Development of a non-linear growth model for predicting temporal evolution of Scenedesmus obliquus with varying irradiance.

In the present study, the effect of irradiance on growth performance of Scenedesmus obliquus was investigated, and various non-linear growth models were evaluated to predict its temporal evolution. This microalga was cultured in a LED-illuminated flat-panel gas-lift photobioreactor operated in batch mode at varying irradiance ranging from 50 to 200 µmol/m2/s keeping all the other physico-chemical parameters constant. When growth data in terms of optical density were fitted in sigmoidal growth models, three non-linear models, namely, Richards model, Gompertz model, and logistic model, were found to be the best fit. Comparing these models based on statistical information, the logistic model could more appropriately and precisely describe algal growth under varying light intensity. Finally, the parameters of the logistic model were determined using regression analysis and were incorporated in the logistic equation to investigate the kinetic characteristics of S. obliquus. The optimum light intensity (Iopt) for growth was found to be 150 µmol/m2/s, at which a maximum specific growth rate (µopt) of 0.35/day was obtained. The model developed was validated experimentally and could successfully explain the photo-inhibition phenomenon occurring at light intensity above 150 µmol/m2/s.

Synthesis, characterization and the release kinetics of antiproliferative agents from polyamidoamine conjugates.

Polyamidoamine conjugates containing curcumin and bisphosphonate were synthesized via a one-pot aqueous phase Michael addition reaction. In the design of the conjugate, bisphosphonate formed an integral part of the polymer carrier backbone. Curcumin was incorporated onto the polyamidoamine backbone via piperazine linker. The conjugates were characterized by Fourier transform spectroscopy, energy-dispersive X-ray analysis, atomic force spectroscopy and nuclear magnetic resonance spectroscopy and it confirmed the successful incorporation of the antiproliferative agents onto the carriers. The weight percentage incorporation of bisphosphonate to the carriers was found to be between 2.56% and 3.34%. The in vitro release studies of curcumin from the polyamidoamine conjugate were performed in dialysis bag at selected pH values. The release of curcumin was significantly slower at pH 7.4 when compared to pH 5.8. The release profiles indicate that the conjugates are more stable at pH 7.4 and are potential sustained drug-delivery systems for combination therapy.

Severe adenomyosis and CA125.

A total of 50 women having uterine enlargement of > 12 weeks' size due to severe adenomyosis with raised CA125, were carefully studied and compared with other cases. The greater the enlargement of the uterus over 240 cm(3) volume or 12 weeks' uterine size due to severe adenomyosis, the greater was the rise of CA125 levels and these were disturbingly high in some cases. However, similarly enlarged uteri due to fibroids did not show a rise in CA125 levels above normal. Raised CA125 thus need not instil anxiety, when the clinical findings are benign.

Dissolvable sodium alginate-based antibacterial wound dressing patches: Design, characterization, and in vitro biological studies.

The treatment of infected wounds in patients with highly sensitive skin is challenging. Some of the available wound dressings cause further skin tear and bleeding upon removal thereby hindering the healing process. In this study, dissolvable antibacterial wound dressing patches loaded with cephalexin monohydrate were prepared from different amounts of sodium alginate (SA) and carboxymethyl cellulose (CMC) by the solvent casting evaporation technique. The patches displayed good tensile strength (3.83-13.83 MPa), appropriate thickness (0.09 to 0.31 mm) and good flexibility (74-98 %) suitable for the skin. The patches displayed good biodegradability and low moisture uptake suitable to prevent microbial invasion on the wound dressings upon storage. The release profile of the drug from the patches was sustained in the range of 47-80 % for 48 h, revealing their capability to inhibit bacterial infection. The biological assay showed that the patches did not induce cytotoxic effects on HaCaT cells, revealing good biocompatibility. The antimicrobial effect of the patches on the different strains of bacteria used in the study was significant. The cell migration (66.7-74.3 %) to the scratched gap was promising revealing the patches' capability to promote wound closure. The results obtained show that the wound dressings are potential materials for the treatment of infected wounds.

Seated reach distance and trunk excursion accurately reflect dynamic postural control in individuals with motor-incomplete spinal cord injury.

