Shreekumar Pillai

Enhanced intracellular translocation and biodistribution of gold nanoparticles functionalized with a cell-penetrating peptide (VG-21) from vesicular stomatitis virus.

Reduced toxicity and ease of modification make gold nanoparticles (GNPs) suitable for targeted delivery, bioimaging and theranostics by conjugating cell-penetrating peptides (CPPs). This study presents the biodistribution and enhanced intracellular uptake of GNPs functionalized with VG-21, a CPP derived from vesicular stomatitis virus glycoprotein (G). Cell penetrating efficiency of VG-21 was demonstrated using CellPPD web server, conjugated to GNPs and were characterized using, UV-visible and FTIR spectroscopy, transmission electron microscopy, dynamic light scattering and zeta potential. Uptake of VG-21 functionalized GNPs (fGNPs) was tested in eukaryotic cell lines, HEp-2, HeLa, Vero and Cos-7, using flow cytometry, fluorescence and transmission electron microscopy (TEM), and inductively coupled plasmon optical emission spectroscopy (ICP-OES). The effects of nanoparticles on stress and toxicity related genes were studied in HEp-2 cells. Cytokine response to fGNPs was studied in vitro and in vivo. Biodistribution of nanoparticles was studied in BALB/c mice using TEM and ICP-OES. VG-21, GNPs and fGNPs had little to no effect on cell viability. Upon exposure to fGNPs, HEp-2 cells revealed minimal down regulation of stress response genes. fGNPs displayed higher uptake than GNPs in all cell lines with highest internalization by HEp-2, HeLa and Cos-7 cells, in endocytotic vesicles and nuclei. Cytokine ELISA showed that mouse J774 cells exposed to fGNPs produced less IL-6 than did GNP-treated macrophage cells, whereas TNF-α levels were low in both treatment groups. Biodistribution studies in BALB/c mice revealed higher accumulation of fGNPs than GNPs in the liver and spleen. Histopathological analyses showed that fGNP-treated mice accumulated 35 ng/mg tissue and 20 ng/mg tissue gold in spleen and liver respectively, without any adverse effects. Likewise, serum cytokines were low in both GNP- and fGNP-treated mice. Thus, VG-21-conjugated GNPs have enhanced cellular internalization and are suitable for various biomedical applications as nano-conjugates.

Bacteria as an agent for change in structural plumage color: correlational and experimental evidence.

Recent studies have documented that a diverse assemblage of bacteria is present on the feathers of wild birds and that uropygial oil affects these bacteria in diverse ways. These findings suggest that birds may regulate the microbial flora on their feathers. Birds may directly inhibit the growth of harmful microbes or promote the growth of other harmless microbes that competitively exclude them. If keratinolytic (i.e., feather‐degrading) bacteria degrade colored feathers, then plumage coloration could reveal the ability of individual birds to regulate microbial flora. We used field‐ and lab‐based methods to test whether male eastern bluebirds (Sialia sialis) with brighter blue structural plumage coloration were better able to regulate their microbial flora than duller males. When we sampled bluebirds in the field, individuals with brighter color had higher bacterial loads than duller individuals. In the lab, we tested whether bacteria could directly alter feather color. We found that keratinolytic bacteria increased the brightness and purity, decreased the ultraviolet chroma, and did not affect the hue of structural color. This change in spectral properties of feathers may occur through degradation of the cortex and spongy layer of structurally colored barbs. These data suggest that bacteria can alter structural plumage color through degradation.

Synthesis of Ag/CNT hybrid nanoparticles and fabrication of their Nylon-6 polymer nanocomposite fibers for antimicrobial applications

