Poly(vinylbenzyl chloride-co-divinyl benzene) polyHIPE monolith-supported o-hydroxynaphthaldehyde propylenediamine Schiff base ligand complex of copper(ii) ions as a catalyst for the epoxidation of cyclohexene.

Poly(vinylbenzyl chloride-co-divinyl benzene)-based polyHIPE monoliths of different porosities were prepared using high-internal-phase emulsions (HIPEs) containing a fixed amount of vinylbenzyl chloride (VBC, 6.0 g, 0.0393 mol) and divinyl benzene (DVB 4.0 g, 0.0308 mol) as the oil phase and different volume ratios of aqueous calcium chloride as the internal phase. Span-80 (2.0 g (4.67 mmol))-stabilized HIPEs were polymerized at 60 °C using potassium persulfate (0.4 g, 1.48 mmol) as the initiator. Upon varying the volume ratio of aqueous calcium chloride from 80 to 90%, the prepared polyHIPE monoliths have shown significant variations in their surface morphology, specific surface area (SA), and pore volumes (V p) as confirmed by scanning electron microscopy (SEM) and a gas adsorption (BET) method. The prepared polyHIPE monoliths were anchored with o-hydroxynaphthaldehyde propylenediamine Schiff base ligand (HNPn) and then loaded with copper(ii) ions (HNPn-Cu) to act as a catalyst. The structural information of unsupported HNPn-Cu complexes was obtained by recording its FT-IR and UV-visible spectra. The amount of copper(ii) ions loaded onto HNPn ligand-anchored polyHIPE monoliths was determined by atomic absorption spectroscopic analysis. In comparison to unsupported HNPn-Cu catalyst, the polyHIPE monolith-supported HNPn-Cu catalyst has shown high catalytic activity (66.8%), product selectivity for epoxycyclohexane (ECH) (94.8%), high turn over number (0.028 mol mol-1 h-1) and low energy of activation (22.4 kJ mol-1) in the epoxidation of cyclohexene in the presence of hydrogen peroxide (H2O2) as an oxidant at 40 °C. The polyHIPE-supported HNPn-Cu catalyst also shows high reuse applications. Studies show that there is sufficient scope to develop polyHIPE monoliths with various properties for specific applications.

Hyaluronic acid-grafted PLGA nanoparticles for the sustained delivery of berberine chloride for an efficient suppression of Ehrlich ascites tumors.

To promote the specific targeting and elimination of CD44-positive cancer cells, berberine chloride (BRB)-encapsulated hyaluronic acid-grafted poly(lactic-co-glycolic acid) copolymer (BRB-d(HA)-g-PLGA) nanoparticles (NPs) were prepared. The targeted action of these NPs was compared to non-targeted BRB-loaded PLGA NPs and bulk BRB. The in vitro studies demonstrated faster release of BRB and increased cytotoxicity of BRB-d(HA)-g-PLGA NPs in Hela and MCF-7 cells in comparison to BRB-PLGA NPs and bulk BRB. The uptake of BRB-d(HA)-g-PLGA NPs was increased in case of MCF-7 cells as compared to HeLa cells owing to the higher expression of CD44 receptors on MCF-7 cells. The CD44 receptor-mediated uptake of these NPs was confirmed through competitive inhibition experiments. The in vitro results were further validated in vivo in Ehrlich Ascites Carcinoma (EAC)-bearing mice. EAC-bearing mice were injected intravenously with these NPs and the results obtained were compared with that of BRB-PLGA NPs and bulk BRB. BRB-d(HA)-g-PLGA NPs were found to significantly enhance apoptosis, sub-G1 content, life span, mean survival time, and ROS levels in EAC cells with subsequent decrease in mitochondrial membrane potential and tumor burden ion tumor-bearing mice. Taking into account the findings of in vitro and in vivo studies, the enhanced and targeted anti-tumor activity of HA-grafted PLGA copolymer-encapsulated NPs of BRB cannot be negated. Therefore, HA-grafted nanoparticle-based delivery of BRB may offer a promising and improved alternative for anti-tumor therapy.

