Search results for: angiotensin converting enzyme inhibition

Authors: Keyur Raval, Steffen Krohn, Bruno Moerschbacher

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Chitin, the biopolymer of the N-acetylglucosamine, is the most abundant biopolymer on the planet after cellulose. Industrially, chitin is isolated and purified from the shell residues of shrimps. A deacetylated derivative of chitin i.e. chitosan has more market value and applications owing to it solubility and overall cationic charge compared to the parent polymer. This deacetylation on an industrial scale is performed chemically using alkalis like sodium hydroxide. This reaction not only is hazardous to the environment owing to negative impact on the marine ecosystem. A greener option to this process is the enzymatic process. In nature, the naïve chitin is converted to chitosan by chitin deacetylase (CDA). This enzymatic conversion on the industrial scale is however hampered by the crystallinity of chitin. Thus, this enzymatic action requires the substrate i.e. chitin to be soluble which is technically difficult and an energy consuming process. We in this project wanted to address this shortcoming of CDA. In lieu of this, we have modeled a fusion protein with CDA and an auxiliary protein. The main interest being to increase the accessibility of the enzyme towards crystalline chitin. A similar fusion work with chitinases had improved the catalytic ability towards insoluble chitin. In the first step, suitable partners were searched through the protein data bank (PDB) wherein the domain architecture were sought. The next step was to create the models of the fused product using various in silico techniques. The models were created by MODELLER and evaluated for properties such as the energy or the impairment of the binding sites. A fusion PCR has been designed based on the linker sequences generated by MODELLER and would be tested for its activity towards insoluble chitin.

Keywords: chitin deacetylase, modeling, chitin binding domain, chitinases

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Authors: Muskan Jain, Baiju Gopal

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Objectives: Intimacy in interpersonal relationships is integral to psychological health and everyday wellbeing; the focus is on intimacy, which can be described as feelings of closeness, connection, and belonging within relationships, which is influenced by an individual's gender identity as well as life experiences. The study aims to explore the experiences of intimacy of the non-binary gender; this marginalized community has increased risks of developing the imposter phenomenon. The study explores the influence of IP on the development and sustenance of intimacy in relationships. Methods: The present study accumulates detailed narratives from 10 non-binary young adults ages 18 to 25 in metropolitan cities of India. Thematic analysis was used for the data analysis. Results: Seven major themes have emerged revolving around internalized criticism and self-depreciating behavior, which causes distance between partners. The four themes that result in the internalization of criticism are lack of social stability, invalidation by social units, adverse life experiences, and estrangement due to gender identity. Three themes that encapsulate major difficulties in relationships are limited self-disclosure, inhibition of physical needs, and fear of taking space. The findings have been critically compared and contrasted with the existing body of literature in the domain, which sets the agenda for further inquiry. Conclusion: It is important for future studies to capture the experiences of non-binary genders in India to provide better therapeutic support in order to assist them in forming meaningful and authentic relationships, thus increasing overall wellbeing.

Keywords: imposter phenomenon, intimacy, internalized criticism, marginalized community

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Authors: Salwa M. Elmesallamy, Ahmed A. Farag, Magd M. Badr, Dalia S. Fathy, Ahmed Bakry, Mona A. El-Etre

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The term green environment is an international trend. It is become imperative to treat the corrosion of steel with a green coating to protect the environment. From the potential adverse effects of the traditional materials.A series of polybenzoxazine/henna composites (PBZ/henna), with different weight percent (3,5, and 7 wt % (of henna), were prepared for corrosion protection of carbon steel. The structures of the prepared composites were verified using FTIR analysis. The mechanical properties of the resins, such as adhesion, hardness, binding, and tensile strength, were also measured. It was found that the tensile strength increases by henna loading up to 25% higher than the tidy resin. The thermal stability was investigated by thermogravimetric analysis (TGA) the loading of lawsone (henna) molecules into the PBZ matrix increases the thermal stability of the composite. UV stability was tested by the UV weathering accelerator to examine the possibility that henna can also act as an aging UV stabilizer. The effect of henna content on the corrosion resistance of composite coatings was tested using potentiostatic polarization and electrochemical spectroscopy. The presence of henna in the coating matrix enhances the protection efficiency of polybenzoxazine coats. Increasing henna concentration increases the protection efficiency of composites. The quantum chemical calculations for polybenzoxazine/henna composites have resulted that the highest corrosion inhibition efficiency, has the highest EHOMO and lowest ELUMO; which is in good agreement with results obtained from experiments.

Keywords: polybenzoxazine, corrosion, green chemistry, carbon steel

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Authors: Monika Szymańska-Czerwińska, Agnieszka Jodełko, Krzysztof Niemczuk

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Q fever (coxiellosis) is an infectious disease of animals and humans causes by C. burnetii and widely distributed throughout the world. Cattle and small ruminants are commonly known as shedders of C. burnetii. The aims of this study were the evaluation of seroprevalence and shedding of C. burnetii in cattle. Genotypes of the pathogen present in the tested specimens were also identified using MLVA (Multiple Locus Variable-Number Tandem Repeat Analysis) and MST (multispacer sequence typing) methods. Sampling was conducted in different regions of Poland in 2018-2021. In total, 2180 bovine serum samples from 801 cattle herds were tested by ELISA (enzyme-linked immunosorbent assay). 489 specimens from 157 cattle herds such as: individual milk samples (n=407), bulk tank milk (n=58), vaginal swabs (n=20), placenta (n=3) and feces (n=1) were subjected to C. burnetii specific qPCR. The qPCR (IS1111 transposon-like repetitive region) was performed using Adiavet COX RealTime PCR kit. Genotypic characterization of the strains was conducted utilizing MLVA and MST methods. MLVA was performed using 6 variable loci. The overall herd-level seroprevalence of C. burnetii infection was 36.74% (801/2180). Shedders were detected in 29.3% (46/157) cattle herds in all tested regions. ST 61 sequence type was identified in 10 out of 18 genotyped strains. Interestingly one strain represents sequence type which has never been recorded previously. MLVA method identified three previously known genotypes: most common was J but also I and BE were recognized. Moreover, a one genotype has never been described previously. Seroprevalence and shedding of C. burnetii in cattle is common and strains are genetically diverse.

