Toxigenic Staphylococcus aureus in some animal-originated food products marketed in Turkey: presence and Public Health concerns.

This study aimed to detect the presence of Staphylococcus aureus in some animal source food (ASF) including emulsified meat products (sausage and salami), dry fermented meat product (soudjouk), semi dry meat product (pastrami) and raw chicken meat. Sixty six  (38.8%) of 170 samples were found to be positive for S. aureus. It was determined that S. aureus was found in 17 (56.6%) salami, 27 (54%) raw chicken meat, 9 (30%) soudjouk, 9 (30%) pastrami, 4 (13.3%) sausage samples. Staphylococcal enterotoxins (SEs) were identified in 5 out of 66 (7.5 %) isolates food matrices including 3 (4.5%) SEA, 2 (3.03%) SEC. The sea and sec genes were detected in 3 (4.5%) of 66 isolates. The results of this study highlight the need to provide suitable control strategies concerning production, sales, and storage to prevent the spread of enterotoxigenic S. aureus isolates in ASF. The key contribution of this study is its revelation of the presence of S. aureus in animal products sold in Turkish local markets, highlighting the potential public health risks associated with animal foods.

Investigation and characterization of Aliarcobacter spp. isolated from cattle slaughterhouse in Türkiye.

Aliarcobacter spp. have been isolated from numerous food products at retail and from animal carcasses and feces at slaughter. The objectives of this study were as follows: (i) to isolate Aliarcobacter species from different slaughterhouses' samples and (ii) to detect genetic diversity, antibiotic resistance, biofilm ability, and putative virulence gene profiles of the isolates. A molecular investigation of antibiotic resistance and virulence factors was also conducted using polymerase chain reaction (PCR). Among 150 samples, a total of 22 (14.6%) Aliarcobacter spp. isolates were obtained, with varying levels of antibiotic resistance observed. The genes tetO, tetW, and gyrA were detected in 0%, 31.8%, and 27.2% of the isolates, respectively. All isolates were resistant to ampicillin, rifampin, and erythromycin, while tetracycline was found to be the most effective antibiotic, with 81.8% of the isolates showing susceptibility to it. All isolates (100%) harbored more than one of the nine putative virulence genes tested, with 18.1% of isolates carrying more than three. Regarding biofilm formation, 7 (31.8%) and 4 (18.1%) isolates were found to form strong and moderate biofilms, respectively, while one (4.5%) isolate was classified as a weak biofilm producer. ERIC-PCR band patterns suggested that the isolated Aliarcobacter spp. from slaughterhouses had different sources of contamination. These findings highlight the potential risk posed by pathogenic and multidrug-resistant Aliarcobacter spp. in food and the need for control measures throughout the food chain to prevent the spread of these strains. The results indicate that foods of animal origin and cattle slaughterhouses are significant sources of antimicrobial resistant Aliarcobacter.

Receptor mediated targeting of EGF-conjugated alginate-PAMAM nanoparticles to lung adenocarcinoma: 2D/3D in vitro and in vivo evaluation.

Carboplatin (cis-diamine (1,1-cyclobutandicarboxylaso)­platinum (II)) is a second-generation antineoplastic drug, which is widely used for chemotherapy of lung, colon, breast, cervix, testicular and digestive system cancers. Although preferred over cisplatin due to the lower incidence of nephrotoxicity and ototoxicity, efficient carboplatin delivery remains as a major challenge. In this study, carboplatin loaded alginate- poly(amidoamine) (PAMAM) hybrid nanoparticles (CAPs) with mean sizes of 192.13 ± 4.15 nm were synthesized using a microfluidic platform, then EGF was conjugated to the surface of CAPs (EGF-CAPs) for the receptor-targeted delivery. Hence, increased FITC+ cell counts were observed in A549 spheroids after EGF-CAP treatment compared to CAP in the 3D cellular uptake study. As such, the cytotoxicity of EGF-CAP was approximately 2-fold higher with an IC50 value of 35.89 ± 10.37 µg/mL compared to the CAPs in A549 spheroids. Based on in vivo experimental animal model, anti-tumor activities of the group treated with CAP decreased by 61 %, whereas the group treated with EGF-CAP completely recovered. Additionally, EGF-CAP application was shown to induce apoptotic cell death. Our study provided a new strategy for designing a hybrid nanoparticle for EGFR targeted carboplatin delivery with improved efficacy both in vitro and in vivo applications.

