinde bu kadar yaygın SYA bulunması, hem amiplerin kendileri hem de taşıdıkları çeşitli bakteriler nedeniyle insan sağlığı açısından önemli risk oluşturmaktadır., Objective: To our knowledge, there is no study dealing with the prevalence of free-living amoebas (FLA) in water sources in Turkey, previous studies were mostly case presentations. The aim of the present study was to investigate the prevalence of FLA from tap water and natural water sources in different parts of the city. Methods: In the study, 250 samples were collected from the city centre, districts and villages. Two litres of water was collected from each source and filtered through a vacuum filtration system. The filter papers were washed in “Page's Amoeba Saline (PAS)” solution and incubated overnight. Filter papers were removed from the tubes and centrifuged; the final pellet was inoculated on non-nutrient agar (NNA) plates. The growth rate of FLA was checked after three days of inoculation and the flagellation test was performed to determine the presence of Naegleria spp. Heat tolerance of isolated strains was checked at 37, 42 and 52°C for the presence of pathogenic Acanthamoeba species. The cyst and trophozoite morphology of amoebas were examined under a light microscope and the genera was identified according to morphotyping keys. Results: FLA were found in 75 (30.0%) of examined water samples. Eleven (4.4%) were identified as Acanthamoeba spp., 25 (10.0%) as Naegleria spp. and 39 (15.6%) as Hartmannella spp. after microscopic examination. Conclusion: Our study revealed that FLA are common inhabitants of household water as they are in the environment, so their own potential risks as well as transferring bacteria as other pathogens is important for human health.