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1.
Cureus ; 16(9): e70084, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39318659

ABSTRACT

INTRODUCTION: Recent revisions of national field triage guidelines recommend the addition of age-specific systolic blood pressure (SBP) measurement for identifying the most severely injured children requiring transport to a trauma center. The purpose of this study was to determine the frequency in which blood pressures are documented by Emergency Medical Service (EMS) providers and the role this measurement has had, among other factors, in triage decisions. METHODS: This is an exploratory descriptive study with a retrospective review from the trauma registry database of all pediatric trauma admissions that arrived by EMS at a level II pediatric trauma center from January 1, 2019 to December 31, 2022. RESULTS: Two hundred ninety-eight patient records of patients aged 0 to 14 were included. EMS providers documented blood pressure in 70.1% of the total sample. A significant difference in the frequency of this documentation was seen between ages zero to nine and = > 10 years (χ2(1,298) = 28.98 p <0.001). No children ages zero to nine years had SBP of < 70 mmHg + (2x age in years) documented by EMS. There were two children aged = > 10 who had a documented SBP < 90 and 12 children with documented EMS heart rate > SBP. CONCLUSION: Many children transported by EMS in this hospital's catchment area did have a field blood pressure measurement documented, but the frequency was significantly less in younger-aged children. The blood pressure measurements of children determined to have severe injuries in the sample did not meet the inclusion criteria for high risk of serious injury by the newly established national guidelines. This suggests other prehospital criteria, such as mechanism of injury or visual cues, prompted EMS to transport these pediatric trauma patients to a regional trauma center for specialized care.

2.
Int J Biol Macromol ; 268(Pt 2): 131847, 2024 May.
Article in English | MEDLINE | ID: mdl-38677678

ABSTRACT

Combining the versatility of electrospinning with the biocompatibility of Polycaprolactone and Collagen, this study aims to create advanced 3D nano scaffolds for effective drug delivery. Ceramic materials like hydroxyapatite (nHAp) are incorporated as bioactive agents in the fibers. Electrospun PCL (Polycaprolactone)/collagen nanofibers and PVA (Poly-vinyl alcohol)/collagen are promising tissue-engineering substitutes with high biocompatibility, low cytotoxicity, and great tensile strength. Small pores in these nanofibers play a major role in drug delivery system. Owing to its short half-life, limited solubility, restricted bioavailability as well as re-crystallization concerns, the application of Cetirizine (CIT) has found little relevance. Electrospun nanofibers impregnated with CIT provide an excellent solution to combat these limitations, yield sustained drug release along with hampering drug re-crystallization. CIT-loaded polyvinyl alcohol (PVA)/collagen (Col) and CIT-loaded PVA/Col/nHAp nanofibers were characterized and further CIT anti-crystallization as well as release behaviors were investigated. FESEM and HRTEM were used to observe the morphology of the as-synthesized nanofibers. FTIR spectroscopy, water contact angle measurement and drug release studies verified the differences in performance of CIT-loaded PVA/Col and PVA/Col/nHAp nanofibers. The release trend of CIT through these as-synthesized nanoscaffolds was analyzed by various kinetic models and exhibited sustained release of CIT for up to 96 h.


Subject(s)
Cetirizine , Collagen , Drug Liberation , Nanofibers , Polyesters , Polyvinyl Alcohol , Tissue Scaffolds , Polyvinyl Alcohol/chemistry , Polyesters/chemistry , Nanofibers/chemistry , Cetirizine/chemistry , Cetirizine/pharmacokinetics , Cetirizine/administration & dosage , Collagen/chemistry , Tissue Scaffolds/chemistry , Kinetics , Tissue Engineering/methods , Drug Delivery Systems
3.
Chembiochem ; 25(11): e202400045, 2024 Jun 03.
Article in English | MEDLINE | ID: mdl-38593270

