Design, synthesis, and molecular docking studies of benzimidazole-1,3,4-triazole hybrids as carbonic anhydrase I and II inhibitors.

In this study, with an aim to develop novel heterocyclic hybrids as potent enzyme inhibitors, we synthesized a series of 10 novel 2-(4-(4-ethyl-5-(2-(substitutedphenyl)-2-oxo-ethylthio)-4H-1,2,4-triazol-3-yl)-phenyl)-5,6-dimethyl-1H-benzimidazole (5a-5j) derivatives and characterized by 1 H-NMR, 13 C-NMR, and HRMS. These compounds were evaluated for their inhibitory activity against hCA I and hCA II. All the compounds exhibited good hCA I and hCA II inhibitory activities with IC50 values in range of 1.288 µM-3.122 µM. Among all these compounds, compound 5e, with an IC50 value of 1.288 µM is the most active against carbonic hCA I. Compound 5h with an IC50 value of 1.532 µM is the most active against carbonic hCA-II. Compounds 5a-5j were also evaluated for their cytotoxic effects on the L929 mouse fibroblast (normal) cell line. The compounds were also analyzed for their antioxidant capacity by TAS, FRAP, and DPPH activity. Enzyme inhibition kinetics showed all compounds 5a-5j to inhibit the enzyme by non-competitive. The most active compound 5e for hCA I and compound 5h for hCA-II were subjected to molecular docking, which revealed their binding interactions with the enzyme's active site, confirming the experimental findings.

Green synthesis, characterization of melatonin-like drug bioconjugated CoS quantum dots and its antiproliferative effect on different cancer cells.

BACKGROUND: Quantum dots are usually particles smaller than 100 nm and have a low toxic effect. This study aimed to bioconjugate the anticancer effective melatonin agonist to quantum dots and demonstrate its effects in two cancer lines. This is the first study that aims to examine the anticancer activity of ramelteon bioconjugation to quantum dots, providing a new perspective on the use of Melatonin and its derivatives in cancer. METHODS AND RESULTS: For this purpose, first of all, cobalt sulfide (CoS) quantum dots were synthesized, bioconjugated and characterized with Punica granatum extract by green synthesis method. The effects of synthesized nanomaterials on neuroblastoma and prostate cancer cells were investigated. It was noted that nanomaterials reduced cell viability by 50% in neuroblastoma and prostate cancer lines at a dose of 50 µg/mL. Ramelteon bioconjugated nanomaterials reduced cancer cell viability by 1.4 times more than free melatonin. CoS quantum dots were determined to double the oxidative stress in the neuroblastoma cell line compared to the control, while no change was observed in prostate cancer. In the gene expression findings, it was observed that CoS nanoparticles caused an increase in the expression levels of apoptosis-related genes in the neuroblastoma cell line and induced key protein expression levels of pathways such as ROR-alpha in the prostate cancer cell line. CONCLUSION: As a result, it was observed that the viability of the neuroblastoma cell line decreased with apoptosis induced by oxidative stress, while this effect was observed in the DU-145 cell line via the ROR-alpha pathway.

The anxiolytic and circadian regulatory effect of agarwood water extract and its effects on the next generation; zebrafish modelling.

Anxiety is one of the most common psychiatric symptoms worldwide. Studies show that there is an increase of >25 % in the prevalence of anxiety with the onset of the COVID-19 pandemic process. Due to the various side effects of drugs used in the treatment of anxiety, interest in natural therapeutic alternatives has increased. Agarwood is a plant used as a natural therapeutic due to its sedative effect as well as many effects such as antioxidant and antibacterial. Although there are many studies with agarwood, comprehensive behavioral studies, including the next generation, are limited. In present study, zebrafish fed with diets containing 10-100 ppm water extract of Agarwood (AWE) for 3 and 8 weeks were exposed to predator stress using Oscar fish in order to test the potential anxiolytic effect of AWE. At the end of the period, zebrafish exposed to predator stress were subjected to anxiety and circadian tests. Histopathological evaluation and immunofluorescent analyzes of BDNF and 5HT4-R proteins were performed in the brains of zebrafish. The effects on the next generation were examined by taking offspring from zebrafish. According to the results, it was observed that AWE had a healing effect on anxiety-like behaviors and on the disrupted circadian rhythm triggered by the predatory stress it applied, especially in the 8 weeks 100 ppm group. Interestingly, it was also found to be effective in offspring of zebrafish fed diets with AWE.