Curcumin loading potentiates the chemotherapeutic efficacy of selenium nanoparticles in HCT116 cells and Ehrlich's ascites carcinoma bearing mice.

The anticancer properties of selenium (Se) and curcumin nanoparticles in solo formulations as well as in combination with other therapeutic agents have been proved time and again. Exploiting this facet of the two, we clubbed their tumoricidal characteristics and designed curcumin loaded Se nanoparticles (Se-CurNPs) to achieve an enhanced therapeutic effect. We evaluated their therapeutic effects on different cancer cell lines and Ehrlich's ascites carcinoma mouse model. In vitro results showed that Se-CurNPs were most effective on colorectal carcinoma cells (HCT116) compared to the other cancer cell lines used and possessed pleiotropic anticancer effects. The therapeutic effect on HCT116 was primarily attributed to an elevated level of autophagy and apoptosis as evident from significant up-regulation of autophagy associated (LC3B-II) and pro-apoptotic (Bax) proteins, down-regulation of anti-apoptotic (Bcl-2) protein and Cytochrome c (cyt c) release from mitochondria along with reduced NFκB signaling and EMT based machineries marked by downregulation of inflammation (NFκB, phospho-NFκB) and epithelial-mesenchymal transition (CD44, N-cadherin) associated proteins. In vivo studies on Ehrlich's ascites carcinoma (EAC) mice model indicated that Se-CurNPs significantly reduced the tumor load and enhanced the mean survival time (days) of tumor-bearing EAC mice.

Does a correlation exist between nasal airway volume and craniofacial morphology: A cone beam computed tomography study.

AIM: To find the correlation between nasal airway volume and the craniofacial morphology using cone beam computed tomography (CBCT). MATERIALS AND METHODS: This study consisted of preorthodontic anonymized CBCT scans of 34 healthy adults in the age span of 18-28 years. The volume was calculated using Dolphin 3D R software 11.5 version using semiautomatic segmentation method to calculate nasal volume after determining the nasal airway boundary. The subjects were grouped according to sagittal skeletal relation, craniofacial width, facial index, and facial form. RESULTS: There was statistically significant correlation between nasal volume and craniofacial width (P = 0.009). CONCLUSION: Nasal volume was correlated only with width of the face and not with width/length ratio of face that could have affected the nasal volume.

Bioreducible polyethylenimine nanoparticles for the efficient delivery of nucleic acids.

Recently, non-viral vectors for nucleic acid delivery have received considerable attention. Among the various non-viral vectors, branched polyethylenimine (bPEI, 25 kDa) has been one of the most widely used carrier systems due to its high transfection efficiency, however, it imparts high cytotoxicity. In this study, we have crosslinked bPEI with a bioreducible linker, 3,3'-dithiodipropionic acid (DTPA), via electrostatic interactions to obtain DTPA crosslinked bPEI (DP) nanoparticles. The crosslinking significantly reduced the cytotoxicity of the nanoparticles. To arrive at the best formulation in terms of nucleic acid transfection, a series of DP nanoparticles were prepared by varying the percentage of crosslinking. The dual action of DTPA, i.e. partial blocking of the charge density as well as crosslinking to convert bPEI into its nanoparticles, did not alter the pDNA condensation ability of the so-formed nanoparticles, rather the strategy favoured the unpackaging of the complexes inside the cells improving the release of pDNA, which resulted in a higher transfection efficiency. All the formulations carried nucleic acids inside the cells and exhibited significantly higher transfection efficiencies than native bPEI and the commercial transfection reagent, Lipofectamine™. Sequential siRNA delivery displayed significant suppression in the target gene expression. All together, the evaluation of the delivery systems demonstrates that the newly synthesized DP NPs are quite promising as non-viral gene carriers.

Biocatalytic and antimicrobial activities of gold nanoparticles synthesized by Trichoderma sp.