ABSTRACT

SUMO (Small Ubiquitin-like Modifiers) proteins are involved in a crucial post-translational modification commonly termed as SUMOylation. In this work, we have investigated the native-state conformational flexibility of human SUMO2 and its interaction with Cu2+ and Zn2+ ions using 15N-1H based 2D NMR spectroscopy. After SUMO1, SUMO2 is the most studied SUMO isoform in humans which shares 45 % and ~80 % similarity with SUMO1 in terms of sequence and structure, respectively. In this manuscript, we demonstrate that compared to SUMO1, several amino acids around the α1-helix region of SUMO2 access energetically similar near-native conformations. These conformations could play a crucial role in SUMO2's non-covalent interactions with SUMO interaction motifs (SIMs) on other proteins. The C-terminal of SUMO2 was found to bind strongly with Cu2+ ions resulting in a trimeric structure as observed by gel electrophoresis. This interaction seems to interfere in its non-covalent interaction with a V/I-x-V/I-V/I based SIM in Daxx protein.


Subject(s)
Copper , Small Ubiquitin-Related Modifier Proteins , Zinc , Humans , Copper/chemistry , Copper/metabolism , Small Ubiquitin-Related Modifier Proteins/metabolism , Small Ubiquitin-Related Modifier Proteins/chemistry , Zinc/chemistry , Zinc/metabolism , Protein Conformation , Nuclear Magnetic Resonance, Biomolecular , Protein Binding
4.
J Funct Biomater ; 15(3)2024 Mar 01.
Article in English | MEDLINE | ID: mdl-38535253

ABSTRACT

Three-dimensional printing (3DP) technology has revolutionized the field of the use of bioceramics for maxillofacial and periodontal applications, offering unprecedented control over the shape, size, and structure of bioceramic implants. In addition, bioceramics have become attractive materials for these applications due to their biocompatibility, biostability, and favorable mechanical properties. However, despite their advantages, bioceramic implants are still associated with inferior biological performance issues after implantation, such as slow osseointegration, inadequate tissue response, and an increased risk of implant failure. To address these challenges, researchers have been developing strategies to improve the biological performance of 3D-printed bioceramic implants. The purpose of this review is to provide an overview of 3DP techniques and strategies for bioceramic materials designed for bone regeneration. The review also addresses the use and incorporation of active biomolecules in 3D-printed bioceramic constructs to stimulate bone regeneration. By controlling the surface roughness and chemical composition of the implant, the construct can be tailored to promote osseointegration and reduce the risk of adverse tissue reactions. Additionally, growth factors, such as bone morphogenic proteins (rhBMP-2) and pharmacologic agent (dipyridamole), can be incorporated to promote the growth of new bone tissue. Incorporating porosity into bioceramic constructs can improve bone tissue formation and the overall biological response of the implant. As such, employing surface modification, combining with other materials, and incorporating the 3DP workflow can lead to better patient healing outcomes.

5.
Curr Hypertens Rev ; 2024 Mar 20.
Article in English | MEDLINE | ID: mdl-38509679

ABSTRACT

Hypertension is one of the primary causes of cardiovascular diseases and death, with a higher prevalence in low- and middle-income countries. The pathophysiology of hypertension remains complex, with 2% to 5% of patients having underlying renal or adrenal disorders. The rest are referred to as essential hypertension, with derangements in various physiological mechanisms potentially contributing to the development of essential hypertension. Hypertension elevates the risk of cardiovascular disease (CVD) events (coronary heart disease, heart failure, and stroke) and mortality. First-line therapy for hypertension is lifestyle change, which includes weight loss, a balanced diet that includes low salt and high potassium intake, physical exercise, and limitation or elimination of alcohol use. Blood pressure-lowering effects of individual lifestyle components are partially additive, enhancing the efficacy of pharmaceutical treatment. The choice to begin antihypertensive medication should be based on the level of blood pressure and the existence of a high atherosclerotic CVD risk. First-line hypertension treatment includes a thiazide or thiazide-like diuretic, an angiotensin-converting enzyme inhibitor or angiotensin receptor blocker, and a calcium channel blocker. Addressing hypertension will require continued efforts to improve access to diagnosis, treatment, and lifestyle interventions.

