Showing papers by "Nalco Holding Company published in 2016"

Delivery of Dual Drug Loaded Lipid Based Nanoparticles across the Blood–Brain Barrier Impart Enhanced Neuroprotection in a Rotenone Induced Mouse Model of Parkinson’s Disease

Abstract: Parkinson’s disease (PD) is the most widespread form of dementia where there is an age related degeneration of dopaminergic neurons in the substantia nigra region of the brain. Accumulation of α-synuclein (αS) protein aggregate, mitochondrial dysfunction, oxidative stress, and neuronal cell death are the pathological hallmarks of PD. In this context, amalgamation of curcumin and piperine having profound cognitive properties, and antioxidant activity seems beneficial. However, the blood–brain barrier (BBB) is the major impediment for delivery of neurotherapeutics to the brain. The present study involves formulation of curcumin and piperine coloaded glyceryl monooleate (GMO) nanoparticles coated with various surfactants with a view to enhance the bioavailability of curcumin and penetration of both drugs to the brain tissue crossing the BBB and to enhance the anti-parkinsonism effect of both drugs in a single platform. In vitro results demonstrated augmented inhibition of αS protein into oligomers and fibril...

Strain and bioprocess improvement of a thermophilic anaerobe for the production of ethanol from wood

Abstract: The thermophilic, anaerobic bacterium Thermoanaerobacterium saccharolyticum digests hemicellulose and utilizes the major sugars present in biomass. It was previously engineered to produce ethanol at yields equivalent to yeast. While saccharolytic anaerobes have been long studied as potential biomass-fermenting organisms, development efforts for commercial ethanol production have not been reported. Here, we describe the highest ethanol titers achieved from T. saccharolyticum during a 4-year project to develop it for industrial production of ethanol from pre-treated hardwood at 51–55 °C. We describe organism and bioprocess development efforts undertaken to improve ethanol production. The final strain M2886 was generated by removing genes for exopolysaccharide synthesis, the regulator perR, and re-introduction of phosphotransacetylase and acetate kinase into the methyglyoxal synthase gene. It was also subject to multiple rounds of adaptation and selection, resulting in mutations later identified by resequencing. The highest ethanol titer achieved was 70 g/L in batch culture with a mixture of cellobiose and maltodextrin. In a “mock hydrolysate” Simultaneous Saccharification and Fermentation (SSF) with Sigmacell-20, glucose, xylose, and acetic acid, an ethanol titer of 61 g/L was achieved, at 92 % of theoretical yield. Fungal cellulases were rapidly inactivated under these conditions and had to be supplemented with cellulosomes from C. thermocellum. Ethanol titers of 31 g/L were reached in a 100 L SSF of pre-treated hardwood and 26 g/L in a fermentation of a hardwood hemicellulose extract. This study demonstrates that thermophilic anaerobes are capable of producing ethanol at high yield and at titers greater than 60 g/L from purified substrates, but additional work is needed to produce the same ethanol titers from pre-treated hardwood.

Overexpression of caspase 7 is ERα dependent to affect proliferation and cell growth in breast cancer cells by targeting p21(Cip).

Abstract: Caspase 7 (CASP7) expression has important function during cell cycle progression and cell growth in certain cancer cells and is also involved in the development and differentiation of dental tissues. However, the function of CASP7 in breast cancer cells is unclear. The aim of this study was to analyze the expression of CASP7 in breast carcinoma patients and determine the role of CASP7 in regulating tumorigenicity in breast cancer cells. In this study, we show that the CASP7 expression is high in breast carcinoma tissues compared with normal counterpart. The ectopic expression of CASP7 is significantly associated with ERα expression status and persistently elevated in different stages of the breast tumor grades. High level of CASP7 expression showed better prognosis in breast cancer patients with systemic endocrine therapy as observed from Kaplan–Meier analysis. S3 and S4, estrogen responsive element (ERE) in the CASP7 promoter, is important for estrogen-ERα-mediated CASP7 overexpression. Increased recruitment of p300, acetylated H3 and pol II in the ERE region of CASP7 promoter is observed after hormone stimulation. Ectopic expression of CASP7 in breast cancer cells results in cell growth and proliferation inhibition via p21Cip reduction, whereas small interfering RNA (siRNA) mediated reduction of CASP7 rescued p21Cip levels. We also show that pro- and active forms of CASP7 is located in the nucleus apart from cytoplasmic region of breast cancer cells. The proliferation and growth of breast cancer cells is significantly reduced by broad-spectrum peptide inhibitors and siRNA of CASP7. Taken together, our findings show that CASP7 is aberrantly expressed in breast cancer and contributes to cell growth and proliferation by downregulating p21Cip protein, suggesting that targeting CASP7-positive breast cancer could be one of the potential therapeutic strategies.