Keywords: Coxiella burnetii, cattle, MST, MLVA, Q fever

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Authors: Linda Chularojmontri, Seewaboon Sireeratawong, Suvara Wattanapitayakul

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Terminalia arjuna Roxb. (family Combretaceae) is commonly known as ‘Sa maw thet’ in Thai. The fruit is used in traditional medicine as natural mild laxatives, carminative and expectorant. Aim of the study: This research aims to study the anti-inflammatory, analgesic and antipyretic activities of Terminalia arjuna extract by using animal models in comparison to the reference drugs. Materials and Methods: The anti-inflammatory study was conducted by two experimental animal models namely ethyl phenylpropionate (EPP)-induced ear edema and carrageenan-induced paw edema. The study of analgesic activity used two methods of pain induction including acetic acid and heat-induced pain. In addition, the antipyretic activity study was performed by induced hyperthermia with yeast. Results: The results showed that the oral administration of Terminalia arjuna extract possessed acute anti-inflammatory effect in carrageenan-induced paw edema. Terminalia arjuna extract showed the analgesic activity in acetic acid-induced writhing response and heat-induced pain. This indicates its peripheral effect by inhibiting the biosynthesis and/or release of some pain mediators and some mechanism through Central nervous system. Moreover, Terminalia arjuna extract at the dose of 1000 and 1500 mg/kg body weight showed the antipyretic activity, which might be because of the inhibition of prostaglandins. Conclusion: The findings of this study indicated that the Terminalia arjuna extract possesses the anti-inflammatory, analgesic and antipyretic activities in animals.

Keywords: analgesic activity, anti-inflammatory activity, antipyretic activity, Terminalia arjuna extract

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Authors: B. Chandar, M. K. Ramasamy, P. Madasamy

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The discovery and development of newer antibiotics are limited with the increase in resistance of such multi-drug resistant bacteria creating the need for alternative new therapeutic agents. The recently discovered New Delhi Metallo-betalactamase-1 (NDM-1), which confers antibiotic resistance to most of the currently available β-lactams, except colistin and tigecycline, is a global concern. Several antibacterial drugs approved are natural products or their semisynthetic derivatives, but plant extracts remain to be explored to find molecules that are effective against NDM-1 bacteria. Therefore, it is necessary to explore the possibility of finding new and effective antibacterial compounds against NDM-1 bacteria. In the present study, we have screened a diverse set South Indian plant species, and report most plant species as a potential source for antimicrobial compounds against NDM-1 bacteria. Ethanol extracts from the leaves of taxonomically diverse South Indian medicinal plants were screened for antibacterial activity against NDM-1 E. coli using streak plate method. Among the plant screened against NDM-1 E. coli, the ethanol extracts from many plant extracts showed minimum bactericidal concentration between 5 and 15 mg /ml and MIC between 2.54 and 5.12 mg/ml. These extracts also showed a potent synergistic effect when combined with antibiotics colistin and tetracycline. Combretum albidum that was effective was taken for further analysis. At 5mg/L concentration, these extracts inhibited the NDM-1 enzyme in vitro, and residual activity for Combretum albidum was 33.09%. Treatment of NDM-1 E. coli with the extracts disrupted the cell wall integrity and caused 89.7% cell death. The plant extract of Combretum albidum that was effective was subjected to fractionation and the fraction was further subjected to HPLC, LC-MS for identification of antibacterial compound.

Keywords: antibacterial activity, combretum albidum, Escherichia coli, NDM-1

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Authors: Kenia Martínez, Geniel Talavera, Juan Alonso

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Moringa oleifera is a plant containing many nutrients that are mostly concentrated within the leaves. Commonly, the separation process of these nutrients involves solid-liquid extraction followed by evaporation and drying to obtain a concentrated extract, which is rich in proteins, vitamins, carbohydrates, and other essential nutrients that can be used in the food industry. In this work, three drying methods were used, which involved very different temperature and pressure conditions, to evaluate the effect of each method on the vitamin C content and the antioxidant efficiency of the extracts. Solid-liquid extractions of Moringa leaf (LE) were carried out by employing an ethanol solution (35% v/v) at 50 °C for 2 hours. The resulting extracts were then dried i) in a convective oven (CO) at 100 °C and at an atmospheric pressure of 750 mbar for 8 hours, ii) in a vacuum evaporator (VE) at 50 °C and at 300 mbar for 2 hours, and iii) in a freeze-drier (FD) at -40 °C and at 0.050 mbar for 36 hours. The antioxidant capacity (EC₅₀, mg solids/g DPPH) of the dry solids was calculated by the free radical inhibition method employing DPPH˙ at 517 nm, resulting in a value of 2902.5 ± 14.8 for LE, 3433.1 ± 85.2 for FD, 3980.1 ± 37.2 for VE, and 8123.5 ± 263.3 for CO. The calculated antioxidant efficiency (AE, g DPPH/(mg solids·min)) was 2.920 × 10^-5 for LE, 2.884 × 10^-5 for FD, 2.512 × 10^-5 for VE, and 1.009 × 10^-5 for CO. Further, the content of vitamin C (mg/L) determined by HPLC was 59.0 ± 0.3 for LE, 49.7 ± 0.6 for FD, 45.0 ± 0.4 for VE, and 23.6 ± 0.7 for CO. The results indicate that the convective drying preserves vitamin C and antioxidant efficiency to 40% and 34% of the initial value, respectively, while vacuum drying to 76% and 86%, and freeze-drying to 84% and 98%, respectively.

Keywords: antioxidant efficiency, convective drying, freeze-drying, Moringa oleifera, vacuum drying, vitamin C content

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Authors: Afsheen Aman, Shah Ali Ul Qader

