Search results for: biochemical leak

Authors: M. Gerasimou, S. Mantzoukis, P. Christodoulou, N. Varsamis, G. Kolliopoulou, N. Zotos

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Purpose: Microbial infection of the blood is a serious condition where bacteria invade the bloodstream and cause systemic disease. In such cases, blood cultures are performed. Blood cultures are a key diagnostic test for intensive care unit (ICU) patients. Material and method: The BacT/Alert system, which measures the production of carbon dioxide with metabolic organisms, is used. The positive result in the BacT/Alert system is followed by culture in the following selective media: Blood, Mac Conkey No 2, Chocolate, Mueller Hinton, Chapman and Sabaureaud agar. Gram staining method was used to differentiate bacterial species. The microorganisms were identified by biochemical techniques in the automated Microscan (Siemens) system and followed by a sensitivity test on the same system using the minimum inhibitory concentration MIC technique. The sensitivity test is verified by a Kirby Bauer-based test. Results: In 2017 the Laboratory of Microbiology received 3347 blood cultures. Of these, 170 came from the ICU. 116 found positive. Of these S. epidermidis was identified in 42, A. baumannii in 27, K. pneumoniae in 12 (4 of these KPC ‘Klebsiella pneumoniae carbapenemase’), S. hominis in 8, E. faecium in 7, E. faecalis in 5, P. aeruginosa in 3, C. albicans in 3, S. capitis in 2, K. oxytoca in 2, P. mirabilis in 2, E. coli in 1, S. intermidius in 1 and S. lugdunensis in 1. Conclusions: The study of epidemiological data and microbial resistance phenotypes is essential for the choice of therapeutic regimen for the early treatment and limitation of multivalent strains, while it is a crucial factor to solve diagnostic problems.

Keywords: blood culture, bloodstream, infection, intensive care unit

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Authors: D. Zahran, M. AlKhazindar, M. Khalil, E. T. A. Sayed

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Streptomyces scabies is a pathogenic actinomycete that infects potato crop causing severe production losses. Actinophages affect the composition and diversity of the bacterial population, thereby, can be used as a biological control. Samples of actinomycetes and phages were collected from different cultivated soils including farms of Faculty of Science, Faculty of Agriculture and different locations in Giza, Egypt. Actinomycetes were identified by using biochemical, morphological tests and molecular studies using 16S rRNA sequencing. Two specific phages (E1 and E2) against Streptomyces scabies and other hosts were isolated. Phages were identified using dilution end point (DEP), longevity in vitro (LIV), thermal inactivation point (TIP), host range and electron microscopy. PhageE1 was characterized by 10-8 (DEP),180 days(LIV), 95°C(TIP), narrow host range and electron microscopy showed ahead (59.9 nm) and neck (10.4nm). On the other hand phageE2 had 10-20 (DEP),180 days(LIV), 90°C(TIP), and the size of head was (67.2 nm) and tail (114nm). Antiviral activity was also studied using different chemicals (NaCL, KCL, CaCL2, BaCL2, CoCL2, AgNO3, ALCL3and HgCL2) with different concentrations and different plant extracts with different concentrations (star anise, tea, tillia, peppermint, ginger, cumin, chamomile, turmeric cinnamon, marjoram and black cumin). Both Phage E1and phage E2 were vulnerable to (cumin, ginger, chamomile, guavas leaves and star anise) but resistant to (Tillie, marjoram, fennelflower seeds, peppermint, and cinnamon).

Keywords: potato scab, actinophages, biological control, electron microscopy, TIP, DEP, LIV, antiviral activity

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Authors: S. H. Atef

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Background: The most common chromosomal abnormalities identified at birth are aneuploidies of chromosome 21, 18, 13, X and Y. Prenatal diagnosis of fetal aneuploidies is routinely done by traditional cytogenetic culture, a major drawback of this technique is the long period of time required to reach a diagnosis. In this study, we evaluated the QF-PCR as a rapid technique for prenatal diagnosis of common aneuploidies. Method:This work was carried out on Sixty amniotic fluid samples taken from patients with one or more of the following indications: Advanced maternal age (3 case), abnormal biochemical markers (6 cases), abnormal ultrasound (12 cases) or previous history of abnormal child (39 cases).Each sample was tested by QF-PCR and traditional cytogenetic. Aneuploidy screenings were performed amplifying four STRs on chromosomes 21, 18, 13, two pseudoautosomal,one X linked, as well as the AMXY and SRY; markers were distributed in two multiplex QFPCR assays (S1 and S2) in order to reduce the risk of sample mishandling. Results: All the QF-PCR results were successful, while there was two culture failures, only one of them was repeated. No discrepancy was seen between the results of both techniques. Fifty six samples showed normal patterns, three sample showed trisomy 21, successfully detected by both techniques and one sample showed normal pattern by QF-PCR but could not be compared to the cytogenetics due to culture failure, the pregnancy outcome of this case was a normal baby. Conclusion: Our study concluded that QF-PCR is a reliable technique for prenatal diagnosis of the common chromosomal aneuploidies. It has the advantages over the cytogenetic culture of being faster with the results appearing within 24-48 hours, simpler, doesn't need a highly qualified staff, less prone to failure and more cost effective.