Antioxidant/protective effects of carob pod (Ceratonia siliqua L.) water extract against deltamethrin-induced oxidative stress/toxicity in zebrafish larvae.

In our study, the antioxidant capacity of carob pods water extract (CPWE) against deltamethrin (DM)-induced oxidative stress, a widely used pesticide around the world, was investigated in vitro and in vivo in a zebrafish model. The in vitro antioxidant capacity of the obtained extract was evaluated with different methods using trolox, BHA and BHT standard antioxidants. For in vivo experiments, 4hpf zebrafish embryos were exposed to 10 ppb and 25 ppb DM for 120 h and the larvae were treated with 1-10 and 100 ppm CPWE for 4 h at 72th hours. According to the results obtained, it has been determined that the exposure of zebrafish to DM during the developmental period causes important body malformations, decrease in survival rate, reduction in eye size, shortening in body length and decrease in locomotor activity in the dark period. In addition, according to the results of whole-mount staining, it was determined that DM caused a significant increase in the amount of free oxygen radicals and apoptotic cells. It was also confirmed by metabolome analysis that CPWE application for 4 h reduced DM-induced toxicity and oxidative stress. As a result, it can be said that CPWE has an important antioxidant capacity in eliminating DM-induced oxidative stress.

Synthesis and Molecular Docking of New N-Acyl Hydrazones- Benzimidazole as hCA I and II Inhibitors.

BACKGROUND: The carbonic anhydrases (CAs) which are found in most living organisms is a member of the zinc-containing metalloenzyme family. The abnormal levels and activities are frequently associated with various diseases therefore CAs have become an attractive target for the design of inhibitors or activators that can be used in the treatment of those diseases. METHODS: Herein, we have designed and synthesized new benzimidazole-hydrazone derivatives to investigate the effects of these synthesized compounds on CA isoenzymes. Chemical structures of synthesized compounds were confirmed by 1H NMR, 13C NMR, and HRMS. The synthetic derivatives were screened for their inhibitory potential against carbonic anhydrase I and II by in vitro assay. RESULTS: These compounds have IC50 values of 5.156-1.684 µM (hCA I) and 4.334-2.188 µM (hCA II). Inhibition types and Ki values of the compounds were determined. The Ki values of the compounds were 5.44 ± 0.14 µM-0.299 ± 0.01 µM (hCA I) and 3.699 ± 0.041 µM-1.507 ± 0.01 µM (hCA II). The synthetic compounds displayed inhibitory action comparable to that of the clinically utilized reference substance, acetazolamide. According to this, compound 3p was the most effective molecule with an IC50 value of 1.684 µM. Accordingly, the type of inhibition was noncompetitive and the Ki value was 0.299 ± 0.01 µM. CONCLUSION: According to the in vitro test results, detailed protein-ligand interactions of the compound 3p, which is more active against hCA I than standard azithromycin (AZM), were analyzed. In addition, the cytotoxic effects of the compounds on the L929 healthy cell line were evaluated.

Effects of Zinc, Copper and Iron Oxide Nanoparticles on Induced DNA Methylation, Genomic Instability and LTR Retrotransposon Polymorphism in Wheat (Triticum aestivum L.).