6.
Chemosphere ; 355: 141791, 2024 May.
Article in English | MEDLINE | ID: mdl-38554868

ABSTRACT

This research investigates into the efficacy of algae and algae-bacteria symbiosis (ABS) in efficiently decolorizing Remazol Red 5B, a prevalent dye pollutant. The investigation encompasses an exploration of the biosorption isotherm and kinetics governing the dye removal process. Additionally, various machine learning models are employed to predict the efficiency of dye removal within a co-culture system. The results demonstrate that both Desmodesmus abundans and a composite of Desmodesmus abundans and Rhodococcus pyridinivorans exhibit significant dye removal percentages of 75 ± 1% and 78 ± 1%, respectively, after 40 min. The biosorption isotherm analysis reveals a significant interaction between the adsorbate and the biosorbent, and it indicates that the Temkin model best matches the experimental data. Moreover, the Langmuir model indicates a relatively high biosorption capacity, further highlighting the potential of the algae-bacteria composite as an efficient adsorbent. Decision Trees, Random Forest, Support Vector Regression, and Artificial Neural Networks are evaluated for predicting dye removal efficiency. The Random Forest model emerges as the most accurate, exhibiting an R2 value of 0.98, while Support Vector Regression and Artificial Neural Networks also demonstrate robust predictive capabilities. This study contributes to the advancement of sustainable dye removal strategies and encourages future exploration of hybrid approaches to further enhance predictive accuracy and efficiency in wastewater treatment processes.


Subject(s)
Water Pollutants, Chemical , Thermodynamics , Coculture Techniques , Adsorption , Water Pollutants, Chemical/analysis , Hydrogen-Ion Concentration , Kinetics
7.
Carbohydr Polym ; 327: 121691, 2024 Mar 01.
Article in English | MEDLINE | ID: mdl-38171696

ABSTRACT

Algal polysaccharides, harnessed for their catalytic potential, embody a compelling narrative in sustainable chemistry. This review explores the complex domains of algal carbohydrate-based catalysis, revealing its diverse trajectory. Starting with algal polysaccharide synthesis and characterization methods as catalysts, the investigation includes sophisticated techniques like NMR spectroscopy that provide deep insights into the structural variety of these materials. Algal polysaccharides undergo various preparation and modification techniques to enhance their catalytic activity such as immobilization. Homogeneous catalysis, revealing its significance in practical applications like crafting organic compounds and facilitating chemical transformations. Recent studies showcase how algal-derived catalysts prove to be remarkably versatile, showcasing their ability to customise reactions for specific substances. Heterogeneous catalysis, it highlights the significance of immobilization techniques, playing a central role in ensuring stability and the ability to reuse catalysts. The practical applications of heterogeneous algal catalysts in converting biomass and breaking down contaminants, supported by real-life case studies, emphasize their effectiveness. In sustainable chemistry, algal polysaccharides emerge as compelling catalysts, offering a unique intersection of eco-friendliness, structural diversity, and versatile catalytic properties. Tackling challenges such as dealing with complex structural variations, ensuring the stability of the catalyst, and addressing economic considerations calls for out-of-the-box and inventive solutions. Embracing the circular economy mindset not only assures sustainable catalyst design but also promotes efficient recycling practices. The use of algal carbohydrates in catalysis stands out as a source of optimism, paving the way for a future where chemistry aligns seamlessly with nature, guiding us toward a sustainable, eco-friendly, and thriving tomorrow. This review encapsulates-structural insights, catalytic applications, challenges, and future perspectives-invoking a call for collective commitment to catalyze a sustainable scientific revolution.


