Search results for: serum

Authors: Hong Yi Huang, Wei Ti Kuo, Yi You Huang

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Non-viral gene carriers composed of biodegradable polymers or lipids have been considered as a safer alternative for gene carriers over viral vectors. We have developed multi-functional nano-micelles for both drug and gene delivery application. Polyethyleneimine (PEI) was modified by grafting stearic acid (SA) and formulated to polymeric micelles (PEI-SA) with positive surface charge for gene and drug delivery. Our results showed that PEI-SA micelles provided high siRNA binding efficiency. In addition, siRNA delivered by PEI-SA carriers also demonstrated significantly high cellular uptake even in the presence of serum proteins. The post-transcriptional gene silencing efficiency was greatly improved by the polyplex formulated by 10k PEI-SA/siRNA. The amphiphilic structure of PEI-SA micelles provided advantages for multifunctional tasks; where the hydrophilic shell modified with cationic charges can electrostatically interact with DNA or siRNA, and the hydrophobic core can serve as payloads for hydrophobic drugs, making it a promising multifunctional vehicle for both genetic and chemotherapy application.

Keywords: polyethyleneimine, gene delivery, micelles, siRNA

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Authors: D. Leskauskaite, I. Jasutiene, M. Kersiene, E. Malinauskyte, P. Matusevicius

Abstract:

In vitro gastro-duodenal digestion model was used to investigate the changes of emulsions under digestion conditions. Oil in water emulsions stabilized by whey proteins (2%) and stabilized by whey proteins (2%) with addition of carboxymethyl cellulose (0.75%) as gelling agent of continuous phase were prepared at pH7. Both emulsions were destabilized under gastric conditions; however the protective role of carboxymethyl cellulose was indicated by recording delay of fat digestibility of this emulsion. In the presence of carboxymethyl cellulose whey proteins on the interfacial surface of droplets were more resistant to gastric degradation causing limited hydrolysis of fat due to the poor acceptability of lipids for the enzymes. Studies of emulsions using in vivo model supported results from in vitro studies. Lower content of triglycerides in blood serum and higher amount of fecal fat of rats were determined when rats were fed by diet containing emulsion made with whey proteins and carboxymethyl cellulose. 

Keywords: Digestibility, emulsions, lipids, rats.

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Authors: Junki Miyamoto, Mariko Kiyomi, Yuuki Nagashio, Takuya Suzuki, Soichi Tanabe

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Anti-allergic effects of royal jelly were evaluated in a human-like mouse model of atopic dermatitis. NC/Nga mice were cutaneously applied with royal jelly for 6 weeks. Royal jelly-treated mice exhibited lower levels of serum total immunoglobulin E in comparison with controls. We found that the treatment decreased (11% to the control) expression of mRNA for aquaporin-3, which is involved in the modulation of epidermal hydration. Microarray analysis revealed more than 10-fold changes in the expression of several genes, such as transglutaminase 2, repetin, and keratins. In normal human epidermal keratinocytes, royal jelly extract suppressed interleukin-8 elevation induced by TNF-α and interferon-γ, suggesting direct anti-inflammatory activity in keratinocytes. Collectively, topical application of royal jelly may be useful for amelioration of lesions and inflammation in atopic dermatitis.

Keywords: Aquaporin 3, immunoglobulin E, NC/Nga, royal jelly.

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Authors: Binnur Sagbas

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Artificial joint replacements such as total knee and total hip prosthesis have been applied to the patients who affected by osteoarthritis. Although different material combinations are used for these joints, biopolymers are most commonly preferred materials especially for acetabular cup and tibial component of hip and knee joints respectively. The main limitation that shortens the service life of these prostheses is wear. Wear is complicated phenomena and it must be considered with friction and lubrication. In this study, micro wave (MW) induced argon+oxygen plasma surface modification were applied on ultra-high molecular weight polyethylene (UHMWPE) and vitamin E blended UHMWPE (VE-UHMWPE) biopolymer surfaces to improve surface wettability and wear resistance of the surfaces. Contact angel measurement method was used for determination of wettability. Ball-on-disc wear test was applied under 25% bovine serum lubrication conditions. The results show that surface wettability and wear resistance of both material samples were increased by plasma surface modification.

Keywords: Artificial joints, plasma surface modification, UHMWPE, vitamin E, wear.

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Authors: Andreea Cârâc, Elena Moroşan, Ana Corina Ioniță, Rica Boscencu, Geta Cârâc

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Although, lanthanum carbonate has not been approved by the FDA for treatment of hyperphosphatemia, we prospectively evaluated the efficacy of the combination of Calcium hydroxyapatite (CHAp) and Lanthanum Carbonate (LaC) for the treatment of hyperphosphatemia on mice. CHAp was prepared by co-precipitation method using Ca(OH)2, H3PO4, NH4OH with calcination at 1200ºC. Lanthanum carbonate was prepared by chemical method using NaHCO3 and LaCl3 at low pH environment, below 4.0. The structures were characterized by FTIR spectra and SEM -EDX analysis. The study group included 16 subjects-mice divided into four groups according to the administered substance: lanthanum carbonate (group A), CHAp (group B), lanthanum carbonate + CHAp (group C) and salt water (group D). The results indicate a phosphate decrease when subjects (mice) were treated with CHAp and lanthanum carbonate (0.5% CMC), in a single dose of 1500 mg/kg. Serum phosphate concentration decreased [(from 4.5 ± 0.8 mg/dL) to 4.05 ± 0.2 mg/dL), P < 0.01] in group A and in group C (to 3.6 ± 0.2 mg/dL) at 12 hours from the administration. The combination of CHAp and lanthanum carbonate is a suitable regimen for hyperphosphatemia treatment because it avoids both the hypercalcemia of CaCO3 and the adverse effects of CHAp.

