Search results for: antioxidant enzyme

Authors: S. S. Singh, A. Bhattacharya, S. Bhattacharya

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The eukaryotic exosome complex plays a pivotal role in RNA biogenesis, maturation, surveillance and differential expression of various RNAs in response to varying environmental signals. The exosome is composed of evolutionary conserved nine core subunits and the associated exonucleases Rrp6 and Rrp44. Rrp6p is crucial for the processing of rRNAs, other non-coding RNAs, regulation of polyA tail length and termination of transcription. Rrp6p, a 3’-5’ exonuclease is required for degradation of 5’-external transcribed spacer (ETS) released from the rRNA precursors during the early steps of pre-rRNA processing. In the parasitic protist Entamoeba histolytica in response to growth stress, there occurs the accumulation of unprocessed pre-rRNA and 5’ ETS sub fragment. To understand the processes leading to this accumulation, we looked for Rrp6 and the exosome subunits in E. histolytica, by in silico approaches. Of the nine core exosomal subunits, seven had high percentage of sequence similarity with the yeast and human. The EhRrp6 homolog contained exoribonuclease and HRDC domains like yeast but its N- terminus lacked the PMC2NT domain. EhRrp6 complemented the temperature sensitive phenotype of yeast rrp6Δ cells suggesting conservation of biological activity. We showed 3’-5’ exoribonuclease activity of EhRrp6p with in vitro-synthesized appropriate RNAs substrates. Like the yeast enzyme, EhRrp6p degraded unstructured RNA, but could degrade the stem-loops slowly. Furthermore, immunolocalization revealed that EhRrp6 was nuclear-localized in normal cells but was diminished from nucleus during serum starvation, which could explain the accumulation of 5’ETS during stress. Our study shows functional conservation of EhRrp6p in E.histolytica, an early-branching eukaryote, and will help to understand the evolution of exosomal components and their regulatory function.

Keywords: entamoeba histolytica, exosome complex, rRNA processing, Rrp6

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Authors: Santram Lodhi, Alok Pal Jain, Awesh K. Yadav, Gopal Rai

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Rhodiola rosea L. (Crassulaceae) is commonly known as golden root or rose root. It is a perennial herbaceous plant and most investigated species of the genus Rhodiola. Rhodiola rosea contains flavonoids, terpenoids, phenylpropanoid glycosides and phenylethanol derivatives in the roots of the plant. The objective of present study was to investigate the protective effect of hydroalcoholic extract from Rhodiola rosea roots in DSS induced colitis in mice. The ulcerative colitis was induced by DSS (3%, w/v) in mice and estimated weight loss and stool consistency. Various parameters including Colon length, spleen weights and ulcer index were also measured. The histological observations were observed by H&E staining. Effect of hydroalcoholic extract on various antioxidant parameter of rat colon such as tissue myeloperoxidase (MPO), reduced GSH, SOD concentrations and lipid peroxidation were determined. Pro-inflammatory mediators, such as tumour necrosis factor-α (TNF-α) and nitric oxide (NO) were determined by ELISA. In DSS induced group, mice body weight decreased gradually as compared to the control group. Redness and edema were observed in the colons intensely and scores representing inflammation in this group. The extract treated showed with tissue levels of TNF-α, IL-6 and MPO activity were significantly (p<0.05) increased. The mice treated with higher doses of hydroalcoholic extract (300 mg/kg) significantly reduced the activity compared with standard drug sulfasalazine (100 mg/kg. B.wt). Conclusion: Results of this study were suggested that the efficacy of hydroalcoholic extract, especially at the higher dose, was similar to that of standard drug, which concerned its potential application as a natural medicine for the treatment of ulcerative colitis.

Keywords: phenylpropanoid, Rhodiola rosea, sulfasalazin, ulcerative colitis

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Authors: Sameh A. S. Thabit Alariqi

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Food is widely packaged with plastic materials to prevent microbial contamination and spoilage. Ionizing radiation is widely used to sterilize the food-packaging materials. Sterilization by γ-radiation causes degradation for the plastic packaging materials such as embrittlement, stiffening, softening, discoloration, odour generation, and decrease in molecular weight. Many antioxidants can prevent γ-degradation but most of them are toxic. The migration of antioxidants to its environment gives rise to major concerns in case of food packaging plastics. In this attempt, we have aimed to utilize synergistic mixtures of stabilizers which are approved for food-contact applications. Ethylene-propylene-diene terpolymer (EPDM) have been melt-mixed with hindered amine stabilizers (HAS), phenolic antioxidants and organo-phosphites (hydroperoxide decomposer). Results were discussed by comparing the stabilizing efficiency of mixtures with and without phenol system. Among phenol containing systems where we mostly observed discoloration due to the oxidation of hindered phenol, the combination of secondary HAS, tertiary HAS, organo-phosphite and hindered phenol exhibited improved stabilization efficiency than single or binary additive systems. The mixture of secondary HAS and tertiary HAS, has shown antagonistic effect of stabilization. However, the combination of organo-phosphite with secondary HAS, tertiary HAS and phenol antioxidants have been found to give synergistic even at higher doses of -sterilization. The effects have been explained through the interaction between the stabilizers. After γ-irradiation, the consumption of oligomeric stabilizer significantly depends on the components of stabilization mixture. The effect of the organo-phosphite antioxidant on the overall stability has been discussed.

Keywords: ethylene-propylene-diene terpolymer, synergistic mixtures, gamma sterilization, gamma stabilization

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Authors: Kadry M. Sadek, Tarek K. Abouzed, Mousa A. Ayoub

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The presence of endocrine-disrupting compounds, such as bisphenol A (BPA), in the environment can cause serious health problems. However, there are controversial opinions. This study investigated the reproductive, metabolic, oxidative and immunologic-disrupting effects of bisphenol A in male rabbits. Rabbits were divided into five groups. The first four rabbit groups were administered oral BPA (1, 10, 50, or 100 mg/kg/day) for ten weeks. The fifth group was administered corn oil as the vehicle. BPA significantly decreased serum testosterone, estradiol and the free androgen index (FAI) and significantly increased sex hormone binding globulin (SHBG) compared with the placebo group. The higher doses of BPA showed a significant decrease in follicular stimulating hormone (FSH) and luteinizing hormone (LH). A significant increase in blood glucose levels was identified in the BPA groups. The non-significant difference in insulin levels is a novel finding. The cumulative testicular toxicity of BPA was clearly demonstrated by the dose-dependent decrease in absolute testes weight, primary measures of semen quality and a significant increase in testicular malonaldehyde (MDA). Moreover, BPA significantly decreased total antioxidant capacity (TAC) and significantly increased immunoglobulin G (IgG) at the highest concentration. Our results suggest that BPA, especially at higher doses, is associated with many adverse effects on metabolism, oxidative stress, immunity, sperm quality and markers of androgenic action. These results may reflect the estrogenic effects of BPA, which we hypothesize could be related, in part, to an inhibitory effect on testicular steroidogenesis. The induction of oxidative stress by BPA may play an additional role in testicular toxicity. These results suggest that BPA poses a threat to endocrine and reproductive functions.

