Can low dose of intratracheal dexmedetomidine be used to attenuate peri-extubation cough? - A prospective, double-blinded, randomized clinical trial.

Background: Peri-extubation cough is an undesirable event during extubation, prevention of which has been studied with multiple drugs, amongst which intravenous dexmedetomidine has emerged as one of the favourable drugs. Intratracheal route is attractive because of its ease of administration, provided it avoids the hypotension and bradycardia that occurs during intravenous bolus administration. There is a paucity of data exploring the utility, doses, and adverse effect of intratracheal dexmedetomidine. Methods: After obtaining ethical committee approval, 60 eligible, consenting adult patients undergoing surgery under general anesthesia in a tertiary teaching hospital were recruited and randomised into three groups-DEX0.3, DEX0.5, and NS. The plan of general anesthesia was standardized. Half an hour prior to extubation, study drug was instilled intratracheally-dexmedetomidine 0.3 mic/kg, 0.5 mic/kg, and NS in groups DEX0.3, DEX0.5, and NS, respectively. 4-point cough score was used to assess extubation response. Hemodynamic response and time to Ramsay sedation score 3 was also recorded. Results: Majority of patients in DEX0.3 (60%) and DEX0.5 (85%) group had no cough (cough score 0), while majority of the patients in the NS group (70%) had either mild or moderate cough (cough score 1, 2). Kruskal Wallis test followed by post-hoc pairwise comparison showed statistically significant difference in 4-point cough score between GroupDEX0.3 and GroupNS (P < 0.001) and between GroupDEX0.5 and GroupNS (P = 0.038). DEX0.5 group, compared to DEX0.3 group, had significantly higher time from reversal to extubation (P < 0.001) and time to achieve Ramsay sedation score of 3 (P < 0.001). Conclusion: We conclude that both 0.3 mic/kg and 0.5 mic/kg of dexmedetomidine when given intratracheally are effective in preventing peri-extubation cough. Further, 0.3 mic/kg dexmedetomidine showed a better recovery profile compared to 0.5 mic/kg dexmedetomidine when administered intratracheally.

Multifunctional applications of natural polysaccharide starch and cellulose: An update on recent advances.

The emergence of clinical complications and therapeutic challenges for treating various diseases necessitate the discovery of novel restorative functional materials. Polymer-based drug delivery systems have been extensively reported in the last two decades. Recently, there has been an increasing interest in the progression of natural biopolymers based controlled therapeutic strategies, especially in drug delivery and tissue engineering applications. However, the solubility and functionalisation due to their complex network structure and intramolecular bonding seem challenging. This review explores the current advancement and prospects of the most promising natural polymers such as cellulose, starch and their derivatives-based drug delivery vehicles like hydrogels, films and composites, in combating major ailments such as bone infections, microbial infections, and cancers. In addition, selective drug targeting using metal-drug (MD) and MD-based polymeric missiles have been exciting but challenging for its application in cancer therapeutics. Owing to high biocompatibility of starch and cellulose, these materials have been extensively evaluated in biomedical and pharmaceutical applications. This review presents a detailed impression of the current trends for the construction of biopolymer-based tissue engineering, drug/gene/protein delivery vehicles.

Ror ß expression in activated macrophages and dental pulp stem cells.

