Search results for: serum protein electrophoresis

Authors: Eizadi Mojtaba

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This study aimed to evaluate the effect of 8 weeks aerobic training on serum osteocalsin as an osteoblasts hormone and cortisol in adult men with asthma. For this purpose, twenty four non-trained adult men with mild to moderate asthma were participated in study voluntarily and divided into exercise (aerobic training, 8 weeks/3 times per week) and control groups by randomly. Pre and post training of serum osteocalsin and cortisol were measured of two groups. Student’s paired ‘t’ test was applied to compare the pre and post training values. A p-value of less than 0.05 was considered to be statistically significant. There were no statistically significant differences with regard to all anthropometrical and biochemical markers between the exercise and control groups at baseline ( P > 0.05 ). Exercise training resulted in a significant increase in serum osteocalsin and decrease in cortisol ( P > 0.05 ), but not in control group. Based on these data, we concluded that aerobic training can be improved Processes of bone formation in asthma patients.

Keywords: osteoblasts, asthma, aerobic exercise, sedentary

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Authors: Babak Moradi, Hassan Zare-Maivan

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A greenhouse investigation has been conducted to study the effect of crude oil on morphology and protein content of Avicennia marina plant. Avicennia marina seeds were sown in different concentrations of the crude oil mixed soil (i.e., 2.5, 5, 7.5, and 10 w/w). Controls and replicates were also set up. Morphological traits were recorded 4 months after plantation. Avicennia marina seedlings could tolerate up to 10% (w/w). Results demonstrated that there was a reduction in plant shoot and root biomass with the increase of crude oil concentration. Plant height, total leaf number and length reduced significantly with increase of crude oil contamination. Investigation revealed that there is a great impact of crude oil contamination on protein content of the roots of the experimental plant. Protein content of roots grown in different concentrations of crude oil were more than those of the control plant. Further, results also showed that protein content was increased with increased concentration of crude oil.

Keywords: Avicennia marina, morphology, oil contamination, protein content

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Authors: Solange Adechian, Michèle Balage, Didier Remond, Carole Migné, Annie Quignard-Boulangé, Agnès Marset-Baglieri, Sylvie Rousset, Yves Boirie, Claire Gaudichon, Dominique Dardevet, Laurent Mosoni

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Studies have shown that timing of protein intake, leucine content, and speed of digestion significantly affect postprandial protein utilization. Our aim was to determine if one can spare lean body mass during energy restriction by varying the quality and the timing of protein intake. Obese volunteers followed a 6-wk restricted energy diet. Four groups were compared: casein pulse, casein spread, milk-soluble protein (MSP, = whey) pulse, and MSP spread (n = 10-11 per group). In casein groups, caseins were the only protein source; it was MSP in MSP groups. Proteins were distributed in four meals per day in the proportion 8:80:4:8% in the pulse groups; it was 25:25:25:25% in the spread groups. We measured weight, body composition, nitrogen balance, 3-methylhistidine excretion, perception of hunger, plasma parameters, adipose tissue metabolism, and whole body protein metabolism. Volunteers lost 7.5 ± 0.4 kg of weight, 5.1 ± 0.2 kg of fat, and 2.2 ± 0.2 kg of lean mass, with no difference between groups. In adipose tissue, cell size and mRNA expression of various genes were reduced with no difference between groups. Hunger perception was also never different between groups. In the last week, due to a higher inhibition of protein degradation and despite a lower stimulation of protein synthesis, postprandial balance between whole body protein synthesis and degradation was better with caseins than with MSP. It seems likely that the positive effect of caseins on protein balance occurred only at the end of the experiment.

Keywords: lean body mass, fat mass, casein, whey, protein metabolism

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Authors: Nilima Barman, M. Atiqul Haque, Debabrata Ghosh

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Background: Zinc, one of the vital micronutrients, has an incredible role in the immune system. Hypozincemia affects host defense by reducing the number of circulating T cells and phagocytosis activity of other cells which ultimately impair cell-mediated immunity 1, 2. The immune system is detrimentally suppressed in multidrug-resistant tuberculosis (MDR-TB) 3, 4, a major threat of TB control worldwide5. As zinc deficiency causes immune suppression, we assume that it might have a role in the development of MDR-TB. Objectives: To estimate the serum zinc level in newly diagnosed multidrug resistant tuberculosis (MDR-TB) in comparison with that of newly diagnosed pulmonary TB (NdPTB) and healthy individuals. Materials and Methods: This study was carried out in the department of Public Health and Informatics, Bangabandhu Sheikh Mujib Medical University, Dhaka in collaboration with National Institute of Diseases of the Chest Hospital (NIDCH), Bangladesh from March’ 2012 to February 2013. A total of 337 respondents, of them 107 were MDR TB patients enrolled from NIDCH, 69 were NdPTB and 161 were healthy adults. All NdPTB patients and healthy adults were randomly selected from Sirajdikhan subdistrict of Munshiganj District. It is a rural community 22 kilometer south from capital city Dhaka. Serum zinc level was estimated by atomic absorption spectrophotometry method from early morning fasting blood sample. The evaluation of serum zinc level was done according to normal range from 70 to120 µgm/dL6. Results: Males were predominant in study groups (p>0.05). Mean (sd) serum zinc levels in MDR-TB, NdPTB and healthy adult group were 65.14 (12.52), 75.22(15.89), and 87.98 (21.80) μgm/dL respectively and differences were statistically significant (F=52.08, P value<0.001). After multiple comparison test (Bonferroni test) significantly lower level of serum zinc was found in MDRTB group than NdPTB and healthy adults (p<.001). Point biserial correlation showed a negative association of having MDR TB and serum zinc level (r= -.578; p value <0.001). Conclusion: The significant low level of serum zinc in MDR-TB patients suggested impaired immune status. We recommended for further exploration of low level of serum zinc as risk factor of MDR TB.