Keywords: bisphenol A, oxidative stress, rabbits, semen quality, steroidogenesis

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Authors: Al-Nouri Doha Mostfa, Al-Khalifa Abdulrahman Salih

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The effects of long-term supplementation with different dietary omega-6/omega-3 (ω-6/ω-3) polyunsaturated fatty acid (PUFAs) ratios on the bone marrow fatty acids level, plasma biomarkers of bone metabolism, and minerals content in bone were evaluated in rabbits. Weanling male and female New Zealand white rabbits were randomly assigned to five groups and fed ad libitum for 100 days on diets containing 70 g/kg different dietary oils which providing the following ω-6/ω-3 ratios: soy bean oil (SBO control, 8.68), sesame oil (SO, 21.75), fish oil (FO, 0.39), DHA algae oil (DHA, 0.63), and DHA and ARA algae oils (DHA/ARA, 0.68). The bone marrow arachidonic (ARA), eicosapentaenoic (EPA), and docosahexaenoic (DHA) fatty acid levels were significantly influenced by and reflected the dietary ω-6/ω-3 ratios fed to rabbits. Rabbits fed on the FO diet maintained a lower ω-6/ω-3 ratio and a higher EPA and DHA levels, those fed on the DHA/ARA diet maintained a lower ω-6/ω-3 ratio and a higher ARA level, while those fed on the SO diet maintained a higher ω-6/ω-3 ratio and a lower ARA level. Plasma alkaline phosphatase (ALP) activity was significantly higher in male and female rabbits fed the DHA/ARA diet compared with those fed the control, SO, FO, or DHA diets. There was a significant main effect of dietary treatment on femur calcium (Ca), phosphorous (P), magnesium (Mg), and zinc (Zn) contents in both genders. This study confirmed that different dietary oil sources with varying ω-6/ω-3 ratios significantly altered the fatty acids level of bone marrow. In addition, the significant elevation in minerals content and the maintenance of optimal Ca/P ratio in bone of DHA/ARA and DHA fed groups beside the significant elevation in ALP activity in the DHA/ARA fed group proved that marine algae oils may be promising dietary sources for promoting bone mineralization and formation, thus improving bone mass during the growth stage.

Keywords: arachidonic (ARA), docosahexaenoic (DHA), eicosapentaenoic (EPA), growing rabbits

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Authors: A. Khan, I. Khan, M. Younus, S. E. Haque, U. Waheed, H. Neubauer, A. A. Anjum, S. A. Muhammad, A. Idrees T. Abbas, S. Raza, M. A. Ali, M. Farooq, M. Mahmood, A. Hussain, H. Danish, U. Tayyab, M. Zafar, M. Aslam.

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Brucellosis is one of the major problems of milk producing animals in our country which deteriorate the health of livestock. It is a disease of zoonotic significance which is capable of producing disease in humans leading to infertility, orchitis, abortions, and synovitis. In this particular study, milk and serum samples of cattle and buffalo (n=402) were collected from different districts of Punjab including Narowal, Gujranwala and Gujrat. Milk samples were analyzed by Milk Ring Test (MRT), while serum samples were tested through Rose Bengal Plate agglutination Test (RBPT) and Indirect Enzyme Linked Immunosorbant Assay (i-ELISA). The sample tested with MRT were 9.5% positive, including cattle 9.6% and buffalo 9.3%. While using the RBPT test for the detection of serum samples and for screening purpose it was observed that 16.4% animals were seropositive, cattle were 18.8% and buffalo were 13.9% seropositive. The higher prevalence of brucellosis indicates the danger of the disease to human population. The serum samples positive by RBPT were further confirmed by the use of most specific and sensitive serological test known as i-ELISA. 11.4% animals were confirmed as seropositive by i-ELISA including cattle 13.5% seropositive and buffalo 9.3%. The results indicated high seroprevalence of brucellosis in cattle as compared to buffalos. Different risk factors were also studied to know the association between disease and their spread. Advanced age, larger herds, history of abortion and pregnancy of the animals is considered to be the important factors for the prevalence and spread of the hazardous zoonotic disease. It is a core issue of developing countries like Pakistan and has major public health impact.

Keywords: humans, bovines, infertility, orchitis, abortions, seroprevalence, brucellosis

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Authors: D. Motlhanka, M. Mine, T. Bagaketse, T. Ngakane

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There is a plethora of evidence from numerous sources that highlights the triumph of naturally derived medicinal plant metabolites with antioxidant capability for repurposed therapeutics. As post-COVID-19 syndromes and non-communicable diseases are on the rise, there is an urgent need to come up with new therapeutic strategies to address the problem. Non-communicable diseases and Post COVID-19 syndromes are classified as socio-economic diseases and are ranked high among threats to health security due to the economic burden they pose to any government budget commitment. Research has shown a strong link between accumulation of free radicals and oxidative stress critical for pathogenesis of non-communicable diseases and COVID-19 syndromes. Botswana has embarked on a robust programme derived from ethno-pharmacognosy and drug repurposing to address these threats to health security. In the current approach, a number of medicinally active plant-derived polyphenolics are repurposed and combined into new medicinal tools to target diabetes, Hypertension, Prostate Cancer and oxidative stress induced Post COVID 19 syndromes such as “brain fog”. All four formulants demonstrated Free Radical scavenging capacities above 95% at 200µg/ml using the diphenylpicryalhydrazyl free radical scavenging assay and the total phenolic contents between 6899-15000GAE(g/L) using the folin-ciocalteau assay respectively. These repurposed medicinal tools offer new hope and potential in the fight against emerging health threats driven by hyper-inflammation and free radical-induced oxidative stress.

