Anju Manuja

Sodium alginate and gum acacia hydrogels of ZnO nanoparticles show wound healing effect on fibroblast cells

An ideal biomaterial for wound dressing applications should possess antibacterial and anti-inflammatory properties without any toxicity to the host cells while providing the maximum healing activity. Zinc oxide nanoparticles (ZnONPs) possess antimicrobial activity and enhance wound healing, but the questions regarding their safety arise before application to the biological systems. We synthesized ZnONPs-loaded-sodium alginate-gum acacia hydrogels (SAGA-ZnONPs) by cross linking hydroxyl groups of the polymers sodium alginate and gum acacia with the aldehyde group of gluteradehyde. Here, we report the wound healing properties of sodium alginate/gum acacia/ZnONPs, circumventing the toxicity of ZnONPs simultaneously. We demonstrated the concentration-dependent zones of inhibition in treated cultures of Pseudomonas aerigunosa and Bacillus cereus and biocompatability on peripheral blood mononuclear/fibroblast cells. SAGA-ZnONPs hydrogels showed a healing effect at a low concentration of ZnONPs using sheep fibroblast cells. Our findings suggest that high concentrations of ZnONPs were toxic to cells but SAGA-ZnONPs hydrogels significantly reduced the toxicity and preserved the beneficial antibacterial and healing effect.

Comparison of cellular schizont, soluble schizont and soluble piroplasm antigens in ELISA for detecting antibodies against Theileria annulata.

The efficacy and suitability of cellular schizont, soluble schizont and soluble piroplasm antigens was compared for detecting antibodies against Theileria annulata. Fifty bovine sera of known identity were evaluated in ELISA using the above mentioned antigens. Antibody titres of 1:100 to 1:51,200 were detected while using soluble piroplasm and cellular schizont antigen in ELISA. The titres ranged between 1:100 to 1:25,600 with the soluble schizont antigen. Soluble piroplasm antigen exhibited the highest antibody titres followed by cellular schizont and soluble schizont antigens. Cellular schizont antigen proved to be better than soluble schizont antigen for detecting anti-schizontal antibodies. Antibody titres obtained by the three antigens exhibited a good linear correlation amongst each other. The study showed that soluble piroplasm and cellular schizont antigens can be used successfully for detecting antibodies against piroplasm and schizont stages of T. annulata, respectively in bovine sera.

Cytotoxicity and genotoxicity of a trypanocidal drug quinapyramine sulfate loaded-sodium alginate nanoparticles in mammalian cells.

We synthesized quinapyramine sulfate loaded-sodium alginate nanoparticles (QS-NPs) to reduce undesirable toxic effects of QS against the parasite Trypanosoma evansi, a causative agent of trypanosomosis. To determine the safety of the formulated nanoparticles, biocompatibility of QS-NPs was determined using Vero, Hela cell lines and horse erythrocytes in a dose-dependent manner. Our experiments unveiled a concentration-dependent safety/cytotoxicity (metabolic activity), genotoxicity (DNA damage, chromosomal aberrations), production of reactive oxygen species and hemolysis in QS-NPs treated cells. Annexin-V propidium iodide (PI) staining showed no massive apoptosis or necrosis. However, at very high doses (more than 300 times than the effective doses), we observed more toxicity in QS-NPs treated cells as compared to QS treated cells. QS-NPs were safe at effective trypanocidal doses and even at doses several times higher than the effective dose.

Stress and its impact on farm animals

This article presents a brief overview of stress and its impact on growth, production, reproduction and disease susceptibility in farm animals. A single measure of stress might not be a reliable indicator and it is usually more informative to combine multiple indicators of stress to assess animal welfare. Popular measures of stress such as alterations in hormonal profiles can be complemented with behavioral and immunological changes. Traditional approaches are insufficient to unravel the role of large number of genes and interconnected genetic pathways leading to multiple responses to stresses. Modern genomic and proteomics methodologies used in identifying the biomarkers and molecular mechanisms of stress response have also been described briefly. With a better understanding of the basic biology, altered physiological processes and the genes/proteins involved in stress adaptability, it would be possible to develop methodologies for selection of animals with better performance and devise strategies for animal welfare.