Keywords: Calcium hydroxyapatite, hyperphosphatemia, lanthanum carbonate, phosphatebinder, structures.

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Authors: M. Petchsangsai, N. Wonglertnirant, T. Rojanarata, P. Opanasopit, T. Ngawhirunpat

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The objective of the present study was to evaluate the potential of hollow microneedles for enhancing the transdermal delivery of Bovine Serum Albumin (MW~66,000 Da)-Fluorescein Isothiocyanate (BSA-FITC) conjugate, a hydrophilic large molecular compound. Moreover, the effect of different formulations was evaluated. The series of binary mixtures composed of propylene glycol (PG) and pH 7.4 phosphate buffer solution (PBS) was prepared and used as a medium for BSA-FITC. The results showed that there was no permeation of BSA-FITC solution across the neonatal porcine skin without using hollow microneedles, whereas the cumulative amount of BSA-FITC released at 8 h through the neonatal porcine skin was about 60-70% when using hollow microneedles. Furthermore, the results demonstrated that the higher volume of PG in binary mixtures injected, the lower cumulative amount of BSA-FITC released and release rate of BSA-FITC from skin. These release profiles of BSA-FITC in binary mixtures were expressed by Fick-s law of diffusion. These results suggest the utilization of hollow microneedle to enhance transdermal delivery of protein and provide useful information for designing an effective hollow microneedle system.

Keywords: Hydrophilic macromolecules, Microneedles, Propylene glycol, Transdermal drug delivery

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Authors: Shanmugasundaram Krishnakumari, Paramasivam Rajeswari, Subramanian Kathiravan

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Mushrooms are a group of fleshy macroscopic fungi. They have been valued throughout the world as both edible and medicine. They are highly nutritious with good amount of quality proteins, vitamins and minerals. An edible mushroom, Calocybe indica was selected to validate its nutritional and medicinal properties. Since tissue damage in hyperglycemia has been related to oxidative stress, we evaluated the enzymatic and non-enzymatic antioxidant status in the serum, liver and kidney since they are the target organs in diabetic complications. From the results, increased oxidative stress and decreased antioxidants might be related to the causation of diabetes mellitus. The treatment in the diabetic rats with the Calocybe indica showed an increase in the antioxidant system and decrease in the production of free radicals. The mushrooms which contain antioxidant phytochemicals has potential free radical scavenging capacity and hence can induce the antioxidant system in the body significantly reduces the generated free radicals thereby maintaining the normal levels of the antioxidants

Keywords: Antioxidants, Calocybe indica, diabetes mellitus, edible mushroom, oxidative stress.

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Authors: J. Suwanprateeb, F. Thammarakcharoen

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Calcium phosphate coating (CaP) has been employed for protein delivery, but the typical direct protein adsorption on the coating led to low incorporation content and fast release of the protein from the coating. By using bovine serum albumin (BSA) as a model protein, rapid biomimetic co-precipitation between calcium phosphate and BSA was employed to control the distribution of BSA within calcium phosphate coating during biomimetic formation on titanium surface for only 6 h at 50oC in an accelerated calcium phosphate solution. As a result, the amount of BSA incorporation and release duration could be increased by using a rapid biomimetic coprecipitation technique. Up to 43 fold increases in the BSA incorporation content and the increase from 6 h to more than 360 h in release duration compared to typical direct adsorption technique were observed depending on the initial BSA concentration used during coprecipitation (1, 10 and 100 μg.ml-1). From x-ray diffraction and Fourier transform infrared spectroscopy studies, the coating composition was not altered with the incorporation of BSA by this rapid biomimetic co-precipitation and mainly comprised octacalcium phosphate and hydroxyapatite. However, the microstructure of calcium phosphate crystals changed from straight, plate-like units to curved, plate-like units with increasing BSA content.

Keywords: Biomimetic, Calcium Phosphate Coating, Protein, Titanium.

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Authors: R. F. Ajayi, U. Sidwaba, U. Feleni, S. F. Douman, E. Nxusani, L. Wilson, C. Rassie, O. Tovide, P. G. L. Baker, S. L. Vilakazi, R. Tshikhudo, E. I. Iwuoha

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Pyrazinamide (PZA) is among the first-line pro-drugs  in the tuberculosis (TB) combination chemotherapy used to treat  Mycobacterium tuberculosis. Numerous reports have suggested that  hepatotoxicity due to pyrazinamide in patients is due to inappropriate  dosing. It is, therefore necessary to develop sensitive and reliable  techniques for determining the PZA metabolic profile of diagnosed  patients promptly and at point-of-care. This study reports the  determination of PZA based on nanobiosensor systems developed  from disuccinimidyl octanedioate modified Cytochrome P450-2E1  (CYP2E1) electrodeposited on gold substrates derivatised with  (poly(8-anilino-1-napthalene sulphonic acid) PANSA/PVP-AgNPs  nanocomposites. The rapid and sensitive amperometric PZA  detection gave a dynamic linear range of 2µM to 16µM revealing a  limit of detection of 0.044µM and a sensitivity of 1.38µA/µM. The  Michaelis-Menten parameters; KM, KM app and IMAX were calculated to  be 6.0µM, 1.41µM and 1.51x10-6 A, respectively, indicating a  nanobiosensor suitable for use in serum.