STUDY DESIGN: To determine whether kinematic measures of reach excursion are valid measures of postural control in individuals with motor-incomplete spinal cord injury (MISCI), seated reach test (SRT; obtained from a wrist marker) values and associated trunk excursion values (obtained from a C7 marker) were compared with center of pressure excursion (COPE). Data were obtained from individuals with MISCI and from non-disabled individuals for each of four directions. To assess the reliability of these measures in subjects with MISCI, these values were collected on two separate days. OBJECTIVE: Dynamic seated postural control is essential for individuals who perform their daily activities from a wheelchair. Although seated reach distance is used as a proxy measure for postural control, it is unknown whether this is an accurate and reliable measure of the limits of stability in individuals with MISCI. RESULTS: In subjects with MISCI, the SRT was significantly correlated with the COPE in three directions of reaching (r ≥ 0.71), with the exception being leftward reaching. Trunk excursion was correlated with COPE in all directions (r ≥ 0.93). In non-disabled individuals, both SRT (r ≥ 0.56) and trunk excursion (r ≥ 0.91) were correlated with COPE for all directions. In individuals with MISCI, there was significant intersession agreement for both reach distance (intraclass correlation coefficient (ICC)≥ 0.78) and trunk excursion (ICC ≥ 0.77) measured in all directions. CONCLUSIONS: Although both SRT and trunk excursion are reasonable reflections of COPE, measurement of postural control based on trunk excursion has advantages for individuals who may have difficulty maintaining arm position during reaching. Reach distance is highly reliable in individuals with MISCI in all directions of reaching.

Pasting properties of hydrothermally treated maize starch with added stearic acid.

The effects of stearic acid addition followed by hydrothermal treatment on the functional properties of maize starch were studied. Addition of stearic acid followed by hydrothermal treatment resulted in non-gelling starch. Starch with stearic acid had significantly (P < 0.05) higher viscosity as compared to heat-moisture treated starch. There was no significant difference on the pasting properties of starch with stearic acid alone and in combination with annealing. Stearic acid addition followed by heat-moisture treatment significantly reduced starch susceptibility to acid hydrolysis as compared to stearic acid alone and heat-moisture treatment alone. These changes related well with the increased amylose lipid complexes and relative crystallinity observed by the DSC and XRD, suggesting that heat-moisture treatment promoted amylopectin side chain crosslinking and amylose-stearic acid complex formation. Stearic acid addition followed by hydrothermal treatment produced a 'clean label' starch that can potentially substitute chemically cross-linked and non-gelling starch in the food industry.

A highly responsive NH3 sensor based on Pd-loaded ZnO nanoparticles prepared via a chemical precipitation approach.

The gas-detecting ability of nanostructured ZnO has led to significant attention being paid to the development of a unique and effective approach to its synthesis. However, its poor sensitivity, cross-sensitivity to humidity, long response/recovery times and poor selectivity hinder its practical use in environmental and health monitoring. In this context, the addition of noble metals, as dopants or catalysts to modify the ZnO surface has been examined to enhance its sensing performance. Herein, we report preparation of Pd-loaded ZnO nanoparticles via a chemical precipitation approach. Various Pd loadings were employed to produce surface-modified ZnO nanostructure sensors, and their resulting NH3 sensing capabilities both in dry and humid environments were investigated. Through a comparative gas sensing study between the pure and Pd-loaded ZnO sensors upon exposure to NH3 at an optimal operating temperature of 350 °C, the Pd-loaded ZnO sensors were found to exhibit enhanced sensor responses and fast response/recovery times. The influence of Pd loading and its successful incorporation into ZnO nanostructure was examined by X-ray diffraction, high resolution-transmission electron microscopy, and X-ray photoelectron spectroscopy. XPS studies demonstrated that in all samples, Pd existed in two chemical states, namely Pd° and Pd2+. The possible sensing mechanism related to NH3 gas is also discussed in detail.

Properties and Characterization of a PLA-Chitin-Starch Biodegradable Polymer Composite.