Nanomaterials with unique and diverse physico-chemical properties are used in plant science since they improve plant growth and development and offer protection against biotic and abiotic stressors. Previous studies have explored the effects of such nanomaterials on different plant mechanisms, but information about the effects of nanomaterials on induced DNA methylation, genomic instability and LTR retrotransposon polymorphism in wheat is lacking. Therefore, the present study highlights the key role of nanoparticles in DNA methylation and polymorphism in wheat by investigating the effects of ZnO, CuO and γ-Fe3O4 nanoparticles (NPs) on mature embryo cultures of wheat (Triticum aestivum L.). Nanoparticles were supplemented with Murashige and Skoog (MS) basal medium at normal (1X), double (2X) and triple (3X) concentrations. The findings revealed different responses to the polymorphism rate depending on the nanoparticle type and concentration. Genomic template stability (GTS) values were used to compare the changes encountered in iPBS profiles. ZnO, CuO and γ-Fe3O4 NPs increased the polymorphism rate and cytosine methylation compared to the positive control while reducing GTS values. Moreover, non-γ-Fe3O4 NPs treatments and 2X ZnO and CuO NP treatments yielded higher polymorphism percentages in both MspI- and HpaII-digested CRED-iPBS assays and were thus classified as hypermethylation when the average polymorphism percentage for MspI digestion was considered. On the other hand, the 3X concentrations of all nanoparticles decreased HpaII and MspI polymorphism percentages and were thus classified as hypomethylation. The findings revealed that MS medium supplemented with nanoparticles had epigenetic and genotoxic effects.

The effects of MgS nanoparticles-Cisplatin-bio-conjugate on SH-SY5Y neuroblastoma cell line.

Magnesium sulfide nanoparticles (MgS NPs) is a nanomaterial that has an important place in diagnosis, treatment, diagnosis, and drug delivery systems. Neuroblastoma, a type of brain cancer, is an extremely difficult cancer to treat with today's treatment options. This study was carried out to determine the cytotoxic, oxidant, and antioxidant effects on the neuroblastoma cancer line (SH-SY5Y cell line) along with the green synthesis and characterization of MgS NPs structures. MgS NPs were synthesized by green synthesis using Na2S and Punica granatum, a cleaner method for toxic effects, and characterized using Scanning Electron Microscopy, Fourier Transform Infrared spectroscopy, X-Ray diffraction methods. In cell culture, SH-SY5Y cells were grown in a suitable nutrient medium under favorable conditions. Five different doses of MgS NPs (10, 25, 50, 75, and 100 µg/mL) were applied to the cell line for 24 h. The analysis of the MgS NPs applications was performed with MTT cytotoxicity test and total oxidant and total antioxidant tests. According to the data obtained, 75 µg/mL MgS NPs application decreased cancer cell viability up to 48.54%. MgS NPs exhibited a dose-dependent effect on the SH-SY5Y cell line. Also, it was determined that MgS NPs increased oxidant activity in neuroblastoma cells, which was compatible with the cytotoxicity test. As a result, MgS NPs exhibited an effective activity on the neuroblastoma cell line. It was clearly seen that NPs obtained by green synthesis prevented the related cancer line from proliferating.

Preparation of CoS nanoparticles-cisplatin bio-conjugates and investigation of their effects on SH-SY5Y neuroblastoma cell line.

Neuroblastoma is one of the most widely seen under the age of 15 tumors that occur in the adrenal medulla and sympathetic ganglia. Cisplatin, an antineoplastic drug, is a Platinum-based compound and is known to inhibit the proliferation of neuroblastoma cells. Effective applications of nanoparticles in biomedical areas such as biomolecular, antimicrobial detection and diagnosis, tissue engineering, theranostics, biomarking, drug delivery, and anti-cancer have been investigated in many studies. This study aims to prepare the bioconjugates of CoS (cobalt sulfide) nanoparticles (NPs) with cisplatin combination groups and to evaluate their effects on the neuroblastoma cell line. Nanoparticle synthesis was done using the green synthesis technique using Punica granatum plant extract. The size and shape of CoS NPs were characterized by SEM, FT-IR, and XRD. Zeta potential was confirmed by the DLS study. For this purpose, the SH-SY5Y neuroblastoma cell line was cultured in a suitable cell culture medium. Cisplatin 5 µg and different concentrations (Cisplatin + CoS NPs bioconjugates (5, 10, 25, 50, 75 µg) doses were applied to SH-SY5Y neuroblastoma cell lines for 24 h. TAC, TOS and MTT tests were performed 24 h after the application. According to the MTT test results, cisplatin and CoS NP combinations reduced the proliferation of neuroblastoma cells by 78 to 57% compared to the cisplatin control. From the findings obtained; the most effective Bio-conjugate group was Cisplatin 5 µg/mL + CoS 75 µg/mL.