AIM: To assess the expression of Retinoic acid-related orphan receptor beta (Ror ß) in human inflamed dental pulp stem cells (hI-DPSCs) and during macrophage phenotypic conversion. METHODOLOGY: Commercially procured THP-1 monocytes conversion to macrophages was judged by their morphology, the percentage of adherent cells and the expression of CD-14 surface marker. THP-1 macrophage cell viability following LPS, IFN-γ/IL-4, IL-13 stimulus was evaluated at 24 and 48h. The phenotypic conversion of macrophages to M1 and M2 was confirmed by flow cytometry and Western blot analysis. Cytokine release following polarization was estimated by the BD cytokine flex kit. The expression of Ror ß in THP-1 macrophages and hI-DPSCs following LPS, IFN-γ/IL-4, IL-13 stimulus was assessed by Western blot analysis. Statistical significance was analysed using one-way Anova followed by Tukey's Post hoc test. RESULTS: THP-1 monocytes pretreated with PMA (100 ng mL-1 ) for 48 h followed by culturing in PMA-free media for another 48 h yielded cells with morphological characteristics similar to macrophages with a high percentage of adherence capability and CD-14 expression. Macrophages treated with LPS 100 ng mL-1 and IFN-γ 20 ng mL-1 or IL-4 20 ng mL-1 had high expression of the respective M1 and M2 CD markers in flow cytometry and Western blot analysis. Cytokine release studies demonstrated the expression of IL-1ß, TNF-α and IL-10 in the M1-polarized macrophages (P < 0.01), whilst TGF- ß levels were seen in the M1 and M2-polarized macrophages. Ror ß expression was upregulated when macrophages and hI-DPSCs were treated with anti-inflammatory cytokines. CONCLUSION: Ror ß was expressed in THP-1 macrophages and hI-DPSCs during their resting stage. Upregulated expression of Ror ß occurred following an anti-inflammatory stimulus.

Facile manufacturing of fused-deposition modeled composite scaffolds for tissue engineering-an embedding model with plasticity for incorporation of additives.

The fused-deposition modeling (FDM) process is carried out at an elevated temperature, preventing the addition of biological factors, drugs, bioactive compounds, etc, during fabrication. To overcome this disadvantage, a 3D interlinked porous polylactic acid (PLA) scaffold was fabricated by FDM, followed by the embedding of a polycaprolactone (PCL) scaffold into the pores of the PLA at room temperature, yielding a PLA-PCL scaffold. In addition, PLA-PCL scaffolds with nanohydroxyapatite (PLA-PCL-nHAP) and multiwalled carbon nanotubes (PLA-PCL-MWCNT) were also fabricated. Here, the FDM-fabricated PLA scaffold functions as the structural component, whereas the embedded PCL scaffold acts as the functional component, which provides a the ability to functionalize the scaffolds with the desired chemical or biological materials. The embedding process is straightforward, cost effective, and does not require sophistication. A mechanical characterization of the scaffolds suggests that the Young's modulus of the PLA-PCL scaffold (16.02 MPa) was higher than that of the FDM-fabricated PLA (9.98 MPa) scaffold, by virtue of embedded PCL matrix. In addition, finite element analysis showed that the von Mises stress on a mandible with scaffolds was 4.04 MPa, whereas for a mandible with a defect, it was 6.7 MPa, confirming the stress distribution efficiency and mechanical stability of these scaffolds. Furthermore, field emission-scanning electron microscope analysis implied the presence of interlinked porous structures with pore diameters of 50 µm to 300 µm. X-ray diffraction results revealed an increased crystallinity (%) in the embedded models (PLA-PCL, PLA-PCL-nHAP and PLA-PCL-MWCNT), compared to a PLA printed scaffold. Additionally, Raman analysis revealed that the embedding process did not cause chemical alterations in the polymeric chains. In vitro analysis with human osteoblasts demonstrated the osteoconductive nature of the scaffold, which supported mineralization. In brief, the advantage of our model is that it helps to overcome the difficulties of manufacturing a filament with the desired additives for FDM, and offers the ability to incorporate the desired concentrations of heat-labile bioactive molecules during the embedding process at ambient temperatures.

Fabrication of tri-layered electrospun polycaprolactone mats with improved sustained drug release profile.

Modulation of initial burst and long term release from electrospun fibrous mats can be achieved by sandwiching the drug loaded mats between hydrophobic layers of fibrous polycaprolactone (PCL). Ibuprofen (IBU) loaded PCL fibrous mats (12% PCL-IBU) were sandwiched between fibrous polycaprolactone layers during the process of electrospinning, by varying the polymer concentrations (10% (w/v), 12% (w/v)) and volume of coat (1 ml, 2 ml) in flanking layers. Consequently, 12% PCL-IBU (without sandwich layer) showed burst release of 66.43% on day 1 and cumulative release (%) of 86.08% at the end of 62 days. Whereas, sandwich groups, especially 12% PCLSW-1 & 2 (sandwich layers-1 ml and 2 ml of 12% PCL) showed controlled initial burst and cumulative (%) release compared to 12% PCL-IBU. Moreover, crystallinity (%) and hydrophobicity of the sandwich models imparted control on ibuprofen release from fibrous mats. Further, assay for cytotoxicity and scanning electron microscopic images of cell seeded mats after 5 days showed the mats were not cytotoxic. Nuclear Magnetic Resonance spectroscopic analysis revealed weak interaction between ibuprofen and PCL in nanofibers which favors the release of ibuprofen. These data imply that concentration and volume of coat in flanking layer imparts tighter control on initial burst and long term release of ibuprofen.