Keywords: Bangladesh, immune status, multidrug-resistant tuberculosis, serum zinc

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Authors: Nnennaya Samuel Okoro

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An experiment was conducted to determine the growth performance, blood parameters and reproductive characteristics of 8-week old male weaner rabbits fed 20% boiled pigeon pea seed meal (PPSM). The study lasted for 16 weeks. Results showed that haematological parameters of the two groups of rabbit bucks were not significantly affected (P>0.05) by the treatment, meaning that the PPSM was adequate for maintaining the blood parameters at the normal levels. The 20% boiled PPSM significantly affected (P<0.05) serum Alanine Aminotransferase (ALT) (67.72±2.5 I.U/I) more than the ALT (57.50±2.02 I.U/I) of the control, which is an indication of liver problem. The globulin level (3.00 ± 0.23 g/d) of the 20% boiled PPSM group was significantly higher than that of the control (2.60±0.06 g/dl), indicating that the test diet did not alter protein metabolism in the rabbits. Boiled pigeon pea seed meal supported organ weight and testicular development in rabbit bucks, suggesting that boiling reduced the level of the anti-nutritional factors in pigeon pea seed meal. Thus, 20% boiled pigeon pea can be included in diets of rabbits without adverse effect on blood parameters and internal organs characteristics.

Keywords: haematology, internal organs, pigeon pea, rabbits, serum biochemistry

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Authors: Singh Baljinder, Sharma Navneet

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Foeniculum vulgare Miller is a biennial medicinal and aromatic plant belonging to the family Apiaceae (Umbelliferaceae). It is a hardy, perennial–umbelliferous herb with yellow flowers and feathery leaves. The aim is to study the control of blood glucose in alloxan induced diabetic mice.Method used for extraction was continuous hot percolation method in which Soxhlet apparatus was used.95%ethanol was used as solvent. Male albino mice weighing about 20-25 g obtained from Guru Angad Dev University of Veterinary Science, Ludhiana were used for the study. Diabetes was induced by a single i.p. injection of 125 mg/kg of alloxan monohydrate in sterile saline (11). After 48 h, animals with serum glucose level above 200 mg/dl (diabetic) were selected for the study. Blood samples from mice were collected by retro-orbital puncture (ROP) technique. Serum glucose levels were determined by glucose oxidase and peroxidase method. Single administration (single dose) of aqueous extract of fennel (25, 50, and 100 mg/kg, p.o.) in diabetic Swiss albino mice, showed reduction in serum glucose level after 45 min. Maximum reduction in serum glucose level was seen at doses of 100 mg/kg. Aqueous extract of fennel in all doses except 25 mg/kg did not cause any significant decrease in blood glucose. It may be said that the aqueous extract of fennel decreased the serum glucose level and improved glucose tolerance owing to the presence of aldehyde moiety. The aqueous extract of fennel has antihyperglycemic activity as it lowers serum glucose level in diabetic mice.

Keywords: Foeniculum vulgare Miller, antihyperglycemic, diabetic mice, Umbelliferaceae

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Authors: Dzatur Rizqi Fathienah Syarifuddin, Isharyah Sunarno, Eddy Hartono, Siti Maisuri T. Chalid

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Introduction: The potential impact of coronavirus disease 2019 (COVID-19) on the health of expectant mothers and fetuses has strained attention. Pregnant women are considered a vulnerable category to respiratory infections. Moreover, several inflammatory cytokines are 2-100 times more abundant in COVID-19 with cytokine storms than in normal individuals; interleukin 6 (IL-6) exhibits much higher elevations. Investigating potential relationships between IL-6 serum levels and the severity of COVID-19 symptoms in pregnant women is the aim of this study. Material and Methods: Sixty-two eligible pregnant women were divided into a positive COVID-19 group (n=31) and a negative COVID-19 group (n=31) in this cross-sectional study. The research subjects were selected using consecutive sampling. The IL-6 was measured from a vein blood specimen using ELISA methods. Results: The COVID-19 positive group had a higher median IL-6 serum level (45.35 (35.15- 153.99) vs. 38.86 ± 11.43 (15.02-59.52), p=0.03) than the negative group. On the other hand, the IL-6 serum level had comparable value according to the COVID-19 symptoms severity (88.35 ± 36.14 ng/mL vs. 51.09 ± 25.48 ng/mL vs. 56.02 ± 33.20 ng/mL in moderate symptoms, mild symptoms, and asymptomatic, respectively; p=0.152). Conclusion: Although the IL-6 serum levels are not related to COVID-19 symptoms severity, an elevated of this biomarker was found in pregnant women with affected diagnoses.