Design of Poly(lactic) acid/gelatin core-shell bicomponent systems as a potential wound dressing material.

The electrospun core-shell nanofiber has great many advantages such as different types of solvents that can be used for changing flexibility, mechanical properties, or surface chemistry of fiber. Hydrophobic Poly(lactic) acid (PLA) and hydrophilic gelatin (Gel) were electrospun by various preparation conditions to design perfect bicomponent PLA:Gel nanofiber in a core-shell structure. Solvent types, the concentration of polymeric components, flow rate, and voltage of the electrospinning process were changed to optimization of nanofiber. According to the SEM images, the best nanofiber structure without beads was obtained at 0.4 ml/h flow rate of PLA solution and 1.2 ml/h flow rate of Gel solution at 45:55 (w:w %) weight ratio of PLA:Gel in trifluoroethanol solvent with a 10 kV voltage at 10 cm distance to the collector. From the TEM images, the existence of the core-shell structure had been proved which all prepared nanofibers with 2,2,2-Trifluoroethanol solvent. Furthermore, contact angle measurements showed a change in wettability when the Gel amount was increased. Therefore, the mildest synthesis conditions were determined for bicomponent PLA:Gel core-shell nanofibers as a potential wound dressing and dual drug carrier materials.

In vitro evaluation and in vivo efficacy studies of a liposomal doxorubicin-loaded glycyrretinic acid formulation for the treatment of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC), more than 800 000 cases reported annually, is the most common primary liver cancer globally. Doxorubicin hydrochloride (Dox-HCl) is a widely used chemotherapy drug for HCC, but efficacy and tolerability are limited, thus critical to develop delivery systems that can target Dox-HCl to the tumour site. In this study, liver-targeting ligand glycyrrhetinic acid (Gly) was conjugated to polyethylene glycol (PEG) via Steglich reaction and incorporated in liposomes, which were then loaded with Dox-HCl by pH gradient method. The optimal formulation Gly-Peg-Dox-ProLP-F6 showed high Dox-HCl encapsulation capacity (90.0%±1.85%), low particle size (120 ± 3.2 nm). Gly-Peg-Dox-ProLP-F6 formulation demonstrated substantially greater toxicity against HCC cells than commercial Dox-HCl formulation (greater against 1.14, 1.5, 1.24 fold against Hep G2, Mahlavu and Huh-7 cells, respectively), but was 1.86-fold less cytotoxic against non-cancerous cell line AML-12. It increased permeability from apical to basolateral (A-B) approximately 2-fold. Gly-Peg-Dox-ProLP-F6 demonstrated superior antitumor efficacy in mouse liver cancer model as evaluated by IVIS. Isolated mouse liver tissue contained 2.48-fold Dox more than Dox-HCl after administration of Gly-Peg-Dox-ProLP-F6, while accumulation in heart tissue was substantially lower. This Gly-Peg-Dox-ProLP-F6 formulation may improve HCC outcomes through superior liver targeting for enhanced tumour toxicity with lower systemic toxicity.

L-glutamic acid-g-poly hydroxyethyl methacrylate nanoparticles: acute and sub-acute toxicity and biodistribution potential in mice.

The aim of this safety study in mice was to determine in vivo toxicity and biodistribution potential of a single and multiple doses of L-glutamic acid-g-p(HEMA) polymeric nanoparticles as a drug delivery system. The single dose did not cause any lethal effect, and its acute oral LD50 was >2.000 mg/kg body weight (bw). Multiple doses (25, 50, or 100 mg/kg bw) given over 28 days resulted in no significant differences in body and relative organ weights compared to control. These results are supported by biochemical and histological findings. Moreover, nanoparticle exposure did not result in statistically significant differences in micronucleus counts in bone marrow cells compared to control. Nanoparticle distribution was time-dependent, and they reached the organs and even bone marrow by hour 6, as established by ex vivo imaging with the IVIS® spectrum imaging system. In conclusion, L-glutamic acid-g-p(HEMA) polymeric nanoparticles appear biocompatible and have a potential use as a drug delivery system.

The effect of Bdellovibrio bacteriovorus containing dressing on superficial incisional surgical site infections experimentally induced by Klebsiella pneumoniae in mice.