Keywords: QF-PCR, traditional cytogenetic fetal aneuploidies, trisomy 21, prenatal diagnosis

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Authors: C. T. Eze, I. N. E. Onwurah

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An important hydrogeologic problem in areas of high faults formations is high environmental health hazard occasioned by microbial and heavy metals contamination of ground waters. Consequently, we examined the microbial load and heavy metals concentration of hospital wastewater discharged into the environment at Park Lane General Hospital Enugu Urban, Nigeria. The microbial counts, characteristics and frequency of occurrences of the isolated microorganisms were determined by cultural, morphological and biochemical characteristics using established procedure while the varying concentrations of the identified heavy metals were determined using the spectrophotometric method. The microbiological analyses showed a mean total aerobic bacteria counts from 13.7 ± 0.65 × 107 to 22.8 ± 1.14 ×1010 CFU/ml, mean total anaerobic bacteria counts from 6.0 ± 1.6 × 103 to 1.7 ± 0.41 ×104 CFU/ml and mean total fungal counts from 0 ± 0 to 2.3 ± 0.16 × 105 CFU/ml. The isolated micro-organisms which included both pathogenic and non-pathogenic organisms were Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Bacillus subtilis, Proteus vulgaris, Klesbsiella pneumonia and bacteriodes sp. The only fungal isolate was Candida albican. The heavy metals identified in the leachate were Arsenic, Cadmium, Lead, Mercury and Chromium and their concentrations ranged from 0.003 ± 0.00082 to 0.14 ± 0.0082 mg/l. These values were above WHO permissible limits while others fall within the limits. Therefore, hospital waste water can pose the environmental health risk when not properly treated before discharge, especially in geologic formations with high fault formations.

Keywords: bacterial isolates, fungal isolates, heavy metals, hospital wastewater, microbial counts

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Authors: Lata Kumari Dhanesh Tiwary, Pradeep Kumar Mishra

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The effluent coming from various industries such as textile, carpet, food, pharmaceutical and many other industries is big challenge due to its recalcitrant and xenobiotiocs in nature. Recently, biodegradation of dye wastewater through biological means was widely used due to eco-friendly and cost effective with the higher percentage of removal of dye from wastewater. The present study deals with the biodegradation and decolourization of Direct Red 23 dye using indigenously isolated bacterial consortium. The bacterial consortium was isolated from soil sample from dye contaminated site near a cluster of Carpet industries of Bhadohi, Uttar Pradesh, India. The bacterial strain formed consortia were identified and characterized by morphological, biochemical and 16S rRNA gene sequence analysis. The bacterial strain mainly Staphylococcus saprophyticus strain BHUSS X3 (KJ439576), Microbacterium sp. BHUMSp X4 (KJ740222) and Staphylococcus saprophyticus strain BHUSS X5 (KJ439576) were used as consortia for further studies of dye decolorization. Experimental investigations were made in a Sequencing Air- lift bioreactor using the synthetic solution of Direct Red 23 dye by optimizing various parameters for efficient degradation of dye. The effect of several operating parameters such as flow rate, pH, temperature, initial dye concentration and inoculums size on removal of dye was investigated. The efficiency of isolated bacterial consortia from dye contaminated area in Sequencing Air- lift Bioreactor with different concentration of dye between 100-1200 mg/l at different hydraulic rate (HRTs) 26h and 10h. The maximum percentage of dye decolourization 98% was achieved when operated at HRT of 26h. The percentage of decolourization of dye was confirmed by using UV-Vis spectrophotometer and HPLC.

Keywords: carpet industry, bacterial consortia, sequencing air-lift bioreactor

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Authors: Lakshmidevi Rajendran, Bharathidasan Kanniappan, Gopi Raja, Muthukumar Karuppan

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This study investigates the feasibility of fed-batch mixotrophic cultivation of microalgae Scenedesmus sp. in a 3-litre airlift photobioreactor under standard operating conditions. The results of this study suggest the algae species may serve as an excellent feed for aquatic species using organic byproducts. Microalgae Scenedesmus sp., was cultured using a synthetic wastewater by stepwise addition of crude glycerol concentration ranging from 2-10g/l under fed-batch mixotrophic mode for a period of 15 days. The attempts were made with the stepwise addition of crude glycerol as a carbon source in the initial growth phase to evade the inhibitory nature of high glycerol concentration on the growth of Scenedesmus sp. Crude glycerol was chosen since it is readily accessible as byproduct from biodiesel production sectors. Highest biomass concentration was achieved to be 2.43 g/l at the crude glycerol concentration of 6g/l after 10 days which is 3 fold times the increase in the biomass concentration compared with the control medium without the addition of glycerol. Biomass growth data obtained for the microalgae Scenedesmus sp. was fitted well with the modified Logistic equation. Substrate utilization kinetics was also employed to model the biomass productivity with respect to the various crude glycerol concentration. The results indicated that the supplement of crude glycerol to the mixotrophic culture of Scenedesmus sp., enhances the biomass concentration, chlorophyll and lutein productivity. Thus the application of fed-batch mixotrophic cultivation with stepwise addition of crude glycerol to Scenedesmus sp., provides a subtle way to reduce the production cost and improvisation in the large-scale cultivation along with biochemical compound synthesis.