This paper presents a comparison on the effects of blending chitin and/or starch with poly(lactic acid) (PLA). Three sets of composites (PLA-chitin, PLA-starch and PLA-chitin-starch) with 92%, 94%, 96% and 98% PLA by weight were prepared. The percentage weight (wt.%) amount of the chitin and starch incorporated ranges from 2% to 8%. The mechanical, dynamic mechanical, thermal and microstructural properties were analyzed. The results from the tensile strength, yield strength, Young's modulus, and impact showed that the PLA-chitin-starch blend has the best mechanical properties compared to PLA-chitin and PLA-starch blends. The dynamic mechanical analysis result shows a better damping property for PLA-chitin than PLA-chitin-starch and PLA-starch. On the other hand, the thermal property analysis from thermogravimetry analysis (TGA) shows no significant improvement in a specific order, but the glass transition temperature of the composite increased compared to that of neat PLA. However, the degradation process was found to start with PLA-chitin for all composites, which suggests an improvement in PLA degradation. Significantly, the morphological analysis revealed a uniform mix with an obvious blend network in the three composites. Interestingly, the network was more significant in the PLA-chitin-starch blend, which may be responsible for its significantly enhanced mechanical properties compared with PLA-chitin and PLA-starch samples.

Application of thin layer activation technique for monitoring corrosion of carbon steel in hydrocarbon processing environment.

Acidic crude oil transportation and processing in petroleum refining and petrochemical operations cause corrosion in the pipelines and associated components. Corrosion monitoring is invariably required to test and prove operational reliability. Thin Layer Activation (TLA) technique is a nuclear technique used for measurement of corrosion and erosion of materials. The technique involves irradiation of material with high energy ion beam from an accelerator and measurement of loss of radioactivity after the material is subjected to corrosive environment. In the present study, TLA technique has been used to monitor corrosion of carbon steel (CS) in crude oil environment at high temperature. Different CS coupons were irradiated with a 13 MeV proton beam to produce Cobalt-56 radioisotope on the surface of the coupons. The corrosion studies were carried out by subjecting the irradiated coupons to a corrosive environment, i.e, uninhibited straight run gas oil (SRGO) containing known amount of naphthenic acid (NA) at high temperature. The effects of different parameters, such as, concentration of NA, temperature and fluid velocity (rpm) on corrosion behaviour of CS were studied.

Life cycle assessment of facile microwave-assisted zinc oxide (ZnO) nanostructures.

The life cycle assessment of several zinc oxide (ZnO) nanostructures, fabricated by a facile microwave technique, is presented. Key synthesis parameters such as annealing temperature, varied from 90°C to 220°C, and microwave power, varied from 110W to 710W, are assessed. The effect of these parameters on both the structural characteristics and the environmental sustainability of the nanostructures is examined. The nanostructures were characterized by means of X-ray diffraction (XRD), focused ion beam scanning electron microscopy (FIB-SEM), ultraviolet-visible spectroscopy (UV-Vis), Photoluminescence (PL) and Brunauer-Emmett-Teller (BET) analysis. Crystalline size was found to be 22.40nm at 110W microwave power, 24.83nm at 310W, and 24.01nm at 710W. Microwave power and synthesis temperature were both directly proportional to the surface area. At 110W the surface area was 10.44m2/g, at 310W 12.88m2/g, and at 710W 14.60m2/g; while it was found to be 11.64m2/g at 150°C and 18.09m2/g at 220°C. Based on these, a life cycle analysis (LCA) of the produced ZnO nanoparticles was carried out, using the ZnO surface area (1m2/g) as the functional unit. It was found that the main environmental weaknesses identified during the production process were; (a) the use of ethanol for purifying the produced nanomaterials and (b) the electricity consumption for the ZnO calcination, provided by South Africa's fossil-fuel dependent electricity source. When the effect of the key synthesis parameters on environmental sustainability was examined it was found that an increase of either microwave power (from 110 to 710W) or synthesis temperatures (from 90 to 220°C), results in higher sustainability, with the environmental footprint reduced by 27% and 41%, respectively. Through a sensitivity analysis, it was observed that an electricity mix based on renewable energy could improve the environmental sustainability of the nanoparticles by 25%.

Triosephosphate isomerase from Plasmodium falciparum: the crystal structure provides insights into antimalarial drug design.