Keywords: Cytochrome P450-2E1, Disuccinimidyl octanedioate, Pyrazinamide, Tuberculosis.

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Authors: Natasha Hussain, Maleeha Aslam, Robina Farooq

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Hepatitis B and hepatitis C are among the most significant hepatic infections all around the world that may lead to hepatocellular carcinoma. This study is first time performed at the blood transfussion centre of Omar hospital, Lahore. It aims to determine the sero-prevalence of these diseases by screening the apparently healthy blood donors who might be the carriers of HBV or HCV and pose a high risk in the transmission. It also aims the comparison between the sensitivity of two diagnostic tests; chromatographic immunoassay – one step test device and Enzyme Linked Immuno Sorbant Assay (ELISA). Blood serum of 855 apparently healthy blood donors was screened for Hepatitis B surface antigen (HBsAg) and for anti HCV antibodies. SPSS version 12.0 and X2 (Chi-square) test were used for statistical analysis. The seroprevalence of HCV was 8.07% by the device method and by ELISA 9.12% and that of HBV was 5.6% by the device and 6.43% by ELISA. The unavailability of vaccination against HCV makes it more prevalent. Comparing the two diagnostic methods, ELISA proved to be more sensitive.

Keywords: ELISA, Sensitivity comparison of diagnostic tests, seroprevalence of Hepatitis B and C

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Authors: K. Mahmoud, T. Shalahmetova, B. Umbayev, Sh. Deraz

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Biochemical investigations were carried out to assess the effect of different exposure regimes of Kazakhstan crude oil (KCO) on hepatic antioxidant defense system in albino rats. Contaminants were delivered under two different dosing regimes, with all treatments receiving the same total contaminant load by the end of the exposure period. Rats in regime A injected with KCO once at a dose of 6 ml/kg bw while in regime B injected multiply at a dose of 1.5 ml/kg bw on day 1, 3, 5 and 8. Antioxidant biomarkers were measured in hepatic tissue after 1, 3, 5 and 8 days. Significant induction was observed in serum aminotransferases (ALT, AST) (p<0.01) and hepatic Glutathione-S-transferase (GST) (p<0.05) in the two exposure regimes, with the majority of significant induction occurring in regime A. Superoxide dismutase (SOD) increased 1-d after injection (p<0.01) but the increase was reduced time dependently thereafter while after 8-d induced again (p<0.01). Malondialdehyde (MDA) significantly induced after 3 and 5-d (p<0.05) in regime A while in regime B was not changed significantly (p>0.05) at short time after exposure. However, there was significant increase after 8-d (p<0.01). Histological examination indicates that crude oil induced pathologic changes from inflammatory cells infiltration to hemorrhage and necrosis of hepatocytes. Acute exposure to crude oil adversely affect hepatic cell so human must avoid such exposure.

Keywords: Kazakhstan crude oil, Antioxidant biomarkers, Histological examination, Dose regime, Rats

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Authors: Gry M, Bergström J, Lengquist J, Lindberg J, Drobin K, Schwenk J, Nilsson P, Schuppe-Koistinen I.

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Sensitive and predictive DILI (Drug Induced Liver Injury) biomarkers are needed in drug R&D to improve early detection of hepatotoxicity. The discovery of DILI biomarkers that demonstrate the predictive power to identify individuals at risk to DILI would represent a major advance in the development of personalized healthcare approaches. In this healthy volunteer acetaminophen study (4g/day for 7 days, with 3 monitored nontreatment days before and 4 after), 450 serum samples from 32 subjects were analyzed using protein profiling by antibody suspension bead arrays. Multiparallel protein profiles were generated using a DILI target protein array with 300 antibodies, where the antibodies were selected based on previous literature findings of putative DILI biomarkers and a screening process using pre dose samples from the same cohort. Of the 32 subjects, 16 were found to develop an elevated ALT value (2Xbaseline, responders). Using the plasma profiling approach together with multivariate statistical analysis some novel findings linked to lipid metabolism were found and more important, endogenous protein profiles in baseline samples (prior to treatment) with predictive power for ALT elevations were identified.

Keywords: DILI, Plasma profiling, PLSDA, Randomforest.