Immunotherapeutic potential of CpG oligodeoxynucleotides in veterinary species

Abstract Innate immunity plays a critical role in host defense against infectious diseases by discriminating between self and infectious non-self. The recognition of infectious non-self involves germ-line encoded pattern recognition receptors (PRRs) that recognize pathogen-associated molecular patterns (PAMPs). The PAMPs are the components of pathogenic microbes which include not only the cell wall constituents but also the unmethylated 2′-deoxy-ribo-cytosine-phosphate-guanosine (CpG) motifs. These CpG motifs present within bacterial and viral DNA are recognized by toll-like receptor 9 (TLR9), and signaling by this receptor triggers a proinflammatory cytokine response which, in turn, influences both innate and adaptive immune responses. The activation of TLR9 with synthetic CpG oligodeoxynucleotides (ODNs) induces powerful Th1-like immune responses. It has been shown to provide protection against infectious diseases, allergy and cancer in laboratory animal models and some domestic animal species. With better understanding of the basic biology and immune mechanisms, it would be possible to exploit the potential of CpG motifs for animal welfare. The research developments in the area of CpG and TLR9 and the potential applications in animal health have been reviewed in this article.

CpG-ODN class C-mediated immunostimulation and its potential against Trypanosoma evansi in equines

Trypanosoma evansi is the causative agent of surra, which is the most common and widespread trypansomal disease. The infection is mainly restricted to animals, but it has also been documented in human. Trypanosomes possess the thick immunogenic surface coat known as variant surface glycoprotein (VSG). The parasite modifies the VSG constantly resulting in continuous antigenic variations and thus evades the host immune response. Due to antigenic variations, vaccination against trypanosomosis is not useful. Therefore, alternate strategies to augment the immune response are required. CpG-ODN class-C has combined immune effects of both A and B classes of CpG-ODN. In this study, we observed that CpG-ODN class-C stimulated horse peripheral blood mononuclear cells (PBMC) induce the expression of interferon-α (IFN-α), tumor necrosis factor-α (TNF-α), IL-12 and nitric oxide (NO) indicating enhanced innate immune response. We have for the first time demonstrated that co-culture of CpG-ODN with T. evansi antigen induces lymphocyte proliferative responses and result in a synergistic effect in eliciting the immune response.

Sodium alginate and gum acacia hydrogels of zinc oxide nanoparticles reduce hemolytic and oxidative stress inflicted by zinc oxide nanoparticles on mammalian cells

Zinc oxide nanoparticles are important nanomaterials currently under research due to their applicability in nanomedicine. Toxicity of ZnO NPs has been extensively studied and has been shown to affect various cell types and animal systems. In this study, we investigated hemolytic potential and oxidative stress inflicted by ZnO NPs and ZnO NPs-loaded-sodium alginate-gum acacia hydrogels on horse erythrocytes and African green monkey kidney (Vero) cells. Our study provides a better understanding of the hemolytic and oxidative effects of interaction of ZnO NPs and ZnO NPs released from polymeric hydrogels with the biological system. Remarkable aggregation of erythrocytes was noted in the higher concentration of ZnO NPs treated erythrocytes as compared to erythrocytes treated with ZnO NPs-loaded hydrogels. ZnO NPs-loaded hydrogels treated Vero cells significantly reduced oxidative stress as evidenced by less malondialdehyde production as compared to that of ZnO NPs treated cells. Normal horse erythrocytes when treated with ZnO NPs in in vitro condition undergo oxidative damage, and contribute in augmenting the toxicity. We demonstrated that polymeric ZnO NPs reduced the undesirable effects provoked by ZnO NPs on mammalian cells.