Subject(s)
Polymers , Sustainable Development , Catalysis , Carbohydrates , Polysaccharides
8.
J Environ Manage ; 347: 119193, 2023 Dec 01.
Article in English | MEDLINE | ID: mdl-37797518

ABSTRACT

From the surface of the earth to the depths of the ocean, microplastics are a hazard for both aquatic and terrestrial habitats. Due to their small size and vast expanse, they can further integrate into living things. The fate of microplastics in the environment depends upon the biotic components such as microorganisms which have potential enzymes to degrade the microplastics. As a result, scientists are interested in using microorganisms like bacteria, fungi, and others to remediate microplastic. These microorganisms release the cutinase enzyme, which is associated with the enzymatic breakdown of microplastics and plastic films. Yet, numerous varieties of microplastics exist in the environment and their contaminants act as a significant challenge in degrading microplastics. The review discusses the cutinases enzyme degradation strategies and potential answers to deal with existing and newly generated microplastic waste - polyethylene (PE), polyethylene terephthalate (PET), poly-ε-caprolactone (PCL), polyurethanes (PU), and polybutylene succinate (PBS), along with their degradation pathways. The potential of cutinase enzymes from various microorganisms can effectively act to remediate the global problem of microplastic pollution.


Subject(s)
Microplastics , Water Pollutants, Chemical , Plastics , Carboxylic Ester Hydrolases/metabolism , Polyethylene Terephthalates
9.
J Environ Manage ; 346: 119004, 2023 Nov 15.
Article in English | MEDLINE | ID: mdl-37734213

ABSTRACT

In the pursuit of effective wastewater treatment and biomass generation, the symbiotic relationship between microalgae and bacteria emerges as a promising avenue. This analysis delves into recent advancements concerning the utilization of microalgae-bacteria consortia for wastewater treatment and biomass production. It examines multiple facets of this symbiosis, encompassing the judicious selection of suitable strains, optimal culture conditions, appropriate media, and operational parameters. Moreover, the exploration extends to contrasting closed and open bioreactor systems for fostering microalgae-bacteria consortia, elucidating the inherent merits and constraints of each methodology. Notably, the untapped potential of co-cultivation with diverse microorganisms, including yeast, fungi, and various microalgae species, to augment biomass output. In this context, artificial intelligence (AI) and machine learning (ML) stand out as transformative catalysts. By addressing intricate challenges in wastewater treatment and microalgae-bacteria symbiosis, AI and ML foster innovative technological solutions. These cutting-edge technologies play a pivotal role in optimizing wastewater treatment processes, enhancing biomass yield, and facilitating real-time monitoring. The synergistic integration of AI and ML instills a novel dimension, propelling the fields towards sustainable solutions. As AI and ML become integral tools in wastewater treatment and symbiotic microorganism cultivation, novel strategies emerge that harness their potential to overcome intricate challenges and revolutionize the domain.

10.
Biochemistry ; 62(5): 1012-1025, 2023 03 07.
Article in English | MEDLINE | ID: mdl-36820504

ABSTRACT

The MID1 TRIM protein is important for ventral midline development in vertebrates, and mutations of its B-box1 domain result in several birth defects. The B-box1 domain of the human MID1 protein binds two zinc atoms and adopt a similar ßßα-RING structure. This domain is required for the efficient ubiquitination of protein phosphatase 2A, alpha4, and fused kinase. Considering the structural similarity, the MID1 B-box1 domain exhibits mono-autoubiquitination activity, in contrast to poly-autoubiquitination observed for RING E3 ligases. To understand its mechanism of action, the interaction of the B-box1 domain with Ube2D1 (UbcH5a, E2), a preferred E2 ligase, is investigated. Using isothermal titration calorimetry, the MID1 RING and B-box1 domains were observed to have similar binding affinities with the Ube2D1 protein. However, NMR 15N-1H Heteronuclear Single Quantum Coherence titration, 15N relaxation data, and High Ambiguity Driven protein-protein DOCKing (HADDOCK) calculations show the B-box1 domain binding on a surface distinct from where RING domains bind. The novel binding interaction shows the B-box1 domain partially overlapping the noncovalent Ube2D1 and a ubiquitin binding site that is necessary for poly-autoubiquitination activity. The B-box1 domain also displaces the ubiquitin from the Ube2D1 protein. These studies reveal a novel binding interaction between the zinc-binding ßßα-fold B-box1 domain and the Ube2D enzyme family and that this difference in binding, compared to RING E3 ligases, provides a rationale for its auto-monoubiquitination E3 ligase activity.