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Authors: Nneka N. Uchegbu

Abstract:

Physical, sensory properties and hypoglycemic effect of crackers produced from sprouted pigeon pea, unripe plantain and brewers’ spent grain fed to diabetic rats were investigated. Different composite flours were used to produce crackers. Physical and sensory properties of the crackers, the blood serum of the rats and changes in the rat body weight were measured. Spread ratio and break strength of the crackers from different flour blends ranges from 7.01 g to 8.51 g and 1.87 g to 3.01 g respectively. The acceptability of the crackers revealed that Sample A (100% wheat crackers) was not significantly (p>0.05) different from Samples C and D. Feeding the rats with formulated crackers caused an increase in the body weight of the rats but a reduced body weight was observed in diabetic rats fed with normal rat feed. The result indicated that cracker produced from the formulated flour blends caused a significant hypoglycemic effect in diabetic rats and led to a reduction of measured biochemical indices. Therefore, this work showed that consumption of crackers from the above formulated flour blend was able to decrease hyperglycemia in diabetic rats.

Keywords: Hypoglycemia, hyperlipidemia, total lipid, triglyceride, total cholesterol.

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Authors: M Nazrul-Hakim, A Yaacob, Y Adam, A Zuraini

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This study examined the toxicological effects and safety of polypeptide k isolated from the seeds of Momordica charantia in laboratory rats. 30 male Sprague Dawley rats (12 weeks old, bodyweight 180-200 g) were randomly divided into 3 groups (1000 mg/kg, 500 mg and 0 mg/kg). Rats were acclimatized to laboratory conditions for 7 days and at day 8 rats were dosed orally with polypeptide k (in 2% DMSO/normal saline) and the controls received the dosed vehicle only. Rats were then observed for 72 hours before sacrificed. Rats were anaesthetized by pentobarbital (50 mg/kg ip) and 2-3.0 mL of blood was taken by cardiac puncture and rats were scarified by anaesthetic overdose. Immediately, organs (heart, lungs, liver, kidneys) were weigh and taken for histology. Organ sections were then evaluated by a histopathologist. Serum samples were assayed for liver functions (ALT and γ-GT) and kidney functions (BUN and creatinine). All rats showed normal behavior after the dosing and no statistical changes were observed in all blood parameters and organ weight. Histological examinations revealed normal organ structures. In conclusion, dosing of rats up to 1000 mg/kg did not have any effects on the rat behavior, liver or kidney functions nor histology of the selected organs.

Keywords: Polypeptide k, safety, histology, toxicology, Momordica charantia

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Authors: Permphan Dharmasaroja

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Mutations of the telomeric copy of the survival motor neuron 1 (SMN1) gene cause spinal muscular atrophy. A deletion of the Eef1a2 gene leads to lower motor neuron degeneration in wasted mice. Indirect evidences have been shown that the eEF1A protein family may interact with SMN, and our previous study showed that abnormalities of neuromuscular junctions in wasted mice were similar to those of Smn mutant mice. To determine potential colocalization between SMN and tissue-specific translation elongation factor 1A2 (eEF1A2), an immunochemical analysis of HeLa cells transfected with the plasmid pcDNA3.1(+)C-hEEF1A2- myc and a new quantitative test of colocalization by intensity correlation analysis (ICA) was used to explore the association of SMN and eEF1A2. Here the results showed that eEF1A2 redistributed from the cytoplasm to the nucleus in response to serum and epidermal growth factor. In the cytoplasm, compelling evidence showed that staining for myc-tagged eEF1A2 varied in synchrony with that for SMN, consistent with the formation of a SMN-eEF1A2 complex in the cytoplasm of HeLa cells. These findings suggest that eEF1A2 may colocalize with SMN in the cytoplasm and may be a component of the SMN complex. However, the limitation of the ICA method is an inability to resolve colocalization in components of small organelles such as the nucleus.

Keywords: Intensity correlation analysis, intensity correlation quotient.

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Authors: Zarmina Gillani, Nuzhat Huma, Aysha Sameen, Mulazim Hussain Bukhari

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Whey is an excellent food ingredient owing to its high nutritive value and its functional properties. However, composition of whey varies depending on composition of milk, processing conditions, processing method, and its whey protein content. The aim of this study was to prepare a whey powder from raw whey and to determine the influence of different processing temperatures (160 and 180 °C) on the physicochemical, functional properties during storage of 180 days and on whey protein denaturation. Results have shown that temperature significantly (P < 0.05) affects the pH, acidity, non-protein nitrogen (NPN), protein total soluble solids, fat and lactose contents. Significantly (p < 0.05) higher foaming capacity (FC), foam stability (FS), whey protein nitrogen index (WPNI), and a lower turbidity and solubility index (SI) were observed in whey powder processed at 160 °C compared to whey powder processed at 180 °C. During storage of 180 days, slow but progressive changes were noticed on the physicochemical and functional properties of whey powder. Reverse phase-HPLC analysis revealed a significant (P < 0.05) effect of temperature on whey protein contents. Denaturation of β-Lactoglobulin is followed by α-lacalbumin, casein glycomacropeptide (CMP/GMP), and bovine serum albumin (BSA).

Keywords: Whey powder, temperature, denaturation, reverse phase – HPLC.