CpG-ODN Class C Mediated Immunostimulation in Rabbit Model of Trypanosoma evansi Infection

CpG oligodeoxynucleotides (CpG-ODN) stimulate immune cells from a wide spectrum of mammalian species. Class C CpG-ODN is relatively stable and has the combined immune effects of both A and B classes of CpG-ODN. Trypanosoma evansi produces the state of immuno-suppression in the infected hosts. The current chemotherapeutic agents against this parasite are limited in number and usually associated with severe side effects. The present work aimed to determine the immunostimulatory effects of CpG-ODN class C in T. evansi infected rabbits. Rabbits inoculated with CpG C and challenged with T. evansi resulted in delayed onset of clinical signs with reduced severity in comparison to that of T. evansi infected rabbits. The treatment also enhanced humoral immune responses. Histopathological findings in liver and spleen revealed enhancement of mononuclear cell infiltration and secondary B cell follicles. These results demonstrate that CpG-ODN class C, has immunostimulatory properties in rabbit model of trypanosomosis. The use of booster doses or sustained delivery of CpG-ODN will further elucidate the prolonged CpG-ODN generated immune responses.

Comparative Analysis of Molecular Structure, Function and Expression of Buffalo (Bubalus bubalis) Toll-Like Receptor 9

Toll-like receptor 9 (TLR9) has been characterized as a receptor that recognizes unmethylated CpG motif and triggers a pro-inflammatory cytokine response that influences both innate and adaptive immunity. Buffalo is an economically important livestock species in many Asian and Mediterranean countries, but there is little information available on its TLR9 structure and response to stimulation with its agonist CpG-ODNs. Hence in this study, we report the analysis of newly sequenced buffalo TLR9 gene fragment. In this study, buffalo TLR9 amino acid sequence revealed close association of TLR9 proteins within other bovines and small ruminants; but high divergence from other species. Multiple alignment of deduced amino acid sequence of Bubalus bubalis TLR9 with other species showed that 156/201 (74.28%) amino acids were conserved in all species. Leucine rich repeat (LRR) motifs in the ectodomain of TLR9 are responsible for molecular recognition of its agonist. The LRR pattern of Bubalus bubalis TLR9 protein was predicted towards N-terminal sequence and was found to be conserved among all species except Rattus norvegicus and Equus caballus. Blast analysis of buffalo TLR9 sequence with single nucleotide polymorphisms (SNPs) database revealed 13 SNPs out of which 7 were cds-synonymous and 6 were of the functional significance. Furthermore, kinetics of TLR9 and proinflammatory IL-1β and TNF-α cytokine expression by buffalo PBMCs influenced by CpG-ODN is also discussed.

Influence of functionalized mesoporous silica in controlling azathioprine drug release and cytotoxicity properties

Abstract The present study is focused on functionalisation of mesoporous silica (or SBA-15) to control azathioprine drug release rate and its toxic effect. The mesoporous silica was functionalised by (γ-chloropropyl)triethoxysilicane and (3-aminopropyl)triethoxysilane (APTES) via hydrothermal process. The azathioprine was loaded into SBA-15 via post impregnation method. Azathioprine-loaded pristine and functionalised SBA-15 samples were characterised using UV–visible spectrophotometry and thermogravimetric analysis to measure the drug loading efficiency. The samples were also characterised by small and wide-angle powder X-ray diffraction, scanning electron microscope, transmission electron microscope, infrared spectroscopy and nitrogen adsorption/desorption analysis to study the structure, morphology, functionalisation and drug loading in detail. The maximum drug loading efficiency of 65(±1)% was achieved. In vitro azathioprine release profiles were studied in phosphate buffered saline (pH 7.4) and results suggested that the drug release amount could be controlled by functionalisation of carrier matrix SBA-15. Azathioprine-loaded APTES-functionalised material revealed lowest release amount of ~64.5% in 60 h. The toxicity of azathioprine was significantly reduced by loading the drug into the mesoporous pristine and functionalised silica. The controlled azathioprine release reduced its repeated administration and can reduce its toxicity and side effects. These outcomes recommend that the functionalised SBA-15 is an advantageous drug carrier for achieving extended release time.