Keywords: drug repurposed plant polyphenolics, free radical damage, non-communicable diseases, post COVID 19 syndromes

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Authors: Elham Shakerian, Reza Afarin

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The hepatic disease causes approximately 2 million deaths/year worldwide. Liver fibrosis is the last stage of numerous chronic liver diseases, and until now there is no definite cure or drug for it. Activation of hepatic stellate cells (HSCs) is the main reason for fibrosis. Transforming growth factor (TGF-β), as a main profibrogenic cytokine, if increased in these cells, leads to liver fibrosis through smad3 signaling pathways and increasing the expressions of Collagen type I and III, and actin-alpha smooth muscle (αSMA) genes. Curcumin (CUR) is a polyphenolic compound and an active ingredient derived from the rhizome of the turmeric plant that exerts effective antioxidant, anti-inflammatory, and antimicrobial activity. It has been shown that daily consumption of curcumin may have a protective effect on the liver against oxidative stress associated with alcohol consumption. In this study, we investigate the role of Curcumin in decreasing HSC activation and treating liver fibrosis. First, the human HSCs were treated with 2 ng/ml of (TGF-β) for 24 hours to become activated, then with Silibinin for 24 hours. Total RNAs were extracted, reversely transcribed into cDNA, Quantitative Real-time PCR, and western blot were performed. The mRNA expression levels of Collagen type I and III, αSMA genes, and the level of smad3 phosphorylation in TGF-β activated human HSCs treated with Curcumin were significantly reduced compared to human HSCs untreated with Curcumin. Curcumin is effective in reducing the expression of fibrogenic genes in the activated human HSCs treated with TGFB through downregulation of the TGF-β/smad3 signaling pathway. Therefore, Curcumin possesses significant antifibrotic properties in hepatic fibrosis

Keywords: hepatic fibrosis, human HSCs, curcumin, fibrogenic genes

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Authors: Gopal Lamichhane, Biswash Sapkota, Grinsun Sharma, Mahendra Adhikari

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Urolithiasis is a condition in which insoluble or less soluble salts like oxalate, phosphate etc. precipitate in urinary tract and causes obstruction in ureter resulting renal colic or sometimes haematuria. It is the third most common disorder of urinary tract affecting nearly 2% of world’s population. Poor urinary drainage, microbial infection, oxalate and calcium containing diet, calciferol, hyperparathyroidism, cysteine in urine, gout, dysfunction of intestine, drought environment, lifestyle, exercise, stress etc. are risk factors for urolithiasis. Wide ranges of treatments are available in allopathic system of medicine but reoccurrence is unpreventable even with the surgical removal of stone or lithotripsy. So, people prefer alternative medicinal systems such as Unani, homeopathic, ayurvedic etc. systems of medicine due to their fewer side effects over allopathic counterpart. Different plants based ethnomedicines are being well established by their continuous effective use in human since long time in treatment of urinary problem. Many studies have scientifically proved those ethnomedicines for antiurolithiatic effect in animal and in vitro model. Plant-based remedies were found to be therapeutically effective for both prevention as well as cure of calcium oxalate urolithiasis. Plants were known to show these effects through a combination of many effects such as antioxidant, diuretic, hypocalciuric, urine alkalinizing effect in them. Berberine, triterpenoids, lupeol are the phytochemicals established for antiurolithiatic effect. Hence, plant-based medicine can be the effective herbal alternative as well as means of discovery of novel drug molecule for curing urolithiatic disorder and should be focused on further research to discover their value in coming future.

Keywords: urolithiasis, herbal medicine, ethnomedicine, kidney stone, calcium oxalate

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Authors: Zenaida M. De Guzman, Davison T. Baldos, Gilberto T. Diano, Jeff Darren G. Valdez, Levelyn Mitos Tolentino, Gina B. Abrera, Ma. Lucia Cobar, Cristina Gragasin

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One of the factors affecting the quality of brown rice is the free fatty acid produced from surface lipids. It is the purpose of the study to determine the effect of gamma radiation, packaging materials and age and variety of paddy on the surface free fatty acid content using two different brown rice variety, namely, RC-160 and SL-7, packed in two different packaging materials, namely, regular polyethylene bag and Super bag irradiated at 0.5 and 1.0 kGy. Brown rice was produced from 2-week old (Lot 1) and two months old paddy (Lot 2) and irradiated at the Co-60 Multipurpose Irradiation Facility, PNRI. The surface Free Fatty Acid (FFA) content was obtained following the AOCS Official Method (1982) with some modifications. The experiment was laid out using Split-Plot Randomized Control Block Design. Analysis of variance (ANOVA) showed that the effects of variety, age of paddy and interactions of both were both significant. The surface FFA of SL-7 variety was found to be significantly higher than the RC-160 variety for all radiation doses. Likewise, Lot 2 was observed to have higher surface FFA than Lot 1 regardless of packaging material and radiation dose. It was observed that the surface FFA of both varieties packed in both packaging materials increased significantly up to the 2nd or 3rd month of storage and remains the same until the 5th month. On the other hand, radiation dose did not significantly affect the surface free fatty acid content for all storage/sampling time while the packaging material significantly interacts with the type of variety and radiation dose. Gamma radiation was proven to have no significant effect on the surface free fatty acid at 0.5 and 1.0 kGy and further analyses are needed to determine the action of gamma radiation to the activity of enzyme (lipase-induced and microbial) responsible for the production of other lipolytic products and the effect of gamma radiation on the integrity of the packaging materials.

Keywords: brown rice, free fatty acid, gamma radiation, polyethylene bag

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Authors: Weaam Aldeeban, Majd Aljamali, Lama A. Youssef

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Background & Objective: Warfarin is considered a problematic drug due to its narrow therapeutic window and wide inter-individual response variations, which are attributed to demographic, environmental, and genetic factors, particularly single nucleotide polymorphism (SNPs) in the genes encoding VKORC1 and CYP2C9 involved in warfarin's mechanism of action and metabolism, respectively. CYP2C9*2rs1799853 and CYP2C9*3rs1057910 alleles are linked to reduced enzyme activity, as carriers of either or both alleles are classified as moderate or slow metabolizers, and therefore exhibit higher sensitivity of warfarin compared with wild type (CYP2C9*1*1). Our study aimed to assess the frequency of *1, *2, and *3 alleles in the CYP2C9 gene in a cohort of Syrian patients receiving a maintenance dose of warfarin for different indications, the impact of genotypes on warfarin dosing, and the frequency of adverse effects (i.e., bleedings). Subjects & Methods: This retrospective cohort study encompassed 94 patients treated with warfarin. Patients’ genotypes were identified by sequencing the polymerase chain reaction (PCR) specific products of the gene encoding CYP2C9, and the effects on warfarin therapeutic outcomes were investigated. Results: Sequencing revealed that 43.6% of the study population has the *2 and/or *3 SNPs. The mean weekly maintenance dose of warfarin was 37.42 ± 15.5 mg for patients with the wild-type allele (CYP2C9*1*1), whereas patients with one or both variants (*2 and/or *3) demanded a significantly lower dose (28.59 ±11.58 mg) of warfarin, (P= 0.015). A higher percentage (40.7%) of patients with allele *2 and/or *3 experienced hemorrhagic accidents compared with only 17.9% of patients with the wild type *1*1, (P = 0.04). Conclusions: Our study proves an association between *2 and *3 genotypes and higher sensitivity to warfarin and a tendency to bleed, which necessitates lowering the dose. These findings emphasize the significance of CYP2C9 genotyping prior to commencing warfarin therapy in order to achieve optimal and faster dose control and to ensure effectiveness and safety.