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Authors: Salim Cerig, Fatime Geyikoglu, Murat Bakir, Suat Colak, Merve Sonmez, Kubra Koc

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Cisplatin (CIS) is one of the most effective an anticancer drug and also toxic to cells by activating oxidative stress. Oleuropein (OLE) has key role against oxidative stress in mammalian cells, but the role of this antioxidant in the toxicity of CIS remains unknown. The aim of the present study was to investigate the efficacy of OLE on CIS-induced liver damages in male rats. With this aim, male Sprague Dawley rats were randomly assigned to one of eight groups: Control group; the group treated with 7 mg/kg/day CIS; the groups treated with 50, 100 and 200 mg/kg/day OLE (i.p.); and the groups treated with OLE for three days starting at 24 h following CIS injection. After 4 days of injections, serum was provided to assess the blood AST, ALT and LDH values. The liver tissues were removed for histological, biochemical (TAC, TOS and MDA) and genotoxic evaluations. In the CIS treated group, the whole liver tissue showed significant histological changes. Also, CIS significantly increased both the incidence of oxidative stress and the induction of 8-hydroxy-deoxyguanosine (8-OH-dG). Moreover, the rats taking CIS have abnormal results on liver function tests. However, these parameters reached to the normal range after administration of OLE for 3 days. Finally, OLE demonstrated an acceptable high potential and was effective in attenuating CIS-induced liver injury. In this trial, the 200 mg/kg dose of OLE firstly appeared to induce the most optimal protective response.

Keywords: Antioxidant response, cisplatin, histology, liver, oleuropein, 8-OhdG.

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Authors: Gouda K. Helal, Marwa I. Shabayek, Heba A. El-Ramly, Heba A. Awida

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The adverse effects of Clindamycin (Clind.) / Ibuprofen (Ibu.) combination on liver, kidney, blood elements and the significances of antioxidants (N-acetylcysteine and Zinc) against these effects were evaluated. The study includes: Group I; control n=30, Group II; patients on Clind.300mg/Ibu.400mg twice daily for a week n=30, Group III; patients on Clind.300mg/Ibu.400mg+Nacetylcysteine 200mg twice daily for a week n=15 and Group IV; patients on Clind.300mg/Ibu.400mg+Zinc50mg twice daily for a week n=15. Serum malondialdehyde (MDA), alanine transferase (ALT), aspartate transferase (AST), γ glutamyl transferase (GGT), creatinine, blood urea nitrogen (BUN) were measured. Applying one way ANOVA followed by Tuckey Kramer post test, Group II showed significant increase in ALT, AST, GGT, BUN and decrease in Hb, RBCs, platelets than Group I. Group III showed significant decrease in ALT, AST, GGT, BUN than Group II. Moreover, Group IV showed significant decrease in ALT, AST, GGT and increase in Hb, RBCs, and platelets than Group II. Conclusively, Adding Zinc or Nacetylcysteine buffer the oxidative stress and improve the therapeutic outcome of Clindamycin/Ibuprofen combination.

Keywords: Clindamycin, Ibuprofen, Adverse effects, Antioxidant, Zinc, N-acetylcysteine.

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Authors: Li Bixia, Ren Shouwen, Fu Yanfeng, Tu Feng, Xiaoming Fang, Xueming Wang

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The utilization of dietary crude fiber in different breed pigs is not the same. Su-shan pigs are a new breed formed by crossing Taihu pigs and Yorkshire pigs. In order to understand the resistance of Su-shan pigs to dietary crude fiber, 150 Su-shan pigs with 60 kg of average body weight and similar body conditions were allocated to three groups randomly, and there are 50 pigs in each group. The percentages of dietary crude fiber were 8.35%, 9.10%, and 11.39%, respectively. At the end of the experiment, 15 pigs randomly selected from each group were slaughtered. The results showed as follows: average daily gain of the 9.10% group was higher than that of the 8.35% group and the 11.39% group; there was a significant difference between the 9.10% group and the 8.35% group (p < 0.05. Levels of urea nitrogen, total cholesterol and high density lipoprotein in the 9.10% group were significantly higher than those in the 8.35% group and the 11.39% group (p < 0.05). Ratios of meat to fat in the 9.10% group and the 11.39% group were significantly higher than that in the 8.35% group (p < 0.05). Lean percentage of 9.10% group was higher than that of 8.35% group and 11.39% group, but there was no significant difference in three groups (p > 0.05). The weight of small intestine and large intestine in the 11.39% group was higher than that in the 8.35% group, and the 9.10% group and the difference reached a significant level (p < 0.05). In conclusion, increasing dietary crude fiber properly could reduce fat percentage, and improve the ratio of meat to fat of finishing Su-shan pigs. The digestion and metabolism of dietary crude fiber promoted the development of stomach and intestine of finishing Su-shan pig.

Keywords: Su-Shan pigs, dietary crude fiber, growth performance, serum biochemical indexes.

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

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

Keywords: Bisphenol A, oxidative stress, rabbits, semen quality, steroidogenesis.

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Authors: Robles F, Uribe A, Guadarrama A , Castellanos L, González C.