The aim of this work was to synthesize gold nanoparticles by Trichoderma viride and Hypocrea lixii. The biosynthesis of the nanoparticles was very rapid and took 10 min at 30 °C when cell-free extract of the T. viride was used, which was similar by H. lixii but at 100 °C. Biomolecules present in cell free extracts of both fungi were capable to synthesize and stabilize the formed particles. Synthesis procedure was very quick and environment friendly which did not require subsequent processing. The biosynthesized nanoparticles served as an efficient biocatalyst which reduced 4-nitrophenol to 4-aminophenol in the presence of NaBH4 and had antimicrobial activity against pathogenic bacteria. To the best of our knowledge, this is the first report of such rapid biosynthesis of gold nanoparticles within 10 min by Trichoderma having plant growth promoting and plant pathogen control abilities, which served both, as an efficient biocatalyst, and a potent antimicrobial agent.

Role of Transrectal Ultrasound in Preoperative Local Staging of Carcinoma Rectum and It's Histopathological Correlation.

A precise knowledge of depth of invasion of tumor is essential for the planning of treatment of rectal cancer. TRUS is a new diagnostic modality that has become useful in determining depth of invasion preoperatively and the presence or absence of metastatic lymph nodes. Our aim was to determine Role of Transrectal Ultrasound in Preoperative Local Staging of Carcinoma Rectum and it's Histopathological Correlation. TRUS was used in preoperative local staging of 30 patients with carcinoma rectum. 25patients underwent APR (abdomino-perineal resection) & 5 underwent AR. (anterior resection). Preoperative TRUS staging was compared with pathological staging obtained from biopsy of resected specimen. In staging depth of invasion of rectal wall (T stage) overall accuracy was 83.3 %, over staged 10 %, under staged in 6.67 % sensitivity was 92.5 %, and specificity was 62.5 %. In staging lymph nodes (N stage) overall accuracy was 76.67 %, sensitivity was 79.31 %, specificity was 87.5 %. TRUS is a safe and accurate preoperative local staging method for assessment of both depth of invasion of rectal wall and presence or absence of metastatic lymph nodes.

Management of odontogenic space infection with microbiology study.

INTRODUCTION: Dental infection has plagued humankind for as long as our civilization has been a fight against microorganisms by man dates back to ancient civilization. The discoveries of antibiotics are encouraging trends towards conquest of the microbial infection. MATERIALS AND METHODS: This study emphasizes the detection of pathogenic microorganisms by microbiological examination and culture of specimens representative of the infection, importance of early and correct diagnosis of infections, prompt treatment and supportive care. RESULTS: The age group most commonly involved was in the third and fourth decades of life. Extraction followed by incision and drainage was done. The most commonly involved space was submandibular followed by buccal space. Thirty isolates were obtained. 43 % of the strains were strict anaerobes and 39 % were aerobes, with mixed growth was seen in 18.52 %. Amongst aerobes alpha hemolytic Streptococcus aureus and Peptostreptococcus as anaerobes were the most predominant followed by Bacteroides and Prevotella. Mixed aerobic and anaerobic isolates were obtained from 18.52 % of total cases. Overall resistance to Penicillin was 22 %, amongst aerobes. CONCLUSION: Amoxicillin and Clavulanic acid combination performed better, as 100 % strains were sensitive to it. The results of this study saw a changing trend in terms of predominance of anaerobic bacteria over aerobic ones.

Galactomannan-PEI based non-viral vectors for targeted delivery of plasmid to macrophages and hepatocytes.