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Dextransucrase [EC 2.4.1.5] is a glucosyltransferase that catalysis the biosynthesis of a natural biopolymer called dextran. It can catalyze the transfer of D-glucopyranosyl residues from sucrose to the main chain of dextran. This unique biopolymer has multiple applications in several industries and the key utilization of dextran lies on its molecular weight and the type of branching. Extracellular dextransucrase from Leuconostoc mesenteroides is most extensively studied and characterized. Limited data is available regarding cell-bound or intracellular dextransucrase and on the characterization of dextran produced by in-vitro reaction of intracellular dextransucrase. L. mesenteroides AA1 is reported to produce extracellular dextransucrase that catalyzes biosynthesis of a high molecular weight dextran with only α-(1→6) linkage. Current study deals with the characterization of an intracellular dextransucrase and in vitro biosynthesis of low molecular weight dextran from L. mesenteroides AA1. Intracellular dextransucrase was extracted from cytoplasm and purified to homogeneity for characterization. Kinetic constants, molecular weight and N-terminal sequence analysis of intracellular dextransucrase reveal unique variation with previously reported extracellular dextransucrase from the same strain. In vitro synthesized biopolymer was characterized using NMR spectroscopic techniques. Intracellular dextransucrase exhibited Vmax and Km values of 130.8 DSU ml-1 hr-1 and 221.3 mM, respectively. Optimum catalytic activity was detected at 35°C in 0.15 M citrate phosphate buffer (pH-5.5) in 05 minutes. Molecular mass of purified intracellular dextransucrase is approximately 220.0 kDa on SDS-PAGE. N-terminal sequence of the intracellular enzyme is: GLPGYFGVN that showed no homology with previously reported sequence for the extracellular dextransucrase. This intracellular dextransucrase is capable of in vitro synthesis of dextran under specific conditions. This intracellular dextransucrase is capable of in vitro synthesis of dextran under specific conditions and this biopolymer can be hydrolyzed into different molecular weight fractions for various applications.

Keywords: characterization, dextran, dextransucrase, leuconostoc mesenteroides

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Authors: Lila Boulekbache-Makhlouf, Fatiha Brahmi

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This study was carried out with the aim of enriching fresh cheese with the food by-products, which are the leaves of shallots and the peels of potatoes. Firstly, the conditions for extracting the total polyphenols using ultrasound are optimized. Then, the contents of total polyphenols PPT , flavonoids and antioxidant activity were evaluated for the extracts obtained by adopting the optimal parameter. On the other hand, we have carried out some physicochemical, microbiological and sensory analyzes of the cheese produced. The maximum total polyphenols value of 70.44 mg GAE gallic acid equivalent / g of dry matter DM of shallot leaves was reached with 40% (v/v) ethanol, an extraction time of 90 min and a temperature of 10 °C. While, the maximum TPP total polyphenols content of potato peels of 45.03 ± 4.16 mg gallic acid equivalent / g of dry matter DM was obtained using an ethanol /water mixture (40%, v/v), a time of 30 min and a temperature of 60 °C and the flavonoid contents were 13.99 and 7.52 QE quercetin equivalent/g dry matter DM, respectively. From the antioxidant tests, we deduced that the potato peels present a higher antioxidant power with the concentration of extracts causing a 50% inhibition IC50s of 125.42 ± 2.78 μg/mL for 2,2-diphényl 1-picrylhydrazyle DPPH, of 87.21 ± 7.72 μg/mL for phosphomolybdate and 200.77 ± 13.38 μg/mL for iron chelation, compared with the results obtained for shallot leaves which were 204.29 ± 0.09, 45.85 ± 3,46 and 1004.10 ± 145.73 μg/mL, respectively. The results of the physicochemical analyzes have shown that the formulated cheese was compliant with standards. Microbiological analyzes show that the hygienic quality of the cheese produced was satisfactory. According to the sensory analysis, the experts liked the cheese enriched with the powder and pieces of the leaves of the shallots.

Keywords: shallots leaves, potato peels, ultrasound extraction, phenolics, cheese

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Authors: Jessica Pinheiro Silva, Brenda Rabello De Camargo, Daniel Gusmao De Morais, Eliane Ferreira Noronha

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The utilization of lignocellulosic biomass as source of polysaccharides for industrial applications requires an arsenal of enzymes with different mode of action able to hydrolyze its complex and recalcitrant structure. Clostridium thermocellum is gram-positive, thermophilic bacterium producing lignocellulosic hydrolyzing enzymes in the form of multi-enzyme complex, termed celulossomes. This complex has several hydrolytic enzymes attached to a large and enzymically inactive protein known as Cellulosome-integrating protein (CipA), which serves as a scaffolding protein for the complex produced. This attachment occurs through specific interactions between cohesin modules of CipA and dockerin modules in enzymes. The present work aims to construct celulosomes in vitro with the structural protein CipA, a xylanase called Xyn10D and a cellulose called CelJ from C.thermocellum. A mini-scafoldin was constructed from modules derived from CipA containing two cohesion modules. This was cloned and expressed in Escherichia coli. The other two genes were cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9 and integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. Purification of each protein is being carried out. Further studies regarding enzymatic activity of the cellulosome is going to be evaluated. The cellulosome built in vitro and composed of mini-CipA, CelJ and Xyn10D, can be very interesting for application in industrial processes involving the degradation of plant biomass.

Keywords: cellulosome, CipA, Clostridium thermocellum, cohesin, dockerin, yeast

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Authors: R. Posia, J. Mistry, K. Kamani

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The health screening of laboratory rodents is essential for ensuring animal health and the validity of biomedical research data. Routine health monitoring is necessary to verify the effectiveness of biosecurity and the specific pathogen free (SPF) status of the colony. The present screening was performed in barrier maintained rat (Rattus norvegicus) colony. Rats were maintained under a controlled environment and strict biosecurity in the facility. The screening was performed on quarterly bases from randomly selected animals from breeding and or maintenance colonies. Selected animals were subject to blood collection under isoflurane anaesthesia. Serum was separated from the collected blood and stored samples at -60 ± 10 °C until further use. A total of 88 samples were collected quarterly bases from animals in a year. In the serological test, enzyme-linked immunosorbent assay (ELISA) was used for screening of serum samples against sialodacryoadenitis virus (SDAV), Sendai virus (SV), and Kilham’s rat virus (KRV). ELISA kits were procured from XpressBio, USA. Test serum samples were run along with positive control, negative control serum in 96 well ELISA plates as per the procedure recommended by the vendor. Test ELISA plate reading was taken in the microplate reader. This screening observed that none of the samples was observed positive for the sialodacryoadenitis virus (SDAV), Sendai virus (SV), and Kilham’s rat virus (KRV), indicating that effectiveness of biosecurity practices followed in the rodent colony. The result of serological screening helps us to declare that our rodent colony is specifically pathogen free for these pathogens.