Keywords: airlift photobioreactor, crude glycerol, microalgae Scenedesmus sp., mixotrophic cultivation, lutein production

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Authors: R. Sharma, S. Kumar, C. Sharma

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A number of toxic chlorophenolic compounds are formed during pulp bleaching. The nature and concentration of these chlorophenolic compounds largely depends upon the amount and nature of bleaching chemicals used. These compounds are highly recalcitrant and difficult to remove but are partially removed by the biochemical treatment processes adopted by the paper industry. Identification and estimation of these chlorophenolic compounds has been carried out in the primary and secondary clarified effluents from the paper mill by GCMS. Twenty-six chorophenolic compounds have been identified and estimated in paper mill waste waters. Electrochemical treatment is an efficient method for oxidation of pollutants and has successfully been used to treat textile and oil waste water. Electrochemical treatment using less expensive anode material, stainless steel electrodes has been tried to study their removal. The electrochemical assembly comprised a DC power supply, a magnetic stirrer and stainless steel (316 L) electrode. The optimization of operating conditions has been carried out and treatment has been performed under optimized treatment conditions. Results indicate that 68.7% and 83.8% of cholorphenolic compounds are removed during 2 h of electrochemical treatment from primary and secondary clarified effluent respectively. Further, there is a reduction of 65.1, 60 and 92.6% of COD, AOX and color, respectively for primary clarified and 83.8%, 75.9% and 96.8% of COD, AOX and color, respectively for secondary clarified effluent. EC treatment has also been found to increase significantly the biodegradability index of wastewater because of conversion of non- biodegradable fraction into biodegradable fraction. Thus, electrochemical treatment is an efficient method for the degradation of cholorophenolic compounds, removal of color, AOX and other recalcitrant organic matter present in paper mill waste water.

Keywords: chlorophenolics, effluent, electrochemical treatment, wastewater

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Authors: Hasan Afarnegan, Ali Shahraki, Jafar Shahraki

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Paraquat is widely used as a strong nitrogen-based herbicide for controlling of weeds in agriculture. This poison is extremely toxic for humans which induces several – organ failure by accumulation in cells and many instances of death occurred due to its poisoning. Paraquat metabolized primarily in the liver. The purpose of this study was to assess the effects of aquatic extract of levisticum officinale on oxidative status and biochemical factors in hepatocytes exposed to paraquat. Our results determined that hepatocytes destruction induced by paraquat is mediated by reactive oxygen species (ROS) production, lipid peroxidation and decrease of mitochondrial membrane potential were significantly (P<0.05) prevented by aquatic extract of Levisicum officinale (100, 200 and 300 µg/ml). These effects of paraquat also prevented via antioxidants and ROS scavengers (α-tocopherol, DMSO, manitol), mitochondrial permeability transition (MPT) pore sealing compound (carnitine).MPT pore sealing compound inhibited the hepatotoxicity, indicating that paraquat induced cell death via mithochondrial pathway. Pretreatment of hepatocytes with aquatic extracts of Levisticum officinale, antioxidants and ROS scavengers also blocked hepatic cell death caused by paraquat, suggesting that oxidative stress may be directly induced decline of mithochondrial membrane potential. In conclusion, paraquat hepatotoxicity can be attributed to oxidative stress and continued by mithochondrial membrane potential disruption. Levisticum officinale aquatic extract, presumably due to its strong antoxidant properties, could protect the destructive effects of paraquat on rat hepatocytes.

Keywords: hepatocyte protection, levisticum officinale, oxidative stress, paraquat

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Authors: K. B. M. Islam, Syeeda Shiraj-Um-Mahmuda, Afroj Jahan, A. A. Bhuiyan

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In Bangladesh, the use of imported probiotics in poultry is gradually being increased. But surprisingly, no probiotic bacteria have been isolated yet in Bangladesh despite the existence of scavenging native poultry as potential source that is seemingly more resistant to GIT infection as well as other diseases. Therefore, the study was undertaken to isolate, identify and characterize the potential probiotic Lactobacillus and Bifidobacteria strains from Bangladeshi indigenous poultry, and to evaluate their suitability to use in poultry industry. Crop and cecal samples from 61 healthy indigenous birds were used to isolate potential probiotics strains following conventional cultural methods. A total of 216 isolates were identified following physical, biochemical and molecular methods that belonged to the genus Lactobacillus and Bifidobacteria. An auto-aggregation test was performed for 180 and 136 isolated lactobacilli and bifidobacteria strains, respectively. Twelve lactobacilli isolates and 7 bifidobacteria isolates were selected because of their convenient aggregation. In vitro tests including antibacterial activity, resistance to low pH, hemolytic activities etc. were performed for evaluation of probiotic potential of each strain. Under the in vitro conditions and with respects to the probiotic traits, three lactobacilli; LS16, LS45, LS133 and two bifidobacteria, BS21 and BS90 were found to be potential probiotic strains. Thus, they are proposed to be evaluated for their in vivo probiotic properties. If the proposed strains are found suitable as the probiotics to be used in commercial poultry industry, it is expected that the local probiotics would be more beneficial and would save the huge amount of money that Bangladesh spends every year for the importation of such materials from abroad.