Intracellular nature and diversified locations of infectious and parasitic diseases such as leishmaniasis, trypanosomiasis, tuberculosis and hepatitis B and C pose a significant global burden and challenge to the scientists working in the area of drug discovery and drug delivery. The macrophages and hepatocytes are considered as potential target sites as they together play an important role in various infectious diseases. The present study scrutinizes the applicability of a natural biopolymer-based chemical vectors, capable of targeting both macrophages and hepatocytes, that can form a complex with plasmid and administer it into cells to produce a desired protein. The investigations were made to develop a novel series of gene carriers by conjugating depolymerized galactomannan (guar gum), a biocompatible polysaccharide with low molecular weight branched PEI (LMWP). A series of conjugates were developed and characterized using physicochemical techniques. All the GP/pDNA complexes showed significantly higher transfection efficiency with GP-3/pDNA, one of the best formulations, showed ~2.0-7.7-folds higher transfection efficacy when compared with the standard transfection reagents. Further, GP-3/pDNA displayed significantly higher target specific transfection efficiency under both in vitro and in vivo conditions. The data demonstrate the potential of GP vectors to deliver nucleic acids simultaneously to macrophages and hepatocytes in gene delivery applications.

Copolymers of covalently crosslinked linear and branched polyethylenimines as efficient nucleic acid carriers.

The present study describes the formation of copolymers of linear and branched PEIs (25 kDa each). These polyethylenimines (bPEI and IPEI) were crosslinked with each other to obtain branched-linear (BL) PEI copolymers using epichlorohydrin as a crosslinker in two steps. First, IPEI was reacted with epichlorohydrin to form IPEI-chlorohydrin (CHL) and subsequently, bPEI was grafted onto CHL in basic medium by in situ generation of epoxy functionalities. The two PEIs were crosslinked by varying the weight ratio of bPEI while keeping the amount of IPEI fixed. The ratio of two PEIs (1:1, 2:1, 3:1, 4:1 and 5:1) and crosslinking percentage of epichlorohydrin (5, 10, 15 and 20%) appeared as the main parameters to have affected the transfection efficiency. The lead conjugate/DNA complex was tested for in vivo transgene expression in Balb/c mice and was found to show maximum expression in the spleen.

Hydrophobic and membrane permeable polyethylenimine nanoparticles efficiently deliver nucleic acids in vitro and in vivo.

Conjugation through primary amines is one of the most commonly used methods to modify cationic vectors for efficient gene delivery. Here, dimethyl suberimidate, a commercially available homobifunctional reagent bearing imidoesters at the termini, has been used to crosslink branched polyethylenimine (bPEI) into its nanoparticles (crosslinked PEI nanoparticles, CLP NPs) specifically through primary amines without altering the total charge on the resulting NPs for interaction with biomolecules and cell membranes. By varying the degree of crosslinking, a small series of CLP NPs was prepared and evaluated for their capability to deliver nucleic acids in vitro and in vivo. Physico-chemical characterization revealed the size of the NPs in the range of ∼152 to 210 nm with zeta potential ∼+35 to +38 mV. The plasmid DNA binding ability of these nanoparticles was examined by mobility shift assay, where the pDNA migration was found to be completely retarded by these NPs at an N/P ratio of 4 (cf. bPEI at N/P 3). In various mammalian cells, CLP/pDNA nanoplexes were not only found to be non-toxic but also exhibited significantly enhanced gene expression with one of the formulations, the CLP3/pDNA nanoplex, displaying the highest transfection efficiency, outperforming native bPEI and the selected commercial transfection reagents both in the presence and absence of serum. Further, the versatility of the vector, CLP3, was demonstrated by sequential delivery of GFP-specific siRNA to HEK293 cells, which resulted in ∼79% suppression of the target gene. Intracellular localization studies showed a significant population of the dual labeled nanoplex (CLP3/pDNA) in the nucleus in just 60 min of incubation. Luciferase reporter gene analysis in Balb/c mice post-intravenous administration of the CLP3/pDNA nanoplex showed the highest gene expression in their spleen. The study suggests that CLP NPs could be used as efficient gene delivery vectors for future gene therapy applications.

Engineered polymer-supported synthesis of 3'-carboxyalkyl-modified oligonucleotides and their applications in the construction of biochips for diagnosis of the diseases.