BACKGROUND: Malaria caused by the parasite Plasmodium falciparum is a major public health concern. The parasite lacks a functional tricarboxylic acid cycle, making glycolysis its sole energy source. Although parasite enzymes have been considered as potential antimalarial drug targets, little is known about their structural biology. Here we report the crystal structure of triosephosphate isomerase (TIM) from P. falciparum at 2.2 A resolution. RESULTS: The crystal structure of P. falciparum TIM (PfTIM), expressed in Escherichia coli, was determined by the molecular replacement method using the structure of trypanosomal TIM as the starting model. Comparison of the PfTIM structure with other TIM structures, particularly human TIM, revealed several differences. In most TIMs the residue at position 183 is a glutamate but in PfTIM it is a leucine. This leucine residue is completely exposed and together with the surrounding positively charged patch, may be responsible for binding TIM to the erythrocyte membrane. Another interesting feature is the occurrence of a cysteine residue at the dimer interface of PfTIM (Cys13), in contrast to human TIM where this residue is a methionine. Finally, residue 96 of human TIM (Ser96), which occurs near the active site, has been replaced by phenylalanine in PfTIM. CONCLUSIONS: Although the human and Plasmodium enzymes share 42% amino acid sequence identity, several key differences suggest that PfTIM may turn out to be a potential drug target. We have identified a region which may be responsible for binding PfTIM to cytoskeletal elements or the band 3 protein of erythrocytes; attachment to the erythrocyte membrane may subsequently lead to the extracellular exposure of parts of the protein. This feature may be important in view of a recent report that patients suffering from P. falciparum malaria mount an antibody response to TIM leading to prolonged hemolysis. A second approach to drug design may be provided by the mutation of the largely conserved residue (Ser96) to phenylalanine in PfTIM. This difference may be of importance in designing specific active-site inhibitors against the enzyme. Finally, specific inhibition of PfTIM subunit assembly might be possible by targeting Cys13 at the dimer interface. The crystal structure of PfTIM provides a framework for new therapeutic leads.

Unusual stability of a multiply nicked form of Plasmodium falciparum triosephosphate isomerase.

BACKGROUND: The limited proteolytic cleavage of proteins can result in distinct polypeptides that remain noncovalently associated so that the structural and biochemical properties of the 'nicked' protein are virtually indistinguishable from those of the native protein. The remarkable observation that rabbit muscle triosephosphate isomerase (TIM) can be multiply nicked by subtilisin and efficiently religated in the presence of an organic solvent formed the stimulus for our study on a homologous system, Plasmodium falciparum triosephosphate isomerase (PfTIM). RESULTS: The subtilisin nicked form of PfTIM was prepared by limited proteolysis using subtilisin and the major fragments identified using electrospray ionization mass spectrometry. The order of susceptibility of the peptide bonds in the protein was also determined. The structure of the nicked form of TIM was investigated using circular dichroism, fluorescence and gel filtration. The nicked enzyme exhibited remarkable stability to denaturants, although significant differences were observed with the wild-type enzyme. Efficient religation could be achieved by addition of an organic cosolvent, such as acetonitrile, in the presence of subtilisin. Religation was also demonstrated after dissociation of the proteolytic fragments in guanidinium chloride, followed by reassembly after removal of the denaturant. CONCLUSIONS: The eight-stranded beta8/alpha8 barrel is a robust, widely used protein structural motif. This study demonstrates that the TIM barrel can withstand several nicks in the polypeptide backbone with a limited effect on its structure and stability.

Flocculation and adsorption properties of biodegradable gum-ghatti-grafted poly(acrylamide-co-methacrylic acid) hydrogels.

This study reports the microwave-assisted synthesis of gum-ghatti (Gg)-grafted poly(acrylamide-co-methacrylic acid) (AAm-co-MAA) hydrogels for the development of biodegradable flocculants and adsorbents. The synthesized hydrogels were characterized using TGA, FTIR and SEM. TGA studies revealed that the synthesized hydrogels were thermally more stable than pristine Gg and exhibited maximum swelling capacity of 1959% at 60°C in neutral pH. The optimal Gg-cl-P(AAm-co-MAA) hydrogel was successfully employed for the removal of saline water from various petroleum fraction-saline emulsions. The maximum flocculation efficiency was achieved in an acidic clay suspension with a 15 mg polymer dose at 40°C. Moreover, the synthesized hydrogel adsorbed 94% and 75% of Pb(2+) and Cu(2+), respectively, from aqueous solutions. Finally, the Gg-cl-P(AAm-co-MAA) hydrogel could be degraded completely within 50 days. In summary, the Gg-cl-P(AAm-co-MAA) hydrogel was demonstrated to have potential for use as flocculants and heavy metal absorbents for industrial waste water treatment.