A review of natural polysaccharides for drug delivery applications: Special focus on cellulose, starch and glycogen.

Natural polysaccharides are renewable with a high degree of biocompatibility, biodegradability, and ability to mimic the natural extracellular matrix (ECM) microenvironment. Comprehensive investigations of polysaccharides are essential for our fundamental understanding of exploiting its potential as bio-composite, nano-conjugate and in pharmaceutical sectors. Polysaccharides are considered to be superior to other polymers, for its ease in tailoring, bio-compatibility, bio-activity, homogeneity and bio-adhesive properties. The main focus of this review is to spotlight the new advancements and challenges concerned with surface modification, binding domains, biological interaction with the conjugate including stability, polydispersity, and biodegradability. In this review, we have limited our survey to three essential polysaccharides including cellulose, starch, and glycogen that are sourced from plants, microbes, and animals respectively are reviewed. We also present the polysaccharides which have been extensively modified with the various types of conjugates for combating last-ditch pharmaceutical challenges.

An inhibitory action of chitosan nanoparticles against pathogenic bacteria and fungi and their potential applications as biocompatible antioxidants.

Chitosan is the second most abundant polymer obtained from the byproduct of seafood. Chitosan and its derivatives and chitosan loaded drugs are the recent area of interest against microbial pathogenesis. The cationic chitosan nanoparticles (ChNPs) interact with the anionic surfaces of the microbial cell membrane, which promotes antimicrobial activity. Although, ChNPs are potential against pathogenic microbes, selection of adaptable, suitable and cost effective synthesis method is much important. In the present study, ChNPs were synthesized adopting ionic gelation using sodium tripolyphosphate as a cross linking agent and characterized by FTIR, DLS, SEM and TEM analysis. ChNPs were investigated for antimicrobial activity against bacterial (Escherichia coli and Staphylococcus aureus) and fungal (Candida albicans) pathogens. ChNPs showed bactericidal activity at the lower minimum inhibitory concentration of about 40-80 µg mL-1. Interestingly, ChNPs exhibits biocompatible antioxidant property by inhibiting DPPH free radicals at 76% and also proven to be a potential candidate against the microbial pathogenesis with an inevitable applications in biomedicine.

Green synthesis of anisotropic zinc oxide nanoparticles with antibacterial and cytofriendly properties.

Zinc oxide nanoparticles (ZnONPs) exhibit abundant biomedical applications. Anisotropic ZnONPs with a defined shape and size were synthesized using Bacillus megaterium (NCIM 2326) cell free extract as a bio-reductant. The study investigated the multidimensional effect of ZnONPs on Helicobacter pylori strains and assessed its biosafety in normal human mesenchymal stem cells (hMSc). The highly stable ZnONPs were produced using B. megaterium and Zinc nitrate as a precursor. The phase of ZnONPs formation and structural characterization were performed by UV- visible (UV-Vis), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD) and Field Emission Scanning electron microscopy (FESEM) analysis. Furthermore, the ZnONPs exhibited higher biocompatibility against human mesenchymal stem cells (hMSC) and proved to be potentially safe in mammalian cells. Corroborating the current investigation, we described the anti-H. Pylori dosage of ZnONPs was safe to hMSC and could efficiently use as nano-antibiotic.

AKT is indispensable for coordinating Par-4/JNK cross talk in p21 downmodulation during ER stress.

