A Highly Efficient and Stable Photocatalyst; N-Doped ZnO/CNT Composite Thin Film Synthesized via Simple Sol-Gel Drop Coating Method.

The thin film of N-doped ZnO/CNT nanocomposite was successfully fabricated on soda lime glass substrate by a simple sol-gel drop-coating method. The structural, morphological, chemical, and optical properties of as prepared samples were characterized by a variety of tools such as X-ray Diffraction (XRD), Field Emission Scanning Electron Microscopy (FE-SEM), Fourier Transform Infrared spectroscopy (FT-IR), and UV-visible spectroscopy. The hexagonal crystalline structure was confirmed from XRD measurement without any other impurity phase detection in samples. The N-doped ZnO/CNT composite showed excellent photo-catalytic activity towards cationic methylene blue (MB) dye degradation with 100% removal rate under UV light irradiation as compared to N-doped ZnO (65%) and pure ZnO (47.36%). The convincing performance has also been observed for the case of visible light irradiation. The enhancement of that photocatalytic activity might be due to narrowing the band gap as well as the reduction of electron-hole pair recombination in ZnO matrix with the incorporation of dopant nitrogen and CNT. It is assumed from the obtained results that N-doped ZnO/CNT nanocomposite thin film can be employed as an economically achievable and ecofriendly method to degrade dye with UV and visible light irradiation. Additionally, density functional theory (DFT) calculations were applied to explore the effect of N-doping on electronic structure of ZnO. The computational study has supported the experimental results of significant band gap contraction, which leads to the maximum absorption towards higher wavelength and no appreciable change of lattice parameters after doping. A conceivable photocatalytic mechanism of N-doped ZnO/CNT nanocomposite has been proposed as well.

Dataset on concentration and enrichment factor of rare earth elements (REEs) in sediments of Linggi River, Malaysia.

This study is on the distribution of rare earth elements (REEs) concentrations in sediments collected from 113 sampling locations of Linggi River. The analysis of sediment samples was performed by Neutron Activation Analysis (NAA) and Inductively Coupled Plasma - Mass spectrometer (ICP-MS). The main compositions of Linggi river sediments were silt > sand > clay. The mean of total concentrations of REEs (ΣREE), light REEs (ΣLREE) and heavy REEs (ΣHREE) in Linggi sediment were 249, 228, and 22.0 mg/kg, respectively. The results of Linggi river sediment were normalised to several reference shale values. REEs of Linggi river sediments were comparable to MUQ reference shale values. Enrichment factors (EF) of mean values indicate Linggi River sediment can be categorised as having minor to moderate enrichment.

Rare earth elements (REEs) as pollution indicator in sediment of Linggi River, Malaysia.

A study was carried out to determine the concentrations of rare earth elements (REEs) in Linggi river sediments collected from 113 sampling locations. The sediment analysis was performed by Neutron activation analysis (NAA) and Inductively coupled plasma - mass spectrometry (ICP-MS). The results of Linggi river sediment were normalized to "recent" reference shale values. The means of total concentrations of REEs (ΣREE), light REEs (ΣLREE) and heavy REEs (ΣHREE) in Linggi sediment were 241.2, 219.2, and 22.0 mg/kg, respectively, which indicates enrichment compared to ΣREE, ΣLREE and ΣHREE reference shale values. Results obtained from enrichment factors (EF) show no enrichment to moderate enrichment of Linggi sediments, indicating the sources of REEs pollution originated from natural and land-based activities. A similar pattern was observed by comparing the REEs values of Linggi sediments to other references shale values. Ce (δCe) and Eu (δEu) anomalies indicate Linggi sediments showed positive anomaly of Ce whilst negative anomaly of Eu.

The sources and ecological risk assessment of elemental pollution in sediment of Linggi estuary, Malaysia.

In this study, concentrations of heavy metals, rare earth elements (REEs), Uranium (U) and Thorium (Th) of the actinide group were determined from Linggi estuary sediment samples by neutron activation analysis (NAA) and inductive coupled plasma - mass spectrometry techniques. The geo-accumulation (Igeo) and ecological risk index (Ri) values were calculated to identify the quality status of Linggi estuary sediments. Results indicated Linggi estuary was polluted by arsenic (As), lead (Pb) and antimony (Sb). REEs, U and Th showed significant increase of concentration in Linggi estuary sediments. Ri of Linggi estuary was categorised as low to considerable ecological risk, which indicates no significant to moderate effect on the majority of the sediment-dwelling organisms. Correlation matrix and principal component analysis assessed pollution sources to be both natural and anthropogenic.