Subject(s)
Microtubule Proteins , Transcription Factors , Ubiquitin-Conjugating Enzymes , Ubiquitin-Protein Ligases , Humans , Amino Acid Sequence , Microtubule Proteins/chemistry , Models, Molecular , Protein Structure, Tertiary , Transcription Factors/chemistry , Ubiquitin/chemistry , Ubiquitin-Conjugating Enzymes/chemistry , Ubiquitin-Protein Ligases/chemistry , Ubiquitination , Zinc/chemistry
11.
Mol Biotechnol ; 64(8): 841-851, 2022 Aug.
Article in English | MEDLINE | ID: mdl-35184267

ABSTRACT

Inadequate rice production worldwide is largely attributed to abiotic and biotic stresses, along with high sensitivity of cultivable plant germplasm. In the field of cereal biotechnology, rice engineering plays an important role in achieving tolerance to such stresses. Plant transformation and selection play crucial role in rice engineering. This review summarized the antibiotic, herbicide and metabolic selection marker genes (SMG) employed in diverse rice engineering studies. These SMGs are no longer required after the transformation has been achieved, hence undesirable at the commercial level. This study also included several strategies employed in rice engineering to eliminate such foreign DNA elements. These include co-transformation, site-specific recombination, transposon and CRISPR base approaches. CRISPR/Cas9 being simple and efficient, is considered a crucial step toward clean gene technology. Further ease and applicability of CRISPR/Cas9 in the embryos directly can help us to modify target genes with efficient marker-free selection in minimum time. Overall, this review summarizes and analyse the recent advances that have enormous potential in rice improvement.


Subject(s)
Oryza , Biotechnology , Cloning, Molecular , Genetic Markers/genetics , Oryza/genetics , Plants, Genetically Modified/genetics
12.
Nutr Cancer ; 74(4): 1489-1496, 2022.
Article in English | MEDLINE | ID: mdl-34309470

ABSTRACT

As the current study reports the utilization of the leaf extract of Catharanthus roseus (C.roseus) for the biological synthesis of zinc oxide nanoparticles (ZnO NPs) because of the importance of the importance of health and environment. Bioinspired synthesis were characterized using Fourier Transform Infrared Spectroscopy (FT-IR), Field Emission-Scanning Electron Microscopy (FE-SEM), Transmission Electron Microscopy (TEM), Energy-Dispersive X-ray Spectroscopy (EDX) and X-Ray diffraction (XRD). XRD and TEM micrograph analysis revealed that the synthesized nanostructures were well-dispersed and spherical with the average particle size in the 18-30 nm range were produced. The FT-IR spectra confirmed presence of phenolic compounds that act as reducing and capping agents. Further, it suggested the possible utilization of hydroxyl groups and amides in the reduction of Zn ions and stablization of ZnO NPs. Zinc oxide nanomaterials are effective in cancer treatments, including the destruction of tumor cells with minimal damage to healthy cells. The toxicity of zinc oxide nanomaterials was checked in vitro in the human breast cancer line MDA-MB-231. Inverse relation of the percentage of viable cells to the concentration of zinc oxide nanomaterials at increasing molar levels was assessed. The cytotoxicity analysis used in the MTT test shows the substantial viable MDA-MB-231-cells despite the increased concentration of exposure to zinc oxide nanomaterials. Reduction in the ratio of viable MDA-MB-231 cells after being exposed to zinc oxide nanomaterials was compared to untreated cancerous cells. The present approach to biosynthesis is quick, inexpensive, eco-friendly, and high-rise stable nanomaterials of zinc oxide with substantial cancer potential. This is the first study that reports molar concentrations (with the lowest concentration of 10 mM) as an anticancer agent for breast cancer and potential clinical uses for synthesized zinc oxide nanomaterials. Thus, C. roseus based synthesized ZnO NPs could be explored not only as environmentally benign method but also as a potential anti-carcinogenic agent.


