Research Journal of Biotechnology

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Research Journal of Biotechnology

Curcumin and Risendronate/hydroxyapatite co-loaded lipid-polymer nanoparticle to enhance the therapeutic efficacy in osteoporosis

Shi Li-Ping, Zhou Jian, Zhao Su-Ping and Wu You-Wei

In this study, a unique lipid polymer nanoparticle loaded with dual drug (CUM and RSN) was formulated to treat osteoporosis. For this purpose, PEG-lecithin coated PLGA NP (LCRHP NPs) was prepared by nanoprecipitation method. The main aim was to increase the stability, sustained release and internalization of nanoformulations in the osteoblast cells. The particles were nanosized indicating its suitability for bone applications. Morphology was investigated by the SEM which showed clear spherical shaped NPs with size less than 200 nm to treat the micropores in the bone. Both the drugs were released in a sustained manner from the nanoparticles throughout the study period. Moreover, a typical biphasic release pattern was observed which could be attributed to the presence of drug on the outer surface as well as on the inner core. Initial fast release of drug will enhance the treatment while slow release from core will maintain the drug reservoir effect in the bone site. Consistently, nanoparticle showed superior killing property and uptake in HFOb 1.19 cells. Most importantly, RSN-HA in the lipid polymer showed strong affinity towards the HA further enhancing the therapeutic efficacy in osteoporosis.

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16S rRNA gene sequencing and culture dependent analysis of bacterial diversity associated with commercially processed salads

Bano Abida and Ali Basharat

The main objective of the present study was to evaluate the biosafety of the raw-eaten processed salads by exploring the general microbial diversity. For this purpose, more than hundred bacterial strains were isolated from the processed salads collected from street restaurants from different localities in Lahore, Pakistan. Sequencing analysis of 16S rRNA gene showed the presence of bacterial strains belonging to the genera of Bacillus, Staphylococcus, Proteus, Macrococcus, Haemophilus, Escherichia, Citrobacter, Enterobacter, Salmonella, Shigella, Klebsiella and Acinetobacter. Antibiotic susceptibility pattern against different antibiotics showed that majority of the strains were resistant against amoxicillin and ampicillin. Strains were also evaluated for biofilm formation, swarming motility and slime production. Maximum biofilm production in single or double cultures was shown by B. cereus C35 and M. caseolyticus C96. In the presence of carbon sources, B. cereus C35 (1% sucrose), B. pumilus C49 (1% fructose) and H. influenzae C410 (3% lactose) showed significant biofilm formation. For swarming motility, P. mirabilis L21 and P. mirabilis B33 were strongly positive. In conclusion, processed salads from different localities of Lahore were inhabited by different potentially pathogenic bacterial strains. E. coli a potential human pathogen was isolated from samples which indicated fecal contamination. Additionally, pathogenic bacteria such as En. cloacae, K. pneumoniae, B. cereus, S. dysentriae, H. influenzae, S. saprophyticus were also identified that make the biosafety of salad sources questionable.

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Toxigenic profiles of diarrheal and emetic toxin in Oceanobacillus sojae and Oceanobacillus oncorhynchi strains isolated from Sichuan Pickle

Xiong H. and Cai T.

The diarrheal and emetic toxin production potential and toxin genes were investigated in twenty nine Oceanobacillus sojae and twenty two Oceanobacillus oncorhynchi from Sichuan Pickle, a typical traditional fermented vegetable product in China. All of O. sojae isolates harbored hblC, nheA and nheC diarrheal toxin genes while only those that possessed the cytK gene (about 86.2%) displayed haemolytic activity on blood agar. In O. oncorhynchi isolates, 59.09% possessed the complete hbl gene (A, C and D), nheB of nhe gene and cytK gene. The hbl and cytK acted together to cause higher haemolytic activity of these O. oncorhynchi isolates (with 15.0 ± 1.0 mm index). In emetic toxin study, all were negative for HEp-2 cell assay. The emetic toxin and toxin gene have not been detected in all Oceanobacillus sojae and twenty two Oceanobacillus oncorhynchi isolates. These results indicated that there is a potential risk of food safety when Sichuan Pickle is contaminated by O. sojae and O. oncorhynchi.