Acylamide and Amine Oxide Derivatives of Linear and Hyperbranched Polyethylenimine. Part 2: Comparison of Gas Kinetic Hydrate Inhibition Performance

Abstract: A series of acylamide and amine oxide derivatives of polyethylenimine, both hyperbranched (HPEI) and linear (LPEI), has been synthesized, and their performance as kinetic hydrate inhibitors (KHIs) with a structure II (SII)-forming natural gas mixture has been investigated in high-pressure steel rocking cell experiments. This is the first time that the KHI performance of linear and hyperbranched polymers with the same functional groups has been compared. The importance of quality control in KHI synthesis is also highlighted. In general, the amine oxides showed better performance than the acylamides when comparing the structurally optimized and best polymers from each class. The amine oxide polymers with best KHI performance were the low-molecular-weight butylated HPEI amine oxides. The performance was similar to those of oligomeric amine oxides reported previously with molecular weights as low as 660 g/mol. This contrasts with other KHI polymer classes, where molecular weights below 1000–1500 g/mol general...

Identification of Novel Sequence Types among Staphylococcus haemolyticus Isolated from Variety of Infections in India.

Abstract: The aim of this study was to determine sequence types of 34 S. haemolyticus strains isolated from a variety of infections between 2013 and 2016 in India by MLST. The MEGA5.2 software was used to align and compare the nucleotide sequences. The advanced cluster analysis was performed to define the clonal complexes. MLST analysis showed 24 new sequence types (ST) among S. haemolyticus isolates, irrespective of sources and place of isolation. The finding of this study allowed to set up an MLST database on the PubMLST.org website using BIGSdb software and made available at http://pubmlst.org/shaemolyticus/. The data of this study thus suggest that MLST can be used to study population structure and diversity among S. haemolyticus isolates.

Sulfitobacter faviae sp. nov., isolated from the coral Faviaveroni.

Abstract: Three closely related, non-sporulating, aerobic, Gram-stain-negative, motile, rod-shaped isolates (S5-53T, S6-62 and S6-64) were obtained from mucus of corals Favia veroni from the Andaman Sea, India. Colonies grown on marine agar were small, circular and cream-coloured. Heterotrophic growth was observed at 10-40 °C and pH 6-10; optimum growth occurred at 25-30 °C and pH 7-8. 16S rRNA gene sequence analysis confirmed the isolates belonged to the genus Sulfitobacter and the three isolates shared more than 99 % pairwise sequence similarity. Strain S5-53T shared highest 16S rRNA gene sequence similarity of 98.43 % with Sulfitobacter dubius KMM 3554T. DNA-DNA relatedness among the three isolates was above 70 % whereas strain S5-53T showed less than 70 % relatedness with the type strains of closely related species. The DNA G+C content of strain S5-53T was 61 mol%. It contained phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and diphosphatidylglycerol as major polar lipids. Predominant fatty acids included C18 : 1ω7c, C18 : 1ω7c 11-methyl, C16 : 0 and C10 : 0 3-OH. Q10 was the major respiratory quinone. Based on this polyphasic analysis, the new isolates (S5-53T, S6-62 and S6-64) are considered to represent a novel species of the genus Sulfitobacter, for which the name Sulfitobacter faviae sp. nov. is proposed. The type strain is S5-53T(=JCM 31093T=LMG 29156T).

Biochemical and molecular characterisation of salt-induced poor grain filling in a rice cultivar.

Abstract: Despite the prevalence of poor grain filling in rice (Oryza sativa L.) under abiotic stress, the reason for this is largely unexplored. Application of 0.75% NaCl to a salt-sensitive rice cultivar at late booting resulted in a >20% yield loss. Spikelets per panicle and the percentage of filled grain decreased significantly in response to NaCl application. The inhibitory effect of NaCl on grain filling was greater in basal than in apical spikelets. Sucrose synthase (SUS) activity was positively correlated with grain weight. The transcript levels of the SUS isoforms differed greatly: the levels of SUS2 increased significantly in response to salt; those of SUS4 decreased drastically. Gene expression studies of starch synthase and ADP-glucose pyrophosphorylase showed that the decreased transcript levels of one isoform was compensated by an increase in those of the other. Salt application also significantly increased the gene expression of the ethylene receptors and the ethylene signalling proteins. The increase in their transcript levels was comparatively greater in basal than in apical spikelets. Significant enhancement in the transcript levels of the ethylene receptors and the increase in the production of ethylene indicated that the salt-induced inhibition of grain filling might be mediated by ethylene. Additionally, the inhibition of chromosomal endoreduplication mediated by decreased transcript levels of B-type cyclin could explain poor grain filling under salt stress. A significant increase in the transcript levels of the ethylene-responsive factors in the spikelets during grain filling in response to salt indicated their possible protective role in grain filling under stress.