Ag-coated CNTs hybrid nanoparticles (Ag/CNTs) were prepared by ultrasonic irradiation of dimethylformamide (DMF) and silver (I) acetate precursors in the presence of CNTs. The morphology of Ag/CNTs was characterized using x-ray diffraction and transmission electron microscopy (TEM) techniques. The Nylon-6 powder and 1 wt% Ag/CNTs mixture was dispersed uniformly using a noncontact spinning technique. The dried mixture was melted in a single screw extrusion machine and then extruded through an orifice. Extruded filaments were later stretched and stabilized by sequentially passing them through a set of tension adjusters and a secondary heater. The Nylon-6/Ag/CNT hybrid polymer nanocomposite (HPNC) fibers, which were of approximately 80 microm size, were tested for their tensile properties. The failure stress and modulus of the extruded HPNC fibers (doped with 1% Ag/CNTs) was about 72.19 % and 342.62% higher than the neat extruded Nylon-6 fiber, respectively. DSC results indicated an increase in the thermal stability and crystallization for HPNC fibers. The antibacterial activity of the Ag-coated CNTs, commercial Ag, neat Nylon-6 and plain CNTs were evaluated. Ag-coated CNTs at 25 microg demonstrated good antimicrobial activity against four common bacterial pathogens as tested by the Kirby-Bauer assay. The mean diameters of the zones of inhibition were 27.9 +/- 6.72 mm, 19.4 +/- 3.64 mm, 21.9 +/- 4.33 mm, and 24.1 +/- 4.14 mm, respectively, for Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli and Salmonella enterica serovar Typhimurium. By comparison, those obtained using the broad spectrum antibiotic amoxicillin-clavulanic acid were 37.7 +/- 2.13 mm, 28.6 +/- 4.27 mm, 22.6 +/- 1.27 mm, and 27.0 +/- 1.41 mm, respectively, for the same strains. The zones of inhibition obtained for Nylon-6 Ag-coated CNT powder at 25 microg were also high, ranging from 15.2 to 25.3 mm in contrast to commercial silver or neat Nylon-6, which did not inhibit the bacterial strains tested. Further, the Nylon-6 nanocomposite fibers infused with Ag/CNTs inhibited bacterial growth by 11-20%. Our results suggest that nylon nanocomposite fibers infused with Ag-coated CNTs have significant antimicrobial activity.

Do feather-degrading bacteria affect sexually selected plumage color?

Models of parasite-mediated sexual selection propose that males with more elaborate sexual traits will have fewer parasites. These models have generally been tested using metazoan or protozoan parasites of the blood, gut, or integument. Fewer studies have examined sexual ornaments in relation to bacterial infections. While most surface bacteria are harmless or beneficial, feather-degrading bacteria may have detrimental effects. In this study, we examined the relationships between overall bacterial load, feather-degrading bacterial load, and sexually selected carotenoid-based plumage color in a wild population of house finches (Carpodacus mexicanus). We found that males with the redder plumage preferred by females had similar overall bacterial loads, but lower feather-degrading bacterial loads, than males with less red plumage. These data suggest that plumage color can signal abundance of feather-degrading bacteria to potential mates. It remains unclear whether feather-degrading bacteria directly or indirectly affect plumage color, but the observed correlations suggest that feather-degrading bacteria may play some role in sexual selection.

Future Prospects for Scaffolding Methods and Biomaterials in Skin Tissue Engineering: A Review

Over centuries, the field of regenerative skin tissue engineering has had several advancements to facilitate faster wound healing and thereby restoration of skin. Skin tissue regeneration is mainly based on the use of suitable scaffold matrices. There are several scaffold types, such as porous, fibrous, microsphere, hydrogel, composite and acellular, etc., with discrete advantages and disadvantages. These scaffolds are either made up of highly biocompatible natural biomaterials, such as collagen, chitosan, etc., or synthetic materials, such as polycaprolactone (PCL), and poly-ethylene-glycol (PEG), etc. Composite scaffolds, which are a combination of natural or synthetic biomaterials, are highly biocompatible with improved tensile strength for effective skin tissue regeneration. Appropriate knowledge of the properties, advantages and disadvantages of various biomaterials and scaffolds will accelerate the production of suitable scaffolds for skin tissue regeneration applications. At the same time, emphasis on some of the leading challenges in the field of skin tissue engineering, such as cell interaction with scaffolds, faster cellular proliferation/differentiation, and vascularization of engineered tissues, is inevitable. In this review, we discuss various types of scaffolding approaches and biomaterials used in the field of skin tissue engineering and more importantly their future prospects in skin tissue regeneration efforts.

Enhanced delivery and expression of a nanoencapsulated DNA vaccine vector for respiratory syncytial virus