Keywords: biosecurity, ELISA, specific pathogen free, serological screening, serum

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Authors: Chika C. Ogueke, Ijeoma M. Agunwah

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The antimicrobial activities and preservative potentials of crude extracts of Alstonia boonei stem bark and Euphorbia hirta leaves were studied. Soxhlet extraction and cold ethanol extraction methods were used for the extraction of the dried and ground plant samples. Well in agar diffusion method was used for the antimicrobial screening at different concentrations of 25mg/ml, 50mg/ml, 100mg/ml and 200mg/ml on E.coli and B.subtilis. The preservative effects of the extracts at 0.1%, 0.2% and 0.3% singly and in combination were determined in minced meat using E. coli and B. subtilis as test isolates. Phytochemical analysis was also conducted on the extracts using standard analytical methods. E.hirta cold and A.boonei cold extracts gave the highest zone of growth inhibition on E. coli and B.substilis with 20mm zone diameter at 200mg/ml concentration. Phytochemical analysis revealed the presence of alkaloids, flavonoids, tannins, saponins and cardiac glycosides. A.boonei at 0.1, 0.2 and 0.3% produced a log cycle reduction on the growth of E.coli. Mixture of A. boonei and E. hirta extracts (1:1) at 0.1% and 0.2% also produced a log cycle reduction on the growth of E.coli and B. subtilis, however the A. boonei extracts had more significant effect on the isolates. The observed antimicrobial activities are attributed to the phytochemicals identified in the extracts. The results reveal the potentials of plant extracts as natural antimicrobial preservatives in minced meat. Thus the crude extracts can act as inhibitors of bacteria in a food system. Upon further purification better results may be obtained.

Keywords: antimicrobial preservative, crude extracts, minced meat, test isolates

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Authors: Camille Charlebois-Plante, Marie-Ève Bourassa, Gaelle Dumel, Meriem Sabir, Louis De Beaumont

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To the extent of our knowledge, there has been little interest in the chronic effects of mild traumatic brain injury (mTBI) on cognition during normal aging. This is rather surprising considering the impacts on daily and social functioning. In addition, sustaining a mTBI during late adulthood may increase the effect of normal biological aging in individuals who consider themselves normal and healthy. The objective of this study was to characterize the persistent neuropsychological repercussions of mTBI sustained during late adulthood, on average 12 months prior to testing. To this end, 35 mTBI patients and 42 controls between the ages of 50 and 69 completed an exhaustive neuropsychological assessment lasting three hours. All mTBI patients were asymptomatic and all participants had a score ≥ 27 at the MoCA. The evaluation consisted of 20 standardized neuropsychological tests measuring memory, attention, executive and language functions, as well as information processing speed. Performance on tests of visual (Brief Visuospatial Memory Test Revised) and verbal memory (Rey Auditory Verbal Learning Test and WMS-IV Logical Memory subtest), lexical access (Boston Naming Test) and response inhibition (Stroop) revealed to be significantly lower in the mTBI group. These findings suggest that a mTBI sustained during late adulthood induces lasting effects on cognitive function. Episodic memory and executive functions seem to be particularly vulnerable to enduring mTBI effects.

Keywords: cognitive function, late adulthood, mild traumatic brain injury, neuropsychology

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Authors: Indu Bhushan Sharma, Rashmi Saraswat

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The thermotolerant, solvent stable and alkalophilic lipase producing bacterial strain was isolated from the water sample of the foothills of Trikuta Mountain in Kakryal (Reasi district) in Jammu and Kashmir, India. The lipase-producing microorganisms were screened using tributyrin agar plates. The selected microbe was optimized for maximum lipase production by subjecting to various carbon and nitrogen sources, incubation period and inoculum size. The selected strain was identified as Bacillus subtilis strain kakrayal_1 (BSK_1) using 16S rRNA sequence analysis. Effect of pH, temperature, metal ions, detergents and organic solvents were studied on lipase activity. Lipase was found to be stable over a pH range of 6.0 to 9.0 and exhibited maximum activity at pH 8. Lipolytic activity was highest at 37°C and the enzyme activity remained at 60°C for 24hrs, hence, established as thermo-tolerant. Production of lipase was significantly induced by vegetable oil and the best nitrogen source was found to be peptone. The isolated Bacillus lipase was stimulated by pre-treatment with Mn2+, Ca2+, K+, Zn2+, and Fe2+. Lipase was stable in detergents such as triton X 100, tween 20 and Tween 80. The 100% ethyl acetate enhanced lipase activity whereas, lipase activity were found to be stable in Hexane. The optimization resulted in 4 fold increase in lipase production. Bacillus lipases are ‘generally recognized as safe’ (GRAS) and are industrially interesting. The inducible alkaline, thermo-tolerant lipase exhibited the ability to be stable in detergents and organic solvents. This could be further researched as a potential biocatalyst for industrial applications such as biotransformation, detergent formulation, bioremediation and organic synthesis.

Keywords: bacillus, lipase, thermotolerant, alkalophilic

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Authors: O. H. Gebril, N. A. Meguid

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Background: Iron had been a focus of interest recently as a main exaggerating factor for oxidative stresses in the central nervous system and a link to various neurological disorders is suspected. Many studies with various techniques showed evidence of disturbed iron-related proteins in the cell in human and animal models of neurodegenerative disorders. Also, linkage to significant pathological changes had been evidenced e.g. apoptosis and cell signaling. On the other hand, the role of iron in neurodevelopmental disorders is still unclear. With increasing prevalence of autism worldwide, some changes in iron parameters and its stores were documented in many studies. This study includes Haemochromatosis HFE gene polymorphisms (p.H63D and p.C282Y) and ferroportin gene (SLC40A1) Q248H polymorphism in autism and control children. Materials and Methods: Whole genome DNA was extracted; p.H63D and p.C282Y genotyping was studied using specific sequence amplification followed by restriction enzyme digestion on a sample of autism patients (25 cases) and twenty controls. Results: The p.H63D is seen more than the C282Y among both autism and control samples, with no significant association of p.H63D or p.C282Y polymorphism and autism was revealed. Also, no association with Q248H polymorphism was evidenced. Conclusion: The study results do not prove the role of cellular iron genes polymorphisms as risk factors for neurodevelopmental disorders, and in turn highlights the specificity of cellular iron related pathways in neurodegeneration. These results demand further gene expression studies to elucidate the main pathophysiological pathways that are disturbed in autism and other neurodevelopmental disorders.