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32bits Arm cortex S3C2440 Microcontroller application for Transcranial Magnetic Stimulation’s Superposition pulse on cell death in ischemia induced rats

Whi-Young Kim

As the pulse train repetition ratio and pulse range are dominant factors, they achieve high treatment and high energy density. Achieving precise control according to purpose of treatment and diagnosis is important. A method for controlling the action time and changing the pulse train repetition range is used mainly through the output pulse of transcranial magnetic stimulation. The 32bits S3C2440 microprocessor was grafted to achieve safe control of circuit operation. Transcranial magnetic stimulation was operated through the power device and control section and research was carried out on the motion and output characteristics of transcranial magnetic stimulation according to the current pulse waveform and pulse train of the stimulation coil. As a result, the transcranial magnetic stimulation output efficiency was reduced and the transcranial magnetic stimulation output showed a regular increase of 50W according to the increase in pulse train repetition rate as pulse repetition rate was increased by 10 Hz from 10Hz to 60 Hz with a condenser charged voltage of 1,800V and 1,900V. This study introduces a treatment Pulse using Transcranial Magnetic Stimulation for changing various bacteria caused by rat head. One can obtain good changing characteristics of various bacteria by adjusting the charging voltage, the treatment pulse forming and the magnetic field inducing time.

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Encapsulation of probiotic bacteria with alginate-starch and evaluation of viability in storage conditions and in food

İşleyen M. Fatih and Çakır İbrahim

Lactobacillus acidophilus KPb4b and Lactobacillus rhamnosus KPb7 bacteria were selected as potential strains for probiotic food development after determining their 16S rRNA sequence and molecular diagnosis. These strains were microencapsulated with 1.5% alginate, 1.5% manucol and 1.5% alginate mixed with 0.5% starch. Coating process was made by using emulsion electrostatic vibration technique with Nisco modular microencapsulation unit Var A. The obtained capsules were between 370 μm and 404 μm in size. Storage stability was determined at temperatures of +24ºC, +5ºC and -18ºC. In addition, the stability of microcapsules was investigated in a model food environment. In terms of storage stability, storing at -18ºC was found to damage the structure of microbeads. It was determined that storage at +5ºC with alginate + starch capsules was the most appropriate temperature and coating material to maintain the viability of the bacteria.

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Biological Properties of 4-Benzyloxybenzaldehyde-4-Methyl-3-Thiosemicarbazone and its Cd(II) Complex

Suvarapu Lakshmi Narayana, Baek Sung-Ok, Vijaya T. and Venkateswarlu N.

The thiosemicarbazones can act as a monodentate ligand that binds to the metal ion through the sulphur atom or as a bidentate ligand that coordinates to the metal ion through the sulphur atom and one of the nitrogen atoms of the hydrazine moiety to form four or five membered chelate rings. The coordination capacity of thiosemicarbazones can be increased due to the presence of aldehydes or ketones containing additional functional group(s) in position(s) suitable for chelation. Apart from their interesting coordination chemistry, thiosemicarbazones have attracted considerable interest because of their potentially beneficial biological activities such as antiviral, antitumor, antimalarial, antifungal and antibacterial activities. The biological activity of thiosemicarbazones and metal-thiosemicarbazones has been receiving considerable attention recently. In the present study we investigated the biological activities of 4-Benzyl -oxybenzaldehyde-4-methyl-3-thiosemicarbazone and its Cd(II) complex. For this concern we evaluate the biological activities such as anti-bacterial activity against two pathogenic gram-positive bacteria such as Bacillus subtilis and Staphylococcus aureus and two gram-negative bacteria such as Escherichia Coli and Pseudomonas aeruginosa anti-fungal activity against Candida albicans and Candida tropicalisand anti-oxidant properties of the both ligand and its Cd(II) complex.

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Simple and rapid method for Rhodamine B determination in beverage samples by using AMWCNT dispersive SPE with icELISA and HPLC

Xixia Liu

Sample pretreatment method is critical for rhodamine B(RB) detection. In this study, the average concentrations required for 50% inhibition concentration (IC50) and the limit of detection (LOD) were 1.78±0.01 and 0.26±0.05 ng/mL respectively (n=6) after optimization of the indirect competitive enzyme-linked immunosorbent assay (icELISA) conditions. The linear response range extended from 0.49ng/mL to 9.56 ng/mL. High Performance Liquid Chromatography (HPLC) analysis showed that the LOD was 0.02 ng/mL, the limit of quantitation was 0.04 ng/mL and the linear response range extended from 0.5ng/mL to 500 ng/mL. The new amino multiwalled carbon nanotubes (AMWCNTs)dispersive solid-phase extraction (d-SPE) for sample pretreatment were established after optimizing the icELISA conditions. The d-SPE method was combined with icELISA and HPLC to determine the RB levels in beverage samples. Results indicated that the corresponding mean recoveries were 73.50%–89.50% and 72.53%–91.57%. Overall, the findings exhibited the potential to develop simple and rapid sample pretreatment commercial kits for RB detection.