Keywords: Bangladeshi poultry, gut microbiota, lactic acid bacteria, scavenging chicken, GIT health

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Authors: Kulvinder Bajwa, Narsi R. Bishnoi

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Finding a sustainable alternative for today’s petrochemical industry is a major challenge facing by researchers, scientists, chemical engineers, and society at the global level. Microorganisms are considered to be sustainable feedstock for 3rd generation biofuel production. In this study, we have investigated the potential of a native bacterial strain isolated from a petrol contaminated site for the production of biodiesel. The bacterium was identified to be Rhodococcus opacus by biochemical test and 16S rRNA. Compositional analysis of bacterial biomass has been carried out by Fourier transform infrared spectroscopy (FTIR) in order to confirm lipid profile. Lipid and biomass were optimized by combination with Box Behnken design (BBD) of response surface methodology. The factors selected for the optimization of growth condition were glucose, yeast extract, and ammonium nitrate concentration. The experimental model developed through RSM in terms of effective operational factors (BBD) was found to be suitable to describe the lipid and biomass production, which indicated higher lipid and biomass with a minimum concentration of ammonium nitrate, yeast extract, and quite higher dose of glucose supplementation. Optimum results of the experiments were found to be 2.88 gL⁻¹ biomass and lipid content 38.75% at glucose 20 gL⁻¹, ammonium nitrate 0.5 gL⁻¹ and yeast extract 1.25 gL⁻¹. Furthermore, GCMS study revealed that Rhodococcus opacus has favorable fatty acid profile for biodiesel production.

Keywords: biofuel, Oleaginious bacteria, Rhodococcus opacus, FTIR, BBD, free fatty acids

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Authors: Larissa M. Vargas, Julia M. Sacchi, Renata O. Pereira, Lucas S. Asano, Iara C. Araújo, Patricia Fiorino, Vera Farah

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The aim of this study was to evaluate the occurrence of subclinical organ injuries induced by high-fat diet. Male wistar rats (n=5/group) were divided in control diet group (CD), commercial rat chow, and hyperlipidic diet (30% lipids) group (HD) administrated during 8 weeks, starting after weaning. All the procedures followed the rules of the Committee of Research and Ethics of the Mackenzie University (CEUA Nº 077/03/2011). At the end of protocol the animals were euthanized by anesthesia overload and the left kidney was removed. Intrarenal lipid deposition was evaluated by histological analyses with oilred. Kidney slices were stained with picrosirius red to evaluate the area of the Bowman's capsule (AB) and space (SB), and glomerular tuft area (GT). The renal expression of sterol regulatory element–binding protein (SREBP-2) was performed by Western Blotting. Creatinine concentration (serum and urine) and lipid profile were determined by colorimetric kit (Labtest). At the end of the protocol there was no differences in body weight between the groups, however the HD showed a marked increase in lipid deposits, glomeruli and tubules, and biochemical analysis for cholesterol and triglycerides. Moreover, in the kidney, the high-fat diet induced a reduction in the AB (13%), GT (18%) and SB (17%) associated with a reduction in glomerular filtration rate (creatinine clearance). The renal SRBP2 expression was increased in HD group. These data suggests that consumption of high-fat diet starting in childhood is associated with subclinical renal damage and function.

Keywords: high-fat diet, kidney, intrarenal lipid deposition, SRBP2

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Authors: Murat Tosunoglu, Cigdem Gul, Nurcihan Hacioglu, Nurdan Tepeova

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Amphibian and Reptilian species are influenced by pollution and habitat destruction. Blood parameters of Amphibia species were particularly affected by the negative environmental conditions. Studied frog samples 36 clinically normal Pelophylax ridibundus individuals were captured along the Biga Stream between April–June 2014. When comparing our findings with the Turkish legislation (Water pollution control regulation), the 1. Locality of the Biga stream in terms of total coliform classified as "high quality water" (Coliform: 866.66 MPN/100 mL), while the 2. locality was a "contaminated water" (Coliform: 53266.66 MPN/100 mL). Blood samples of the live specimens were obtained in the laboratory within one day of their capture. The blood samples were taken from the etherized frogs by means of ventriculus punctures, via heparinized hematocrit capillaries. Hematological and biochemical analyses based on high quality water and contaminated water, respectively, are as follows: Red blood cell count (444210.52-426846.15 per cubic millimeter of blood), white blood cell count (4215.78-4684.61 per cubic millimeter of blood), hematocrit value (29.25-29.43 %), hemoglobin concentration (7.76-7.22 g/dl), mean corpuscular volume (637.64-719.99 fl), mean corpuscular hemoglobin (184.78-174.75 pg), mean corpuscular hemoglobin concentration (29.44-24.82 %), glucose (103.74-124.13 mg/dl), urea (87.68-81.72 mg/L), cholesterol (148.20-197.39 mg/dl), creatinine (0.29-0.28 mg/dl), uric acid (10.26-7.55 mg/L), albumin (1.13-1.39 g/dl), calcium (11.45-9.70 mg/dl), triglyceride (135.23-155.85 mg/dl), total protein (4.26-3.73 g/dl), phosphorus (6.83-17.86 mg/dl), and magnesium (0.95-1.06 mg/dl). The some hematological parameters in P. ridibundus specimens are given for the first time in this study. No water quality dependent variation was observed in clinic hematology parameters measured.