UNLABELLED This study evaluated the efficiency of chitosan-encapsulated DNA-based respiratory syncytial virus (RSV) vaccine. Antigenic regions of RSV F, M2, and G genes were cloned into the human cytomegalovirus promoter-based constitutive expression vector, resulting in a DNA vaccine vector named DR-FM2G. This vector was used to formulate DNA-chitosan nanoparticles (DCNPs) using a complex coacervation process that yielded an encapsulation efficiency of 94.7%. The DCNP sizes ranged from 80 to 150 nm with uniform size distribution and spherical shape. DNA release was between 50% and 60% when DCNPs were incubated with similar gastrointestinal fluid (pH 2), whereas 21% to 25% of DNA was released from DCNPs in 30 minutes at pH 10. Differential scanning calorimetry showed DCNPs to be more stable than naked DNA or chitosan, offering protection from DNA degradation by nucleases. DCNPs were not toxic to cells when used at concentrations < or =400 microg/mL. Immunohistochemical and real-time polymerase chain reaction results showed a higher level of RSV protein expression in mouse tissues given when DCNPs were injected intravenously as compared with naked DNA. FROM THE CLINICAL EDITOR This study evaluated the efficiency of chitosan-encapsulated DNA-based respiratory syncytial virus (RSV) vaccine, showing a higher level of RSV protein expression in mouse tissues given when DCNPs were injected intravenously as compared with naked DNA.

Recent Advances in Diagnosis, Prevention, and Treatment of Human Respiratory Syncytial Virus

Human respiratory syncytial virus (RSV) is a common cause of respiratory infection in infants and the elderly, leading to significant morbidity and mortality. The interdisciplinary fields, especially biotechnology and nanotechnology, have facilitated the development of modern detection systems for RSV. Many anti-RSV compounds like fusion inhibitors and RNAi molecules have been successful in laboratory and clinical trials. But, currently, there are no effective drugs for RSV infection even after decades of research. Effective diagnosis can result in effective treatment, but the progress in both of these facets must be concurrent. The development in prevention and treatment measures for RSV is at appreciable pace, but the implementation into clinical practice still seems a challenge. This review attempts to present the promising diverse research approaches and advancements in the area of diagnosis, prevention, and treatment that contribute to RSV management.

Advances in Skin Regeneration Using Tissue Engineering

Tissue engineered skin substitutes for wound healing have evolved tremendously over the last couple of years. New advances have been made toward developing skin substitutes made up of artificial and natural materials. Engineered skin substitutes are developed from acellular materials or can be synthesized from autologous, allograft, xenogenic, or synthetic sources. Each of these engineered skin substitutes has their advantages and disadvantages. However, to this date, a complete functional skin substitute is not available, and research is continuing to develop a competent full thickness skin substitute product that can vascularize rapidly. There is also a need to redesign the currently available substitutes to make them user friendly, commercially affordable, and viable with longer shelf life. The present review focuses on providing an overview of advances in the field of tissue engineered skin substitute development, the availability of various types, and their application.

Toxicity Issues Related to Biomedical Applications of Carbon Nanotubes

Carbon Nanotubes (CNTs) have emerged as a potential candidate for application in targeted drug delivery, cancer treatment, gene therapy and diagnostics. This review critically evaluates the biomedical potential of CNTs, and possible toxicological implications. Success of CNTs in other fields has not yet been translated to the biomedical field due to the lack of consistent data on toxicity, variation of toxicity due to characteristics such as shape and size along with the method of preparation, and limited control over functionalized CNTs behavior. Thus there is a lack of a predictable toxicity pattern of CNTs. This review summarizes the key findings on the toxicity of CNTs and the role of functionalization with hydrophilic moieties to reduce their toxicity, and increase cell penetrability for drug delivery and gene therapy applications. Caution is urged when handling this ‘wonder material’ but the immense potential for its commercial utilization and clinical trials infuses hope for future biomedical applications.

Direct Electrochemistry of Glucose Oxidase at a Gold Electrode Modified with Graphene Nanosheets

In this work, we report the direct electrochemistry of glucose oxidase (GOD) observed at a gold electrode modified with graphene nanosheets. Initially, graphene nanosheets were synthesized and conjugated to the enzyme GOD and immobilized on to a gold electrode surface. Cyclic voltammetry was then performed using Gold-Graphene-GOD modified electrodes in a pH 7.2 phosphate buffered saline (PBS). A pair of well-defined redox peaks was obtained for GOD with the reduction peak centered at +180 mV and a peak separation of 70 mV in PBS under physiological conditions. Moreover, the electron transfer rate of GOD redox reaction was greatly enhanced and the peak potential was found to be pH dependent at the graphene-GOD surface. Further, the performance of the Gold-Graphene-GOD was found to be stable and excellent under physiological conditions indicating the possibility of employing this platform for real time analysis. The observed results indicated that the 2D-graphene holds great promise for conjugation ability with a variety of enzymes. Further, our results also confirmed that graphene is capable of holding the enzyme GOD in a favorable position and retains its original structure and functionality that are essential for biosensing.