Bdellovibrio bacteriovorus is a bacterial agent that stands out for its ability to act as a predator against gram-negative bacteria and has found application against antibiotic-resistant pathogens. The aim of this study is to determine the efficacy of Bdellovibrio bacteriovorus against antibiotic-resistant pathogens, particularly those causing infections in surgical incision sites. A total of 6 experimental groups were created in mice, and surgical area infections were initiated with Klebsiella pneumoniae in incision sites. The effects of antibiotics and Bdellovibrio bacteriovorus alone or in combination were compared to the control group. In the Bdellovibrio bacteriovorus treatment group, edema and redness were observed in all mice at 24th hours, in 20% of mice at 48th hours, and in none at the 72 nd h. A significant difference was observed in the Bdellovibrio bacteriovorus treatment groups in reducing Klebsiella pneumoniae burden in the incision area compared to antibiotics alone or Bdellovibrio bacteriovorus + antibiotics, (p < 0.001). Likewise, cytokine level determinations indicated that B. bacteriovorus applications generated a therapeutic response without inducing an inflammatory response.

Arcobacter species isolated from various seafood and water sources; virulence genes, antibiotic resistance genes and molecular characterization.

Arcobacter spp. has gained clinical significance as an emerging diarrheagenic pathogen associated with water reservoirs in recent years. The complete clinical significance of Arcobacter remains rather speculative due to the virulence and antibiotic susceptibility of individual strains. This study aimed to assess the prevalence of Arcobacter spp. in fish, water, and shellfish. A total of 150 samples were collected from the Adana, Kayseri and Kahramanmaras provinces in Turkey. Arcobacter spp. was isolated from 32 (21%) of the 150 samples. The most prevalent species was A. cryaerophilus, 17 (56%), A. butzleri 13 (37%) and A. lacus 2 (6%). As a result, the ratios of the mviN, irgA, pldA, tlyA and hecA target genes were found as 17 (51%), 1 (3%), 7 (23%), 7 (23%), 1 (3%), respectively. While bla OXA-61, tetO and tetW were positive in all isolates, were found as mcr1/2/6, mcr3/7, and mcr5, genes %37.5, %25, and %34.3, respectively. Although in A. butzleri was found 10 (58%), 1 (3%), 3 (43%), 2 (28%) (mviN, irgA, pldA, and tlyA, respectively) virulence genes 7 (42%), 4 (57%), 5 (72%), 1 (3%) was found (mviN, irgA, tlyA, and hecA, respectively) virulence genes in A. cryoaerophilus. Moreover, was found for the mcr 1/2/6 7 (58%) genes, for the mcr 3/7 genes 3 (38%) in A. butzleri. In A. cryoaerophilus was found for the mcr 1/2/6 genes 5 (42%), for the mcr 3/7 genes 5 (62%), and for the mcr 5 gene 10 (100%). Thus, the current study indicated that the existence of Arcobacter spp. isolated from fish and mussel samples may pose a potential risk to public health.

Synthesis, characterization, thermokinetic analysis and biological application of novel allyl glucosamine based glycopolymers.

The synthesis of glycopolymers by copolymerising an allyl glucosamine (AG) monomer with co-monomers methyl methacrylate (MMA), acrylonitrile (AN) and 2-hydroxyethyl methacrylate (HEMA) was investigated via free-radical polymerisation of 2,2-azobisisobutyronitrile (AIBN) in dimethylformamide (DMF). Three new copolymers, poly(AG-co-MMA), poly(AG-co-AN) and poly(AG-co-HEMA), were obtained. The chemical structures of the glycopolymers were analysed using 1H-NMR, 13C-NMR and FTIR. The thermal properties and degradation kinetics of the three glycopolymers were examined by thermogravimetric (TG) analysis at different heating rates. The effects of different co-monomers on the copolymerisation yield, thermal properties and biological activities of the resulting glycopolymers were investigated. The activation energies of the decomposition stages were calculated using the Flynn-Wall-Ozawa (FWO) and Kissinger methods. Furthermore, the biological activity of AG monomers and glycopolymers was studied and compared to chitosan. Poly(AG-co-HEMA) had the most significant effect on MCF-7 cell viability, and all glycopolymers have a low toxic effect profile on MCF-7 cell lines.