Scaffold-free and scaffold-assisted 3D culture enhances differentiation of bone marrow stromal cells

Abstract: 3D cultures of stem cells can preserve differentiation potential or increase the efficiency of methods that induce differentiation. Mouse bone marrow-derived stromal cells (BMSCs) were cultured in 3D as scaffold-free spheroids or "mesoid bodies" (MBs) and as aggregates on poly(lactic) acid microspheres (MB/MS). 3D cultures demonstrated viable cells, interaction on multiple planes, altered cell morphology, and the formation of structures similar to epithelial cell bridges. Cell proliferation was limited in suspension cultures of MB and MB/MS; however, cells regained proliferative capacity when transferred to flat substrates of tissue culture plates (TCPs). Expanded as monolayer, cells retained expression of Sca-1 and CD44 stem cell markers. 3D cultures demonstrated enhanced potential for adipogenic and osteogenic differentiation showing higher triglyceride accumulation and robust mineralization in comparison with TCP cultures. Enhanced and efficient adipogenesis was also observed in 3D cultures generated in a rotating cell culture system. Preservation of multilineage potential of BMSC was demonstrated in 5-azacytidine treatment of 3D cultures and TCP by expression of cardiac markers GATA4 and ACTA1 although functioning cardiomyocytes were not derived.

Identification and functional characterization of the miRNA-gene regulatory network in chronic myeloid leukemia lineage negative cells.

Abstract: Chronic myeloid leukemia (CML) is maintained by leukemic stem cells (LSCs) which are resistant to the existing TKI therapy. Hence a better understanding of the CML LSCs is necessary to eradicate these cells and achieve complete cure. Using the miRNA-gene interaction networks from the CML lin(-) cells we identified a set of up/down-regulated miRNAs and corresponding target genes. Association studies (Pearson correlation) from the miRNA and gene expression data showed that miR-1469 and miR-1972 have significantly higher number of target genes, 75 and 50 respectively. We observed that miR-1972 induces G2-M cell cycle arrest and miR-1469 moderately arrested G1 cell cycle when overexpressed in KCL22 cells. We have earlier shown that a combination of imatinib and JAK inhibitor I can significantly bring down the proliferation of CML lineage negative cells. Here we observed that imatinib and JAK inhibitor I combination restored the expression pattern of the down-regulated miRNAs in primary CML lin(-) cells. Thus effective manipulation of the deregulated miRNAs can restore the miRNA-mRNA networks that can efficiently inhibit CML stem and progenitor cells and alleviate the disease.

Psychrobacter pocilloporae sp. nov., isolated from a coral, Pocillopora eydouxi.

Abstract: Two closely related aerobic, Gram-stain-negative, rod-shaped bacteria (S6-60T and S6-67) were isolated from the mucus of the coral, Pocillopora eydouxi, from the Andaman Sea, India. Heterotrophic growth on marine agar was observed at 4–37 °C and at pH 6.5–10.0; optimum growth occurred at 25–30 °C and at pH 7–9. 16S rRNA gene sequence analysis confirmed that the isolates belong to the genus Psychrobacter ; the two isolates shared more than 99.5 % pairwise sequence similarity. Strain S6-60T showed a maximum 16S rRNA similarity of 98.92 % with Psychrobacter pacificensis DSM 23406T. DNA–DNA homology between the two isolates, S6-60T and S6-67, was above 90 %, whereas strain S6-60T showed less than 70 % homology with closely related type species. The DNA G+C content was 47.7 mol%. It contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phospholipid as the major polar lipids. C10 : 0, C12 : 0 3OH, C16 : 0, C18 : 1ω9c, C17 : 1ω8c and C16 : 1ω7c were found to be the predominant fatty acids. Based on a polyphasic analysis, the isolates (S6-60T and S6-67) represent a novel species of the genus Psychrobacter for which the name Psychrobacter pocilloporae sp. nov. is proposed with S6-60T(=JCM 31058T=LMG 29157T) as the type strain.

Phenotypic and genetic characteristics of Vibrio cholerae O1 carrying Haitian ctxB and attributes of classical and El Tor biotypes isolated from Silvassa, India.