Keywords: Pelophylax ridibundus, hematological parameters, biochemistry, freshwater quality

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Authors: Mojtaba Mirmontazemi, Habibollah Dadgar

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Bone is the most common metastasis site in some advanced malignancies, such as prostate and breast cancer. Bone metastasis generally indicates fewer prognostic factors in these patients. Different radiological and molecular imaging modalities are used for detecting bone lesions. Molecular imaging including computed tomography, magnetic resonance imaging, planar bone scintigraphy, single-photon emission tomography, and positron emission tomography as noninvasive visualization of the biological occurrences has the potential to exact examination, characterization, risk stratification and comprehension of human being diseases. Also, it is potent to straightly visualize targets, specify clearly cellular pathways and provide precision medicine for molecular targeted therapies. These advantages contribute implement personalized treatment for each patient. Currently, NaF PET/CT has significantly replaced standard bone scintigraphy for the detection of bone metastases. On one hand, 68Ga-PSMA PET/CT has gained high attention for accurate staging of primary prostate cancer and restaging after biochemical recurrence. On the other hand, FDG PET/CT is not commonly used in osseous metastases of prostate and breast cancer as well as its usage is limited to staging patients with aggressive primary tumors or localizing the site of disease. In this article, we examine current studies about FDG, NaF, and PSMA PET/CT images in bone metastases diagnostic utility and assess response to treatment in patients with breast and prostate cancer.

Keywords: skeletal metastases, fluorodeoxyglucose, sodium fluoride, molecular imaging, precision medicine, prostate cancer (68Ga-PSMA-11)

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Authors: Farideh K. Khosroushahi, Serkan Erdal, Mucip Geni̇şel

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Soil salinity is one of the major abiotic stress factors that restricts arable land and reduces crop productivity worldwide. High salt concentration adversely affects plant growth and development inducing water deficit, ionic toxicity, nutrient imbalance, and lead to oxidative stress. Although the stimulating role of mammalian sex hormones on various biological and biochemical processes under normal and stress condition have been proven, there is no study regarding with these hormone's effect on modulation of the alternative respiration pathway and AOX gene expression. In this study, changes in alternative respiration pathway in leaves of maize seedlings under salinity and the possible modulating effect of estrogen on these changes were investigated. Maize seedlings were grown in a hydroponic media for 11 days and then were exposed to salt stress for 3 days after being sprayed estrogen. The data obtained from oxygen consumption revealed that salt stress elevated cellular respiration value in the leaves. In addition, a marked increase was observed at alternative respiration level in salt-stressed seedlings. Compared to salt application alone, supplementation with estrogen resulted in a significant rise in alternative oxidase (AOX) activities. Similarly, while salt stress caused to rise in expressions of AOX gene compared to control seedlings, estrogen application resulted in further activation of these genes’ expression compared to stressed-seedlings alone. These data revealed that mitigating role of estrogen against the detrimental effects of salt stress is linked to modulation of alternative respiration pathway.

Keywords: alternative oxidase, estrogen, Ssalt stress, AOX, maize

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Authors: Preeyarach Whisetkhan, Malai Taweechotipatr, Ulisa Pachekrepapol

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Elevated level of blood cholesterol or hypercholesterolemia may lead to atherosclerosis and poses a major risk for cardiovascular diseases. Probiotics play a crucial role in human health, and probiotic bacteria that possesses bile salt hydrolase (BSH) activity can be used to lower cholesterol level of the host. The aim of this study was to investigate whether lactic acid bacteria (LAB) isolated from traditional Thai fermented foods were able to exhibit bile salt hydrolase activity and their use in fermented milk. A total of 28 isolates were tested for BSH activity by plate method on MRS agar supplemented with 0.5% sodium salt of taurodeoxycholic acid and incubated at 37°C for 48 h under anaerobic condition. The results showed that FN1-1 and FN23-3 isolates possessed strong BSH activity. FN1-1 and FN23-3 isolates were then identified for phenotype, biochemical characteristics, and genotype (16S rRNA sequencing). FN1-1 isolate showed 99.92% similarity to Lactobacillus pentosus DSM 20314(T), while FN23-3 isolate showed 99.94% similarity to Enterococcus faecium CGMCC1.2136 (T). Lactobacillus pentosus FN1-1 and Enterococcus faecium FN23-3 were tolerant of pH 3-4 and 0.3 and 0.8% bile. Bacterial count and pH of milk fermented with Lactobacillus pentosus FN1-1 at 37°C and 43°C were investigated. The results revealed that Lactobacillus pentosus FN1-1 was able to grow in milk, which led to decrease in pH level. Fermentation at 37°C resulted in faster growth rate than at 43 °C. Lactobacillus pentosus FN1-1 was a candidate probiotic to be used in fermented milk products to reduce the risk of high-cholesterol diseases.

Keywords: probiotics, lactic acid bacteria, bile salt hydrolase, cholesterol

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Authors: Omar Alhamd, Peter A. Thomas, Trevor J. Greenhough, Annette K. Shrive

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The Pseudomonas syringae species complex includes many plant pathogenic strains with highly specific interactions with varied host species and cultivars. The rapid spread of these bacteria over the last ten years has become a cause for concern. Nanoparticles have previously shown promise in microbiological action. We have therefore investigated in vitro and in vivo the effects of different types and sizes of nanoparticles in order to provide quantitative information about their effect on the bacteria. The effects of several different nanoparticles against several bacteria strains were investigated. The effect of NP on bacterial growth was studied by measuring the optical density, biochemical and nutritional tests, and transmission electron microscopy (TEM) to determine the shape and size of NP. Our results indicate that their effects varied, with either a negative or a positive impact on both bacterial and plant growth. Additionally, the methods of exposure to nanoparticles have a crucial role in accumulation, translocation, growth response and bacterial growth. The results of our studies on the behaviour and effects of nanoparticles in model plants showed. Cerium oxide (CeO₂) and silver (Ag) NP showed significant antibacterial activity against several pathogenic bacteria. It was found that titanium nanoparticles (TiO₂) can have either a negative or a positive impact, according to concentration and size. It is also thought that environmental conditions can have a major influence on bacterial growth. Studies were therefore also carried out under some environmental stress conditions to test bacterial survival and to assess bacterial virulence. All results will be presented including information about the effects of different nanoparticles on Pseudomonas syringae bacteria.