Keywords: warfarin, CYP2C9, polymorphisms, Syrian, hemorrhage

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Authors: Samy S. Eleawa, Mahmoud A. Alkhateeb, Fahaid H. Alhashem, Ismaeel bin-Jaliah, Hussein F. Sakr, Hesham M. Elrefaey, Abbas O. Elkarib, Mohammad A. Haidara, Abdullah S. Shatoor, Mohammad A. Khalil

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This effect of Resveratrol (RES) against CdCl2- induced toxicity in the rat testes was investigated. Seven experimental groups of adult male rats were formulated as follows: A) Controls + NS, B) Control+ vehicle (saline solution of hydroxypropyl cyclodextrin), C) RES treated, D) CdCl2 +NS, E) CdCl2+ vehicle, F) RES followed by CdCl2 and M) CdCl2 followed by RES. At the end of the protocol, serum levels of FSH, LH, and testosterone were measured in all groups. Testicular levels of TBARS and Super Oxide Dismutase (SOD) activity were also measured. Epidydidimal semen analysis was performed and testicular expression of Bcl-2, p53 and Bax were assessed by RT-PCR. Also, histopathological changes of testes were examined microscopically and described. Pre and Post administration of RES in cadmium chloride-intoxicated rats improved semen parameters including count, motility, daily sperm production and morphology, increased serum concentrations of gonadotropins and testosterone, decreased testicular lipid peroxidation and increased SOD activity. Not only RES attenuated cadmium chloride induced testicular histopathology but was also able to protect against the onset of cadmium chloride testicular toxicity. Cadmium chloride downregulated the anti-apoptotic gene Bcl2 and upregulated the expression of both pro-apoptotic genes p53 and Bax. Resveratrol protected from and partially reversed cadmium chloride testicular via upregulation of Bcl2 and down regulation of p53 and Bax gene expression. Antioxidant activity of RES protects against cadmium chloride testicular toxicity and partially reverses its effect via upregulation of BCl2 and downregulation of p53 and Bax expression. These findings have far reaching implications on subfertility and impotency frequently seen in hypertensive as well as metabolic syndrome patients.

Keywords: resveratrol, cadmium, infertility, sperm, testis, metabolic syndrome

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Authors: M. Zeitoun, K. Abdallah, M. Rashwan

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Introduction: A growing body of evidence indicates that apelin, a relatively recent member of the adipokines family, has a potential anti-atherogenic effect. An association between low serum apelin state and coronary artery disease (CAD) was previously reported; however, the relationship between apelin and the atherosclerotic burden was unclear. Objectives: Our aim was to explore the correlation of serum apelin level with coronary calcium score (CCS) as a quantitative marker of coronary atherosclerosis. Methods: This observational cross-sectional study enrolled 100 consecutive subjects referred for cardiac multi-detector computed tomography (MDCT) for assessment of CAD (mean age 54 ± 9.7 years, 51 male and 49 females). Clinical parameters, glycemic and lipid profile, high sensitivity CRP (hsCRP), homeostasis model assessment of insulin resistance (HOMA-IR), serum creatinine and complete blood count were assessed. Serum apelin levels were determined using a commercially available Enzyme Immunoassay (EIA) Kit. High-resolution non-contrast CT images were acquired by a 64-raw MDCT and CCS was calculated using the Agatston scoring method. Results: Forty-three percent of the studied subjects had positive coronary artery calcification (CAC). The mean CCS was 79 ± 196.5 Agatston units. Subjects with detectable CAC had significantly higher fasting plasma glucose, HbA1c, and WBCs count than subjects without detectable CAC (p < 0.05). Most importantly, subjects with detectable CAC had significantly lower serum apelin level than subjects without CAC (1.3 ± 0.4 ng/ml vs. 2.8 ± 0.6 ng/ml, p < 0.001). In addition, there was a statistically significant inverse correlation between serum apelin levels and CCS (r = 0.591, p < 0.001); on multivariate analysis this correlation was found to be independent of traditional cardiovascular risk factors and hs-CRP. Conclusion:To the best of our knowledge, this is the first report of an independent association between apelin and CCS in patients with suspected CAD. Apelin emerges as a possible novel biomarker for CAD, but this result remains to be proved prospectively.

Keywords: HbA1c, apelin, adipokines, coronary calcium score (CCS), coronary artery disease (CAD)

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Authors: Valiollah Babaeipour, Mahdi Rahaie

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food supplements are rich in specific nutrients and bioactive compounds that eliminate free radicals and improve cellular metabolism. The major bioactive compounds are found in bran and cereal sprouts. Secondary metabolites of these microorganisms have antioxidant properties that can be used alone or in combination with chemotherapy and radiation therapy to treat cancer. Biologically active compounds such as benzoquinone derivatives extracted from fermented wheat germ extract (FWGE) have several positive effects on the overall state of human health and strengthen the immune system. The present work describes the discontinuous fermentation of raw wheat germ for FWGE production through the simultaneous culture process using the probiotic strains of Saccharomyces cerevisiae, Lactobacillus plantarum, and the possibility of using solid waste. To increase production efficiency, first to select important factors in the optimization of each fermentation process, using a factorial statistical scheme of stirring fraction (120 to 200 rpm), dilution of solids to solvent (1 to 8-12), fermentation time (16 to 24 hours) and strain to wheat germ ratio (20% to 50%) were studied and then simultaneous culture was performed to increase the yields of 2 and 6 dimethoxybenzoquinone (2,6-DMBQ). Since 2 and 6 dimethoxy benzoquinone were fermented as the main biologically active compound in wheat germ extract, UV-Vis analysis was performed to confirm the presence of 2 and 6 dimethoxy benzoquinone in the final product. In addition, 2,6-DMBQ of some products was isolated in a non-polar C-18 column and quantified using high performance liquid chromatography (HPLC). Based on our findings, it can be concluded that the increase of 2 and 6 dimethoxybenzoquinone in the simultaneous culture of Saccharomyces cerevisiae - Lactobacillus plantarum compared to pure culture of Saccharomyces cerevisiae (from 1.89 mg / g) to 28.9% (2.66 mg / g) Increased.