Keywords: iron, neurodevelopmental, oxidative stress, haemohromatosis, ferroportin, genes

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Authors: Carlo Stephen O. Moneva, Sharon Rose M. Tabugo

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The present study aims to assess polymorphism in the prolactin (C576A) gene and determine the influence of different prolactin (PRL) genotypes to milk yield performance in crossbred Anglo-Nubian dairy goats raised from Awang, Opol, Misamis Oriental and Talay, Dumaguete City, Negros Oriental. Genomic DNA was extracted from hair follicles and Polymerase Chain Reaction – Restriction Fragment Length Polymorphism (PCR-RFLP) was performed for the genotyping of the C576A polymorphism located in exon 5 of goats’ prolactin gene using Eco241 restriction enzyme. Genotypic and allelic frequencies of 0.56 for AA, 0.44 for AB, 0.78 for A, and 0.22 for B were recorded. Observed heterozygosity values were higher than the expected heterozygosity. All populations followed the Hardy–Weinberg principle at p>0.05, except for dairy goats from Farm A located in Opol, Misamis Oriental. A two-way factorial (2 x 4) in a Randomized Complete Block Design was used to be able to evaluate the relationship between genotypes and milk yield performance. PRL genotypes and parity were used as main factors and farm as the blocking factor. AB genotype goats produced significantly higher average daily milk yield and total milk production than AA genotype (p<0.05), an indication that the polymorphism in the caprine PRL (C576A) gene influenced milk yield performance in the population of crossbred Anglo-Nubian goats from Opol, Misamis Oriental and Dumaguete City, Negros Oriental. However, these results have to be validated in other dairy goat breeds.

Keywords: polymorphism, prolactin, milk yield, Anglo-Nubian, PCR-RFLP

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Authors: Bercem Kiran-Yildirim, Volker Gaukel

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Ice recrystallization (IR) which occurs especially during frozen storage is an undesired process due to the possible influence on the quality of products. As a result of recrystallization, the total volume of ice remains constant, but the size, number, and shape of ice crystals change. For instance, as indicated in the literature, the size of ice crystals in ice cream increases due to recrystallization. This results in texture deterioration. Therefore, the inhibition of ice recrystallization is of great importance, not only for food industry but also for several other areas where sensitive products are stored frozen, like pharmaceutical products or organs and blood in medicine. Ice-binding proteins (IBPs) have the unique ability to inhibit ice growth and in consequence inhibit recrystallization. This effect is based on their ice binding affinity. In the presence of IBP in a solution, ice crystal growth is inhibited during temperature decrease until a certain temperature is reached. The melting during temperature increase is not influenced. The gap between melting and freezing points is known as thermal hysteresis (TH). In literature, the TH activity is usually investigated under laboratory conditions in IBP buffer solutions. In product applications (e.g., food) there are many other solutes present which may influence the TH activity. In this study, a subset of IBPs, so-called antifreeze proteins (AFPs), is used for the investigation of the influence of sucrose solution concentration on the TH activity. For the investigation, a polarization microscope (Nikon Eclipse LV100ND) equipped with a digital camera (Nikon DS-Ri1) and a cold stage (Linkam LTS420) was used. In a first step, the equipment was established and validated concerning the accuracy of TH measurements based on literature data.

Keywords: ice binding proteins, ice crystals, sucrose solution, thermal hysteresis

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Authors: Ga-Young Lee, Hyun-Man Kim

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The modulation of the cell-cell junction is critical in epithelial-mesenchymal transition during tumorigenesis. As RhoA activity is known to be up-regulated to dissociate cell-cell junction by contracting acto-myosin complex in various cancer cells, the present study investigated if RhoA activity was also associated with the disruption of the cell-cell junction of oral cancer cells. We studied SCC-25 cells which are established from oral squamous cell carcinoma if their E-cadherin junction (ECJ) was under control of RhoA. Interestingly, development of ECJ of SCC-25 cells depended on the amount of fibronectin (FN) coated on the culture dishes. Seeded cells promptly aggregated to develop ECJ on the substrates coated with a low amount of FN, whereas they were retarded in the development of ECJ on the substrates coated with a high amount of FN. However, it was an unexpected finding that total RhoA activity was lower in the dissociated cells on the substrates of high FN than in the aggregated cells on the substrates of low FN. Treating the dissociated cells on the substrates of high FN with LPA, a RhoA activator, promoted the development to ECJ. In contrast, treating the aggregated cells on the substrates of low FN with Clostridium botulinum C3, a toxin decreasing RhoA activity, dissociated cells concomitant with the disruption of ECJ. Genetical knockdown of RhoA expression by transfecting RhoA siRNA also down-regulated the development of ECJ in SCC-25 cells. Furthermore, PMA, an activator of protein kinase C (PKC), down-regulated the development of ECJ junction of SCC-25 cells on the substrates coated with low FN. In contrast, GO6976, a PKC inhibitor, up-regulated the development of ECJ of SCC-25 cells with the activation of RhoA on the substrates coated with high FN. In conclusion, in the present study, we demonstrated unexpected results that the activation of RhoA promotes the development of ECJ, whereas the inhibition of RhoA retards the development of ECJ in SCC-25 cells.

Keywords: E-cadherin junction, oral squamous cell carcinoma, PKC, RhoA, SCC-25

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Authors: Gajanan V. Mane, Rashmi More, Mahesh S. Sonawane, Tushar Lodha, Rohit Sharma

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The diversity of fungi isolated from two different termite species viz., Odontoterms assmuthi and O. abesus was investigated by dilution- plate method, combined with morphological characteristics and sequencing of internal transcribed spacer region. In total, ninety-six fungi were isolated and purified, out of which 69 isolates were obtained from O. assmuthi belonging to 18 genera and 31 species, whereas 27 isolates were obtained from O. abesus belonging to 15 genera and 17 species. The fungal strains were screened for laccase, amylase, cellulase and pectinase enzymes production. Twenty-seven strains were positive for laccase, 59 strains were positive for amylase, 71 strains were positive for cellulase and 72 strains were positive for pectinase enzymes. The antimicrobial activities of the isolated fungi were tested by the dual plate culture method against standard pathogens. Bioactive secondary metabolites were identified by HPLC and LCMS. Four isolates viz., Penicillium goetzii MG 57, Epicoccum sp. MG 39, Penicillium tanzanicum MG 30, Aspergillus polyporicola MG 54, showed positive antimicrobial activity against standard pathogens, Streptococcus pneumonia MCC 2425, Staphylococcus aureus MCC 2408, Pseudomonas aeruginosa MCC 2080, Escherichia coli MCC 2412, Enterococcus faecalis MCC 2409, Klebsiella pneumonia MCC 2451, Micrococcus luteus MCC 2155 and Candida albicans MCC 1151. In conclusion, the study showed that the insect gut harbor fungal diversity, which is futuristic with biotechnological potential and could be a good source of enzymes and antibiotics.