The double-edged role of p21 to command survival and apoptosis is emerging. The current investigation highlights ER stress-mediated JNK activation that plausibly triggers cell death by attenuating endogenous p21 level. Here, we demonstrated that ER stress activator 3-AWA diminishes the p21 levels in cancer cells by averting the senescent phenotype to commence G2/M arrest. In essence, the deceleration in p21 level occurs through ER stress/JNK/Caspase-3 axis via activation/induction of proapoptotic Par-4 and inhibition of AKT. The molecular dynamics studies identified important interactions, which may be responsible for the AKT inhibition and efficacy of 3-AWA towards AKT binding pocket. Interestingly, the p21 deceleration was rescued by incubating the cells with 3-AWA in the presence of an ER stress inhibitor, Salubrinal. Furthermore, we demonstrated that p21 expression decreases solitarily in Par-4+/+ MEFs; albeit, ER stress-induced JNK activation was observed in both Par-4+/+ and Par-4-/- MEFs. Par-4 knockdown or overexpression studies established that ectopic Par-4 along with ER stress are not sufficient to downregulate p21 in PC-3 cells but are adequate for DU-145 cells and that the ER stress inflicted activation of JNK, inhibition of AKT and Par-4 induction are all crucial to p21 downmodulation by 3-AWA. By using isogenic cell lines, such as HCT-116 p53+/+ and HCT-116 p53-/-, we found that deceleration in p21 expression due to ER stress is p53 independent. Moreover, in orthotopic carcinogen-induced rat colorectal carcinoma model, we found that 3-AWA inhibits colorectal tumor growth and formation of colorectal polyps at a tolerable dose, similar to the first-line drug for colorectal cancer-5-fluorouracil.

Synthesis and characterization of polyethylene glycol (PEG) coated Fe3O4 nanoparticles by chemical co-precipitation method for biomedical applications.

Polyethylene glycol (PEG) coated Fe3O4 nanoparticles were synthesized by chemical co-precipitation method. With polyethylene glycol (PEG) as a stabilizer and dispersant. The X-ray diffraction and selected area electron diffraction (SAED) results show that the cubic inverse spinel structure of pure phase polycrystalline Fe3O4 was obtained. The scanning electron microscopy (SEM) and field emission transmission electron microscopy (FE-TEM) results exhibited that the resulted Fe3O4 nanoparticles were roughly spherical in shape with narrow size distribution and homogenous shape. Fourier transform infrared spectroscopy (FT-IR) results suggested that PEG indicated with Fe3O4 via its carbonyl groups. Results of vibrating sample magnetometer (VSM) indicated that the prepared Fe3O4 nanoparticles exhibit superparamagnetic behavior and high saturation magnetization at room temperature. Such Fe3O4 nanoparticles with favorable size and tunable magnetic properties are promising biomedical applications.

In ovo delivery of Newcastle disease virus conjugated hybrid calcium phosphate nanoparticle and to study the cytokine profile induction.

In this report, the hybrid calcium phosphate (CaP) nanoparticles were synthesized and functionalized with Newcastle disease virus (NDV). These nanoparticles were synthesized by a combination of co-precipitation and polymerization process and functionalized with amino propyl triethoxy silane before coupling to NDV. The 5-dimethylthiazol-2-yl-2, 5-diphenyltetrazolium bromide (MTT) assay of chicken spleen cells incubated with these nanoparticles indicated that, these particles did not exert any significant cytotoxicity. The effects of hybrid CaP nanoparticles on cell cycle were assayed using a flow cytometer. The results demonstrated that the cell viability and proliferation capacity of spleen cells were not affected by hybrid CaP nanoparticles compared with their control cells. The hybrid CaP nanoparticles were characterized by scanning/transmission electron microscopy (SEM/TEM); Fourier transformed infrared spectroscopy (FTIR), X-ray diffraction patterns (XRD), Raman spectroscopy and energy-dispersive X-ray spectroscopy (EDX). These methods revealed that NDV was successfully conjugated on nanoparticles. The ability of the hybrid CaP nanoparticles to induce different cytokine mRNAs in the spleen cells of 18-day old embryonated chicken eggs (ECEs) was studied by quantitative real time polymerase chain reaction (qRT-PCR). NDV conjugated particles induced a high expression of Th1 cytokines such as interferon (IFN)-α, tumor necrosis factor (TNF)-α of and Th2 cytokines, interleukin (IL) 6 and IL-10. Uncoupled NDV induced only Th1 cytokines, IFN-α, INF-γ and TNF-α. The hybrid particles alone did not induce any cytokines. This confirmed that nanoparticle coupling could induce differential cytokine profiles and hence can be used as an alternate strategy to direct favorable immune responses in animals or chickens using appropriate vaccination carrier.