Keywords: wheat germ, FWGE, saccharomyces cerevisiae, lactobacillus plantarum, co-culture, 2, 6-DMBQ

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Authors: Waleed Aly Sayed Ahmed, Eman Kishk, Tahani Shams

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Aim: To study the effects of co-administration of phosphodiesterase-5 inhibitor (PDE-5) and sodium selenite against the damage induced by ovarian ischemia-reperfusion in rats. Materials and Methods: A total of forty-two sexually mature, virgin, female rats were divided randomly into six groups of seven each: sham group (C), ischemia group (I), ischemia/reperfusion group (I/R), ischemia/reperfusion plus 1.4mg/kg sildenafil (I/R+S) group, ischemia/reperfusion plus 0.2mg/kg selenium (I/R+Se) group and ischemia/reperfusion plus combination of sildenafil and selenium (I/R+S+Se) group. In ischemia group (I), rats were exposed to ischemia for 3 hours (h). In ischemia/reperfusion group (I/R), rats were exposed to ischemia for 3 h followed by 6 h of reperfusion. Treated groups received 1.4mg/kg sildenafil or 0.2 mg/kg selenium or both 30 min before reperfusion. Both ovaries were surgically removed carefully. One ovary was examined for histopathological changes and the other was subject to biochemical analysis including malondialdehyde (MDA), catalase (CAT) and glutathione peroxidase (GPx). Results: Assessment of ovarian tissue damage using a scoring system showed marked vascular congestion, interstitial edema, leukocyte infiltration, hemorrhage, and follicular degeneration in ischemia and ischemia/reperfusion groups. Tissue damage score for I, IR and all treated groups were significantly higher than those of the sham group (p<0.001), while tissue damage score decreased significantly in I/R+S and I/R+Se groups compared to I/R group (p<0.05), and notably, the difference was highly significant in I/R+S+Se group (p<0.001). There was significant increase in MDA levels and reduction in activities of CAT and GPx in I/R group compared to the sham group (p < 0.05). In I/R+S and I/R+Se groups, MDA was significantly decreased compared to the I/R group (p<0.05) and the difference was highly significant with co-administration of sildenafil and selenium (p<0.001). CAT and GPx were higher in all treated groups compared to I/R group (p<0.05). Conclusion: The co-administration of sildenafil citrate and selenium are highly protective against damage induced by ovarian ischemia/reperfusion in rats.

Keywords: phosphodiesterase-5 inhibitor, sildenafil, antioxidant, selenium, ovarian ischemia

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Authors: Quyen Vu Dang Ha, Oanh Truong Thi, Thuoc Tran Linh, Kent Carpenter, Thinh Doan Vu, Binh Dang Thuy

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Enzyme restriction site associated DNA (EzRAD) has recently emerged as a promising genomic approach for exploring fish genetic diversity on a genome-wide scale. This is a simplified method for genomic genotyping in non-model organisms and applied for SNPs discovery in the population genetics of freshwater and marine fish in the South of Vietnam. The observations of regional-scale differentiation of commercial freshwater fish (smallscale croakers Boesemania microlepis) and marine fish (emperor Lethrinus lentjan) are clarified. Samples were collected along Hau River and coastal area in the south and center Vietnam. 52 DNA samples from Tra Vinh, An Giang Province for Boesemania microlepis and 34 DNA samples of Lethrinus lentjan from Phu Quoc, Nha Trang, Da Nang Province were used to prepare EzRAD libraries from genomic DNA digested with MboI and Sau3AI. A pooled sample of regional EzRAD libraries was sequenced using the HiSeq 2500 Illumina platform. For Boesemania microlepis, the small scale population different from upstream to downstream of Hau river were detected, An Giang population exhibited less genetic diversity (SNPs per individual from 14 to 926), in comparison to Tra Vinh population (from 11 to 2172). For Lethrinus lentjan, the result showed the minor difference between populations in the Northern and the Southern Mekong River. The numbers of contigs and SNPs vary from 1315 to 2455 and from 7122 to 8594, respectively (P ≤ 0.01). The current preliminary study reveals regional scale population disconnection probably reflecting environmental changing. Additional sampling and EzRad libraries need to be implemented for resource management in the Mekong Delta.

Keywords: Boesemania microlepis, EzRAD, Lethrinus lentjan, SNPs

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Authors: Szu-Chieh Yu, Pei-Chin Chiand, Chih-Yi Lin, Yi-Wen Chien

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UV radiation from sunlight cause numbers of acute and chronic skin damage which can result in inflammation, immune changes, physical changes and DNA damage that facilitates skin aging and the development of skin carcinogenesis. Reactive oxygen species (ROS) are generated by excessive solar UV radiation, resulting in oxidative damage to cellar components, proteins, lipids, and nucleic acids. Thus, antioxidation plays an important role that protects skin against ROS-induced injury. Safflower (Carthamus tinctorius L.) is an important Chinese medicine contained abundance flavones and hydroxysafflor yellow A (HSYA) which is main active ingredient. HSYA is part of quinochalcone and has unique structures of hydroxy groups that provided the antioxidant effect. In this study, the aim was to investigate the protective role of HYSA in human keratinocytes (HaCaT) against UVA-induced oxidative damage and the possible mechanism. The HaCaT cells were UVA-irradiated and the effects of HYSA on cell viability, reactive oxygen species generation, DNA fragmentation and lipid peroxidation were measured. The mRNA expression of matrix metalloproteinase Ι (MMP Ι), cyclooxygenase-2 (COX-2) were determined by RT-PCR. In this study, UVA exposure lead to decrease in cell viability and increase in reactive oxygen species generation in HaCaT cells. HYSA could effectively increase the viability of HaCaT cells after UVA exposure and protect them from UVA-induced oxidative stress. Moreover, HYSA can reduce inflammation through inhibition the mRNA expression of MMP Ι and COX-2. Our results suggest that HSYA can act as a free radical scavenger while keratinocytes were photodamaged. HYSA could be a useful natural medicine for the protection of epidermal cells from UVA-induced damage and will be developed into products for skin care.

Keywords: HaCaT keratinocytes, hydroxysafflor yellow A (HSYA), MMP Ι, oxidative stress

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Authors: Ibtissem El Ouar, Cornelia Braicu, Dalila Naimi, Alexendru Irimie, Ioana Berindan-Neagoe

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Helix aspersa, 'the garden snail' is a big land snail widely found in the Mediterranean countries, it is one of the most consumed species in the west of Algeria. It is commonly used in zootherapy to purify blood and to treat cardiovascular diseases and liver problems. The aim of our study is to investigate, the antitumor activity of an aqueous extract from Helix aspersa prepared by the traditional method on Hs578T; a triple negative breast cancer cell line. Firstly, the free radical scavenging activity of H. aspersa extract was assessed by measuring its capability for scavenging the radical 2,2-diphenyl-1-picrylhydrazyl (DPPH), as well as its ability to reduce ferric ion by the FRAP assay (ferric reducing ability). The cytotoxic effect of H. aspersa extract against Hs578T cells was evaluated by the MTT test (3-(4,5- dimethylthiazl-2-yl)-2,5- diphenyltetrazolium bromide)) while the mode of cell death induced by the extract has been determined by fluorescence microscopy using acredine orange/ethidium bromide (AO/EB) probe. The level of TNFα has also measured in cell medium by ELISA method. The results suggest that H. aspersa extract has an antioxidant activity, especially at high concentrations, it can reduce DPPH radical and ferric ion. The MTT test shows that H. aspersa extract has a great cytotoxic effect against breast cancer cells, the IC50 value correspond of the dilution 1% of the crude extract. Moreover, the AO/EB staining shows that TNFα induced necrosis is the main form of cell death induced by the extract. In conclusion, the present study may open new perspectives in the search for new natural anticancer drugs.