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The main objective of this study was to demonstrate that differentiation of infected and vaccinated animals (DIVA) strategy using different ELISA tests is possible when a subunit vaccine (Haemagglutinin protein) is used to prevent Avian influenza. Special emphasis was placed on the differentiation in the serological response to different components of the AIV (Nucleoprotein, Neuraminidase, Haemagglutinin, Nucleocapsid) between chickens that were vaccinated with a whole virus kill vaccine and recombinant vaccine. Furthermore, the potential use of this DIVA strategy using ELISA assays to detect Neuraminidase 1 (N1) was analyzed as strategy in countries where the field virus is H5N1 and the vaccine used is formulated with H5N2. Detection of AIV-s antibodies to any component in serum was negative for all animals on the study days 0-13. At study day 14 the titers of antibodies against Nucleoprotein (NP) and Nucleocapsid (NC) rose in the experimental groups vaccinated with Volvac® AI KV and were negatives during all the trial in the experimental groups vaccinated with a subunit H5; significant statistically differences were observed between these groups (p < 0.05). The seroconversion either Haemagglutinin or Neuraminidase was evident after 21 days post-vaccination in the experimental groups vaccinated with the respective viral fraction. Regarding the main aim of this study and according with the results that were obtained, use a combination of different ELISA test as a DIVA strategy is feasible when the vaccination is carry out with a subunit H5 vaccine. Also is possible to use the ELISA kit to detect Neuraminidase (either N1 or N2) as a DIVA concept in countries where H5N1 is present and the vaccination programs are done with H5N2 vaccine.

Keywords: Avian Influenza Virus, "DIVA concept", ELISAassay, subunit H5 vaccine.

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Authors: N. Baspinar, A. Basoglu, O. Ozdemir, C. Ozel, F. Terzi, O. Yaman

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Current research is targeting new molecular mechanisms that underlie non-alcoholic fatty liver disease (NAFLD) and associated metabolic disorders like non-alcoholic steatohepatitis (NASH). Forty New Zealand White rabbits have been used and fed a high protein (HP) and energy diet based on grains and containing 11.76 MJ/kg. Boron added to 3 experimental groups’ drinking waters (30 mg boron/L) as boron compounds. Biochemical analysis including boron levels, and nuclear magnetic resonance (NMR) based metabolomics evaluation, and mRNA expression of peroxisome proliferator-activated receptor (PPAR) family was performed. LDLcholesterol concentrations alone were decreased in all the experimental groups. Boron levels in serum and feces were increased. Content of acetate was in about 2x higher for anhydrous borax group, at least 3x higher for boric acid group. PPARα mRNA expression was significantly decreased in boric acid group. Anhydrous borax attenuated mRNA levels of PPARγ, which was further suppressed by boric acid. Boron supplementation decreased the degenerative alterations in hepatocytes. Except borax group other boron groups did not have a pronounced change in tubular epithels of kidney. In conclusion, high protein and energy diet leads hepatocytes’ degenerative changes which can be prevented by boron supplementation. Boric acid seems to be more effective in this situation.

Keywords: High protein and energy diet, boron, metabolomic, transcriptomic.

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Authors: S. Shalaweh, P. Bouic, F. Weitz, R. Henkel

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More than 3000 plants of notable phyto-therapeutic value grow in South Africa; these include Cissampelos capensis, commonly known in Afrikaans as dawidjie or dawidjiewortel. C. capensis is the most significant and popular medicinal plant used by the Khoisan as well as other rural groups in the Western region of South Africa. Its rhizomes are traditionally used to treat male fertility problems. Yet, no studies have investigated the effects of this plant or its extracts on human spermatozoa. Therefore, this study aimed at investigating the effects of C. capensis rhizome extract (CRE) fractions on ejaculated human spermatozoa in vitro. Spermatozoa from a total of 77 semen samples were washed with human tubular fluid medium supplemented with bovine serum albumin (HTF-BSA) and incubated for 2 hours with 20 μg/ml progesterone (P4) followed by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200 μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH, F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A sample without addition of CRE fractions served as control. Samples were analyzed for sperm motility, reactive oxygen species (ROS), DNA-fragmentation, acrosome reaction and capacitation. Results showed that F1 resulted in significantly higher values for ROS, capacitation and hyper-activation compared to F2, F3, and F4 with P4-stimulated samples generally having higher values. No significant effect was found for the other parameters. In conclusion, alkaloids present in F1 of CRE appear to have triggered sperm intrinsic ROS production leading to sperm capacitation and acrosome reaction induced by P4.

Keywords: Capacitation, acrosome reaction, Cissampelos capensis, DNA fragmentation, ROS.

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Authors: Duane Barker, John Holliday

Abstract:

In a 10-week (May – August, 2008) Phase I trial, 840, 1+ rainbow trout, Oncorhynchus mykiss, received a commercial oral immunomodulator, Fin Immune™, at four different dosages (0, 10, 20 and 30 mg g-1) to evaluate immune response and growth. The overall objective of was to determine an optimal dosage of this product for rainbow trout that provides enhanced immunity with maximal growth and health. Biweekly blood samples were taken from 10 randomly selected fish in each tank (30 samples per treatment) to evaluate the duration of enhanced immunity conferred by Fin-Immune™. The immunological assessment included serum white blood cell (lymphocyte, neutrophil) densities and blood hematocrit (packed cell volume %). Of these three variables, only lymphocyte density increased significantly among trout fed Fin- Immune™ at 20 and 30 mg g-1 which peaked at week 6. At week 7, all trout were switched to regular feed (lacking Fin-Immune™) and by week 10, lymphocyte levels decreased among all levels but were still greater than at week 0. There was growth impairment at the highest dose of Fin-Immune™ tested (30 mg g-1) which can be associated with a physiological compensatory mechanism due to a dose-specific threshold level. Thus, our main objective of this Phase I study was achieved, the 20 mg g-1 dose of Fin-Immune™ should be the most efficacious (of those we tested) to use for a Phase II disease challenge trial.