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Characterization of bacterial and yeast populations in fermentation of finger millet (Eleusine coracana)

Ramakrishnan Sudha Rani, Chelliah Ramachandran and Antony Usha

Fermented finger millet porridge (koozh) is widely consumed traditional food in the southern parts of India. Indigenous microflora in koozh has not been studied in detail. In this study, Lactic acid bacteria (LAB) and yeast strains from koozh were isolated, identified and characterized based on phenotype and sequence of 16S and 18S ribosomal RNA. Three different strains were isolated from koozh. Among them, two were bacterial (Enterococcus faecalis and Lactobacillus casei) and one was yeast strain (Saccharomyces cerevisiae). L.casei is the dominant LAB followed by E.faecalis in the initial 18 h of fermentation. Yeast dominated (S.cerevisiae) at later stages till 48 h of fermentation that leads to softening of the product. The antibiotic sensitivity of LAB, E.faecalis and S.cerevisiae isolates were determined against seven different antibiotics with three different modes of actions. It was revealed that KS9 and KS4 strains showed inhibitory activity against the antibiotics but KS10 showed resistance. The results demonstrated that LAB species and yeast were dominant during fermentation. LAB and yeast affected the aroma and flavour of koozh. The characterization of the microbial population in the traditional fermented food and their activity in the presence of antibiotics were demonstrated.

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Optimization in pre-term birth classification with velocity based mutation of particle swarm optimizations

Thomas J. and Kulanthaivel G.

Preterm birth has become a usual happening in spite of recent advancement in monitoring the fetal in mother’s womb. In this research work we have proposed a novel algorithm PSO_DE where we have varied the velocity of every particles in the swarm by mutating the existing velocity with respect to addressing the current environment by selecting randomly the velocity of current generation individuals. The particle swarm optimization algorithm simulates the social behaviorisms of certain organisms in search of food. Evolutionary algorithm evolves better and better in each evolution to give a most optimized solution. The proposed novel algorithm provides a dynamism in searching the solution space which is the back bone of the evolutionary algorithmic approach for faster convergence of the solution being more precise in accuracy. This approach has proved to be very effective in improving the performance of predicting with an accuracy of 0.99935 in pre-term birth classification which helps medical practitioners to develop better diagnostic model.

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Acanthus ilicifolius Linn. enhances glucose uptake in L6 cell lines promising for an antidiabetic potential in streptozotocin induced type II diabetic rats

Mahalingam Gayathri and G.A. Gayathri

The aim of this study was to explore in vitro glucose uptake activity in L6 cell lines and anti-diabetic activity of methanolic leaf extract of Acanthus ilicifolius (MLEA) in streptozotocin (STZ) induced type II diabetic rats. In vitro cytotoxicity and glucose uptake assay were carried out in L6 skeletal muscle cells using standard protocols. In vivo acute toxicity study was performed for 14 days. Type 2 diabetes was confirmed after 4 days of single intraperitoneal injection of STZ (40mg/kg bw) in albino Wistar rats. MLEA (100, 200 mg/kg bw) and metformin (500 mg/kg bw) were administered orally for 28 days. Body weight and glucose level from tail vein were monitored at regular interval. After 48 days, overnight fasted rats were sacrificed and blood was collected for further biochemical analysis. MLEA did not show any mortality up to 3000 mg/kg bw. Diabetic rats showed significant changes in fasting blood glucose, HbA1C, when compared with diabetic control group. There was significant elevation in body weight of diabetic treated rats. Thus, it was concluded that MLEA triggers glucose uptake in L6 cell lines and possesses antidiabetic activity in STZ induced diabetic rats.