Keywords: interleukin-6, pregnancy, COVID-19, several inflammatory

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Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

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Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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Authors: R. Charoen, S. Tipkanon, W. Savedboworn, N. Phonsatta, A. Panya

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Rice bran protein hydrolysates (RBPH) were prepared from defatted rice bran of two different Thai rice cultivars (Plai-Ngahm-Prachinburi; PNP and Khao Dok Mali 105; KDM105) using an enzymatic method. This research aimed to optimize enzyme-assisted protein extraction. In addition, the functional properties of RBPH and their stabilities to environmental stresses including pH (3 to 8), ionic strength (0 mM to 500 mM) and the thermal treatment (30 °C to 90 °C) were investigated. Results showed that enzymatic process for protein extraction of defatted rice bran was as follows: enzyme concentration 0.075 g/ 5 g of protein, extraction temperature 50 °C and extraction time 4 h. The obtained protein hydrolysate powders had a degree of hydrolysis (%) of 21.05% in PNP and 19.92% in KDM105. The solubility of protein hydrolysates at pH 4-6 was ranged from 27.28-38.57% and 27.60-43.00% in PNP and KDM105, respectively. In general, antioxidant activities indicated by total phenolic content, FRAP, ferrous ion-chelating (FIC), and 2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) of KDM105 had higher than PNP. In terms of functional properties, the emulsifying activity index (EAI) was was 8.78 m²/g protein in KDM105, whereas PNP was 5.05 m²/g protein. The foaming capacity at 5 minutes (%) was 47.33 and 52.98 in PNP and KDM105, respectively. Glutamine, Alanine, Valine, and Leucine are the major amino acid in protein hydrolysates where the total amino acid of KDM105 gave higher than PNP. Furthermore, we investigated environmental stresses on the stability of 5% oil in water emulsion (5% oil, 10 mM citrate buffer) stabilized by RBPH (3.5%). The droplet diameter of emulsion stabilized by KDM105 was smaller (d < 250 nm) than produced by PNP. For environmental stresses, RBPH stabilized emulsions were stable at pH around 3 and 5-6, at high salt (< 400 mM, pH 7) and at temperatures range between 30-50°C.

Keywords: functional properties, oil in water emulsion, protein hydrolysates, rice bran protein

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Authors: Habib Aghdam Shahryar

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The aim of the present study was investigation on the effect of intraperipheral (IP) injection of ghrelin on serum insulin and glucose levels in native turkey. Seventy-two 28 day (d)-old native turkey were assigned into three treatments and four replicate for 40 days experimental rearing period: group 1, intact without any injection, group 2, injected 50 ng ghrelin/kg body weight (BW), and group 3, injected 100 ng ghrelin/kg BW. Intraperitoneal injection of rat ghrelin was conducted on d 28 and before the onset of the experimental rearing period. Blood samples were taken 12 hr after injection and 40 days after injection. The result showed glucose concentrations have been affected by administered ghrelin and significant between groups (P<0.01). Injection of ghrelin at G 100 increased glucose level of serum in 12 hr after injection and 40 days after injection (276.6 and 260.0 mg/dl, respectively). Also, by increasing the dose of injected ghrelin, insulin levels than the control group showed an increase (P < 0.001). This study suggests roles of ghrelin in serum biochemical regulation may show a different effect of ghrelin on blood parameters in avian species.

Keywords: ghrelin injection, insulin, glucose, turkey

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Authors: Ezeugwunne Ifeoma Priscilla, Charles Chinedum Onyenekwe, Joseph Eberendu Ahaneku, Rosemary Adanma Analike, Adesuwa Peace Eidangbe

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This study was designed to assess the effect of malaria parasitaemia on serum reproductive hormone levels of asymptomatic HIV adult subjects. A total of 271 participants aged between 17 and 58 ears were conveniently recruited. 135 asymptomatic HIV-infected subjects participated in the study; 67 of them had malaria parasitaemia. 136 HIV seropositive control subjects, 68 of them had malaria parasitaemia. Blood samples were collected from the participants for the determination of HIV status by immunoassay and immunochromatography. Enzyme-linked immunosorbent assay (ELISA) was used to assay for serum LH, FSH, Estrogen, testosterone, progesterone, prolactin, and PSA levels, CD4+T cell counts by Cyflow method, thick and thin films determination of malaria parasitaemia count and density by WHO. Student's t-tests and ANOVA were used to compare means. P<0.05 was considered statistically significant. The results showed significant differences in serum levels of LH, FSH, PSA, estrogen, progesterone, and testosterone amongst the groups at P<0.05, respectively. The serum levels of LH, FSH, and PSA were significantly higher in malaria-infected asymptomatic HIV subjects than in asymptomatic HIV subjects with malaria parasitaemia (P<0.05 in each case). Also, the serum levels of LH, FSH, PSA, estrogen, and progesterone were significantly higher in malaria-infected asymptomatic HIV subjects compared with malaria-infected HIV seronegative subjects (P<0.05, respectively). The mean MP counts and MP density were significantly higher in asymptomatic HIV subjects compared to HIV seronegative subjects (P<0.05, in each case). The mean serum levels of testosterone were significantly lower in both malaria-infected and malaria uninfected HIV seronegative subjects (P<0.05, in each case). In conclusion, Malaria and HIV co-infection might increase the burden of hypogonadism as well as primary testicular failure, hyperprogesteronaemia, elevated levels of estrogen, and PSA in adult males asymptomatic HIV subjects.