Keywords: termites, fungi, its, enzyme, antimicrobial activity

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Authors: Irma Gabriela Lopez-Moreno, Elva Perez-Luque, Herlinda Aguilar-Zavala

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Introduction: Type 2 Diabetes Mellitus (T2DM) is characterized by combination of insulin resistance and deterioration of insulin secretion. Sfrp5 is a protein that antagonizes Wnt5a proteins by preventing it from reaching its receptor and activating the Wnt/β-catenin signaling pathway, this pathway is one of the most important regulators of adipogenesis. Although metformin decreases glucose levels its mechanisms of action are not fully known but it has been implicated in the inhibition of the Wnt/β-catenin signaling pathway. Objective: The objective was evaluating the effects of metformin on serum levels of Sfrp5 in patients with T2DM treated and not treated with metformin. Methods: Two groups of patients were selected: one group of T2DM patients treated with metformin (n = 35) and another group of subjects with recent diagnosis of T2DM untreated (n = 35) with a mean age of 48 ± 9 years. In these subjects anthropometric measures were taken as weight, height, waist and hip circumference, were calculated the percentage of body fat, visceral fat and muscle mass. In addition, were measured glucose levels, lipid profile, adiponectin and Sfrp5. Results: Sfrp5 were higher in metformin-treated patients compared to the untreated group (19.9 vs 13.6 ng/mL p < 0.001), a negative correlation was found between Sfrp5 levels and total cholesterol levels (r= -0.25, p = 0.03) and percentage of visceral fat (r = -0.26, p = 0.03) and a positive correlation with HDL cholesterol levels (r = 0.31, p = 0.01) and adiponectin (r=0.65, p = < 0.001). Conclusions: The findings show that metformin consumption increased levels of Sfrp5, which may lead to a decrease in the activation of the WNT/β-catenin pathway impacting on adipogenesis.

Keywords: adiponectin, diabetes, metformin, Sfrp5

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Authors: Divya Vohora, Md. Jamir Anwar

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Antiepileptic drugs (AEDs)-induced adverse consequences on bone are now well recognized. Despite this, there is limited data on the effect of anti-osteoporotic therapies on AEDs-induced bone loss. Both phenytoin (PHT) and sodium valproate (SVP) inhibit human aromatase enzyme and stimulate microsomal catabolism of oestrogens. Estrogen deficiency states are known to reduce the deposition of transforming growth factor-β (TGF-β3), a bone matrix protein, having anti-osteoclastic property. Thus, an attempt was made to investigate the effect of raloxifene, a selective oestrogen receptor modulator, in comparison with CVD supplementation, on PHT and SVP-induced alterations in bone in mice. Further, the effect of raloxifene on seizures and on the antiepileptic efficacy of AEDs was also investigated. Swiss strains of female mice were treated with PHT (35 mg/kg, p.o.) and SVP (300 mg/kg, p.o.) for 120 days to induce bone loss as evidenced by reduced bone mineral density (BMD) and altered bone turnover markers in lumbar bones (alkaline phosphatase, tartarate resistant acid phosphatase, hydroxyproline) and urine (calcium). The bone loss was accompanied by reduced serum estradiol levels and bone TGF-β3 content. Preventive and curative treatment with raloxifene ameliorated bony alterations and was more effective than CVD. Deprived estrogen levels (that in turn reduced lumbar TGF-β3 content) following PHT and SVP, thus, might represent one of the various mechanisms of AEDs-induced bone loss. Raloxifene preserved the bony changes without interfering with their antiepileptic efficacy, and hence raloxifene could be a potential therapeutic option in the management of PHT and SVP-induced bone disease if clinically approved.

Keywords: antiepileptic drugs, osteoporosis, raloxifene, TGF-β3

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Authors: Boris Ershov

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Depression is the most common mood disorder experienced by patients who have sustained a traumatic brain injury (TBI) and is associated with poorer cognitive functional outcomes. However, in some cases, similar cognitive impairments can also be observed in depression. There is not enough information about the features of the cognitive deficit in patients with TBI in relation to patients with depression. TBI patients without depressive symptoms (TBInD, n25), TBI patients with depressive symptoms (TBID, n31), and 28 patients with bipolar II disorder (BP) were included in the study. There were no significant differences in participants in respect to age, handedness and educational level. The patients clinical status was determined by using Montgomery–Asberg Depression Rating Scale (MADRS). All participants completed a cognitive battery (The Brief Assessment of Cognition in Affective Disorders (BAC-A)). Additionally, the Rey–Osterrieth Complex Figure (ROCF) was used to assess visuospatial construction abilities and visual memory, as well as planning and organizational skills. Compared to BP, TBInD and TBID showed a significant impairments in visuomotor abilities, verbal and visual memory. There were no significant differences between BP and TBID groups in working memory, speed of information processing, problem solving. Interference effect (cognitive inhibition) was significantly greater in TBInD and TBID compared to BP. Memory bias towards mood-related information in BP and TBID was greater in comparison with TBInD. These results suggest that depressive symptoms are associated with impairments some executive functions in combination at decrease of speed of information processing.

Keywords: bipolar II disorder, depression, neurocognitive deficits, traumatic brain injury

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Authors: Haile F. Darge, Hsieh C. Tsai

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The tumor microenvironment affects the therapeutic outcomes of cancer disease. In a malignant tumor, overexpression of vascular endothelial growth factor (VEGF) provokes the production of pathologic vascular networks. This results in a hostile tumor environment that hinders anti-cancer drug activities and profoundly fuels tumor progression. In this study, we develop a strategy of sequential sustain release of the anti-angiogenic drug: Bevacizumab(BVZ), and anti-cancer drug: Doxorubicin(DOX) which had a synergistic effect on cancer treatment. Poly (D, L-Lactide)- Poly (ethylene glycol) –Poly (D, L-Lactide) (PDLLA-PEG-PDLLA) thermo-sensitive hydrogel was used as a vehicle for local delivery of drugs in a single platform. The in vitro release profiles of the drugs were investigated and confirmed a relatively rapid release of BVZ (73.56 ± 1.39%) followed by Dox (61.21 ± 0.62%) for a prolonged period. The cytotoxicity test revealed that the copolymer exhibited negligible cytotoxicity up to 2.5 mg ml-1 concentration on HaCaT and HeLa cells. The in vivo study on Hela xenograft nude mice verified that hydrogel co-loaded with BVZ and DOX displayed the highest tumor suppression efficacy for up to 36 days with pronounce anti-angiogenic effect of BVZ and with no noticeable damage on vital organs. Therefore, localized co-delivery of anti-angiogenic drug and anti-cancer drugs by the hydrogel system may be a promising approach for enhanced chemotherapeutic efficacy in cancer treatment.