Keywords: Blood Lymphocyte, Neutrophil Response of Cultured Rainbow Trout, Oncorhynchus mykiss, Oral Immunomodulator – 'Fin-ImmuneTM'.

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Authors: Maryam Nazari, Majid Karandish, Alihossein Saberi

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Obesity as a global health challenge motivates pharmaceutical industries to produce anti-obesity drugs. However, effectiveness of these agents is remained unclear. Because of popularity of dietary supplements, the aim of this study was tp investigate the effects of Conjugated Linoleic Acid (CLA) and L-carnitine (LC) on serum glucose, triglyceride, cholesterol and weight changes in diet induced obese rats. 48 male Wistar rats were randomly divided into two groups: Normal fat diet (n=8), and High fat diet (HFD) (n=32). After eight weeks, the second group which was maintained on HFD until the end of study, was subdivided into four categories: a) 500 mg Corn Oil (as control group), b) 500 mg CLA, c) 200 mg LC, d) 500 mg CLA+ 200 mg LC.All doses are planned per kg body weights, which were administered by oral gavage for four weeks. Body weights were measured and recorded weekly by means of a digital scale. At the end of the study, blood samples were collected for biochemical markers measurement. SPSS Version 16 was used for statistical analysis. At the end of 8^th week, a significant difference in weight was observed between HFD and NFD group. After 12 weeks, LC significantly reduced weight gain by 4.2%. Trend of weight gain in CLA and CLA+LC groups was insignificantly decelerated. CLA+LC reduced triglyceride level significantly, but just CLA had significant influence on total cholesterol and insignificant decreasing effect on FBS. Our results showed that an obesogenic diet in a relative short time led to obesity and dyslipidemia which can be modified by LC and CLA to some extent.

Keywords: Conjugated linoleic acid, high fat diet, L-carnitine, obesity.

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Authors: Rohayu Izanwati M. R., Muhamad Ridhwan M. R., Abbe Maleyki M. J., Ahmad Zubaidi A. L., Zahri M. K.

Abstract:

PPARs function as regulators of lipid and lipoprotein metabolism. The aim of the study was to compare the lipid profile between two phases of fasting and to examine the frequency and relationship of peroxisome proliferator-activated receptor, PPARα gene polymorphisms to lipid profile in fasting respondents. We conducted a case-control study protocol, which included 21 healthy volunteers without gender discrimination at the age of 18 years old. 3 ml of blood sample was drawn before the fasting phase and during the fasting phase (in Ramadhan month). 1ml of serum for the lipid profile was analyzed by using the automated chemistry analyser (Olympus, AU 400) and the data were analysed using the Paired T-Test (SPSS ver.20). DNA was extracted and PCR was conducted utilising 6 sets of primer. Primers were designed within 6 exons of interest in PPARα gene. Genetic and metabolic characteristics of fasting respondents and controls were estimated and compared. Fasting respondents were significantly have lowered the LDL levels (p=0.03). There were no polymorphisms detected except in exon 1 with 5% of this population study respectively. The polymorphisms in exon 1 of the PPARα gene were found in low frequency. Regarding the 1375G/T and 1386G/T polymorphisms in the exon 1 of the PPARα gene, the T-allele in fasting phase had no association with the decreased LDL levels (Fisher Exact Test). However this association is more promising when the sample size is larger in order to elucidate the precise impact of the polymorphisms on lipid profile in the population. In conclusion, the PPARα gene polymorphisms do not appear to affect the LDL of fasting respondents.

Keywords: Fasting, LDL, Peroxisome proliferator activated receptor alpha (PPAR-α), Polymorphisms.

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Authors: A. Asambe, A. K. B. Sackey, L. B. Tekdek

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A serosurveillance study was conducted to detect the presence of antibodies to African swine fever virus (ASFV) and Classical swine fever virus in pigs sampled from piggeries and Makurdi central slaughter slab in Benue State, Nigeria. 416 pigs from 74 piggeries across 12 LGAs and 44 pigs at the Makurdi central slaughter slab were sampled for serum. The sera collected were analysed using Indirect Enzyme Linked Immunosorbent Assay (ELISA) test kit to test for antibodies to ASFV, while competitive ELISA test kit was used to test for antibodies to CSFV. Of the 416 pigs from piggeries and 44 pigs sampled from the slaughter slab, seven (1.7%) and six (13.6%), respectively, tested positive to ASFV antibodies and was significantly associated (p < 0.0001). Out of the 12 LGAs sampled, Obi LGA had the highest ASFV antibody detection rate of (4.8%) and was significantly associated (p < 0.0001). None of the samples tested positive to CSFV antibodies. The study concluded that antibodies to CSFV were absent in the sampled pigs in piggeries and at the Makurdi central slaughter slab in Benue State, while antibodies to ASFV were present in both locations; hence, the need to keep an eye open for CSF too since both diseases may pose great risk in the study area. Further studies to characterise the ASFV circulating in Benue State and investigate the possible sources is recommended. Routine surveillance to provide a comprehensive and readily accessible data base to plan for the prevention of any fulminating outbreak is also recommended.