Assessment of toxic elements in sediments of Linggi River using NAA and ICP-MS techniques.

Fourteen sediment samples were collected along Linggi River, Malaysia. Neutron activation analysis (NAA) and inductively coupled plasma-mass spectrometry (ICP-MS) techniques were used in the determination of toxic element contents. The results showed that As, Cd and Sb concentrations were higher at all sampling stations, with enrichment factor values ranging from 17.7 to 75.0, 2.1 to 19.5 and 6.6 to 28.4, respectively. Elements of Pb and Zn) were also enriched at most of the sampling stations whilst Cu, Cr and Ni were shown as background levels. The sediment of Linggi River can be categorised as low (<8.0) to very high degree of contamination (>32.0). The mean concentrations of elements viz. Cd, Cr, Ni, Pb, Sb and Zn were lower than the threshold effect level (TEL) of FSQGs values except for As. The concentration of As (arsenic) was higher than PEL and PEC of FSQGs values.

Multivariate analysis for source identification of pollution in sediment of Linggi River, Malaysia.

Rapid socioeconomic development in the Linggi River Basin has contributed to the significant increase of pollution discharge into the Linggi River and its adjacent coastal areas. The toxic element contents and distributions in the sediment samples collected along the Linggi River were determined using neutron activation analysis (NAA) and inductively coupled plasma-mass spectrometry (ICP-MS) techniques. The measured mean concentration of As, Cd, Pb, Sb, U, Th and Zn is relatively higher compared to the continental crust value of the respective element. Most of the elements (As, Cr, Fe, Pb, Sb and Zn) exceeded the freshwater sediment quality guideline-threshold effect concentration (FSQG-TEC) value. Downstream stations of the Linggi River showed that As concentrations in sediment exceeded the freshwater sediment quality guideline-probable effect concentration (FSQG-PEC) value. This indicates that the concentration of As will give an adverse effect to the growth of sediment-dwelling organisms. Generally, the Linggi River sediment can be categorised as unpolluted to strongly polluted and unpolluted to strongly to extremely polluted. The correlation matrix of metal-metal relationship, principle component analysis (PCA) and cluster analysis (CA) indicates that the pollution sources of Cu, Ni, Zn, Cd and Pb in sediments of the Linggi River originated from the industry of electronics and electroplating. Elements of As, Cr, Sb and Fe mainly originated from motor-vehicle workshops and metal work, whilst U and Th originated from natural processes such as terrestrial runoff and land erosion.

Distribution of Heavy Metals in Core Marine Sediments of Coastal East Malaysia by Instrumental Neutron Activation Analysis and Inductively Coupled Plasma Spectroscopy.

Fifty-five core marine sediments from three locations at South China Sea and one location each at Sulu Sea and Sulawesi Sea of coastal East Malaysia were analyzed for heavy metals by instrumental neutron activation analysis and inductively coupled plasma mass spectroscopy. The enrichment factor and the modified degree of contamination were used to calculate the anthropogenic and pollution status of the elements in the samples. The enrichment factor of As, Cd, Cr, Cu, Ni, Pb, and Zn varied from 0.42-4.26, 0.50-2.34, 0.31-0.82, 0.20-0.61, 0.91-1.92, 0.23-1.52, and 0.90-1.28, respectively, with the modified degree of contamination values below 0.6. Comparative data showed that coastal East Malaysia has low levels of contamination.

RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake.

In the paralytic disease botulism, the botulinum neurotoxin (BoNT) passes through the bloodstream to reach and inactivate neuromuscular junctions. Monoclonal antibodies (mAbs) may be useful BoNT countermeasures, as mAb combinations can rapidly clear BoNT from the blood circulation. We have previously shown that the BoNT-neutralizing potency of mAbs can be improved through red blood cell (RBC) immunoadherence. For example, a fusion protein (FP) that adheres biotinylated mAbs to the RBC surface enabled a pair of mAbs to neutralize 5000 LD50 BoNT/A in the mouse protection assay. Here, we added two mAbs to that combination, creating a 4-mAb:FP complex that neutralized 40,000 LD50 BoNT/A in vivo, and analyzed functional correlates of neutralization. The FP enhanced potency of BoNT/A immune complexes, providing the greatest magnitude of benefit to the 4-mAb combination. RBC binding of a BoNT/A complexed with 4-mAb:FP exhibited a bi-phasic clearance process in vivo. Most of the complexes were cleared within five minutes; the rest were cleared gradually over many hours. Peritoneal macrophages showed better uptake of the 4-mAb complex than the 3-mAb complex, and this was not affected by the presence of the FP. However, the addition of RBCs to the 4-mAb:FP BoNT/A doubled macrophage uptake of the complexes. Lastly, the 4-mAb:FP BoNT/A complex synergistically induced M2 macrophage polarization, as indicated by IL-10 expression, whether or not RBCs were present. RBC-targeted immunoadherence through the FP is a potent enhancer of mAb-mediated BoNT/A neutralization in vivo, and can have positive effects on BoNT/A sequestration, immune complex uptake, and macrophage activation.