Abstract: Vibrio cholerae O1 biotype El Tor, the causative agent of the seventh pandemic, has recently been replaced by strains carrying classical and Haitian ctxB in India, Haiti and other parts of the world. We conducted phenotypic and genetic tests to characterize V. cholerae O1 isolated between 2012 and 2014 from Silvassa, India, to examine the presence of virulence and regulatory genes, seventh pandemic marker, ctxB type and biofilm formation and to study genomic diversity. Of the 59 V. cholerae O1, eight isolates belong to El Tor prototype, one to classical prototype and the remaining isolates have attributes of both classical and El Tor biotypes. PCR and ctxB gene sequencing revealed the presence of classical ctxB in four strains and Haitian ctxB in 55 isolates; indicating that isolates were either an El Tor or hybrid variant. All isolates carried virulence, regulatory, adherence, Vibrio seventh pandemic pathogenicity island I and seventh pandemic group-specific marker VC2346, in addition to tcpAET and rstRET, the features of seventh pandemic strains, and produced cholera toxin and biofilm. PFGE analysis showed that the majority of isolates are clonal and belong to fingerprint pattern A; however, pattern B is unrelated and patterns C and D are distinct, suggesting considerable diversity in the genomic content among them. These data thus show that isolates from Silvassa are genetically diverse and that Haitian ctxB and hybrid phenotypes are undergoing global dissemination.

Multidrug-resistant Staphylococcus haemolyticus isolates from infected eyes and healthy conjunctivae in India.

Abstract: This study aimed to determine the presence of antibiotic resistance genes (ARGs), SCCmec elements and genetic relatedness among Staphylococcus haemolyticus isolated from patients with a variety of eye infections (n=11) and from healthy conjunctiva (n=7). Minimum inhibitory concentrations were determined for 14 antimicrobials according to BSAC guidelines. PCR was used to identify the presence of mecA, mecC, SCCmec type and ARGs. Sequencing was used to determine mutations in gyrA, gyrB, topoisomerase IVA and IVB genes. Genetic relatedness was determined by PFGE. Of the 18 isolates, 17 showed resistance to at least one antibiotic, but none showed resistance to vancomycin or rifampicin. Ten isolates were oxacillin-resistant and carried the mecA gene, eight of which belonged to SCCmec type V. The presence of non-mec SCC elements in two meticillin-susceptible isolates and untypeable SCC elements in meticillin-resistant isolates suggests the involvement of S. haemolyticus in the diversification of SCC elements. Sequence analysis revealed point mutations in gyrA (Ser-84→Leu) and topoisomerase IVA genes (Ser-80→Leu) in 13 isolates, and additional variation in the QRDR (Asp-84→Asn) of two isolates, showing good correlation between mutations in gyrA and topoisomerase IV genes and the level of resistance to fluoroquinolones. PFGE analysis showed distinct pulsotypes forming two major clusters, indicating the existence of diversity among isolates, irrespective of the source of isolation. This study suggests that S. haemolyticus isolates from infected eyes and healthy conjunctivae invariably carried ARGs and SCCmec elements and showed diversity in their genomic content, irrespective of the source of isolation.

S95C substitution in CuZn-SOD of Ipomoea carnea: impact on the structure, function and stability

Abstract: Superoxide dismutase (SOD) in general is a unique homo-dimeric enzyme that can scavenge toxic superoxide radicals by dismutation reaction. In IcSOD (Ipomoea carnea SOD), the presence of cysteine (Cys) plays an essential role in protein behaviour. This study analysed the role of Cys in modulating the stability and kinetic properties of IcSOD. To investigate the significance of the dimeric structure in modulating the structure/function relationship of CuZn-SODs, we have substituted a conserved serine by cysteine (Ser95Cys) in Ipomoea carnea CuZn-SOD. The results demonstrate that this mutation leads to an increase in dimeric strength, as reflected by size exclusion chromatography, differential scanning calorimetry, and high-temperature circular dichroism spectroscopy measurements. The mutant form, as compared to the native enzyme, shows a relatively low tendency to form aggregates but encountered a reduction in both dismutase and peroxidase activities. This study provides new mechanistic insight into the role of free cysteine in CuZn-SODs and such mutation may be used to increase dimeric strength. Protein docking and molecular dynamics simulations further demonstrate that Ser95Cys substitution in Ipomoea carnea CuZn-SOD leads to the creation of a new subunit interface resulting in increased dimeric strength of the protein.