Keywords: malaria parasitaemia, HIV, CD4, reproductive hormones

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Authors: A. M. Gadanya, B. A. Ahmad, U. Maigatari

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Effect of oral administration of Gadagi tea on superoxide dismutase activity was assessed on twenty (20) male subjects (aged 21-40years). Ten (10) male non Gadagi tea consumers (aged 20-26 years), were used as control. Blood samples were collected from the subjects and analysed for serum superoxide dismutase activity using R&D Enzyme Linked Immunosorbent Assay method (ELISA). The subjects were grouped into four based on age i.e group I (21-25 years), group II (26-30 years), and also based on duration of the tea consumption, i.e group A (5-9 years) , group B (10-14 years), group C (15-19 years) and group D (20-24 years). The subjects in group I (0.12 U mg-l +0.05), group II (0.11 U mg-l +0.01), group III (0.14 U mg-l +0.08) and group IV (0.17 U mg-l +0.11) showed increased activity of serum superoxide dismutase when compared with the control subjects (0.88 U mg-l +0.02) (P<0.05). There was no statistical significant difference in superoxide dismutase activity within the case groups (P<0.05), based on age and duration of consumption of the tea. Thus, Gadagi tea consumption could increase serum superoxide dismutase activity in humans.

Keywords: “Gadagi” tea, Serum, Superoxide dismutase, Humans.

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Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: Gospodinka Prakova, Pavlina Gidikova, Gergana Sandeva, Kamelia Haracherova, Emil Slavov

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Neopterin was demonstrated to be a sensitive marker of cell-mediated immune reactions which plays a key role in the interaction of monocyte / macrophage activation. The purpose of this work was to investigate changes in serum neopterin in workers exposed to different composition of mineral dust. Material and Methods: Serum neopterin was studied in 193 exposed workers, divided into three groups, depending on the mineral dust and content of the quartz in the respirable fraction. The I-st group-coal dust containing less than 2% free crystalline silica (n=44), II-nd group-coal dust containing over 2% free crystalline silica (n=94) and the III-rd group-mixed dust with corundum and carborundum (n=55). The control group was composed of 21 individuals without exposure to dust. Serum neopterin was investigated by Elisa method in ng/ml according to the instructions of the manufacturer. Results and Discussion: It was found significantly higher level of serum neopterin in exposed workers of mineral dust (2,10 ± 0,62 ng / ml), compared with that of the control group (1,10 ± 0,85 ng/ml; p < 0,05). Neopterin levels in workers exposed to coal dust (1,87 ± 0,42 ng / ml-I-st and 3,32 ± 0,77 ng / ml-II-nd group) were significantly higher compared with those exposed to a mixed dust (1,31±0,68 mg / ml-third) and control group (p < 0,05). No significant difference in serum neopterin when exposed to a mixed dust composed of corundum and carborundum (III-rd) and a control group. Conclusion: The results of this study indicate activates a cell-mediated immune response when exposed to a mineral dust. The level of that activation depends mainly on the composition of the dust and is significantly highest in workers exposed to coal dust.

Keywords: mineral dust, neopterin, occupational exposure, respirable crystalline silica

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Authors: B. Nageshwar Rao

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Urinary tract infection (UTI) represents the most commonly acquired bacterial infection worldwide, with substantial morbidity, mortality, and economic burden. The objective of the study is to characterize the microbial profiles of uropathogenic in the obstetric population by RTPCR. Study design: An observational cross-sectional study was performed at a single tertiary health care hospital among 50 pregnant women with UTIs, including asymptomatic and symptomatic patients attending the outpatient department and inpatient department of Obstetrics and Gynaecology.Methods: Serotyping and genes detection of various uropathogens were studied using RTPCR. Pulse filed gel electrophoresis methods were used to determine the various genetic profiles. Results: The present study shows that CsgD protein, involved in biofilm formation in Escherichia coli, VIM1, IMP1 genes for Klebsiella were identified by using the RTPCR method. Our results showed that the prevalence of VIM1 and IMP1 genes and CsgD protein in E.coli showed a significant relationship between strong biofilm formation, and this may be due to the prevalence of specific genes. Finally, the genetic identification of RTPCR results for both bacteria was correlated with each other and concluded that the above uropathogens were common isolates in producing Biofilm in the pregnant woman suffering from urinary tract infection in our hospital observational study.

Keywords: biofilms, Klebsiella, E.coli, urinary tract infection

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Authors: Yana Posmitnaya, Galina Rudnitskaya, Tatyana Lukashenko, Anton Bukatin, Anatoly Evstrapov