Treatment of drug-induced gingival overgrowth by full-mouth disinfection: A non-surgical approach.

BACKGROUND: Drug-induced gingival overgrowth is a common finding in the modern era. These gingival overgrowths are usually treated by various modalities namely substitution of drugs, surgical, and non-surgical treatment. The recent concept mainly involves full-mouth scaling and root planing (the entire dentition in two visits within 24 hours, i.e., two consecutive days) followed by chair side mouth rinsing by the patient with a 0.2% chlorhexidine solution for 2 minutes and brushing the tongue of the patient with 1% chlorhexidine gel. This is followed by an additional subgingival irrigation (three times, repeated within 10 minutes) of all pockets with a 1% chlorhexidine gel. MATERIALS AND METHODS: Twenty patients between the ages of 20 and 50 years with drug-induced gingival overgrowth were treated using the full-mouth disinfection approach. The patients were evaluated at 3 months and 6 months after therapy. The data obtained for plaque index, bleeding on probing index, probing pocket depth, and gingival overgrowth scores were tabulated and compared statistically using the one sample unpaired t test. STATISTICAL ANALYSIS: Statistically significant difference (P < 0.05) was found in PI GBI, PPD, and GO score between baseline, 3 months, and 6 months. RESULTS: All clinical parameters improved significantly after therapy without the need of further surgical treatment. CONCLUSIONS: Full-mouth disinfection might be a beneficial treatment concept in patients with drug-induced gingival overgrowth, thus decreasing the need for surgical therapy.

Oral findings in a patient with Sebaceous Nevi - A Case Report.

The presentation herewith as scripted is to describe a case with Nevus sebaceous with oral manifestations. Nevus Sebaceous or Jadassohn's nevus is an epidermal nevus with predominant sebaceous glands seen histologically. Reports of oral involvement have been few ranging from papillomatous growths of the tongue, gingiva, palate to dental abnormalities such as anodontia and dysodontia. The present case describes a nevus sebaceous present on the right half of the face and neck, showing intraoral papillomatous growth on the lateral part of the tongue on the right side. The patient was healthy and did not report involvement of any other organ systems. Intraoral involvement may be seen in patients with Nevus Sebaceous, hence proper screening is important. In patients presenting with large nevi on the head and neck such as ours, involvement of other systems such as ocular, neurologic and oral lesions may be seen, therefore screening of such patients is of importance. Patients with nevus sebaceous may be predisposed to the occurrence of tumours. Therefore, careful screening of such patients is necessary. How to cite this article: Baliga V, Gopinath VP, Baliga S, Chandra U. Oral findings in a patient with Sebaceous Nevi - A Case Report. J Int Oral Health 2013; 5(5):139-42.

Intraoral manifestations in a patient with Epidermal nevus syndrome.

AIM: To report rare findings of oral and periodontal manifestations in a patient with Epidermal nevus syndrome (ENS). BACKGROUND: The ENS describes the rare association of an epidermal nevus with abnormalities of central nervous system,ocular and skeletal abnormalities. Reports of oral involvement have been few. Also, most of the intraoral lesions have been reported in patients with nevi that do not fulfill the criteria for the diagnosis of ENS. CASE DESCRIPTION: This report describes a case of ENS that, in addition to cutaneous manifestations showed skeletal involvement and intraoral manifestations such as the extension of the nevi on the face intraorally involving the labial mucosa, hypoplasia, hypodontia of teeth and severe periodontal destruction. CONCLUSION: Patients with extensive epidermal nevi and systemic abnormalities should be suspected of having the ENS. Evaluation and management of patients with ENS requires a multidisciplinary team approach involving the dermatologist, pediatrician, ophthalmologist, neurologist, genetist, plastic surgeon and orthopedic services. Although uncommonly described in association with ENS, significant intraoral lesions do occur. Periodontal manifestations as in our patient, which to our knowledge has not been described in association with ENS so far, may also be present. CLINICAL RELEVANCE: Alteration of the response of periodontal tissues to dental plaque in the presence of certain systemic diseases has been reported, but not in association with ENS. Severe periodontal destruction due to exaggerated response to dental plaque was seen in the present case. Hence, emphasis on oral hygiene maintenance in such patients is essential. Patients with ENS must be evaluated periodically as they show a persistent predisposition for the development of tumors.