Distribution of Trace Elements in Core Marine Sediments of Coastal East Malaysia by Instrumental Neutron Activation Analysis.

A study was carried out on the distribution and enrichment of trace elements in the core marine sediments of East Malaysia from three stations at South China Sea and one station each at Sulu Sea and Sulawesi Sea. Five stations of sediment cores were recovered and the vertical concentration profiles of six elements namely Br, Cs, Hf, Rb, Ta, and V were determined using the instrumental neutron activation analysis. The enrichment factor, geoaccumulation index and the modified degree of contamination were used to calculate the anthropogenic and pollution status of the elements in the samples. Except for Cs and Hf, which by the enrichment factor are categorized from minimum enrichment to moderate enrichment in all stations and for V and Rb in Sulu Sea and Sulawesi Sea, which are categorized minimum enrichment, other elements are found to be no enrichment at all stations. The geoaccumulation index of Hf in one station shows moderately polluted and for other elements are unpolluted. However, the modified degree values of all samples are less than 1, suggesting very low contamination of elements found in all the stations.

Rare earth elements in core marine sediments of coastal East Malaysia by instrumental neutron activation analysis.

A study was carried out on the concentration of REEs (Dy, Sm, Eu,Yb, Lu, La and Ce) that are present in the core marine sediments of East Malaysia from three locations at South China Sea and one location each at Sulu Sea and Sulawesi Sea. The sediment samples were collected at a depth of between 49 and 109 m, dried, and crushed to powdery form. The entire core sediments prepared for Instrumental Neutron Activation Analysis (INAA) were weighted approximately 0.0500 g to 0.1000 g for short irradiation and 0.1500 g to 0.2000 g for long irradiation. The samples were irradiated with a thermal neutron flux of 4.0×10(12) cm(-2) s(-1) in a TRIGA Mark II research reactor operated at 750 kW. Blank samples and standard reference materials SL-1 were also irradiated for calibration and quality control purposes. It was found that the concentration of REEs varies in the range from 0.11 to 36.84 mg/kg. The chondrite-normalized REEs for different stations suggest that all the REEs are from similar origins. There was no significant REEs contamination as the enrichment factors normalized for Fe fall in the range of 0.42-2.82.

Mechanisms of enhanced neutralization of botulinum neurotoxin by monoclonal antibodies conjugated to antibodies specific for the erythrocyte complement receptor.

Immune complexes formed between monoclonal antibodies (mAbs) and toxins can neutralize toxicity in vivo by multiple mechanisms. Toxin sequestration and clearance by mAbs may be improved by enhancing their ability to bind to red blood cells (RBCs) through immune adherence. This can be achieved by converting the mAbs to heteropolymers (HPs), which are antigen-specific mAbs cross-linked to mAbs targeting the complement receptor (CR1), a protein that is expressed on the surface of RBCs in primates and mediates delivery of complement C3b-containing immune complexes to tissue macrophages. Conversion of mAbs to HPs has been shown to enhance clearance of multivalent antigens from the blood circulation, but the interaction of HPs with monovalent toxins has not been examined. Using botulinum neurotoxin (BoNT) as a model system, we studied the effect of conversion of a pair of BoNT-specific mAbs into HPs on toxin neutralization and handling in vivo. Two HPs given in combination had 166-fold greater potency than un-modified mAbs, neutralizing 5000 LD50 BoNT, when tested in transgenic mice expressing human CR1 on RBC membranes. Improvement required adherence of BoNT to the RBC in vivo and 2 HPs, rather than an HP+mAb pair. The HP pair bound BoNT to RBCs in the circulation for 2h, in comparison to BoNT-neutralizing anti-serum, which induced no detectable RBC binding. HP pairs exhibited enhanced uptake by peritoneal macrophages in vitro, compared to pairs of mAbs or mAb+HP pairs. In a post-exposure therapeutic model, HPs gave complete protection from a lethal BoNT dose up to 3h after toxin exposure. In a pre-exposure prophylaxis model, mice given HP up to 5 days prior to BoNT administration were fully protected from a lethal BoNT dose. These studies elucidate general mechanisms for the neutralization of toxins by HP pairs and demonstrate the potential utility of HPs as BoNT therapeutics.