Keywords: plant microbiome, nanoparticles, 16S rRNA gene sequencing, bacterial survival

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Authors: Mubashir Hussain, Mu Lv, Xiaohan Dong, Zhiyang Li, Bin Liu, Nongyue He

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Detection and classification of microbes have a vast range of applications in biomedical engineering especially in detection, characterization, and quantification of bacterial contaminants. For identification of pathogens, different techniques are emerging in the field of biomedical engineering. Latest technology uses light scattering, capable of identifying different pathogens without any need for biochemical processing. Bacterial Pathogens Identification System (BPIS) which uses a laser beam, passes through the sample and light scatters off. An assembly of photodetectors surrounded by the sample at different angles to detect the scattering of light. The algorithm of the system consists of two parts: (a) Library files, and (b) Comparator. Library files contain data of known species of bacterial microbes in the form of binned plots, while comparator compares data of unknown sample with library files. Using collected data of unknown bacterial species, highest voltage values stored in the form of peaks and arranged in 3D histograms to find the frequency of occurrence. Resulting data compared with library files of known bacterial species. If sample data matching with any library file of known bacterial species, sample identified as a matched microbe. An experiment performed to identify three different bacteria particles: Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli. By applying algorithm using library files of given samples, results were compromising. This system is potentially applicable to several biomedical areas, especially those related to cell morphology.

Keywords: microbial identification, laser scattering, peak identification, binned plots classification

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Authors: Marija Knez, James C.R. Stangoulis, Manja Zec, Zoran Pavlovic, Jasmina D. Martacic, Mirjana Gurinovic, Maria Glibetic

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Background: Zinc is an essential micronutrient for humans with important physiological functions. A sensitive and specific biomarker for assessing Zn status is still needed. Objective: The major aim of this study was to examine if the changes in the content of plasma phospholipid LA, DGLA and LA: DGLA ratio can be used to efficiently predict the dietary Zn intake and plasma Zn status of humans. Methods: The study was performed on apparently healthy human volunteers. The dietary Zn intake was assessed using 24h recall questionnaires. Plasma phospholipid fatty acid analysis was done by gas chromatography and plasma analysis of minerals by atomic absorption spectrometry. Biochemical, anthropometrical and hematological parameters were assessed. Results: No significant relationship was found between the dietary and plasma zinc status (r=0.07; p=0.6). There is a statistically significant correlation between DGLA and plasma Zn (r=0.39, p=0.00). No relationship was observed between the linoleic acid and plasma Zn, while there was a significant negative correlation between LA: DGLA ratio and plasma Zn status (r=-0.35, p=0.01). Similarly, there were statistically significant difference in DGLA status (p=0.004) and LA: DGLA ratio (p=0.042) between the Zn formed groups. Conclusions: This study is an initial step in evaluating LA: DGLA ratio as a biomarker of Zn status in humans. The results are encouraging as they show that concentration of DGLA is decreased and LA: DGLA ratio increased in people with lower dietary Zn intake. However, additional studies are needed to fully examine the sensitivity of this biomarker.

Keywords: dietary Zn intake Zinc, fatty acid composition, LA: DGLA, healthy population, plasma Zn status, Zn biomarker

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Authors: Devendra Sillu, Shekhar Agnihotri

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The 'nano-biocatalyst' utilizes an ordered assembling of enzyme on to nanomaterial carriers to catalyze desirable biochemical kinetics and substrate selectivity. The current study describes an inter-disciplinary approach for converting agriculture waste, sugarcane bagasse into D-glucose exploiting halloysite nanotubes (HNTs) decorated cellulase enzyme as nano-biocatalytic system. Cellulase was successfully immobilized on HNTs employing polydopamine as an eco-friendly crosslinker while iron oxide nanoparticles were attached to facilitate magnetic recovery of material. The characterization studies (UV-Vis, TEM, SEM, and XRD) displayed the characteristic features of both cellulase and magnetic HNTs in the resulting nanocomposite. Various factors (i.e., working pH, temp., crosslinker conc., enzyme conc.) which may influence the activity of biocatalytic system were investigated. The experimental design was performed using Response Surface Methodology (RSM) for process optimization. Analyses data demonstrated that the nanobiocatalysts retained 80.30% activity even at elevated temperature (55°C) and excellent storage stabilities after 10 days. The repeated usage of system revealed a remarkable consistent relative activity over several cycles. The immobilized cellulase was employed to decompose agro-waste and the maximum decomposition rate of 67.2 % was achieved. Conclusively, magnetic HNTs can serve as a potential support for enzyme immobilization with long term usage, good efficacy, reusability and easy recovery from solution.