Keywords: breast cancer, Helix aspersa, Hs578t cell line, necrosis

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Authors: Mohammad Karam, Abdulmalik Alsaif, Abdulrahman Al-Naseem, Amrit Hayre, Abdurrahman Al Jabbouri, Ahmad Aldubaikhi, Narvair Kahlar, Salem Al-Mutairi

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Purpose: To compare the outcomes of mycophenolate mofetil (MMF) versus methotrexate (MTX) in non-infectious ocular inflammatory disease (NIOID). Methods: A systematic review and meta-analysis were performed as per the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) Guidelines and an electronic search was conducted to identify all comparative studies of MMF versus MTX in NIOID. Treatment results and side effects were primary outcome measures. Secondary outcome measures included visual acuity and resolution of macular oedema. Fixed and random-effects models were used for the analysis. Results: Four studies enrolling 905 patients were identified. There was no significant difference between MMF and MTX groups in overall treatment success (Odds Ratio [OR] = 0.97, P = 0.96) and failure (OR = 0.86, P = 0.85) of NIOID. Although treatment success of uveitis showed no significant difference for anterior and intermediate uveitis cases (OR = 2.33, P = 0.14), MTX showed a significantly improved effect in cases involving posterior uveitis and panuveitis (OR = 0.41, P = 0.003). However, the median dose required for treatment success was lower for MTX whereas MMF was associated with a faster median time to treatment success. Further to this, MMF showed a reduced rate of side effects when compared to MTX, but MTX failed to reach statistical significance, most notably for liver enzyme elevation (OR = 0.65, P = 0.16), fatigue (OR = 0.84, P = 0.49) and headache (OR = 0.81, P = 0.37). For secondary outcomes, no significant difference was noted in visual acuity and resolution of macular edema. Conclusions: MMF is comparable to MTX in the treatment of NIOID as there was no significant difference in the outcome of treatment success and side effect profiles.

Keywords: Mycophenolate mofetil, methotrexate, non-infectious ocular inflammation, uveitis, scleritis

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Authors: Tahira Hyder, Saima Quraeshi, Zohaib Akram

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Salvadora Persica (SP) is known to have anti-inflammatory, antioxidant, anti-coagulant and anti-bacterial properties that may provide therapeutic benefits in the treatment of chronic periodontitis (CP). The current clinical trial was designed to investigate the clinical and anti-microbial effects of SP gel as an adjunct to scaling and root planning (SRP) in subjects with generalized CP. Sixty-six subjects with CP were randomized allocated into two groups: SRP + SP gel (test group) and SRP only (control group). Clinical parameters (periodontal pocket depth, gingival recession, clinical attachment level, bleeding score and plaque score) were recorded at baseline before SRP and at 6 weeks. At baseline and 6 weeks subgingival plaque samples were collected and periodontopathogen Porphyromonas Gingivalis (Pg) quantified using Real-time Polymerase Chain Reaction (RT-PCR). Both therapies reduced the mean periodontal pocket depth (PPD), plaque score (PS) and bleeding score (BOP) and improved the mean clinical attachment level (CAL) between baseline and 6 weeks. In subjects receiving adjunctive SP gel a statistically significant improvement was observed in BOP at follow-up compared to control group (15.01±3.47% and 22.81±6.81% respectively, p=0.001), while there was no statistically significant difference in periodontal pocket depth, gingival recession, clinical attachment level and plaque score between both groups. The test group displayed significantly greater Pg reduction compared to the control group after 6 weeks. The current study establishes that local delivery of SP gel into periodontal pocket in CP stimulated a significant reduction in bacteria Pg level and an improvement in gingival health, as evident from a reduced bleeding score, when used as an adjunct to SRP.

Keywords: miswak, scaling and root planing, porphyromonas gingivalis, chronic periodontitis

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Authors: Rajesh S. Mony, Vaidyaratnam Oushadhasala

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An attempt is made in this study to evolve a drug development modality based on classical Ayurvedic knowledge base as well as on modern scientific methodology. The present study involves (a) identification of a specific ailment condition, (b) the selection of a polyherbal formulation, (c) deciding suitable extraction procedure, (d) confirming the efficacy of the combination by in-vitro trials and (e) fixing up the recommended dose. The ailment segment selected is arthritic condition. The selected herbal combination is Kunturushka, Vibhitaki, Guggulu, Haridra, Maricha and Nirgundi. They were selected as per Classical Ayurvedic references, Authentified as per API (Ayurvedic Pharmacopeia of India), Extraction of each drug was done by different ratios of Hydroalcoholic menstrums, Invitro assessment of each extract after removing residual solvent for anti-Inflammatory, anti-arthritic activities (by UV-Vis. Spectrophotometer with positive control), Invitro assessment of each extract for COX enzyme inhibition (by UV-Vis. Spectrophotometer with positive control), Selection of the extracts was made having good in-vitro activity, Performed the QC testing of each selected extract including HPTLC, that is the in process QC specifications, h. Decision of the single dose with mixtures of selected extracts was made as per the level of in-vitro activity and available toxicology data, Quantification of major groups like Phenolics, Flavonoids, Alkaloids and Bitters was done with both standard Spectrophotometric and Gravimetric methods, Method for Marker assay was developed and validated by HPTLC and a good resolved HPTLC finger print was developed for the single dosage API (Active Pharmaceutical Ingredient mixture of extracts), Three batches was prepared to fix the in process and API (Active Pharmaceutical Ingredient) QC specifications.

Keywords: drug development, antiinflammatory, quality stardardisation, planar chromatography

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Authors: Dawid Sołoducha, Tomasz Borowski, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

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Many studies report varied effects of the magnetic field in medicine, but applications are still missing. Also, essential oils (EOs) were historically used in healing therapies, food preservation and the cosmetic industry due to their wound healing and antioxidant properties and antimicrobial activity. Unfortunately, the chemical characterization of EOs activates its antibacterial action only at a fairly high concentration. They can cause skin reactions, e.g., irritation (irritant contact dermatitis) or allergic contact dermatitis; therefore, they should always be used with caution. However, the administration of EOs to achieve the desired antimicrobial activity and stability with long-term medical usage in low concentration is challenging. The aim of this work was to investigate the antimicrobial activity of commercial Pinus sylvestris L. essential oil from Polish company Avicenna-Oil® under Rotating Magnetic Field (RMF) at f = 1 – 50 Hz. The novel construction of the magnetically assisted self-constructed reactor (MAP) was applied for this study. The chemical composition of essential pine oil was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Different concentrations of pine oil was prepared: 100% 50%, 25%, 12.5% and 6.25%. The disc diffusion and MIC test were done. To examine the effect of essential pine oil and rotating magnetic field RMF on antibacterial performance agar plate method was used. Pine oil consist of α-pinene (28.58%), β-pinene (17.79%), δ-3-carene (14.17%) and limonene (11.58%). The present study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of pine oil. We have shown that the rotating magnetic fields (RMF) at a frequency, f, between 25 Hz to 50 Hz, increase the antimicrobial efficiency of oil at lower than 50% concentration. The new method can be applied in many fields e.g. aromatherapy, medicine as a component of dressing, or as food preservatives.