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An important area of biological and medical research is the study of genetic mutations and polymorphisms that can alter gene function and cause inherited diseases and other diseases. The following methods to analyse DNA fragments are used: capillary electrophoresis and electrophoresis on microfluidic chip (MFC), mass spectrometry with electrophoresis on MFC, hybridization assay on microarray. Electrophoresis on MFC allows to analyse small volumes of samples with high speed and throughput. A soft lithography in polydimethylsiloxane (PDMS) was chosen for operative fabrication of MFCs. A master-form from silicon and photoresist SU-8 2025 (MicroChem Corp.) was created for the formation of micro-sized structures in PDMS. A universal topology which combines T-injector and simple cross was selected for the electrophoretic separation of the sample. Glass K8 and PDMS Sylgard® 184 (Dow Corning Corp.) were used for fabrication of MFCs. Electroosmotic flow (EOF) plays an important role in the electrophoretic separation of the sample. Therefore, the estimate of the quantity of EOF and the ways of its regulation are of interest for the development of the new methods of the electrophoretic separation of biomolecules. The following methods of surface modification were chosen to change EOF: high-frequency (13.56 MHz) plasma treatment in oxygen and argon at low pressure (1 mbar); 1% aqueous solution of polyvinyl alcohol; 3% aqueous solution of Kolliphor® P 188 (Sigma-Aldrich Corp.). The electroosmotic mobility was evaluated by the method of Huang X. et al., wherein the borate buffer was used. The influence of physical and chemical methods of treatment on the wetting properties of the PDMS surface was controlled by the sessile drop method. The most effective way of surface modification of MFCs, from the standpoint of obtaining the smallest value of the contact angle and the smallest value of the EOF, was the processing with aqueous solution of Kolliphor® P 188. This method of modification has been selected for the treatment of channels of MFCs, which are used for the separation of mixture of oligonucleotides fluorescently labeled with the length of chain with 10, 20, 30, 40 and 50 nucleotides. Electrophoresis was performed on the device MFAS-01 (IAI RAS, Russia) at the separation voltage of 1500 V. 6% solution of polydimethylacrylamide with the addition of 7M carbamide was used as the separation medium. The separation time of components of the mixture was determined from electropherograms. The time for untreated MFC was ~275 s, and for the ones treated with solution of Kolliphor® P 188 – ~ 220 s. Research of physical-chemical methods of surface modification of MFCs allowed to choose the most effective way for reducing EOF – the modification with aqueous solution of Kolliphor® P 188. In this case, the separation time of the mixture of oligonucleotides decreased about 20%. The further optimization of method of modification of channels of MFCs will allow decreasing the separation time of sample and increasing the throughput of analysis.

Keywords: electrophoresis, microfluidic chip, modification, nucleic acid, polydimethylsiloxane, soft lithography

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Authors: El-Sohaimy Sobhy A., Androsova Natalia, Toshev Abuvali Djabarovec

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The conditions of the isolation of proteins from the hemp seeds were optimized in the current work. Moreover, the physicochemical and functional properties of hemp protein isolate were evaluated for its potential application in food manufacturing. The elastin protein is the most predominant protein in the protein profile with a molecular weight of 58.1 KDa, besides albumin, with a molecular weight of 31.5 KDa. The FTIR spectrum detected the absorption peaks of the amide I in 1750 and 1600 cm⁻¹, which pointed to C=O stretching while N-H was stretching at 1650-1580 cm⁻¹. The peak at 3250 was related to N-H stretching of primary aliphatic amine (3400-3300 cm⁻¹), and the N-H stretching for secondary (II) amine appeared at 3350-3310 cm⁻¹. Hemp protein isolate (HPI) was showed high content of arginine (15.52 g/100 g), phenylalanine+tyrosine (9.63 g/100 g), methionine + cysteine (5.49 g/100 g), leucine + isoleucine (5.21 g/100 g) and valine (4.53 g/100 g). It contains a moderate level of threonine (3.29 g/100 g) and lysine (2.50 g/100 g), with the limiting amino acid being a tryptophan (0.22 g/100 g HPI). HPI showed high water-holding capacity (4.5 ± 2.95 ml/g protein) and oil holding capacity (2.33 ± 1.88 ml/g) values. The foaming capacity of HPI was increased with increasing the pH values to reach the maximum value at pH 11 (67.23±3.20 %). The highest emulsion ability index of HPI was noted at pH 9 (91.3±2.57 m2/g) with low stability (19.15±2.03).

Keywords: Cannabis sativa ssp., protein isolate, isolation conditions, amino acid composition, chemical properties, functional properties

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Authors: Zhaojun Zheng, Yuanfa Liu, Jiaxin Li, Jinwei Li, Yong-jiang Xu, Chen Cao

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Black bean is an excellent protein source for preparing hydrolysates, which attract much attention due to their biological activity. The objective of this study was to characterize the physicochemical and antioxidant properties of black bean protein, hydrolyzed by ficin, bromelain or alcalase until 300 min of hydrolysis. Results showed that bromelain and alcalase hydrolysates possessed a higher degree of hydrolysis (DH) than that of ficin, thereby presenting different ultraviolet absorption, fluorescence intensity, and circular dichroism. Moreover, all hydrolysates possessed the capacity to scavenge DPPH radical with the lowest IC₅₀ of 21.11 µg/mL, as well as to chelate ferrous ion (Fe²⁺) with the IC₅₀ values ranging from 6.82 to 30.68 µg/mL. Intriguingly, the oxidation of linoleic acid, sunflower oil, and sunflower oil-in-water emulsion was remarkedly retarded by the three selected protein hydrolysates, especially by bromelain-treated protein hydrolysate, which might attribute to their high hydrophobicity and emulsifying properties. These findings can provide strong support for black bean protein hydrolysates to be employed in food products acting as natural antioxidant alternatives.