Antiviral drugs and plasma therapy used for Covid-19 treatment: a nationwide Turkish algorithm.

The Coronavirus outbreak described as COVID-19 is an insidious and enormous biohazard which began to be noticed in November 2019. When the virus was determined to cause serious upper respiratory tract infections resulting in death, pandemics were declared in the world. As of today, the number of cases exceeded 221 thousand people in Turkey, the number of patients who died had reached 5526. In more than 200 countries around the world, 15.1 million people fight the disease, while the number of people recovered is over 9.134 million, and the number of deaths has exceeded 620 thousand. The top 5 countries in the world are USA, Brazil, Russia, India and Spain. The countries with the highest number of cases after America (approximately 4 million 28 thousand) are Brazil (approximately 2 million 166 thousand), India (about 1 million 195 thousand), Russia (approximately 789 thousand), South Africa (approximately 382 thousand). In addition, the number of deaths and cases caused by Covid 19 continues to increase day by day. In this review, it was aimed to discuss that Covidien-19 against antiviral drugs used in the struggle across the globe and plasma treatment options about the current state of knowledge and Turkey algorithm by comparing the therapeutic treatment options.

Immobilization of Purified Pectin Lyase from Pseudomonas putida onto Magnetic Lily Flowers (Lilium candidum L.) Nanoparticles and Applicability in Industrial Processes.

Pectinases are an important class of enzymes distributed in many higher plants and microorganisms. One of these enzymes is pectin lyase which has an important role in industrial applications such as clarification of fruit juices. Pectin lyase was purified with 73% yield from Pseudomonas putida bacteria and was 220.7-fold using three phase precipitation technique. Molecular weight of purified pectin lyase was determined as 32.88 kDa with SDS-polyacrylamide gel electrophoresis. The pectin lyase was immobilized covalently via the L-glutaraldehyde spacer to the cellulosic structures of lily flowers (Lilium candidum L.). The immobilized enzyme was then magnetized by modifying with γ-Fe3O4 nanoparticles and determined the most appropriate immobilization conditions as pH 6 and 30 °C. Purified pectin lyase was connected to magnetized support material after 60 min at the rate of 86.4%. The optimum pH and temperatures for the free and immobilized pectin lyase was found to be 6.0 and 40 °C. pH and thermal stabilities of the free and immobilized pectin lyase enzyme have been preserved at high-low temperatures and pH. The structural characterization of the immobilized pectin lyase was performed by SEM, FT-IR, and XRD chromatographic analyses and it was observed that the support materials structure was appropriated to immobilization with pectin lyase and to modify with Fe3O4 nanoparticles.

In silico Molecular Docking and ADME Studies of 1,3,4-Thiadiazole Derivatives in Relation to in vitro PON1 Activity.

BACKGROUND: Paraoxonase 1 (PON1) is a paraoxonase, arylesterase and lactonase associated with protection of lipoproteins and cell membranes against oxidative modification. OBJECTIVE: Based on antioxidative properties of PON1 and significance of 1,3,4-thiadiazoles in pharmaceutical chemistry, herein we aimed to evaluate the potentials of 1,3,4-thiadiazole derivatives as PON1 activators. METHODS: 2-[[5-(2,4-Difluoro/dichlorophenylamino)-1,3,4-thiadiazol-2-yl]thio]acetophenone derivatives (1-18) were in vitro evaluated for their activator effects on PON1 which was purified using ammonium sulfate precipitation (60-80%) and DEAE-Sephadex anion exchange chromatography. Molecular docking studies were performed for the detection of affinities of all compounds to the active site of PON1. Moreover, Absorption, Distribution, Metabolism and Excretion (ADME) properties of all compounds were also in silico predicted. In silico molecular docking and ADME studies were carried out according to modules of Schrodinger's Maestro molecular modeling package. RESULTS: All compounds, particularly compounds 10, 13 and 17, were determined as promising PON1 activators and apart from compound 1, all of them were detected in the active site of PON1. Besides, ADME results indicated that all compounds were potential orally bioavailable drug-like molecules. CONCLUSION: PON1 activators, compounds 10, 13 and 17 stand out as potential drug candidates for further antioxidant studies and these compounds can be investigated for their therapeutic effects in many disorders such as atherosclerosis, diabetes mellitus, obesity, chronic liver inflammation and many more.