Keywords: halloysite nanotubes, enzyme immobilization, cellulase, response surface methodology, magnetic recovery

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Authors: Gauri Shankar Shah

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Objectives: The Type I diabetes mellitus in children is frequently a missed diagnosis and children presents in emergency with diabetic ketoacidosis having significant morbidity and mortality. The present study was done to find out the clinical presentation and outcome at a tertiary-care centre. Methods: This was retrospective analysis of data of Type I diabetes mellitus reporting to our centre during last one year (2012-2013). Results: There were 12 patients (8 males) and the age group was 4-14 years (mean ± 3.7). The presenting symptoms were fever, vomiting, altered sensorium and fast breathing in 8 (66.6%), 6 (50%), 4 (33.3%), and 4 (33.3%) cases, respectively. The classical triad of polyuria, polydypsia, and polyphagia were present only in two patients (33.2%). Seizures and epigastric pain were found in two cases each (33.2%). The four cases (33.3%) presented with diabetic ketoacidosis due to discontinuation of insulin doses, while 2 had hyperglycemia alone. The hemogram revealed mean hemoglobin of 12.1± 1.6 g/dL and total leukocyte count was 22,883.3 ± 10,345.9 per mm3, with polymorphs percentage of 73.1 ± 9.0%. The mean blood sugar at presentation was 740 ± 277 mg/ dl (544–1240). HbA1c ranged between 7.1-8.8 with mean of 8.1±0.6 %. The mean sodium, potassium, blood ph, pCO2, pO2 and bicarbonate were 140.8 ± 6.9 mEq/L, 4.4 ± 1.8mEq/L, 7.0 ± 0.2, 20.2 ± 10.8 mmHg, 112.6 ± 46.5 mmHg and 9.2 ± 8.8 mEq/L, respectively. All the patients were managed in pediatric intensive care unit as per our protocol, recovered and discharged on intermediate insulin given twice daily. Conclusions: Thus, it shows that these patients have uncontrolled hyperglycemia and often presents in emergency with ketoacidosis and deranged biochemical profile. The regular administration of insulin, frequent monitoring of blood sugar and health education are required to have better metabolic control and good quality of life.

Keywords: type I diabetes mellitus, hyperglycemia, outcome, glycemic control

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Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provides better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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Authors: Abigail Tan, Heng Liang Tan, Vanessa Ding, James Hui, Eng Hin Lee, Andre Choo

Abstract:

Introduction: Comparative oncology investigates the similarities in spontaneous carcinogenesis between humans and animals, in order to identify treatments that can benefit these patients. Companion animals (CA), like canines and felines, are of special interest when it comes to studying human cancers due to their exposure to the same environmental factors and develop tumours with similar features. The purpose of this study is to explore the cross-reactivity of monoclonal antibodies (mAbs) across cancers in humans and CA. Material and Methods: A panel of CA mAbs generated in the lab was screened on multiple human cancer cell lines through flow cytometry to identify for positive binders. Shortlisted candidates were then characterised by biochemical and functional assays e.g., antibody-drug conjugate (ADC) and western blot assays, including glycan studies. Results: Candidate mAb KP52 was generated from whole-cell immunisation using feline mammary carcinoma. KP52 showed strong positive binding to human cancer cells, such as breast cancer and ovarian cancer. Furthermore, KP52 demonstrated strong killing ( > 50%) as an ADC with Saporin as the payload. Western blot results revealed the molecular weight of the antigen targets to be approximately 45kD and 50kD under reduced conditions. Glycan studies suggest that the epitope is glycan in nature, specifically an O-linked glycan. Conclusion: Candidate mAb KP52 has a therapeutic potential as an ADC against feline mammary cancer, human ovarian cancer, human mammary cancer, human pancreatic cancer, and human gastric cancer.

Keywords: ADC, comparative oncology, mAb, therapeutic

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Authors: D. Shehu, Z. Alias

Abstract:

Glutathione S-transferases (GSTs) are family of enzymes that function in the detoxification of variety of electrophilic substrates. In the present work, we report a novel zeta-like GST (designated as KKSG9) from the biphenyl/polychlorobiphenyl degrading organism Acidovorax sp. KKS102. KKSG9 possessed low sequence similarity but similar biochemical properties to zeta class GSTs. The gene for KKSG9 was cloned, purified and biochemically characterized. Functional analysis showed that the enzyme exhibits wider substrate specificity compared to most zeta class GSTs by reacting with 1-chloro-2,4-dinitrobenzene (CDNB), p-nitrobenzyl chloride (NBC), ethacrynic acid (EA), hydrogen peroxide, and cumene hydroperoxide (CuOOH). The enzyme also displayed dehalogenation function against dichloroacetate (a common substrate for zeta class GSTs) in addition to permethrin, and dieldrin. The functional role of Tyr12 was also investigated by site-directed mutagenesis. The mutant (Y12C) displayed low catalytic activity and dehalogenation function against all the substrates when compared with the wild type. Kinetic analysis using NBC and GSH as substrates showed that the mutant (Y12C) displayed a higher affinity for NBC when compared with the wild type, however, no significant change in GSH affinity was observed. These findings suggest that the presence of tyrosine residue in the motif might represent an evolutionary trend toward improving the catalytic activity of the enzyme. The enzyme as well could be useful in the bioremediation of various types of organochlorine pollutants.