Subject(s)
Breast Neoplasms , Catharanthus , Metal Nanoparticles , Nanoparticles , Zinc Oxide , Anti-Bacterial Agents , Breast Neoplasms/drug therapy , Female , Green Chemistry Technology/methods , Humans , Metal Nanoparticles/chemistry , Microbial Sensitivity Tests , Nanoparticles/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Plant Leaves , Spectroscopy, Fourier Transform Infrared , Zinc Oxide/chemistry
13.
Int J Biol Macromol ; 151: 204-211, 2020 May 15.
Article in English | MEDLINE | ID: mdl-32061854

ABSTRACT

Metal ions like Cu2+ and Zn2+ have been shown to impact protein misfolding pathways in neurodegenerative proteinopathies like Alzheimer's and Parkinson's. Also, due to their strong interaction with Ubiquitin, they interfere in degradation of misfolded proteins by impairing the ubiquitin-proteasome system (UPS). In this work, we have studied the interaction of these metal ions with a small Ubiquitin like post-translation modifier SUMO1, which is known to work co-operatively with Ubiquitin to regulate UPS system. Between Cu2+ and Zn2+, the former binds more strongly with SUMO1 as determined using fluorescence spectroscopy. SUMO1 aggregates, forming trimer and higher oligomers in presence of Cu2+ ions which were characterized using gel electrophoresis, Bradford assay, and transmission electron microscopy. Chemical shift analysis using 15N/1H based NMR spectroscopy revealed that SUMO1 retains its structural fold in its trimeric state. Cu2+ induced paramagnetic quenching and Zn2+ induced chemical shift perturbation of 15N-1H cross-peaks were used to identify their respective binding sites in SUMO1. Binding sites so obtained were further validated with molecular dynamics studies. Our findings provide structural insights into the SUMO1-Cu2+/Zn2+ interaction, and its impact on aggregation of SUMO1 which might affect its ability to modify functions of target proteins.


Subject(s)
Binding Sites , Copper/chemistry , Ions , SUMO-1 Protein/chemistry , Zinc/chemistry , Amino Acid Sequence , Molecular Dynamics Simulation , Protein Binding , Protein Stability , Protein Structure, Secondary , Recombinant Proteins , SUMO-1 Protein/metabolism , Spectrum Analysis , Structure-Activity Relationship
14.
Int J Biol Macromol ; 134: 536-546, 2019 Aug 01.
Article in English | MEDLINE | ID: mdl-31100392

ABSTRACT

The present work aims to characterize thermo and alkali stable ß-mannanase (ManSS11) from newly isolated Klebsiella pneumoniae SS11 and its food stain (mannan based) removal efficiency. The enzyme, ManSS11 was stable over a wide range of pH and temperature. The highest activity of ManSS11 was observed at pH 9.0 and temperature, 70 °C, with t1/2 of 135.91 min at the same temperature while, >70% of its initial activity was retained at pH 7.0-10.6. It was purified to 5.50-fold homogeneity with a final recovery of 9.6% and a specificity of 7573.57 U/mg protein. Purified ManSS11 was visible as a single protein band with a molecular weight of ~45 kDa. The kinetic parameters of Km, Vmax and kcat were 1.66 mg/mL, 833.33 µmolmL-1 min-1 and 1190.47 s-1 respectively. The compatibility of ß-mannanase with different detergents together with wash performance test confirmed its potential usability in laundry sector. Wash performance analysis confirmed that the enzyme displayed great efficiency in the removal of stains caused by mannan containing foods.


Subject(s)
Alkalies/chemistry , Thermodynamics , beta-Mannosidase/chemistry , Chemical Phenomena , Coloring Agents , Enzyme Activation , Enzyme Stability , Food Handling/methods , Hydrogen-Ion Concentration , Kinetics , Molecular Weight , Substrate Specificity , Temperature , beta-Mannosidase/isolation & purification
15.
Proteins ; 87(8): 658-667, 2019 08.
Article in English | MEDLINE | ID: mdl-30958586