Keywords: antioxidant activity, black bean protein hydrolysate, emulsion physicochemical properties, sunflower oil

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Authors: Neslihan Taskale Karatug, Mustafa Akcelik

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Some literature data show that misL protein play a role on host immune response formed against Salmonella Typhimurium. The aim of the present study is to learn the role of the protein in S. Typhimurium pathogenicity. To describe certain functions of the protein, primarily recombinant misL protein was produced and purified. PCR was performed using a primer set targeted to passenger domain of the misL gene on S. Typhimurium LT2 genome. Amplicon and pet28a vector were enzymatically cleaved with EcoRI and NheI. The digested DNA materials were purified with High Pure PCR Product Purification Kit. The ligation reaction was achieved with the pure products. After preparation of competent Escherichia coli Dh5α, ligation mix was transformed into the cell by electroporation. To confirm the existence of insert gene, recombinant plasmid DNA of Dh5α was isolated with high pure plasmid DNA kit. Proved the correctness of recombinant plasmid was electroporated to BL21. The cell was induced by IPTG. After induction, the presence of recombinant protein was checked by SDS-PAGE. The recombinant misL protein was purified using HisPur Ni-NTA spin colon. The pure protein was shown by SDS-PAGE and western blot immünoassay. The concentration of the protein was measured BCA Protein Assay kit. In the wake of ligation with digested products (2 kb misL and 5.4 kb pet28a) visualised on gel size of the band was about 7.4 kb and was named as pNT01. The pNT01 recombinant plasmid was transformed into Dh5α and colonies were chosen in selective medium. Plasmid DNA isolation from them was carried out. PCR was achieved on the pNT01 to check misL and 2 kb band was observed on the agarose gel. After electroporation of the plasmid and induction of the cell, 68 kDa misL protein was seen. Subsequent to the purification of the protein, only a band was observed on SDS-PAGE. Association of the pure protein with anti-his antibody was verified by the western blot assay. The concentration of the pure misL protein was determined as 345 μg/mL. Production of polyclonal antibody will be achieved by using the obtained pure recombinant misL protein as next step. The role of the protein will come out on the immune system together some assays.

Keywords: cloning, Escherichia coli, recombinant protein purification, Salmonella Typhimurium

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Authors: Masoumeh Hosseini

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Obesity is known to be associated with cardiovascular diseases and metabolic syndrome. This study aimed to assess the effect of a long term aerobic training program on serum ghrelin in obese men. For this purpose, twenty four sedentary adult obese men aged 30-40 years and body mass index 30-36 kg/m2 were participated in this study and divided randomly into exercise (3 months aerobic training, 3 times/weekly) or control (no training) groups. Serum ghrelin and cardiovascular risk factor (TG, TC, LDL, and HDL) were measured before and after treatment. Anthropometrical markers were measured at two occasions. Data were analyzed by independent-paired T-test. Significance was accepted at P < 0.05. Aerobic training resulted in significant decrease in serum ghrelin and TG in exercise group. All anthropometrical markers decreased significantly in exercise group but not in control subjects. Based on these data, it is concluded that weight loss by aerobic training can be affect serum ghrelin in obese subject, although some cardiovascular risk factor remained without changed.

Keywords: aerobic training, homeostasis, lipid profile, obesity

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Authors: Mohammad Hossein Harirchian, Siavash Babaie, Nika keshtkaran, Sama Bitarafan

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Background: Fatigue is a common complaint of multiple sclerosis (MS) patients, adversely affecting their quality of life. There is a lot of evidence showing that Carnitine deficiency is linked to fatigue development and severity in some conditions. This study aimed to compare the levels of Free L-Carnitine (FLC) between MS patients and healthy people and evaluate its association with the severity of fatigue. Methods: This case-control study included 30 patients with relapsing-remitting MS (RRMS) in 2 sex-matched equal-number groups according to the presence or absence of fatigue and 30 sex-matched healthy people in the control group. In addition, between two patient groups, we compared Serum level of FLC between the patient and healthy group. Fatigue was scored using two valid questionnaires of fatigue Severity Scale (FSS) and Modified Fatigue Impact Scale (MFIS). In addition, association between Serum level of FLC and fatigue severity was evaluated in MS patients. Results: There was no significant difference in serum levels of FLC between MS patients and healthy people. The patients with fatigue had a significantly lower FLC (mg/dl) value than patients without fatigue (22.53 ± 15.84 vs. 75.36 ± 51.98, P < 0.001). The mean value of FSS and MFIS in patients with fatigue were 48.80±8.55 and 62.87 ± 13.63, respectively, which was nearly two-fold higher than group without fatigue (P < 0.001). There was a negative correlation between the serum level of FLC and fatigue severity scales (Spearman rank correlation= 0.76, P < 0.001). Conclusion: We showed healthy people and MS patients were not different in levels of FLC. In addition, patients with lower serum levels of FLC might experience more severe fatigue. Therefore, this could clarify that supplementation with L-Carnitine might be considered as a complementary treatment for MS-related fatigue.

Keywords: fatigue, multiple sclerosis, L-carnitine, modified fatigue impact scale

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Authors: Alok Raghav, Jamal Ahmad

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Background: Serum albumin glycation and advanced glycation end products (AGE) formation correlates in diabetes and its associated complications. Extensive modified human serum albumin is used to study the biochemical, electrochemical and functional properties in hyperglycemic environment with relevance to diabetes. We evaluate Spectroscopic, side chain modifications, amino acid analysis, biochemical and functional group properties in four glucose modified samples. Methods: A series four human serum albumin samples modified with glucose was characterized in terms of amino acid analysis, spectroscopic properties and side chain modifications. The diagnostic technique employed incorporates UV Spectroscopy, Fluorescence Spectroscopy, biochemical assays for side chain modifications, amino acid estimations. Conclusion: Glucose modified human serum albumin confers AGE formation causes biochemical and functional property that depend on the reactivity of glucose and its concentration used for in-vitro glycation. A biochemical and functional characterization of modified albumin in-vitro produced AGE product that will be useful to interpret the complications and pathophysiological significance in diabetes.