Modified Asphaltene Capillary Deposition Unit: A Novel Approach to Inhibitor Screening

Abstract: Asphaltene deposition in capillaries is a tool that has been used in an attempt to better understand asphaltene deposition in the field. However, data reproducibility and inhibitor ranking present some challenges with this technique. An improved asphaltene capillary deposition unit and a novel experimental protocol were developed to address these problems and are presented here. Using untreated Gulf of Mexico oil, the current study generated reproducible amounts of asphaltene deposit inside the capillary. It further identified the fact that residual oil inside the capillary tube can be a limitation to inhibitor selection. Evaluating the amount of asphaltene depositing in the capillary as a function of time proved successful in addressing this issue and led to inhibitor performance differentiation.

Axial stress localization facilitates pressure propagation in gelled pipes

Abstract: Paraffin wax-oil gels are unique rheological fluids which undergo shear degradation starting at a deformation (shear strain level) of approximately 1%. Flow commencement in pipelines filled with wax-oil gels is a complex hydrodynamic process involving propagation of acoustic, diffusive, and rheological degradation pressure wave fronts. Dynamic simulation informed by qualified rheological relations provides useful insight into the physical nature of these flow processes. Eulerian simulations are presented which emulate known physical phenomena and essential characteristics of wax-gel flow dynamics. A constitutive rheological equation set accounts for deformation-driven reduction in yield stress and viscosity terms. No explicit time-dependent rheological parameters are utilized in the equations. Rheological yielding alters the nature of the dominant pressure wave from inherently diffusive towards self-sharpening. Axial stress localization effectively sequentializes the gel breakage process, quantified by reduced length of the pressure wave-front zone. Ultimately, axial stress localization allows flow in longer pipe segments, albeit with a concomitant time delay. Viscous behavior and yielding degradation behavior are shown to account for upward and downward concavity in transient axial pressure profiles, respectively. Overall, a unique synergy between gel compressibility and gel degradation is revealed. Deformation-coupled interaction between compressibility and degradation allows pressure propagation and subsequent sustained flow through a gel material which is otherwise immobile in the incompressible case.

Otosclerosis Associated with a De Novo Mutation -832G > A in the TGFB1 Gene Promoter Causing a Decreased Expression Level

Abstract: Otosclerosis (OTSC) is defined by abnormal bone remodeling in the otic capsule of middle ear which leads to conductive hearing loss. In our previous study, we have identified a de novo heterozygous mutation −832G > A in the promoter of TGFB1 in an otosclerosis patient. In the present study, we progressively screened this mutation in a cohort of 254 cases and 262 controls. The family members of the patient positive for −832G > A variation were also screened and found inheritance of this variation only to her daughter. Interestingly, this variation is associated with a decreased level of the TGFB1 transcript in the patient compared to her parents and controls. In silico analysis of this mutation predicted the altered binding of two transcription factors v-Myb and MZF1 in the mutated promoter sequence. Further, functional analysis of this mutation using in vitro luciferase and electrophoretic mobility shift assays revealed that this variation is associated with decreased gene expression. In conclusion, this study established the fact that TGFB1 mutation −832G > A altered the TGFB1 promoter activity, which could affect the susceptibility to otosclerosis development. Further, systemic analysis of TGFB1 gene sequence and expression analysis of this gene might reveal its precise role in the pathogenesis of otosclerosis.

Draft Genome Sequence of Chelatococcus sambhunathii Strain HT4T (DSM 18167T) Isolated from a Hot Spring in India

Abstract: The moderately thermophilic bacterium Chelatococcus sambhunathii strain HT4T was isolated from hot spring sediment. Based upon the draft genome sequence, the genome is 4.4 Mb and encodes 4,147 proteins.

Draft Genome Sequence of Comamonas thiooxydans Strain S23T (DSM 17888T), a Thiosulfate-Oxidizing Bacterium Isolated from a Sulfur Spring in India

Abstract: The genus Comamonas contains species isolated from various environments, such as termite guts, wetlands, activated sludge, soil, humans, and fresh water. Here, we report the draft genome sequence of Comamonas thiooxydans strain S23T capable of oxidizing thiosulfate under mixotrophic growth conditions. Based upon draft genome sequencing, the genome is 5.3 Mb and encodes 4,767 proteins. The Comamonas thiooxydans whole-genome sequence will help understand the metabolic diversity in sulfur oxidation pathways.

Draft Genome Sequence of Anoxybacillus suryakundensis Strain JS1T (DSM 27374T) Isolated from a Hot Spring in Jharkhand, India.

Abstract: Anoxybacillus suryakundensis strain JS1T, a facultative anaerobic, moderately thermophilic, alkalitolerant bacterium, was isolated from a hot spring. The estimated genome is 2.6 Mb and encodes 2,668 proteins.