Biofilm formation and antibiotic resistance profiles of water-borne pathogens.

Water sources (surface water, drinking water, rivers, and ponds) are significant reservoirs for transmitting antibiotic-resistant bacteria. In addition, these waters are an important public health problem because they are suitable environments for transferring antibiotic resistance genes between bacterial species. Our study aimed to assess the prevalence of Extended-spectrum beta-lactamase (ESBL) producing isolates in water samples, the susceptibility of the isolates to the specified antibiotics, the determination of biofilm ability, antibiotic resistance genes, and the molecular typing of the isolates. For this purpose, Polymerase chain reaction (PCR) and Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analyses were used. Out of 70 isolates, 15 (21%) were ESBL producing, and sent for the MALDI-TOF analysis, where Escherichia coli, Acinetobacter calcoaceticus, Enterobacter bugandensis, Acinetobacter pittii, Pseudomonas aeruginosa, Acinetobacter junii, Pseudomonas oleovorans, and Enterobacter ludwigigii were identified. Moreover, colistin resistance genes (mcr 1/2/6, mcr 4, mcr 5, mcr 3/7, and mcr 8), ESBL-encoding genes (blaSHV, blaTEM, and blaCTX-M) and carbapenemase genes (blaNDM, blaOXA-48, and blaKPC) using molecular analysis (PCR) were confirmed. The colistin resistance gene was detected at 80% (12/15) in the isolates obtained. The distribution of these isolates according to resistance genes was found as mcr 1/2/6 4 (20%), mcr 3/7 3 (13%), and mcr 5 (40%). Additionally, the isolates harbored blaSHV(6.6%) and blaTEM (6.6%) genes. However, blaNDM, blaOXA-48, blaKPC, and blaCTX-M genes were not detected in any isolates. According to the Congo red agar method, seven (46.6%) isolates showed negative biofilm ability, and eight (53.3%) showed moderate biofilm ability. However, the microplate method detected weak biofilm in 53.3% of the isolates. In conclusion, this study provides evidence for the existence of multidrug-resistant bacteria that co-exist with mcr and ESBL genes in water sources. These bacteria can migrate to other environments and pose increasing threats to public health.

Profile of Aliarcobacter spp. from edible giblets: Genetic diversity, antibiotic resistance, biofilm formation.

Aliarcobacter spp. are recognized as emerging foodborne pathogens and consumption of foods contaminated with them can be a hazard to human and animal health. This study was conducted to investigate the prevalence of Aliarcobacter spp. in edible internal organs of different animal species from retail markets and giblet sellers. Additionally, this study was focused on the antimicrobial resistance, virulence profiles, biofilm-forming capabilities, and phylogenetic relationships of obtained isolates. A total of 270 samples were analyzed from which, 28 (10.4 %) were isolated as Aliarcobacter spp. by conventional methods. Within the 28 Aliarcobacter spp. isolates, 17 (60.7 %) were identified as A. butzleri, 10 (35.7 %) were A. cryaerophilus and one (3.5 %) was A. skirrowii by PCR method. The disc diffusion method showed that the highest resistance rate of Aliarcobacter spp. was seen against oxacillin (78.5 %), and 20 (71.4 %) out of the 28 isolates exhibited multidrug resistance (MDR). Out of the 28 isolates, mviN, pldA, tlyA, and hecB virulence genes were detected in 85.7 %, 46.4 %, 46.4 %, and 3.5 %, respectively, but irgA, Cj1349, ciaB, cadF, and hecA genes were not detected. According to the microplate test, 27 (96.4 %) isolates had weak biofilm ability while one A. cryaerophilus isolate (3.6 %) exhibited strong biofilm formation. ERIC-PCR band patterns suggested that isolated Aliarcobacter spp. from giblets, have different contamination sources. The presence of pathogenic and multidrug-resistant Aliarcobacter spp. in food poses a potential risk to public health and control measures throughout the food chain are necessary to prevent the spread of these strains.

Escherichia coli serogroups in slaughterhouses: Antibiotic susceptibility and molecular typing of isolates.