Molecular Docking and Interactions of Pueraria Tuberosa with Vascular Endothelial Growth Factor Receptors.

Pueraria tuberosa is known for its therapeutic potentials in cardiovascular disorders, but its effect in angiogenesis has not been studied so far. In this study, a computational approach has been applied to elucidate the role of the phytochemicals in inhibition of angiogenesis through modulation of vascular endothelial growth factor receptors: Vascular endothelial growth factor receptor-1 and vascular endothelial growth factor receptor-2, major factors responsible for angiogenesis. Metabolite structures retrieved from PubChem and KNApSAcK - 3D databases, were docked using AutoDock4.2 tool. Hydrogen bond and molecular docking, absorption, distribution, metabolism and excretion and toxicity predictions were carried out using UCSF Chimera, LigPlot(+) and PreADMET server, respectively. From the docking analysis, it was observed that puerarone and tuberostan had significant binding affinity for the intracellular kinase domain of vascular endothelial growth factor receptors-1 and vascular endothelial growth factor receptor-2 respectively. It is important to mention that both the phytochemicals shared similar interaction profile as that of standard inhibitors of vascular endothelial growth factor receptors. Also, both puerarone and tuberostan interacted with Lys861/Lys868 (adenosine 5'-triphosphate binding site of vascular endothelial growth factor receptors-1/vascular endothelial growth factor receptors-2), thus providing a clue that they may enforce their inhibitory effect by blocking the adenosine 5'-triphosphate binding domain of vascular endothelial growth factor receptors. Moreover, these molecules exhibited good drug-likeness, absorption, distribution, metabolism and excretion properties without any carcinogenic and toxic effects. The interaction pattern of the puerarone and tuberostan may provide a hint for a novel drug design for vascular endothelial growth factor tyrosine kinase receptors with better specificity to treat angiogenic disorders.

Kinetic release studies of nitrogen-containing bisphosphonate from gum acacia crosslinked hydrogels.

Natural polymer hydrogels are useful for controlling release of drugs. In this study, hydrogels containing gum acacia were synthesized by free-radical polymerization of acrylamide with gum acacia. The effect of gum acacia in the hydrogels on the release mechanism of nitrogen-containing bisphosphonate (BP) was studied at pH 1.2 and 7.4. The hydrogels exhibited high swelling ratios at pH 7.4 and low swelling ratios at pH 1.2. The release study was performed using UV-Visible spectroscopy via complex formation with Fe(III) ions. At pH 1.2, the release profile was found to be anomalous while at pH 7.4, the release kinetic of BP was a perfect zero-order release mechanism. The hydrogels were found to be pH-sensitive and the release profiles of the BP were found to be influenced by the degree of crosslinking of the hydrogel network with gum acacia. The preliminary results suggest that these hydrogels are promising devices for controlled delivery of bisphosphonate to the gastrointestinal region.

Disulfide engineering at the dimer interface of Lactobacillus casei thymidylate synthase: crystal structure of the T155C/E188C/C244T mutant.

The crystal structure of a covalently cross-linked Lactobacillus casei thymidylate synthase has been determined at 2.8 A resolution. The sites for mutation to achieve the bis-disulfide linked dimer were identified using the disulfide modeling program MODIP. The mutant so obtained was found to be remarkably thermostable. This increase in stability has been reasoned to be entirely a consequence of the covalent gluing between the two subunits.

Unusual genome organisation in Entamoeba histolytica leads to two overlapping transcripts.

We have isolated homologs of the mini chromosome maintenance (MCM) gene family from the parasitic protozoan Entamoeba histolytica. The full length genomic and cDNA clones for the Eh MCM3 gene have been characterised. The Eh MCM3 gene is much smaller than the Saccharomyces cerevisiae MCM3 gene and other eukaryotic homologs of the MCM3/P1 family. The predicted Eh Mcm3 protein was 597 amino acids long and showed 37 and 46% positional identity with the Sc Mcm3 and the mouse P1 homologs respectively. While proceeding along the chromosome from the Eh MCM3 gene, we have identified a homolog (Eh PAK) of the murine p21 activated kinase (Rn KPAK), or S. cerevisiae STE20. Eh PAK lies 126 bp upstream of the Eh MCM3 gene. The predicted Eh p21 activated kinase protein was 459 amino-acids long and showed 33% positional identity with the murine p21 activated kinase and its yeast homolog Ste20. Analysis of cDNA and genomic sequences shows that the 3' untranslated region (UTR) of the Eh PAK mRNA and the 5' UTR of the Eh MCM3 mRNA are transcribed from a common 40 bp genomic segment. This is the first report of an amoeba gene being physically linked to a second gene such that their transcripts are overlapping and there is no non-transcribed intergenic region between the two genes. Primer extension studies have confirmed that unlike most E. histolytica genes, which have short 5' UTRs, the Eh MCM3 mRNA has a 126 bp long 5' UTR and the Eh PAK mRNA has a 265 bp long 5' UTR.