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An effective protocol for regenerating mature Pinus massoniana L. trees by tissue culture

Yao Rui-Ling and Wang Yin

The propagation of adult pine trees by tissue culture has been studied for a few decades but problems related to the juvenile to adult phase change of trees have limited the practical applications of these tissue culture procedures. This study describes a micropropagation protocol for the in vitro propagation of mature Pinus massoniana trees. In this study, nodal segments were used as the explants for in vitro regeneration of adult P. massoniana trees (25-30 years old). The sampled explants were cultured on four media (DCR, SH, GD and modified MS (MMS)) supplemented with three cytokinins (BA, KT and ZT) at three different concentrations (5, 10 and 20 μM). The evaluated culture media and cytokinin treatments showed significant differences in organogenic responses. The highest organogenic response was obtained with nodal segments cultured on MMS media and by explants cultured on medium supplemented with 10 μM zeatin (ZT). A successful and efficient protocol is first developed for the micropropagation of adult P. massoniana trees by biotechnological approach.

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A New Spectrophotometric Assay Method for omega-Transaminase using 2,4-Dinitrophenylhydrazine

Jeon Hyunwoo, Ravikumar Yuvaraj, Nadarajan Saravanan Prabhu and Yun Hyungdon

A new spectrophotometric assay method using 2,4-dinitrophenylhydrazine was developed to determine the activity of -transaminase. 2,4-Dinitrophenylhydrazine reacts with ketone produced during -transaminase reaction to form a orange color hydrazone which can be easily quantified using UV/Vis spectrophotometer at 450 nm. The results obtained by spectrophotometric assay with 2,4-dinitrophenylhydrazine and conventional HPLC analysis agreed well. -Transaminase reactions were analyzed in 96-well microplates using the 2,4-dinitrophenylhydrazine, leading to quite simple, but agreeable rapid determination of the reaction profile and apparent enantioselectivity of the enzyme.

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Comparing the clinical outcomes of shoulder hemiarthroplasty versus locking plate fixation for the treatment of complex proximal humeral fractures in elderly patients

Gu Jun, Cai Weihua, Ni Yingjie, Sun Jianfei, Fang Jiahu and Yang Zibin

The aim of this study was to compare the clinical efficacy and complications of hemiarthroplasty versus locking plate fixation for the treatment of complex 3- or 4-part proximal humeral fracture (PHF) in the elderly population. A total of 156 elderly patients at age of > 65 years with complex PHF who underwent shoulder hemiarthroplasty or locking plate fixation during the period from January 2006 to June 2014 were enrolled in the study including 74 patients undergoing hemiarthroplasty and 82 patients with locking plate fixation. The clinical outcomes and complications were retrospectively evaluated and compared. Hemiarthroplasty had a shorter duration of operation (74 ± 18 vs. 86 ± 20 min, P = 0.003), less intraoperative blood loss (135 ± 40 vs. 320 ± 45 ml, P = 0.001) and higher Neer score (91.2 ± 12.5 vs. 77.0 ± 9.2, P = 0.001) and constant-murley score (86.7 ± 12.8 vs. 74.5 ± 15.1, P = 0.002). There was no significant difference in the incidence of postoperative complications between the two treatment groups (P > 0.05). It is concluded that both hemiarthroplasty and locking plate fixation are effective for complex PHF in the elderly patients. However, hemiarthroplasty should be given a high priority if patient’s demand for joint function is not strict and the economic condition is permitted while locking plate fixation is preferentially applied if the preoperative activity is fine, or the patients’ functional requirement is high.

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Morphological traits alteration of mutant common turf grass (Cynodon dactylon) induced by gamma ray irradiation

Azahar Mohd Abdul Halim Bin Baharun, Juraimi Abdul Shukor, Yusof Mohd Rafii, Harun Abdul Rahim, Kamaluddin Md and Alam Md Amirul

The experiment was conducted to study Cynodon dactylon morphological improvement and breeding by induced mutation using gamma ray irradiation at Universiti Putra Malaysia (UPM) and Malaysian Institute of Nuclear Technology Research (MINT) center. C. dactylon is a widely used turf in Malaysia especially for golf course and football field. However, its coarse leaf texture and long internodes are undesirable for good quality golf field. In this purpose mutagenesis by gamma ray irradiation was employed using 0, 20, 40, 60, 80, 100, 120 and 140 Gy to treat 30 single node stolons per treatment. Dosages of 90 Gy were determined as LD50 for the radio sensitivity test. Survival rate of C. dactylon stolon was greatly reduced when irradiated with higher dosages. This experiment was repeated using LD50 on 1500 single node stolons. Twenty two (22) morphological mutants were identified and evaluated. Most mutants were semi-dwarf type with reduced internode length and leaf blade length. The altered morphological traits were stable after third cutting back (M1V3) shown by their morphological performance. Mutation breeding is effective in improving C. dactylon when easily recognized cultivars are needed.

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