This study aimed to investigate the contamination of carcasses and slaughterhouse environment with Escherichia coli O157:H7 and non-O157 serogroups (O45:H2, O103:H2, O121:H19, O145:H28, O26:H11, O111:H8). For this purpose, a total of 150 samples (30 carcasses, 30 shredding units, 30 knives, 30 slaughterhouse waste water and 30 wall surfaces) were collected from 5 different slaughterhouses in Kayseri, Turkey. The conventional and molecular methods were performed in order to detect Escherichia coli and its serogroups. Of the 150 samples, 55 (36%) were found to be contaminated with E. coli. Among isolates, E. coli serogroup (O157:H7) were detected in 2 (11%) carcass and 2 (11%) wastewater samples. None of the E. coli isolates harbored tested genes (stx1, stx2, eaeA, and hylA). Effective infection control measures and antibiotic stewardship programs should be adopted to limit the spread of multidrug-resistant bacteria. It was also deduced that these isolates resistance to different antibiotics could be hazardous for public health.

Prevalence and molecular characterization of Listeria monocytogenes isolated from wastewater of cattle slaughterhouses in Turkey.

AIM: The study aimed to investigate the role of cattle slaughterhouse wastewater as a possible source for the environmental distribution of Listeria monocytogenes. METHODS AND RESULTS: Listeria spp. isolation was performed by collecting 117 wastewater samples from four different cattle slaughterhouses in Turkey. Species-specific identification was performed biochemically, and L. monocytogenes isolates were confirmed with polymerase chain reaction (PCR). In all, 71 (62.2%) of the wastewater samples were found to be positive for Listeria spp., and 17 (14.9%) of these samples were contaminated with L. monocytogenes. Pulsed field gel electrophoresis (PFGE) analysis revealed that all L. monocytogenes isolates were of different pulsotypes and isolates belonged to seven different phylogenetic clusters. Multiplex PCR analysis for genoserotypes and lineage determination showed that the isolates were divided into genoserotypes IVb and IIc in Lineages I and II. Also, it has been investigated with SYBR-Green Real-time PCR whether the L. monocytogenes isolates harboured virulence genes (hly, sigB, plcA, plcB, inlA, inlB, inlC and inlJ), and it was found that all isolates were substantially positive. Antibiotic resistance profiles and MIC values of the isolates were determined, and all L. monocytogenes isolates were found susceptible to ampicillin. In contrast, two isolates were resistant to meropenem and erythromycin, and three isolates were resistant to trimethoprim/sulfamethoxazole. CONCLUSION: L. monocytogenes, which pose a threat to public health and resists to antibiotics effectively used in treatments, can environmentally spread via wastewater of cattle slaughterhouses. The wastewater of the food industry, which has rich microbiota, should be treated carefully, and possible environmental contamination should be prevented. SIGNIFICANCE AND IMPACT OF STUDY: This is the first study that investigates the molecular characterization of L. monocytogenes isolated from cattle slaughterhouse wastewater and the findings represent the importance of cattle wastewater in the epidemiology of L. monocytogenes in Turkey.

Mycotoxigenic and phylogenetic perspective to the yeasts and filamentous moulds in mould-matured Turkish cheese.

This study was conducted to determine the diversity of yeasts and filamentous moulds in mould-matured cheese (MMC) consumed in Turkey. Overall, 120 samples were collected from 12 different geographical locations between March 2016 and April 2017. The morphological observation was applied in combination with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and molecular analyses to determine yeasts and filamentous moulds in the cheeses. High-performance liquid chromatography (HPLC) technique was used to evaluate the ability of mycotoxins production of fungal isolates and the presence of mycotoxins in cheese samples. A total of 241 fungi (81 filamentous moulds and 160 yeast) were recovered, and Penicillium roqueforti and Debaryomyces hansenii were the most frequently isolated species in all cheese samples. The rep-PCR results indicated a high level of genetic diversity among fungal isolates, regardless of isolation source or geographical origin. Filamentous mould strains isolated from MMC were found to synthesize at least one mycotoxin (Aflatoxin B1, B2, G1 and G2, citrinine, cyclopiazonic acid, mycophenolic acid, ochratoxin A, penicillic acid and roquefortine C). Although mycotoxin producing ability was observed from all isolates, none of the cheese samples were found positive for these mycotoxins. AFM1 was detected in 8 (6.6%) MMC samples from which 2 (1.6%) were above the legal limits (0.05 µg/kg) set by the Turkish Food Codex (TFC) and European Commission (EC). In conclusion, Turkish MMCs were found to be contaminated with toxigenic fungi, so a potential public health risk, while low, exists. Therefore, the selection of nontoxigenic filamentous mould strains for cheese manufacturing and control of the ripening conditions is a critical need to ensure the quality and safety of Turkish MMC.