Keywords: anti-angiogenesis, chemotherapy, controlled release, thermo-sensitive hydrogel

Procedia PDF Downloads 134

Authors: Vineeta Kaushik, Manisha Goel

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Chaperones are proteins that help other proteins fold correctly, and are found in all domains of life i.e., prokaryotes, eukaryotes and archaea. Various comparative genomic studies have suggested that the archaeal protein folding machinery appears to be highly similar to that found in eukaryotes. In case of protein folding; slow rotation of peptide prolyl-imide bond is often the rate limiting step. Formation of the prolyl-imide bond during the folding of a protein requires the assistance of other proteins, termed as peptide prolyl cis-trans isomerases (PPIases). Cyclophilins constitute the class of peptide prolyl isomerases with a wide range of biological function like protein folding, signaling and chaperoning. Most of the cyclophilins exhibit PPIase enzymatic activity and play active role in substrate protein folding which classifies them as a category of molecular chaperones. Till date, there is not very much data available in the literature on archaeal cyclophilins. We aim to compare the structural and biochemical features of the cyclophilin protein from within the three domains to elucidate the features affecting their stability and enzyme activity. In the present study, we carry out in-silico analysis of the cyclophilin proteins to predict their conserved residues, sites under positive selection and compare these proteins to their bacterial and eukaryotic counterparts to predict functional divergence. We also aim to clone and express these proteins in heterologous system and study their biophysical characteristics in detail using techniques like CD and fluorescence spectroscopy. Overall we aim to understand the features contributing to the folding, stability and dynamics of the archaeal cyclophilin proteins.

Keywords: biophysical characterization, x-ray crystallography, chaperone-like activity, cyclophilin, PPIase activity

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Authors: Turan Yaman, Zabit Yener, Ismail Celik

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The aim of this study was to investigate the antioxidant properties and protective role of honey, considered a part of traditional medicine, against carcinogen chemical aflatoxin (AF) exposure in rats, which were evaluated by histopathological changes in liver and kidney, measuring level of serum marker enzymes [aspartate aminotransferase (AST), alanin aminotransferase (ALT), gamma glutamil transpeptidase (GGT)], antioxidant defense systems [Reduced glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione-S-transferase (GST) and catalase (CAT)], and lipid peroxidation content in liver, erythrocyte, brain, kidney, heart and lungs. For this purpose, a total of eighteen healthy Sprague-Dawley rats were randomly allocated into three experimental groups: A (Control), B (AF-treated) and C (AF+honey-treated). While rats in group A were fed with a diet without AF, B, and C groups received 25 µg of AF/rat/day, where C group additionally received 1 mL/kg of honey by gavage for 90 days. At the end of the 90-day experimental period, we found that the honey supplementation decreased the lipid peroxidation and the levels of enzyme associated with liver damage, increased enzymatic and non-enzymatic antioxidants in the AF+honey-treated rats. Hepatoprotective and nephroprotective effects of honey is further substantiated by showing almost normal histological architecture in AF+honey-treated group, compared to degenerative changes in the liver and kidney of AF-treated rats. Additionally, honey supplementation ameliorated antioxidant defense systems and lipid peroxidation content in other tissues of AF+honey-treated rats. In conclusion, the present study indicates that honey has a hepatoprotective and nephroprotective effect in rats with experimental aflatoxicosis due to its antioxidant activity.

Keywords: aflatoxicosis, honey, histopathology, malondialdehyde, antioxidant, rat

Procedia PDF Downloads 334

Authors: Shih-Huang Pan, Tsuyoshi Miyazaki, Minoru Otani, Santhanamoorthi Nachimuthu, Jyh-Chiang Jiang

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Ammonia (NH₃) is crucial in renewable energy and agriculture, yet its traditional production via the Haber-Bosch process faces challenges due to the inherent inertness of nitrogen (N₂) and the need for high temperatures and pressures. The electrocatalytic nitrogen reduction (ENRR) presents a more sustainable option, functioning at ambient conditions. However, its advancement is limited by selectivity and efficiency challenges due to the competing hydrogen evolution reaction (HER). The critical roles of protonation of N-species and HER highlight the necessity of selecting optimal catalysts and solvents to enhance ENRR performance. Notably, transition metal oxides, with their adjustable electronic states and excellent chemical and thermal stability, have shown promising ENRR characteristics. In this study, we use density functional theory (DFT) methods to investigate the ENRR mechanisms on IrO₂ (110), a material known for its tunable electronic properties and exceptional chemical and thermal stability. Employing the constant electrode potential (CEP) model, where the electrode - electrolyte interface is treated as a polarizable continuum with implicit solvation, and adjusting electron counts to equalize work functions in the grand canonical ensemble, we further incorporate the advanced 3D Reference Interaction Site Model (3D-RISM) to accurately determine the ENRR limiting potential across various solvents and pH conditions. Our findings reveal that the limiting potential for ENRR on IrO₂ (110) is significantly more favorable than for HER, highlighting the efficiency of the IrO₂ catalyst for converting N₂ to NH₃. This is supported by the optimal *NH₃ desorption energy on IrO₂, which enhances the overall reaction efficiency. Microkinetic simulations further predict a promising NH₃ production rate, even at the solution's boiling point¸ reinforcing the catalytic viability of IrO₂ (110). This comprehensive approach provides an atomic-level understanding of the electrode-electrolyte interface in ENRR, demonstrating the practical application of IrO₂ in electrochemical catalysis. The findings provide a foundation for developing more efficient and selective catalytic strategies, potentially revolutionizing industrial NH₃ production.