Evidence that botulinum toxin receptors on epithelial cells and neuronal cells are not identical: implications for development of a non-neurotropic vaccine.

Botulinum toxin typically interacts with two types of cells to cause the disease botulism. The toxin initially interacts with epithelial cells in the gut or airway to undergo binding, transcytosis, and delivery to the general circulation. The toxin then interacts with peripheral cholinergic nerve endings to undergo binding, endocytosis, and delivery to the cytosol. The receptors for botulinum toxin on nerve cells have been identified, but receptors on epithelial cells remain unknown. The initial toxin binding site on nerve cells is a polysialoganglioside, so experiments were performed to determine whether polysialogangliosides are also receptors on epithelial cells. A series of single mutant and dimutant forms of the botulinum toxin type A binding domain (HC50) were cloned and expressed. One of these (dimutant HC50 A(W1266L,Y1267S)) was shown to have lost its ability to bind nerve cells (phrenic nerve-hemidiaphragm preparation), yet it retained its ability to bind and cross human epithelial monolayers (T-84 cells). In addition, the wild-type HC50 and the dimutant HC50 displayed the same ability to undergo binding and transcytosis (absorption) in a mouse model. The fact that the dimutant retained the ability to cross epithelial barriers but did not possess the ability to bind to nerve cells was exploited to create a mucosal vaccine that was non-neurotropic. The wild-type HC50 and non-neurotropic HC50 proved to be comparable in their abilities to: 1) evoke a circulating IgA and IgG response and 2) evoke protection against a substantial challenge dose of botulinum toxin.

An adenoviral vector-based mucosal vaccine is effective in protection against botulism.

A replication-incompetent adenoviral vector encoding the heavy chain C-fragment (H(C)50) of botulinum neurotoxin type C (BoNT/C) was evaluated as a mucosal vaccine against botulism in a mouse model. Single intranasal inoculation of the adenoviral vector elicited a high level of H(C)50-specific IgG, IgG1 and IgG2a in sera and IgA in mucosal secretions as early as 2 weeks after vaccination. The antigen-specific serum antibodies were maintained at a high level at least until the 27th week. Immune sera showed high potency in neutralizing BoNT/C as indicated by in vitro toxin neutralization assay. The mice receiving single dose of 2 x 10(7) p.f.u. (plaque-forming unit) of adenoviral vector were completely protected against challenge with up to 10(4) x MLD(50) of BoNT/C. The protective immunity showed vaccine dose dependence from 10(5) to 2 x 10(7) p.f.u. of adenoviral vector. In addition, animals receiving single intranasal dose of 2 x 10(7) p.f.u. adenoviral vector could be protected against 100 x MLD(50) 27 weeks after vaccination. Animals with preexisting immunity to adenovirus could also be vaccinated intranasally and protected against lethal challenge with BoNT/C. These results suggest that the adenoviral vector is a highly effective gene-based mucosal vaccine against botulism.

Protective immunity against botulism provided by a single dose vaccination with an adenovirus-vectored vaccine.

Botulinum neurotoxins cause botulism, a neuroparalytic disease in humans and animals. We constructed a replication-incompetent adenovirus encoding a synthesized codon-optimized gene for expression of the heavy chain C-fragment (H(C)50) of botulinum neurotoxin type C (BoNT/C). This recombinant human serotype 5 adenoviral vector (Ad5) was evaluated as a genetic vaccine candidate against botulism caused by BoNT/C in a mouse model. A one-time intramuscular injection with 10(5) to 2 x 10(7)pfu of adenoviral vectors elicited robust serum antibody responses against H(C)50 of BoNT/C as assessed by ELISA. Immune sera showed high potency in neutralizing the active BoNT/C in vitro. After a single dose of 2 x 10(7)pfu adenoviral vectors, the animals were completely protected against intraperitoneal challenge with 100 x MLD(50) of active BoNT/C. The protective immunity appeared to be vaccine dose-dependent. The anti-toxin protective immunity could last for at least 7 months without a booster injection. In addition, we observed that pre-existing immunity to the wild-type Ad5 in the host had no significant influence on the protective efficacy of vaccination. The data suggest that an adenovirus-vectored genetic vaccine is a highly efficient prophylaxis candidate against botulism.