Thymoquinone Glucuronide Conjugated Magnetic Nanoparticle for Bimodal Imaging and Treatment of Cancer as a Novel Theranostic Platform.

BACKGROUND: Theranostic oncology combines therapy and diagnosis and is a new field of medicine that specifically targets the disease by using targeted molecules to destroy the cancerous cells without damaging the surrounding healthy tissues. OBJECTIVE: We aimed to develop a tool that exploits enzymatic TQ release from glucuronide (G) for the imaging and treatment of lung cancer. We added magnetic nanoparticles (MNP) to enable magnetic hyperthermia and MRI, as well as 131I to enable SPECT imaging and radionuclide therapy. METHODS: A glucuronide derivative of thymoquinone (TQG) was enzymatically synthesized and conjugated with the synthesized MNP and then radioiodinated with 131I. New Zealand white rabbits were used in SPECT and MRI studies, while tumor modeling studies were performed on 6-7- week-old nude mice utilized with bioluminescence imaging. RESULTS: Fourier-transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) spectra confirmed the expected structures of TQG. The dimensions of nanoparticles were below 10 nm and they had rather polyhedral shapes. Nanoparticles were radioiodinated with 131I with over 95% yield. In imaging studies, in xenograft models, tumor volume was significantly reduced in TQGMNP-treated mice but not in non-treated mice. Among mice treated intravenously with TQGMNP, xenograft tumor models disappeared after 10 and 15 days, respectively. CONCLUSION: Our findings suggest that TQGMNP in solid, semi-solid and liquid formulations can be developed using different radiolabeling nuclides for applications in multimodality imaging (SPECT and MRI). By altering the characteristics of radionuclides, TQGMNP may ultimately be used not only for diagnosis but also for the treatment of various cancers as an in vitro diagnostic kit for the diagnosis of beta glucuronidase-rich cancers.

Visualisation of real-time oral biodistribution of fluorescent labeled self-microemulsifying drug delivery system of olmesartan medoxomil using optical imaging method.

In the present study, the biodistribution of self-microemulsifying drug delivery system of hydrophobic olmesartan medoxomil (OM-SMEDDS) was determined by labeling with a fluorescent dye VivoTag®680 XL and Xenolight® DiR. Labeled OM-SMEDDS and control dye solution administered orally to mice; real-time dynamic biodistributions over 7 h were determined by 2D-fluorescent imaging to verify their anatomic location. Fluorescent Emissions by Vivotag 680® XL and Xenolight® DiR labeled OM-SMEDDS emitted 2 to 24 times stronger emission than control dye administered group. To further confirm the results, organs were removed and examined using the same technique at the end of 7 h. VivoTag®680XL and Xenolight® DiR emitted 4 and 1.7 times stronger emission respectively than control dye administered mice in ex-vivo organ imaging studies. This study showed that OM-SMEDDS can be succesfully labeled with fluorescent dye and tracked with optical imaging method for the visualisation of biodistribution of drugs and is also useful for enhanced bioavailability.

Enterocytozoon bieneusi in raw milk of cattle, sheep and water buffalo in Turkey: Genotype distributions and zoonotic concerns.

Raw milk is a continued threat to public health due to possible contamination with zoonotic pathogens. Enterocytozoon bieneusi is one of the most prevalent pathogenic fungi in a wide range of vertebrate hosts, causing diarrheal disease. Although there has been some evidence, the role and potential risk of raw milk of dairy animals in the transmission dynamics of E. bieneusi is not clear. Therefore, we aimed to determine the occurrence and genotypes of E. bieneusi in raw milk of dairy animals in several farms of the Central Anatolia Region. We also investigated if there is a relation between the presence of E. bieneusi and mastitis. Genomic DNAs from a total of 450 raw milk including 200, 200 and 50 samples from cattle, sheep and water buffalo respectively were analyzed using nested PCR, targeting the internal transcribed spacer of E. bieneusi. Totally milk samples of 9 (4.5%) dairy cattle, 36 (18.0%) sheep, and 1 (2.0%) water buffalo were PCR-positive. A significant relationship was determined between mastitis and the presence of E. bieneusi. Sequence analysis revealed the presence of eight genotypes: two known (ERUSS1, BEB6) and six novel genotypes (named as TREb1 to TREb6). The genotype ERUSS1 and BEB6 were the most common genotypes, found in all cattle and sheep farms. Phylogenetic analysis clustered all the identified genotypes in Group 2. This study provides novel findings that contribute to the transmission dynamics and molecular epidemiology of E. bieneusi. Our study also highlighted the potential risk of raw milk for public health with respect to microsporidia infections.