An engineered polymer support 5 has been prepared for the solid-phase assembly of 3'-carboxyalkyl-modified oligonucleotides using commonly available reagents. A two-step deprotection procedure resulted in the quantitative cleavage of oligonucleotides from the support and removal of the protecting groups from phosphodiesters and exocyclic amino groups of the nucleic bases. The fully deprotected oligomers, obtained in high yield, were desalted and analyzed on RP-HPLC. After characterization by MALDI-TOF, these carboxyalkylated oligonucleotides were immobilized onto the epoxy-functionalized glass microslides to prepare biochips. The performance of these biochips was evaluated under different sets of conditions and then successfully validated by the detection of base mismatches and human infectious disease, bacterial meningitis, caused by N. meningitidis.

Polyglutamic acid-based nanocomposites as efficient non-viral gene carriers in vitro and in vivo.

A series of polyethylenimine (PEI) and γ-polyglutamic acid (PGA) nanocomposites (PPGA) was prepared and evaluated in terms of their cell viability and transfection efficiency in vitro and in vivo. On complexion with pDNA, the positively charged PPGA/DNA nanocomposites resulted in a higher level of in vitro reporter gene transfection (2.7-7.9-fold) as compared to native PEI, and selected commercial reagents and >95% cell viability in HEK293, HeLa and HepG2 cell lines. Further, PPGA-5 nanocomposite (the best working system in terms of transfection efficiency among the series) was found to efficiently transfect primary mouse keratinocytes up to 22% above the control level. PPGA-5, when tested for in vivo cytotoxicity in Drosophila, did not induce any stress in the exposed larvae in comparison with control. In vivo gene expression using PPGA-5 showed the highest transfection efficiency in spleen of mouse closely followed by heart tissues after intravenous injection through tail vein. Besides, these nanocomposites also delivered siRNA efficiently into mammalian cells, resulting in ∼ 80% suppression of EGFP expression. These results together demonstrated the potential of the projected nanocomposites for in vivo gene delivery.

Gellan gum-PEI nanocomposites as efficient gene delivery agents.

Of the non-viral vectors, a cationic polymer like PEI is an attractive candidate which however, has been negatively impacted due to its marked toxicity. An anionic sugar polymer gelan gum (GG) has been introduced into PEI system to increase transfection efficiency with minimal toxicity. We showed that one of the synthesized (GP1-GP6) GG-PEI nanocomposites (NCs), GP3, exhibited negligible toxicity in in vitro (primary keratinocytes, HEK293, HeLa and HepG2 cells) and in vivo (Drosophila melanogaster) as compared to PEI or lipofectamin. GP3-pDNA complex was found to be transfected efficiently in the above cells as confirmed by FACS analysis (72.0 + 5.5%) while lipofectamine showed only 12.4 + 3.5% efficiency. GP3 mediated GFP specific siRNA delivery resulted in the knockdown of the GFP expression by approximately 77% and JNK (60%). In vivo gene expression studies in mice revealed reporter gene expression in spleen. The study demonstrates that GG blended PEI NCs hold promise for future applications in gene delivery both in vitro and in vivo.

In vitro and in vivo evaluation of linear polyethylenimine nanoparticles.

bPEI (polyethylenimine, 25 kDa, gold standard) is highly effective in transfection efficiency owing to its high buffering capacity, however, cytotoxicity limits its use in in vivo applications. We hypothesized that partial conversion of secondary amines in IPEI to tertiary amines, while preserving the overall number of amines, would result in improved buffering capacity, which may, in turn, improve transfection efficiency of the resulting nanoparticles with cell viability comparable to that of native IPEI. IPEI was crosslinked with BDE to obtain a series of IPEI nanoparticles (LPN-1 to LPN-8) which were obtained in approximately 80-85% yield. These particles were relatively non-toxic in vitro and in vivo. In vivo gene expression studies using LPN-5 in Balb/c mice through i.v. injection showed maximum expression of the reporter gene in the spleen. These results demonstrate the potential of these particles as efficient transfection reagents.