Keywords: glycation, diabetes, human serum albumin, biochemical and electrochemical characterization

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Authors: Baigts-Allende Diana, Klojdová Iveta, Kozlu Ali, Metri-ojeda Jorge

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Emulsion gels are constituted by a colloidal system (emulsion) stabilized by a polymeric gel matrix. These systems are more homogeneous and stable than conventional emulsions and can behave as either gel-like or soft-solid. Protein-based emulsion gels (PEG) have been used as carrier systems of bioactive compounds and as food structuring to improve the texture and consistency, mainly in producing low-fat content products. This work studied the effect of protein: polysaccharide ratio 0.75:1.25, 1:1, and 1.25:0.75 (levels -1, 0, and +1) and pH values (2-9) on the stability of protein-based emulsion gels using soy protein isolate and sodium alginate. Protein emulsion capacity was enhaced with increased pH (6,7,8 and 9) compared to acid pH values. The smaller particle size for PEG was at pH 9 (~23µm); however, with increasing protein ratio (level +1), higher particle size was observed (~23µm). The same trend was observed for rheological measurements; the consistency index (K) increased at pH 9 for level -1 (1.17) in comparison to level +1 (0.45). The studied PEG showed good thermal stability at neutral and pH 9 (~98 %) for all biopolymer ratios. Optimal conditions in pH and biopolymer ratios were determined for PEG using soy protein and sodium alginate ingredients with potential use in elaborating stable systems for broad application in the food sector.

Keywords: emulsion gels, food structuring, biopolymers, food systems

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Authors: M. Titaouine, T. Meziane, K. Deghnouche

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The purpose of this study was to determine reference serum biochemistry values from dromedary (Camelus dromedarius) in Algeria and to evaluate potential sources of physiological variability such as the sex, age and season on serum data. Usual serum biochemistry values were determined in blood samples from 26 apparently healthy dromedaries, 11 males and 15 females, divided into 3 lots (ender 4years), (between 5 and 10 years), (up 10 years). Parametric reference ranges and physiological variations are determined for calcium (Ca), organic phosphate (P), magnesium (Mg), natrium (Na), potassium (K), iron (Fe), glucose, triglycerides (TG), cholesterol, urea, creatinine, total proteins and albumin. The results demonstrate: * Values which agreed with literature * Significant statistically differences (Anova test, p < 0.05) depending on: -the sex for Na, glucose, TG, cholesterol, urea, creatinine, albumin, -the age for Ca, P, K, Mg, glucose, TG, b and g globulin, -and season for Fe, urea, total proteins, TG, cholesterol and glucose. These reference ranges for serum biochemical analysis can be used for metabolic and nutritional disorders detection in dromedary.

Keywords: age, biochemistry, dromadery, season, sex

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Authors: Suchao Donpudsa, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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A crustin gene, LV-SWD1, previously found in the hemocyte cDNA library of Litopenaeus vannamei, contains the open reading frames of 288 bp encoding a putative protein of 96 amino acid residues. The putative signal peptides of the LV-SWD1 were identified using the online SignalP 3.0 with predicted cleavage sites between Ala24-Val25, resulting in 72 residue mature protein with calculated molecular mass of 7.4 kDa and predicted pI of 8.5. This crustin contains a Arg-Pro rich region at the amino-terminus and a single whey acidic protein (WAP) domain at the carboxyl-terminus. In order to characterize their properties and biological activities, the recombinant crustin protein was produced in the Escherichia coli expression system. Antimicrobial assays showed that the growth of Bacillus subtilis was inhibited by this recombinant crustin with MIC of about 25-50 µM.

Keywords: crustin, single whey acidic protein, Litopenaeus vannamei, antimicrobial activity

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Authors: Olawuyi Sikiru Owolabi

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A research was carried out to assess the biochemical effects of crude oil contaminated cat fish on selected rat kidney function tests. Thirty-six (36) albino rats (rattus novergicus) were grouped into six (6) of (6) in each group. The rats in group one served as control and they were placed on feed formulated with catfish cultured in borehole water while those ones from group 2 to group 6 were placed on feed formulated with catfish exposed to various concentrations of crude oil (0.1%,0.25%,0.5%,0.75% and 1% respectively).The results obtained showed that there was a significant increase in serum concentration of creatinine, Urea, sodium and potassium ions in the kidney of experimental rats when compared with the control. This may be interpreted to mean possible adverse effects on the kidney. Several studies have been done especially on the biological effects of crude oil in fish. These include Direct Lethal Toxicity, Sub-Lethal disruption of physiological and behavioral activities, interference with feeding and reproduction, direct coating or tainting of fish, effect of entry of hydrocarbons into the food web as well as alteration of biological habitat. The present study attempts to assess the effects of crude oil contaminated diet on rat kidney by carrying out some kidney function tests like determination of serum sodium and potassium ions by flame photometry method, determination of serum urea and determination of serum creatinine.