Keywords: rotating magnetic field, pine oil, antimicrobial activity, Escherichia coli

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Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa

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Authors: Ahmad Ali Shahid, Mukhtar Ahmed

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The demand for long staple fiber having better strength and length is increasing with the introduction of modern spinning and weaving industry in Pakistan. Work on gene discovery from developing cotton fibers has helped to identify dozens of genes that take part in cotton fiber development and several genes have been characterized for their role in fiber development. Sucrose synthase (SuS) is a key enzyme in the metabolism of sucrose in a plant cell, in cotton fiber it catalyzes a reversible reaction, but preferentially converts sucrose and UDP into fructose and UDP-glucose. UDP-glucose (UDPG) is a nucleotide sugar act as a donor for glucose residue in many glycosylation reactions and is essential for the cytosolic formation of sucrose and involved in the synthesis of cell wall cellulose. The study was focused on successful Agrobacterium-mediated stable transformation of SuS gene in pCAMBIA 1301 into cotton under a CaMV35S promoter. Integration and expression of the gene were confirmed by PCR, GUS assay, and real-time PCR. Young leaves of SuS overexpressing lines showed increased total soluble sugars and plant biomass as compared to non-transgenic control plants. Cellulose contents from fiber were significantly increased. SEM analysis revealed that fibers from transgenic cotton were highly spiral and fiber twist number increased per unit length when compared with control. Morphological data from field plants showed that transgenic plants performed better in field conditions. Incorporation of genes related to cotton fiber length and quality can provide new avenues for fiber improvement. The utilization of this technology would provide an efficient import substitution and sustained production of long-staple fiber in Pakistan to fulfill the industrial requirements.

Keywords: agrobacterium-mediated transformation, cotton fiber, sucrose synthase gene, staple length

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Authors: Dawang D. N., Dasat G. S., Nden D.

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Endophyte means “in the plant” are referred to all microorganisms that live in the internal tissues of stems, petioles, roots and leaves of plants causing no apparent symptoms of disease. Secondary metabolites from fungal endophytes have an enormous potential applications as antioxidant, antimicrobial, anticancer and antidiabeties. Thus, this study aimed to determine the antimicrobial activity of these metabolites against some clinical isolates. The fungi were subjected to fermentation medium and the metabolites were extracted using ethyl acetate. The fungal extracts showed both antibacterial and antifungal activities with maximum zone of inhibition diameter of 10.5mm against Aspergillus fumigatus. Staphylococcus aureus was inhibited by all the five crude extracts with inhibition zone diameter of 4mm. Endophytic fungal crude extract2 (EDF2) exhibited antimicrobial effect against all the test organisms used, EDF4 was active against all test organisms except on Penicillium chrysogenum and Klebsiella pneumoniae. Antibacterial standard of ciprofloxacin which is 15mm is comparable to the effect of endophytic extract of EDF1 and EDF2. Klebsiella pneumoniae was resistant to EDF4 and EDF5. EDF3 showed a wide range of antimicrobial activity against all the test organisms used. The highest inhibition zone diameter of 10.50mm recorded against Aspergillus fumigatus is comparable to antifungal standard of fluconazole (15.5mm). The result of this study suggests that endophytic fungi associated with the roots of Irish potato could be a promising source of novel bioactive compounds of pharmaceutical and industrial importance.

Keywords: endophyte, fungal extract, antimicrobial, potato

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Authors: Mattea Carmen Castrovilli, Antonella Cartoni

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Electrospray ionisation (ESI), a well-established technique widely used to produce ion beams of biomolecules in mass spectrometry (ESI-MS), can be used for ambient soft landing of enzymes on a specific substrate. In this work, we show how the ambient electrospray deposition (ESD) technique can be successfully exploited for manufacturing a promising, green-friendly electrochemical amperometric laccase-based biosensor with unprecedented reuse and storage performance. These biosensors have been manufactured by spraying a laccase solution of 2μg/μL at 20% of methanol on a commercial carbon screen printed electrode (C-SPE) using a custom ESD set-up. The laccase-based ESD biosensor has been tested against catechol compounds in the linear range 2-100 μM, with a limit of detection of 1.7 μM, without interference from cadmium, chrome, arsenic, and zinc and without any memory effects, but showing a matrix effect in lake and well water. The ESD biosensor shows enhanced performances compared to the ones fabricated with other immobilization methods, like drop-casting. Indeed, it retains 100% activity up to two months of storage at ambient conditions without any special care and working stability up to 63 measurements on the same electrode just prepared and 20 on a one-year-old electrode subjected to redeposition together with a 100% resistance to use of the same electrode in subsequent days. The ESD method is a one-step, environmentally friendly method that allows the deposition of the bio-recognition layer without using any additional chemicals. The promising results in terms of storage and working stability also obtained with the more fragile lactate oxidase enzyme suggest these improvements should be attributed to the ESD technique rather than to the bioreceptor, highlighting how the ESD could be useful in reducing pollution from disposable devices. Acknowledgment: The understanding at the molecular level of this promising biosensor by using different spectroscopies, microscopies and analytical techniques is the subject of our PRIN 2022 project ESILARANTE.