Synthesis, molecular modeling, in vivo study and anticancer activity against prostate cancer of (+) (S)-naproxen derivatives.

In this study, (S)-naproxen thiosemicarbazides (3a-d), 1,2,4-triazoles (4a-c), triazole-thioether hybride compounds (5a-p) were synthesized and their structures (3a, 3d, 4a and 5a-p) were confirmed by FT-IR, 1H NMR,13C NMR, HR-Mass spectra and elemental analysis. These compounds are designed to inhibit methionine amino peptidase-2 (MetAP2) enzyme in prostate cancer. These compounds (3d, 5a-p) evaluated against androgen-independent prostate adenocarcinoma (PC-3, DU-145) and androgen-dependent prostate adenocarcinoma (LNCaP) cell lines by using MTS method. Compounds 5a, 5b, 5d and 5e showed 14.2, 5.8, 10.8 and 8.4 µM anticancer activity against PC-3 cell lines, compounds 5e, 5g and 5n presented anticancer activity against DU-145 cell lines 18.8, 12.25 and 10.2 µM, and compounds 5g, 5m and 5n exhibited anticancer activity against LNCaP cell lines 12.25, 22.76 and 2.21 µM, respectively. Consequently, of these results, compounds 5e and 5n showed the highest activities against androgen dependent and independent prostate cancer cell lines, so these compounds could be potent small molecules against prostate cancer. Furthermore, mitogen-activated protein kinase (MAPK) pathway activation, AKT (protein kinase B) phosphorylation and androgen receptor activation of compound 5n (SGK636) were investigated in LNCaP cells by using Western blot method. Compound 5n (SGK636) was also tested against mRNA expression analysis of the Bax, Bcl-2, Caspase 3, Caspase 9 by using real-time PCR analysis. Compound 5n was given to nude male mice with cancer in comparison to the control group. Compound 5n was found to reverse the malignant phenotype in the nude male mice, whereas the prostate cancer progressed in the control group. Analysis of some blood parameters in the study showed that they were within the normal values with respect to the control. The blood values of the animals treated according to the control group also exhibited compliance with the blood limit values. Molecular docking and dynamics simulation of compound 5n binding to Methionine Aminopeptidase 2 (MetAP2) enzyme rationalized its potential activity. In addition, inhibition assay MetAP2 enzyme of compound 5n was evaluated. Taken together, we suggest compound 5n to be a potential candidate for prostate cancer therapy.

[Virulence Genes, Antibiotic Susceptibility Profiles and Molecular Characterization of Campylobacter Species Isolated from Different Slaughterhouses]. / Farkli Kesimhanelerden Izole Edilen Campylobacter Türlerinin Virülans Genleri, Antibiyotik Duyarlilik Profilleri ve Moleküler Karakterizasyonu.