Green synthesis of Ce2O3 NPs and determination of its antioxidant activity.

In this study, the authors presented synthesis of ceria nanoparticles (NPs) by the bio-reduction method and their antioxidative activity. Aqueous extract of Euphorbia (Euphorbia amygdaloides) was used as reducing and stabilising agents. They used aqueous extract of Euphorbia (E. amygdaloides) as reducing and stabilising agent. Ultraviolet-visible (UV-vis) absorption spectroscopy was used to monitor the quantitative formation of ceria NPs. They also addressed the characteristics of the obtained ceria NPs using scanning electron microscopy (SEM), X-ray diffraction (XRD) and transmitting electron microscope (TEM). The synthesised cerium (III) oxide (Ce2O3) NPs were initially noted through visual colour change from colourless pale yellow cerium (III) to light yellow cerium (IV) and further confirmed the band at 345 nm employing UV-vis spectroscopy. The average diameter of the prepared NPs was about 8.6-10.5 nm. In addition, the synthesised Ce2O3 NPs were tested for antioxidant and anti-bacterial activities using ferric reducing antioxidant power, cupric reducing antioxidant capacity, ferrous ions chelating activity, superoxide the anion radical scavenging and 2, 2'-azinobis 3-ethylbenzothiazol to-6-sulphonic acid scavenging activity. It could be concluded that Euphorbia (E. amygdaloides) extract can be used efficiently in the production of potential antioxidant and anti-bacterial Ce2O3 NPs for commercial applications.

Potential inhibitors of human carbonic anhydrase isozymes I and II: Design, synthesis and docking studies of new 1,3,4-thiadiazole derivatives.

In the last years, inhibition of carbonic anhydrase (CA) has emerged as a promising approach for pharmacologic intervention in a variety of disorders such as glaucoma, epilepsy, obesity, and cancer. As a consequence, the design of CA inhibitors (CAIs) is a highly dynamic field of medicinal chemistry. Due to the therapeutic potential of thiadiazoles as CAIs, new 1,3,4-thiadiazole derivatives were synthesized and investigated for their inhibitory effects on hCA I and hCA II. Although the tested compounds did not carry a sulfonamide group, an important pharmacophore for CA inhibitory activity, it was a remarkable finding that most of them were more effective on hCAs than acetazolamide (AAZ), the reference agent. Among these compounds, N'-((5-(4-chlorophenyl)furan-2-yl)methylene)-2-((5-(phenylamino)-1,3,4-thiadiazol-2-yl)thio)acetohydrazide (3) was found to be the most effective compound on hCA I with an IC50 value of 0.14nM, whereas N'-((5-(2-chlorophenyl)furan-2-yl)methylene)-2-((5-(phenylamino)-1,3,4-thiadiazol-2-yl)thio)acetohydrazide (1) was found to be the most potent compound on hCA II with an IC50 value of 0.15nM. According to molecular docking studies, all compounds exhibited high affinity and good amino acid interactions similar to AAZ on the both active sites of hCA I and hCA II enzymes.

Green synthesis and characterisation of platinum nanoparticles using quail egg yolk.

Nanotechnology is extensively used in all parts today. Therefore, nano synthesis is also significant in all explored areas. The results of studies conducted have revealed that nanoparticle synthesis is performed by using both chemical and physical methods. It is well known that these syntheses are carried out at high charge, pressure and temperature in harsh environments. Therefore, this study investigated green synthesis method that sustains more mild conditions. In this study, quail egg yolk having high vitamin and protein content was prepared for green synthesis reaction and used for the synthesis of platinum nanoparticles in the reaction medium. Reaction situations were optimised as a function of pH, temperature, time and concentration by using quail egg yolk. The results showed that the highest platinum nanoparticles were synthesised at 20°C and pH6.0 for 4h. Also, optimal concentration of metal ions was established as 0.5mM. The synthesised platinum nanoparticles were characterised by using UV spectrum, X-ray diffraction and scanning electron microscope.