Keywords: reuse, storage performance, immobilization, electrospray deposition, biosensor, laccase, catechol detection, green chemistry

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Authors: Alma Gómez-Loredo, José González-Valdez, Jorge Benavides, Marco Rito-Palomares

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Fucoxanthin is a carotenoid that exerts multiple beneficial effects on human health, including antioxidant, anti-cancer, antidiabetic and anti-obesity activity; making the development of a whole process for its production and recovery an important contribution. In this work, the lab-scale production and purification of fucoxanthin in Isocrhysis galbana have been studied. In batch cultures, low light intensities (13.5 μmol/m2s) and bubble agitation were the best conditions for production of the carotenoid with product yields of up to 0.143 mg/g. After fucoxanthin ethanolic extraction from biomass and hexane partition, further recovery and purification of the carotenoid has been accomplished by means of alcohol – salt Aqueous Two-Phase System (ATPS) extraction followed by an ultrafiltration (UF) step. An ATPS comprised of ethanol and potassium phosphate (Volume Ratio (VR) =3; Tie-line Length (TLL) 60% w/w) presented a fucoxanthin recovery yield of 76.24 ± 1.60% among the studied systems and was able to remove 64.89 ± 2.64% of the carotenoid and chlorophyll pollutants. For UF, the addition of ethanol to the original recovered ethanolic ATPS stream to a final relation of 74.15% (w/w) resulted in a reduction of approximately 16% of the protein contents, increasing product purity with a recovery yield of about 63% of the compound in the permeate stream. Considering the production, extraction and primary recovery (ATPS and UF) steps, around a 45% global fucoxanthin recovery should be expected. Although other purification technologies, such as Centrifugal Partition Chromatography are able to obtain fucoxanthin recoveries of up to 83%, the process developed in the present work does not require large volumes of solvents or expensive equipment. Moreover, it has a potential for scale up to commercial scale and represents a cost-effective strategy when compared to traditional separation techniques like chromatography.

Keywords: aqueous two-phase systems, fucoxanthin, Isochrysis galbana, microalgae, ultrafiltration

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Authors: Satender Saraswat, Dharmendra Prasad Singh, Rajesh Kumar Verma, Swati Sarswat

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Background and Purpose: The predominant cause of candidiasis was Candida albicans which has shifted towards non-Candida albicans Candida (NCAC) (Candida species other than the C. albicans). NCAC, earlier considered non-pathogenic or minimally virulent, are now considered a primary cause of morbidity and mortality in immunocompromised. With the NCAC spp. gaining weightage in the clinical cases, this study was conducted to determine the prevalence of NCAC spp. in different clinical specimens and to assess a few of their virulence factors. Material and Methods: Routine samples for bacterial culture and sensitivity, showing colony characteristics like Candida on Blood Agar and microscopic features resembling Candida spp. were processed further. Candida isolates were tested for chlamydospore formation, biochemical tests including sugar fermentation and sugar assimilation tests, and growth at 42oC, colony colour on HiCrome™ Candida Differential Agar, HiCandida Identification Kit and VITEK-2 Compact. Virulence factors like adherence to buccal epithelial cells (ABEC), biofilm formation, hemolytic activity, and production of coagulase enzyme were also tested. Results: Mean age of the patients was 38.46 with a male-female ratio of 1.36:1. 137 Candida isolates were recovered. 45.3% isolates were isolated from urine, 19.7% from vaginal swabs and 13.9% from oropharyngeal swabs. 55 (40.1%) isolates of C. albicans and 82 (59.9%) of NCAC spp. were identified, with C. tropicalis (23.4%) in NCAC. C. albicans (3; 50%) was the commonest species in cases of candidemia. Haemolysin production (85.5%) and ABEC (78.2%) were the major virulence factors in C. albicans. C. tropicalis (59.4%) and C. dubliniensis (50%) showed maximum ABEC. Biofilm forming capacity was higher in C. tropicalis (78.1%) than C. albicans (67%). Conclusion: This study suggests varied prevalence and virulence based on geographical locations, even within a subcontinent. It clearly demarcates the emergence of NCAC and their predominance in different body fluids. Identification of Candida to species level should become a routine in all the laboratories.

Keywords: ABEC, NCAC, non-Candida albicans Candida, Vitek-2TM compact

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Authors: Samira Bensoltane, Smina Ait Hamlet, Samti Meriem, Semmasel Asma

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Living organisms in the soil are subject to regular fluctuations of abiotic parameters, as well as a chemical contamination of the environment due to human activities. They are subject to multiple stressors they face. The aim of our work was to study the effects of insecticide: thiamethoxam (neonicotinoid), and the potential reversibility of the effects by an antioxidant: ginger on a bioindicator species in ecotoxicology, the land snail Helix aspersa. The effects were studied by a targeted cell approach of evaluating the effect of these molecules on tissue and cellular aspect of hepatopancreas through histological study. Treatment with thiamethoxam concentrations 10, 20, and 40 mg/l shows signs of inflammation even at low concentrations and from the 5th day of treatment. Histological examination of the hepatopancreas of snails treated with thiamethoxam showed significant changes from the lowest concentrations tested , note intertubular connective tissue enlargement, necrosis deferent types of cells (cells with calcium , digestive, excretory) , also damage acini, alteration of the apical membrane and lysis of the basement membrane in a dose- dependent manner. After 10 days of treatment and with 40 mg/l, the same changes were observed with a very advanced degeneration of the wall of the member that could be confused with the cell debris. For cons, the histological study of the hepatopancreas in Helix aspersa treated with ginger for a period of 15 days after stopping treatment with thiamethoxam has shown a partial regeneration of hepatopancreatic tissue snails treated with all concentrations of thiamethoxam and especially in the intertubular connective tissue of the wall and hepatopancreatic digestive tubules. Finally, we can conclude that monitoring the effect of the insecticide thiamethoxam showed significant alterations, however, treatment with ginger shows regeneration of damaged cells themselves much sharper at low concentration (10 mg/L).

Keywords: Helix aspersa, insecticides, thiamethoxam, ginger, hepatopancreas

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Authors: Aastha Agarwal, Veena Sharma

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N-nitrosopyrollidine or NPYR is a tobacco-specific nitrosamine which upon intoxicated causes abnormal production of Reactive Oxygen Species disrupt the endogenous antioxidant system. The study was designed to evaluate the histological changes in lung tissue of Mus musculus in NPYR administered lungs and effect of isolated flavonoid 3,6-dihydroxy-(3’,4’,7’-trimethoxyphenyl)-chromen-4-one-7-glucoside (ITC) from experimental plant Indigofera tinctorial. Post treatment with isolated compound significantly restored the abnormal symptoms and changes in pulmonary tissue. Transverse section of mouse lung in control animals appeared as a thin lace. Histologically, most of the lung was arranged as alveoli which were thin walled structures made up of single layered squamous epithelial cells. In the transverse section of lung at 100 X will clearly show the component of alveoli, surround by a thin layer of connective tissue and blood vessels. Smaller bronchioles were lined by cuboidal epithelial cells while larger bronchioles were lined by ciliated columnar epithelium layer while in NPYR intoxicated lungs signs of vast pulmonary damages and carcinogenesis as alveolar damage, necrosis, DADs or defused alveolar damages hyperplasia, metaplasia, dysplasia and next stage of carcinogenesis were revealed. Treatment with ITC showed the significant positive changes in the lung tissue due to the side hydroxyl and methoxy groups in its structure which help in combating oxidative injuries and give protection from the free radicals generated during the metabolism of NPYR in body. Thus, histopathological analysis confirms the development of the cancerous conditions in the lung tissue in mice model and the protective effects of ITC.

Keywords: flavonoid, histopathology, Indigofera tinctoria, lung

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