The aim of this study was to investigate the frequency of Campylobacter species, to detect the antibiotic resistance profiles and the virulence genes and to determine the clonal proximity of the isolates in the samples of cutting board, slaughterhouse waste water, wall, knife and carcass from three different slaughterhouses in Kayseri region. For this purpose, a total of 150 samples, 10 of each from knife, wall, cutting board, carcass smear sample and slaughterhouse wastewater were collected from each of the three types of slaughterhouses in 2018 in Kayseri. For the isolation of the Campylobacter species, following preenrichment, the suspensions were inoculated onto modified charcoal cefoperazone desoxycholate (CCD) agar and were incubated at 37°C under microaerophilic condition for 48-72 hours. Suspicious colonies with gray-white color were recovered and subjected to phenotypical (Gram staining, oxidase, catalase test, and motion test) tests. Multiplex polymerase chain reaction (mPCR) was used for the molecular identification of the Campylobacter species. Antimicrobial susceptibilities of the isolates identified at the species level were detected by using the disk diffusion test and antibiotic gradient test. Virulence genes (iam, cadF, cdtA, flaA, ceuE, cdtC, cdtB and virB11) among the isolates were evaluated by PCR. The molecular typing of the isolates determined at species level was performed by Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR). In the study, 17 (11.3%) of the 150 samples taken from the slaughterhouse were found to be suspicious in terms of Campylobacter spp. and as a result of phenotypic identification tests, all of the isolates were verified as Campylobacter spp.. As a result of mPCR; eight of the isolates were identified as Campylobacter jejuni, eight as Campylobacter fetus and one as Campylobacter coli. The isolation of the Campylobacter species from different sources was found to be higher in slaughterhouse wastewater than those of others (p<0.001) and the difference in the proportional distribution of the Campylobacter species obtained from various sources was statistically significant (p<0.05). As a result of the disk diffusion test, while, all C.jejuni isolates were resistant to ciprofloxacin, 87.5%, 25%, 25% and 12.5% of C.jejuni isolates were resistant to enrofloxacin, neomycin, amoxicillin/clavulanic acid, and erythromycin, respectively. In addition, 25%, 25% and 12.5% of C.fetus isolates were resistant to amoxicillin/clavulanic acid, neomycin and gentamicin, respectively. C.coli isolate was not resistant to any of the antibiotics tested. Antibiotic gradient test results were found to be compatible with the disc diffusion test results. One of the virulence genes examined, virB11, was not detected in any of the isolates. Moreover, iam gene was not present in C.fetus and C.coli isolates, but only in one C.jejuni isolate. The flaA gene was detected in six C.jejuni isolates. C.coli isolate and seven C.jejuni and seven C.fetus isolates were positive in terms of the cdtC gene. The cdtA, cdtB, ceuE and cadF genes were found to be positive in all C.jejuni isolates. All isolates analyzed in the study demonstrated different ERIC-PCR profiles. In conclusion, it was shown that Campylobacter strains isolated from slaughterhouses were resistant to the most of the current antibiotics. Moreover, the presence of highly virulent Campylobacters in the slaughterhouse environment threatens public health due to the risk of contamination of the humans via carcasses and foods. Therefore, it is recommended that strict hygiene rules should be followed to reduce Campylobacter species contamination in slaughterhouses.

Synthesis, molecular modeling, in vivo study, and anticancer activity of 1,2,4-triazole containing hydrazide-hydrazones derived from (S)-naproxen.

A new series of 1,2,4-triazole containing hydrazide-hydrazones derived from (S)-naproxen ( 7a-m) was synthesized in this study. The structures of these compounds were characterized by spectral (Fourier-transform infrared spectroscopy, 1 H-nuclear magnetic resonance (NMR), 13 C-NMR, and high-resolution electron ionization mass spectrometry) methods. Furthermore, molecular modeling of these compounds was studied on human methionine aminopeptidase-2. All synthesized compounds were screened for anticancer activity against three prostate cancer cell lines (PC3, DU-145, and LNCaP) using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium colorimetric method. Compound 7a showed the best activity against the PC3, DU-145 and LNCaP cancer cell lines with IC50 values of 26.0, 34.5, and 48.8 µM, respectively. Compounds 7b, 7k, and 7m showed anticancer activity against cancer cell lines PC3 and DU-145 with IC50 values of 43.0, 36.5, 29.3 µM and 49.8, 49.1, 31.6 µM, respectively. Compounds 7f and 7g showed anticancer activity against PC3 cells with IC50 values of 43.4 and 34.5 µM, respectively. To assess the biodistribution in mice of IRDye800, dye-labeled compound 7a or 100 µM of free dye was injected intravenously into the mice's tail. In vivo images were taken with in vivo imaging system spectrum device at 60, 120, 180, 240, 300, and 360 min after injection. At the end of 360 min, ex vivo studies were carried out to determine in which organs the dye was accumulated in the urogenital system. Ex vivo studies showed that the accumulation of compound 7a in the prostate is greater than that of the free dye, and it is concluded that compound 7a may be promising for the treatment of prostate cancer.