Keywords: density functional theory, electrocatalyst, nitrogen reduction reaction, electrochemistry

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Authors: A. A. M. Azoddein, Y. Nuratri, A. B. Bustary, F. A. M. Azli, S. C. Sayuti

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Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage in 4oC without vacuum. Polyethylene glycol (PEG) pre-treated freeze dry cells and broth pre-treated freeze dry cells after freeze-dry recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introduce freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 56.78% and 17.91%. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks were 26.35% and 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been growth in agar. Result from this study may be beneficial and useful as initial reference before commercialize freeze-dried P. putida.

Keywords: Pseudomonas putida, freeze-dry, PEG, tween80/Sucrose, mercury, cell viability

Procedia PDF Downloads 355

Authors: Kamran Jawed, Anu Jose Mattam, Zia Fatma, Saima Wajid, Malik Z. Abdin, Syed Shams Yazdani

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Worldwide demand of natural and sustainable fuels and chemicals have encouraged researchers to develop microbial platform for synthesis of short chain fatty acids as they are useful precursors to replace petroleum-based fuels and chemicals. In this study, we evaluated the role of fatty acid synthesis and β-oxidation cycle of Escherichia coli to produce butyric acid, a 4-carbon short chain fatty acid, with the help of three thioesterases, i.e., TesAT from Anaerococcus tetradius, TesBF from Bryantella formatexigens and TesBT from Bacteroides thetaiotaomicron. We found that E. coli strain transformed with gene for TesBT and grown in presence of 8 g/L glucose produced maximum butyric acid titer at 1.46 g/L, followed by that of TesBF at 0.85 g/L and TesAT at 0.12 g/L, indicating that these thioesterases were efficiently converting short chain fatty acyl-ACP intermediate of fatty acid synthesis pathway into the corresponding acid. The titer of butyric acid varied significantly depending upon the plasmid copy number and strain genotype. Deletion of genes for fatty acyl-CoA synthetase and acyl-CoA dehydrogenase, which are involved in initiating the fatty acid degradation cycle, and overexpression of FadR, which is a dual transcriptional regulator and exerts negative control over fatty acid degradation pathway, reduced up to 30% of butyric acid titer. This observation suggested that β-oxidation pathway is working synergistically with fatty acid synthesis pathway in production of butyric acid. Moreover, accelerating the fatty acid elongation cycle by overexpressing acetyl-CoA carboxyltransferase (Acc) and 3-hydroxy-acyl-ACP dehydratase (FabZ) or by deleting FabR, the transcription suppressor of elongation, did not improve the butyric acid titer, rather favored the long chain fatty acid production. Finally, a balance between cell growth and butyric acid production was achieved with the use of phosphorous limited growth medium and 14.3 g/L butyric acid, and 17.5 g/L total free fatty acids (FFAs) titer was achieved during fed-batch cultivation. We have engineered an E. coli strain which utilizes the intermediate of both fatty acid synthesis and degradation pathway, i.e. butyryl-ACP and -CoA, to produce butyric acid from glucose. The strategy used in this study resulted in highest reported titers of butyric acid and FFAs in engineered E. coli.

Keywords: butenoic acid, butyric acid, Escherichia coli, fed-batch fermentation, short chain fatty acids, thioesterase

Procedia PDF Downloads 371

Authors: Mukunda Upreti, Jill Storry, Rafael Björk, Emilie Louvet, Johan Normark, Sven Bergström

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Borrelia duttonii and most other relapsing fever species are Gram-negative bacteria which cause a blood borne infection characterized by the binding of bacterium to erythrocytes. The bacteria associate with two or more erythrocytes to form clusters of cells into rosettes. Rosetting is a major virulence factor and the mechanism is believed to facilitate persistence of bacteria in the circulatory system and the avoidance of host immune cells through masking or steric hindrance effects. However, the molecular mechanisms of rosette formation are still poorly understood. This study aims at determining the molecules involved in the rosette formation phenomenon. Fractionated serum, using different affinity purification methods, was investigated as a rosetting agent and IgG and at least one other serum components were needed for rosettes to form. An IgG titration curve demonstrated that IgG alone is not enough to restore rosette formation level to the level whole serum gives. IgG hydrolysis by IdeS ( Immunoglobulin G-degrading enzyme of Streptococcus pyogenes) and deglycosylation using N-Glycanase proved that the whole IgG molecule regardless of saccharide moieties is critical for Borrelia induced rosetting. Complement components C3 and C4 were also important serum molecules necessary to maintain optimum rosetting rates. The deactivation of complement network and serum depletion with C3 and C4 significantly reduced the rosette formation rate. The dependency of IgG and complement components also implied involvement of the complement receptor (CR1). Rosette formation test with Knops null RBC and sCR1 confirmed that CR1 is also part of Borrelia induced rosette formation.

Keywords: complement components C3 and C4, complement receptor 1, Immunoglobulin G, Knops null, Rosetting

Procedia PDF Downloads 322

Authors: Leila Karshenas, Hamid Reza Khodaei, Behnaz Mahdavi

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We know that nitric oxide (NO) plays an important role in all sexual activities of animals. It is made in body from NO synthase enzyme and L-arginin molecule. NO can bound with sulfur-iron complexes and because production of steroid sexual hormones is related to enzymes which have this complex, NO can change the activity of these enzymes. NO affects many cells including endothelial cells of veins, macrophages and mast cells. These cells are found in testis leydig cells and therefore are important source of NO in testis tissue. Minimizing damages to sperm at the time of sperm freezing and thawing is really important. The goal of this study was to determine the function of NO before freezing and its effects on quality and viability of sperms after thawing and incubation. 4 Holstein bulls were selected from the age of 4, and artificial insemination was done for 3 weeks (2 times a week). Treatments were 0, 10, 50 and 100 nm of sodium nitroprusside (SNP). Data analysis was performed by SAS98 program. Also, mean comparison was done using Duncan's multiple ranges test (P<0.05). Concentrations used was found to increase motility and viability of spermatozoa at 1, 2 and 3 hours after thawing significantly (P<0.05), but there was no significant difference at zero time. SNP levels reduced the amount of lipid peroxidation in sperm membrane, increased acrosome health and improved sample membranes especially in 50 and 100 nm treatments. According to results, adding SNP to semen diluents increases motility and viability of spermatozoa. Also, it reduces lipid peroxidation in sperm membrane and improves sperm function.

Keywords: sperm motility, nitric oxide, lipid peroxidation, spermatozoa

Procedia PDF Downloads 361