Clinical and radiographic evaluation of regenerative potential of GTR membrane (Biomesh®) along with alloplastic bone graft (Biograft®) in the treatment of periodontal intrabony defects.

BACKGROUND AND OBJECTIVES: The primary goal of periodontal therapy is to restore the tooth supporting tissues lost due to periodontal disease. The aim of the present study was to compare the efficacy of combination of GTR membrane and alloplastc bone graft with open flap debridement (OFD) in treatment of periodontal intrabony defects. METHODS: Twenty paired intrabony defects were surgically treated using split mouth design. The defects were randomly assigned to treatment with OFD, GTR membrane+bone graft (test) or OFD alone (control). The clinical efficacy of two treatment modalities was evaluated at 6 months postoperatively by clinical, radiographical parameters. The measurements included probing pocket depth (PD), clinical attachment level (CAL), gingival recession (GR), bone fill (BF), bone density (BD). RESULTS: The mean reduction in PD at 0 to 6 months was 3.20±0.82 mm and CAL gain of 3.10±1.51 mm occurred in the GTR membrane+bone graft (test) group; corresponding values for OFD (control) were 2.10±0.63 mm and 1.90±0.57 mm. Similar pattern of improvement was observed when radiographically postoperative evaluation was made. All improvement in different parameters was statistically significant (p<0.01). CONCLUSION: Treatment with a combination of collagen membrane and bone graft led to a significantly more favorable clinical outcome in intrabony defects as compared to open flap debridement alone.

Extracellular biosynthesis of silver nanoparticles using Bacillus sp. GP-23 and evaluation of their antifungal activity towards Fusarium oxysporum.

In last few decades nanoparticles have attracted and emerged as a field in biomedical research due to their incredible applications. The current research was focused on extracellular synthesis of silver nanoparticles (AgNPs) using cell free culture supernatant of strain GP-23. It was found that the strain GP-23 belonged to Bacillus species by 16S rRNA sequence analysis. Biosynthesis of AgNPs was achieved by addition of culture supernatant with aqueous silver nitrate solution, after 24 h it turned to brown color solution with a peak at 420 nm corresponding to the Plasmon absorbance of AgNPs by UV-Vis Spectroscopy. The nanoparticles were characterized by FTIR, XRD, HRTEM, EDX and AFM. The synthesized nanoparticles were found to be spherical in shape with size in the range of 7-21 nm. It was stable in aqueous solution for five months period of storage at room temperature under dark condition. The biosynthesized AgNPs exhibited strong antifungal activity against plant pathogenic fungus, Fusarium oxysporum at the concentration of 8 µg ml(-1). The results suggest that the synthesized AgNPs act as an effective antifungal agent/fungicide.

Synthesis of anisotropic silver nanoparticles using novel strain, Bacillus flexus and its biomedical application.

Synthesis of metallic nanoparticles has attracted by bacterial based production and alternative to physical and chemical approaches. The present work was focused to nominate a bacterial strain for synthesis of potential silver nanoparticles. The target was achieved by screening of 127 isolates from silver mining wastes. A strain designated S-27 found to be a potential candidate for rapid synthesis of silver nanoparticles among tested microorganisms. It was subjected to molecular characterization by 16S rDNA sequence analysis. It was found that S-27 belonging to Bacillus flexus. Synthesis of silver nanoparticles was achieved by addition of culture supernatants with aqueous silver nitrate solution, immediately it turns to brown colour solution showed a peak at 420 nm corresponding to the plasmon absorbance of silver nanoparticles by UV-vis spectroscopy. Various instrumentation techniques, such as AFM, FESEM, XRD and FTIR, were adopted to characterize the synthesized nanoparticles. Anisotropic nanoparticles, such as spherical and triangular shaped nanoparticles, have been synthesized and sizes were found to be 12 and 65 nm, respectively. It was stable in aqueous solution in five months period of storage at room temperature in the dark. Synthesized nanoparticles showed efficacy on antibacterial property against clinically isolated multi-drug resistant (MDR) microorganisms. It is suggested that biogenic synthesis of nanoparticles have wide-application in medicine and physical chemistry and it can produce with eco-friendly, easy downstream processing and rapid scale-up processing.