Keywords: African swine fever, classical swine fever, piggery, slaughter slab, surveillance.

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Authors: Siobhan O’Shea, Sangeetha Vijaysri Nair, Hee Cheol Kim, Charles Thomas Nugent, Cheuk Yan William Tong, Sam Douthwaite, Andrew Worlock

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The Aptima® HIV-1 Quant Dx Assay is a fully automated assay on the Panther system. It is based on Transcription- Mediated Amplification and real time detection technologies. This assay is intended for monitoring HIV-1 viral load in plasma specimens and for the detection of HIV-1 in plasma and serum specimens. Nine-hundred and seventy nine specimens selected at random from routine testing at St Thomas’ Hospital, London were anonymised and used to compare the performance of the Aptima HIV-1 Quant Dx assay and Roche COBAS® AmpliPrep/COBAS® TaqMan® HIV-1 Test, v2.0. Two-hundred and thirty four specimens gave quantitative HIV-1 viral load results in both assays. The quantitative results reported by the Aptima Assay were comparable to those reported by the Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, v2.0 with a linear regression slope of 1.04 and an intercept on -0.097. The Aptima assay detected HIV-1 in more samples than the COBAS assay. This was not due to lack of specificity of the Aptima assay because this assay gave 99.83% specificity on testing plasma specimens from 600 HIV-1 negative individuals. To understand the reason for this higher detection rate a side-by-side comparison of low level panels made from the HIV-1 3rd international standard (NIBSC10/152) and clinical samples of various subtypes were tested in both assays. The Aptima assay was more sensitive than the COBAS assay. The good sensitivity, specificity and agreement with other commercial assays make the HIV-1 Quant Dx Assay appropriate for both viral load monitoring and detection of HIV-1 infections.

Keywords: HIV viral load, Aptima, Roche, Panther system.

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Authors: Yemisi Rufina Alli Smith, Isaac Gbadura Adanlawo

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Liver disorders are one of the major problems of the world. Despite its frequent occurrence, high morbidity and high mortality, its medical management is currently inadequate. This study was designed to evaluate the hepatoprotective effect of saponin extract of the root of Garcinia kola on the integrity of the liver of paracetamol induced wistar albino rats. Twenty five (25) male adult wistar albino rats were divided into five (5) groups. Group I was the Control group that received distilled water only, group II was the negative control that received 2 g/kg of paracetamol on the 13th day, and group III, IV and V were pre-treated with 100, 200 and 400mg/kg of the saponin extract before inducing the liver damage on the 13th day with 2 g/kg of paracetamol. Twenty four (24) h after administration, the rats were sacrificed and blood samples were collected. The serum Alanine Transaminase (ALT), Aspartate Transaminase (AST), Alkaline Phosphatase (ALP) activities, Bilirubin and conjugated bilirubin, glucose and protein concentrations were evaluated. The liver was fixed immediately in Formalin and was processed and stained in Haematoxylin and Eosin (H&E). Administration of saponin extract from the root of Garcinia kola significantly decreased paracetamol induced elevated enzymes in the test group. Also histological observations showed that saponin extract of the root of Garcinia kola exhibited a significant liver protection against the toxicant as evident by the cells trying to return to normal. Saponin extract from the root of Garcinia kola indicated a protection of structural integrity of the hepatocytic cell membrane and regeneration of the damaged liver.

Keywords: Garcinia kola, Hepatoprotective, paracetamol, Saponin.

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Authors: Vladyslav Povoroznyuk, Galina Dubetska, Roksolana Povoroznyuk

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In recent years, the problem of hyperuricemia is getting a particular importance due to its increased incidence in the world population. The aim of this study was to determine uriс acid level in blood serum, incidence of hyperuricemia among women in postmenopausal period and their association with body mass index and some components of metabolic syndrome (triglyceride, cholesterol, systolic and diastolic pressure). We examined 412 women in postmenopausal period. They were divided in to the following groups: I group (BMI = 18,5-24,9), II group (BMI = 25,0-29,9), III group (BMI = 30,0-34,9), IV group (BMI > 35). We determined uric acid level among women during postmenopausal period depending on their body mass index. The higher level of uric acid was found in patients with the maximal body mass index (BMI > 35). In the I group it was 277,52 ± 8,40; in the II group – 286,81 ± 7,79; in the III group – 291,81 ± 7,56; in the IV group – 327,17 ± 12,17. Incidence of hyperuricemia among women in the I group was 10,2%, in the II group – 15,9%; in the III group – 21,2%, in the IV group – 34,2%. We found an interdependence between an uric acid level and BMI in the examined women (r = 0,21, p < 0,05). We determined that the highest level of triglyceride (F = 18,62, p < 0,05), cholesterol (F = 3,64, p < 0,05), atherogenic coefficient (F = 22,64, p < 0,05), systolic (F = 10,5, p < 0,05) and diastolic pressure (F = 4,30, p < 0,05) was among women with hyperuricemia. It was an interdependence between an uric acid level and triglyceride (r = 0,26, p < 0,05), atherogenic coefficient (r = 0,24, p < 0,05) among women in postmenopausal period.

Keywords: Hyperuricemia, uric acid, body mass index, metabolic syndrome, triglyceride, cholesterol, atherogenic coefficient, systolic and diastolic pressure, women.

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