Removing Trypan blue dye using nano-Zn modified Luffa sponge.

This study has presented specific features that are examined to remove the Trypan blue dye from the waste using Luffa sponge (LS) and modified Luffa sponge with zinc nanoparticles (ZnNPs). Peroxidase enzyme was obtained from Euphorbia amygdaloides plant and it was used with the green synthesis of Zn nanoparticles. Luffa sponge was used to be a support material for immobilized nanoparticles and it also used in remediation work. The obtained membrane forms, fibrous materials, (LS, ZnNPs-LS) were characterized with SEM and XRD. LS and ZnNPs-LS were employed as adsorbent to be used for the removal of Trypan blue dye from aqueous via batch studies. Measurements were made for the equilibrium, pH, temperature, concentration of dye with UV-visible spectrometer (590nm; for Trypan blue dye). The optimum removal of Trypan blue dye was found at pH7, the equilibrium was attained within 30min. The thermodynamic properties ΔG0, ΔH0, and ΔS0 showed that adsorption of Trypan blue dye onto LS and ZnNPs-LS were spontaneous and endothermic. The equilibrium isotherm data were analyzed using Langmuir and Freundlich models and the sorption process was described by the Langmuir isotherm with maximum monolayer adsorption capacity of 45.32 and 47.3mg/g for LS and LS-ZnNPs at 303±1°K, respectively.

Synthesis and evaluation of new benzodioxole-based dithiocarbamate derivatives as potential anticancer agents and hCA-I and hCA-II inhibitors.

In the current work, new benzodioxole-based dithiocarbamate derivatives were synthesized via the reaction of N-(1,3-benzodioxol-5-ylmethyl)-2-chloroacetamide with appropriate sodium salts of N,N-disubstituted dithiocarbamic acids. These derivatives were evaluated for their cytotoxic effects on A549 human lung adenocarcinoma and C6 rat glioma cell lines. N-(1,3-Benzodioxol-5-ylmethyl)-2-[4-(4-nitrophenyl)-1-piperazinylthiocarbamoylthio]acetamide (10) can be identified as the most promising anticancer agent against C6 cell line due to its notable inhibitory effect on C6 cells with an IC50 value of 23.33 ± 7.63 µg/mL when compared with cisplatin (IC50 = 19.00 ± 5.29 µg/mL). On the other hand, compound 10 did not show any significant cytotoxic activity against A549 cell line. The compounds were also tested for their in vitro inhibitory effects on hCA-I and hCA-II. Generally, the tested compounds were more effective on CAs than acetazolamide, the reference agent. Among these compounds, N-(1,3-benzodioxol-5-ylmethyl)-2-[(morpholinyl)thiocarbamoylthio]acetamide (3) and N-(1,3-benzodioxol-5-ylmethyl)-2-[(thiomorpholinyl)thiocarbamoylthio]acetamide (4) were found to be the most effective compounds on hCA-I with IC50 values of 0.346 nM and 0.288 nM, and hCA-II with IC50 values of 0.287 nM and 0.338 nM, respectively.

Purification and characterization of an extracellular cellulase from Anoxybacillus gonensis O9 isolated from geothermal area in Turkey.

In the present study, cellulase was purified and characterized from Anoxybacillus gonensis (Gen bank Number: KM596794) which was isolated and characterized from Agri Diyadin Hot spring. It was found to synthesize cellulase which had a wide range of industrial applications. Twenty four-hour-cultured bacteria induced cellulase production and specific activities during the purification steps were 1.47, 81.06 and 109.4 EU mg(-1) protein at crude extract, ammonium sulphate precipitated and DEAE-Sephadex purification steps. The highest enzyme activity was observed at 50°C and the optimum range of pH was 3-10. Molecular weight of enzyme was determined approximately 40kDa. The kinetic parameters of cellulase against carboxymethylcellulose (CMC) were 153.4 pmol min(-1) mg for Vmax and 0.46mM for Km. Among effectors of the enzyme, Zn2+, Ca2+, Co2+ and EDTA decreased enzyme activity.