Keywords: Acidovorax sp. KKS102, bioremediation, glutathione s-transferase, site-directed mutagenesis, zeta

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Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

Abstract:

PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provide better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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Authors: Rozeena Shaikh, Syed M Shahid, Jamil Ahmad, Qaisar Mansoor, Muhammad Ismail, Abid Azhar

Abstract:

Introduction: Diabetes mellitus (DM) is a prevalent non-communicable disease worldwide. In most high-income countries as well as middle-income and low- income countries. DM is among the top causes of deaths. DM may lead to many vascular complications like hypertension, nephropathy, retinopathy, neuropathy, and foot. Diabetic nephropathy (DN) characterized by persistent albuminuria is a leading cause of end stage renal failure (ESRF). Pathogenesis of diabetic nephropathy is implicated by the polymorphisms in genes encoding the components of reninangiotensin- aldosteron system (RAAS) which include angiotensinogen (AGT), angiotensin-II receptor and particularly angiotensin converting enzyme (ACE) gene. Method: Study subjects include 110 control, 110 patients with DM without hypertension, 110 patients with DM with hypertension and 110 patients with DN. Blood samples were collected for Biochemical analysis and PCR and sequencing for the specific region of both genes. Results: The frequency of DD genotype and D allele of ACE (I/D) was significantly (p<0.05) high in DM normotensive, DM hypertensive and DN patients when compared to control. The ACE G2350A genotypes and allele frequencies were significantly different (p<0.05) in DM hypertensive patients as compared to control and DN, while no difference was observed between DM normotensive and DN when compared to control. The genotypes and alleles of AGT (M268T) polymorphism were significantly different (p<0.05) in DM normotensive, DM hypertensive and DN when compared to control. Conclusion: The DD genotype and D allele of ACE (I/D), GG genotype and G allele of ACE (G2350A) and the TT genotype and T allele of AGT (M268T) polymorphism have shown a significant difference in genotype and allele frequencies between controls and patients.

Keywords: genetic variations, ACE, AGT, diabetes mellitus, diabetic nephropathy, Pakistan

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Authors: Rafael Zhindon Almeida

Abstract:

Surface water quality is an important concern for the evaluation and prediction of water quality conditions. The objective of this study is to develop a statistical model that can accurately predict the water quality of the El Macho estuary in the city of Machala, El Oro province. The methodology employed in this study is of a basic type that involves a thorough search for theoretical foundations to improve the understanding of statistical modeling for water quality analysis. The research design is correlational, using a multivariate statistical model involving multiple linear regression and principal component analysis. The results indicate that water quality parameters such as fecal coliforms, biochemical oxygen demand, chemical oxygen demand, iron and dissolved oxygen exceed the allowable limits. The water of the El Macho estuary is determined to be below the required water quality criteria. The multiple linear regression model, based on chemical oxygen demand and total dissolved solids, explains 99.9% of the variance of the dependent variable. In addition, principal component analysis shows that the model has an explanatory power of 86.242%. The study successfully developed a statistical model to evaluate the water quality of the El Macho estuary. The estuary did not meet the water quality criteria, with several parameters exceeding the allowable limits. The multiple linear regression model and principal component analysis provide valuable information on the relationship between the various water quality parameters. The findings of the study emphasize the need for immediate action to improve the water quality of the El Macho estuary to ensure the preservation and protection of this valuable natural resource.

Keywords: statistical modeling, water quality, multiple linear regression, principal components, statistical models

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Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

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Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

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Authors: Vineeta Kaushik, Manisha Goel

Abstract:

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

Abstract:

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

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Authors: Nalan Kaya, Elif Erdem, Mehmet Ali Kisacam, Gonca Ozan, Enver Ozan

Abstract:

Tobacco smokers continuously inhale thousands of carcinogens and free radicals. It is estimated that about 1017 oxidant molecules are present in each puff of tobacco smoke. It is known that smoking has adverse effects on the structure and functions of the liver. Ellagic acid (EA) has antioxidant, antiapoptotic, anticarcinogenic, antibacterial and antiinflammatory effects. The aim of our study was to investigate the possible protective effect of ellagic acid against tobacco smoke-mediated oxidative stress in the rat liver. Twenty-four male adult (8 weeks old) Spraque-Dawley rats were divided randomly into 4 equal groups: group I (control), group II (tobacco smoke), group III (tobacco smoke + corn oil) and group IV (tobacco smoke + ellagic acid). The rats in group II, III and IV, were exposed to tobacco smoke 1 hour twice a day for 12 weeks. In addition to tobacco smoke exposure, 12 mg/kg ellagic acid (dissolved in corn oil), was applied to the rats in group IV by oral gavage. An equal amount of corn oil used in solving ellagic acid was applied to the rats by oral gavage in group III. At the end of the experimental period, rats were decapitated, and liver tissues were removed. Histological and biochemical analyzes were performed. Sinusoidal dilatation, inflammatory cell infiltration in portal area, increased Kuppfer cells were examined in tobacco smoke group and tobacco smoke+ corn oil groups. The results, observed in tobacco smoke and tobacco smoke+corn oil groups, were found significantly decreased in tobacco smoke+EA group. Group-II and group-III MDA levels were significantly higher, and GSH activities were not different than group-I. Compared to group-II, group-IV MDA level was decreased, and GSH activities was increased significantly. The results indicate that ellagic acid could protect the liver tissue from the tobacco smoke harmful effects.

Keywords: ellagic acid, liver, rat, tobacco smoke

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