Biosynthesis of silver nanoparticles from Tribulus terrestris and its antimicrobial activity: a novel biological approach.

In the recent decades, increased development of green synthesis of nanoparticles is inevitable because of its incredible applications in all fields of science. There were numerous work have been produced based on the plant and its extract mediated synthesis of nanoparticles, in this present study to explore that the novel approaches for the biosynthesis of silver nanoparticles using plant fruit bodies. The plant, Tribulus terrestris L. fruit bodies are used in this study, where the dried fruit body extract was mixed with silver nitrate in order to synthesis of silver nanoparticles. The active phytochemicals present in the plant were responsible for the quick reduction of silver ion (Ag(+)) to metallic silver nanoparticles (Ag(0)). The reduced silver nanoparticles were characterized by Transmission Electron Microscope (TEM), Atomic Force Microscope (AFM), XRD, FTIR, UV-vis spectroscopy. The spherical shaped silver nanoparticles were observed and it was found to be 16-28 nm range of sizes. The diffraction pattern also confirmed that the higher percentage of silver with fine particles size. The antibacterial property of synthesized nanoparticles was observed by Kirby-Bauer method with clinically isolated multi-drug resistant bacteria such as Streptococcus pyogens, Pseudomonas aeruginosa, Escherichia coli, Bacillus subtilis and Staphylococcus aureus. The plant materials mediated synthesis of silver nanoparticles have comparatively rapid and less expensive and wide application to antibacterial therapy in modern medicine.

Molecular cloning and tissue-specific expression of Toll-like receptor 5 gene from turkeys.

Toll-like receptors (TLRs), a family of transmembrane and cytosolic proteins, detect microbial patterns, initiating innate immune responses in various organisms. Although they are abundant, genetic characterization and functional differences of TLRs in economically important avian species such as chickens and turkeys have not been investigated in detail. In this study, the putative TLR5 coding region from turkey genome was sequenced, and its homology to other vertebrate species was analyzed. Secondary structure analysis revealed protein motifs typical of the chicken TLR5 protein structure, with 97% amino acid identity between them. mRNA expression profiling in adult turkeys revealed abundant TLR5 expression in a broad range of tissues. Stimulation with the TLR5 ligand flagellin resulted in the production of the inflammatory mediators interleukin (IL)-1beta, IL-6, and nitric oxide in peripheral blood mononuclear cells. To our knowledge, this is the first complete turkey TLR5 coding DNA sequence reported in sequence databases.

Rapid detection of Newcastle disease virus replication in embryonated chicken eggs using quantitative real time polymerase chain reaction.

Newcastle disease virus (NDV), an avian paramyxovirus, is an economically important disease of poultry globally. Rapid methods to detect and differentiate the virus are important to curtail the spread of this virus. Nucleic acid based detection methods are routinely employed for diagnosis that suffer from the disadvantage of failure to discriminate viable virus and non-infectious genome. However, virus isolation remains the gold standard for diagnosis of field outbreaks. The sensitivity of virus isolation was combined with nucleic acid based detection methods so that the time taken for confirmatory diagnosis could be considerably reduced while increasing sensitivity. Quantitative real time reverse transcription polymerase chain reaction (qRT-PCR) and conventional RT-PCR techniques were compared for the detection of NDV genome replication in 9-11-day-old embryonated chicken eggs (ECE) using the nucleoprotein (NP) gene of the virus as a target. The results suggest that at least two to fourfold increase in cycle threshold (C(t)) values over the baseline C(t) value of samples lacking infectious virus, would indicate live NDV replication. The limit of detection of NDV replication using qRT-PCR was 1×10(4.0) mean embryo infective doses (EID(50)). The earliest time point when live virus replication was detectable by qRT-PCR or RT-PCR was 30h post-inoculation in ECE.