Heterogeneity of DNA content and expression of cell cycle genes in axenically growing Entamoeba histolytica HM1:IMSS clone A.

The cell division cycle of Entamoeba histolytica was studied using multi-parametric flow cytometry in asynchronous and partially synchronised cells. Dynamic changes in the DNA synthesis and DNA content of axenically growing trophozoites were observed by using 5-bromo-2'-deoxyuridine (BrdU) uptake and DNA specific fluorochromes. It was observed that DNA synthesis in these cells continues beyond the typical S-phase stop point when DNA duplication is complete. Asynchronously growing E. histolytica cells could be synchronised by serum starvation followed by serum re-addition. BrdU incorporation in synchronised cells showed that cell synchrony is maintained for at least one generation time, in which the G1 phase lasts for 2-3 h and the S-phase lasts for 5-6 h. Analysis of our results revealed that E. histolytica trophozoites, growing in axenic medium, are made up of a heterogenous population of euploid and polyploid cells. The number of polyploid cells increases with age of the cells in culture. Expression of putative cell cycle and signal transduction markers was studied using specific antibodies and changes in their expression levels have been correlated with changes in the DNA content. Based upon our results we could identify G1, S and G2 phases of the cell cycle of E. histolytica and also predict the mechanism underlying the generation of polyploidy in these cells, which may have significant effects on its biology and pathogenesis.

An electrospray ionization mass spectrometry investigation of 1-anilino-8-naphthalene-sulfonate (ANS) binding to proteins.

The binding of 1-anilino-8-naphthalene-sulfonic acid (ANS) to various globular proteins at acidic pH has been investigated by electrospray ionization mass spectrometry (ESI-MS). Maximal ANS binding is observed in the pH range 3-5. As many as seven species of dye-bound complexes are detected for myoglobin. Similar studies were carried out with cytochrome c, carbonic anhydrase, triosephosphate isomerase, lysozyme, alpha-lactalbumin, and bovine pancreatic trypsin inhibitor (BPTI). Strong ANS binding was observed wherever molten globule states were postulated in solution. ANS binding is not observed for lysozyme and BPTI, which have tightly folded structures in the native form. Alpha-lactalbumin, which is structurally related to lysozyme but forms a molten globule at acidic pH, exhibited ANS binding. Reduction of disulfide bonds in these proteins leads to the detection of ANS binding even at neutral pH. Binding was suppressed at very low pH (<2.5), presumably due to neutralization of the charge on the sulfonate moiety. The distribution of the relative intensities of the protein bound ANS species varies with the charge state, suggesting heterogeneity of gas phase conformations. The binding strength of these complexes was qualitatively estimated by dissociating them using enhanced nozzle skimmer potentials. The skimmer voltages also affected the lower and higher charge states of these complexes in a different manner.

Metabolic enzymes as potential drug targets in Plasmodium falciparum.

Plasmodium falciparum causes the most severe form of malaria that is fatal in many cases. Emergence of drug resistant strains of P. falciparum requires that new drug targets be identified. This review considers in detail enzymes of the glycolytic pathway, purine salvage pathway, pyrimidine biosynthesis and proteases involved in catabolism of haemoglobin. Structural features of P. falciparum triosephosphate isomerase which could be exploited for parasite specific drug development have been highlighted. Utility of P. falciparum hypoxanthine-guanine-phosphoribosyltransferase, adenylosuccinate synthase, dihydroorotate dehydrogenase, thymidylate synthase-dihydrofolate reductase, cysteine and aspartic proteases have been elaborated in detail. The review also briefly touches upon other potential targets in P. falciparum.