Purification and characterization of [Formula: see text]-mannanase from Bacillus pumilus (M27) and its applications in some fruit juices.

Thermo alkaline mannanase was purified from the bacteria of Bacillus pumilus (M27) using the techniques of ammonium sulphate precipitation, DEAE-Sephadex ion exchange chromatography and Sephacryl S200 gel filtration chromatography with 111-fold and 36 % yield. It was determined that the enzyme had 2 sub-units including 35 kDa and 55 kDa in gel filtration chromatography and SDS-PAGE electrophoresis systems. The optimum pH and temperature was determined as 8 and 60 °C, respectively. It was also noticed that the enzyme did not lose its activity at a wide interval such as pH 3-11 and at high temperatures such as 90 °C. Additionally, the effects of some metal ions on the mannanase enzyme activity. Moreover, the clarifying efficiency of purified mannanase enzyme with some fruit juices such as orange, apricot, grape and apple was also investigated. Enzymatic treatment was carried out with 1 mL L(-1) of purified mannanase for 1 h at 60 °C. It was determined that the highest pure enzyme was efficient upon clarifying the apple juice at 154 % rate.

Synthesis and evaluation of new thiadiazole derivatives as potential inhibitors of human carbonic anhydrase isozymes (hCA-I and hCA-II).

2-[[5-(2,4-Difluoro/dichlorophenylamino)-1,3,4-thiadiazol-2-yl]thio] acetophenone derivatives (3a--s) were designed as human carbonic anhydrase isozymes (hCA-I and hCA-II) inhibitors and synthesized. hCA-I and hCA-II were purified from erythrocyte cells by the affinity chromatography. The inhibitory effects of 18 newly synthesized acetophenones on hydratase activity of these isoenzymes were studied in vitro. The average IC50 values of the new compounds for hydratase activity ranged from 0.033 to 0.14 µM for hCA-I and from 0.030 to 0.11 µM for hCA-II. Among the newly synthesized compounds, 2-[[5-(2,4-dichlorophenylamino)-1,3,4-thiadiazol-2-yl]thio]-4'-bromoacetophenone (3n) can be considered as a promising hCA-II inhibitor owing to its selective and potent inhibitory effect on hCA-II.

Purification of Chitinase enzymes from Bacillus subtilis bacteria TV-125, investigation of kinetic properties and antifungal activity against Fusarium culmorum.

BACKGROUND: Chitin is the main structural component of cell walls of fungi, exoskeletons of insects and other arthropods and shells of crustaceans. Chitinase enzyme is capable of degrading chitin, and this enzyme can be used as a biological fungicide against phytopathogenic fungi, as well as an insecticide against insect pests. METHODS: In this study, 158 isolates, which were derived from bacteria cultures isolated from leaves and root rhizospheres of certain plants in Turkey, were selected after confirming that they are not phytopathogenic based on the hypersensitivity test performed on tobacco; and antifungal activity test was performed against Fusarium culmorum, which is a pathogenic fungi that cause decomposition of roots of vegetables. Accordingly, chitinase enzyme activity assay was performed on 31 isolates that have an antifungal activity, and among them the isolate of Bacillus subtilis TV-125 was selected, which has demonstrated the highest activity. RESULTS: Chitinase enzyme was purified by using ammonium sulphate and DEAE-sephadex ion exchange chromatography. Ammonium sulphate precipitation of chitinase enzyme from Bacillus subtilis TV-125 isolate was performed at maximum range of 0-20%, and 28.4-fold purification was obtained with a 13.4% of yield. Optimum activity of the purified enzyme was observed at pH 4.0 and at 50°C of temperature. In addition, it was identified that Bacillus subtilis TV-125A isolate retains 42% of its activity at 80°C temperature. CONCLUSION: In the last phase of the study, chitinase enzyme purified from Bacillus subtilis TV-125A was tested on four fungal agents, although all the results were positive, it was particularly effective on F. culmorum according to the findings.