Keywords: crude oil, serum urea, creatinine, wistar rats

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Authors: Ali Golestani, Vahid Naseri, Hossein Taheri

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Recently, some of studies examined the effect of exercise on neurotrophic factors influencing the growth, protection, plasticity and function in central and peripheral nerve cells. The aim of this study was to investigate the effects of continuous and interval aerobic exercises with moderate intensity on serum levels of glial cell line-derived neurotrophic factor (GDNF) and aerobic capacity in obese children. 21 obese students with an average age of 13.6 ± 0.5 height 171 ± 5 and BMI 32 ± 1.2 were divided randomly to control, continuous aerobic and interval aerobic groups. Training protocol included continuous or interval aerobic exercises with moderate intensity 50-65%MHR, three times per week for 10 weeks. 48 hours before and after executing of protocol, blood samples were taken from the participants and their GDNF serum levels were measured by ELISA. Aerobic power was estimated using Shuttle-run test. T-test results indicated a small increase in their GDNF serum levels, which was not statistically significant (p =0.11). In addition, the results of ANOVA did not show any significant difference between continuous and interval aerobic training on the serum levels of their GDNF but their aerobic capacity significantly increased (p =0.012). Although continuous and interval aerobic exercise improves aerobic power in obese children, they had no significant effect on their serum levels of GDNF.

Keywords: aerobic power, continuous aerobic training, glial cell line-derived neurotrophic factor (GDNF), interval aerobic training, obese children

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Authors: Joan O. Ogundele, Aladesanmi A. Oshodi, Adekunle I. Amoo

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Abstract In-vitro multi-enzyme protein digestibility (IVMPD) and some anti-nutritional factors (ANF) of five melon (egusi) seed flours (MSF) and their protein isolates (PI) were carried out. Their PI have potentials comparable to that of soya beans. It is important to know the IVMPD and ANF of these protein sources as to ensure their safety when adapted for use as alternate protein sources to substitute for cow milk, which is relatively expensive in Nigeria. Standard methods were used to produce PI of Citrullus colocynthis, Citrullus vulgaris, African Wine Kettle gourd (Lageneria siceraria I), Basket Ball gourd (Lagenaria siceraria II) and Bushel Giant Gourd (Lageneria siceraria III) seeds and to determine the ANF and IVMPD of the MSF and PI unheated and at 37oC. Multi-enzymes used were trypsin, chymotrypsin and peptidase. IVMPD of MSF ranged from (70.67±0.70) % (C. vulgaris) to (72.07± 1.79) % (L.siceraria I) while for their PI ranged from 74.33% (C.vulgaris) to 77.55% (L.siceraria III). IVMPD of the PI were higher than those of MSF. Heating increased IVMPD of MSF with average value of 79.40% and those of PI with average of 84.14%. ANF average in MSF are tannin (0.11mg/g), phytate (0.23%). Differences in IVMPD of MSF and their PI at different temperatures may arise from processing conditions that alter the release of amino acids from proteins by enzymatic processes. ANF in MSF were relatively low, but were found to be lower in the PI, therefor making the PI safer for human consumption as an alternate source of protein.

Keywords: Anti-nutrients, Enzymatic protein digestibility, Melon (egusi)., Protein Isolates.

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Authors: Amit Chaudhary, B. S. Yadav, P. K. Maurya, A. M., S. Srivastava, S. Singh, A. Mani

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Cold shock protein A (CspA) is major cold inducible protein present in Escherichia coli. The protein is involved in stabilizing secondary structure of RNA by working as chaperone during cold temperature. Two RNA binding motifs play key role in the stabilizing activity. This study aimed to investigate implications of low temperature on structure and RNA binding activity of E. coli CspA. Molecular dynamics simulations were performed to compare the stability of the protein at 37°C and 10 °C. The protein was mutated at RNA binding motifs and docked with RNA to assess the stability of both complexes. Results suggest that CspA as well as CspA-RNA complex is more stable at low temperature. It was also confirmed that RNP1 and RNP2 play key role in RNA binding.

Keywords: CspA, homology modelling, mutation, molecular dynamics simulation

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Authors: Francesco Angelucci, Katerina Veverova, Alžbeta Katonová, Lydia Piendel, Martin Vyhnalek, Jakub Hort

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Alzheimer's disease (AD) is a central nervous system (CNS) disease characterized by loss of memory, cognitive functions and neurodegeneration. Plasmin is an enzyme degrading many plasma proteins. In the CNS, plasmin may reduce the accumulation of A, and have other actions relevant to AD pathophysiology. Brain plasmin synthesis is regulated by two enzymes: one activating, the tissue plasminogen activator (tPA), and the other inhibiting, the plasminogen activator inhibitor-1 (PAI-1). We investigated whether tPA and PAI-1 serum levels in AD and amnestic mild cognitive impairment (aMCI) patients are altered compared to cognitively healthy controls. Moreover, we examined the PAI-1/tPA ratio in these patient groups. 40 AD, 40 aMCI and 10 healthy controls were recruited. Venous blood was collected and PAI-1 and tPA serum concentrations were quantified by sandwich ELISAs. The results showed that PAI-1 levels increased in AD and aMCI patients. This increase negatively correlated with cognitive deficit measured by MMSE. Similarly, the ratio between tPA and PAI-1 gradually increases in aMCI and AD patients. This study demonstrates that AD and aMCI patients have altered PAI-1 serum levels and PAI-1/tPA ratio. Since these enzymes are CNS regulators of plasmin, PAI-1 serum levels could be a marker reflecting a cognitive decline in AD.

Keywords: Alzheimer disease, amnestic mild cognitive impairment, plasmin, tissue-type plasminogen activator

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