ABSTRACT

Structural heterogeneity in the native-state ensemble of dSmt3, the only small ubiquitin-like modifier (SUMO) in Drosophila melanogaster, was investigated and compared with its human homologue SUMO1. Temperature dependence of amide proton's chemical shift was studied to identify amino acids possessing alternative structural conformations in the native state. Effect of small concentration of denaturant (1M urea) on this population was also monitored to assess the ruggedness of near-native energy landscape. Owing to presence of many such amino acids, especially in the ß2 -loop-α region, the native state of dSmt3 seems more flexible in comparison to SUMO1. Information about backbone dynamics in ns-ps timescale was quantified from the measurement of 15 N-relaxation experiments. Furthermore, the noncovalent interaction of dSmt3 and SUMO1 with Daxx12 (Daxx729 DPEEIIVLSDSD740 ), a [V/I]-X-[V/I]-[V/I]-based SUMO interaction motif, was characterized using Bio-layer Interferometery and NMR spectroscopy. Daxx12 fits itself in the groove formed by ß2 -loop-α structural region in both dSmt3 and SUMO1, but the binding is stronger with the former. Flexibility of ß2 -loop-α region in dSmt3 is suspected to assist its interaction with Daxx12. Our results highlight the role of native-state flexibility in assisting noncovalent interactions of SUMO proteins especially in organisms where a single SUMO isoform has to tackle multiple substrates single handedly.


Subject(s)
Drosophila Proteins/chemistry , Drosophila melanogaster/chemistry , Repressor Proteins/chemistry , SUMO-1 Protein/chemistry , Adaptor Proteins, Signal Transducing/chemistry , Adaptor Proteins, Signal Transducing/metabolism , Animals , Drosophila Proteins/metabolism , Drosophila melanogaster/metabolism , Humans , Models, Molecular , Nuclear Magnetic Resonance, Biomolecular , Nuclear Proteins/chemistry , Nuclear Proteins/metabolism , Protein Conformation , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Repressor Proteins/metabolism , SUMO-1 Protein/metabolism , Small Ubiquitin-Related Modifier Proteins
16.
Langmuir ; 35(11): 4085-4093, 2019 03 19.
Article in English | MEDLINE | ID: mdl-30810316

ABSTRACT

The development of new strategies for thermal stability and storage of enzymes is very important, considering the nonretention of catalytic activity by enzymes under harsh conditions of temperature. Following this, herein, a new approach based on the interfacial adsorption of lysozyme (LYZ) at nanointerfaces of ionic liquid (IL)-based microemulsions, for enhanced thermal stability of LYZ, is reported. Microemulsions (MEs) composed of dialkyl imidazolium-based surface active ILs (SAILs) as surfactants, ILs as the nonpolar phase, and ethylene glycol (EG) as the polar phase, without any cosurfactants, have been prepared and characterized in detail. Various regions corresponding to polar-in-IL, bicontinuous, and IL-in-polar phases have been characterized using conductivity measurements. Dynamic light scattering (DLS) measurements have provided insights into the size distribution of microdroplets, whereas temperature-dependent DLS measurements established the thermal stability of the MEs. Nanointerfaces formed by SAILs with EG in thermally stable reverse MEs act as fluid scaffolds to adsorb and provide thermal stability, up to 120 °C, to LYZ. Thermally treated LYZ upon extraction into a buffer shows enzyme activity owing to negligible change in the active site of LYZ, as marked by retention of microenvironment of Trp residues present in the active site of LYZ. The present work is expected to establish a new platform for the development of novel nanointerfaces utilizing biobased components for other biomedical applications.


Subject(s)
Ionic Liquids/chemistry , Nanotechnology , Temperature , Emulsions/chemistry , Enzyme Stability , Muramidase/chemistry , Particle Size , Surface Properties
17.
Int J Biol Macromol ; 123: 446-456, 2019 Feb 15.
Article in English | MEDLINE | ID: mdl-30439429

ABSTRACT

The proteins secreted by bacteria contribute to immune mediated gastric inflammation and epithelial damage; thus aid bacterial invasion in host tissue, and may also interact with host proteins, conspirating a mechanism against host-immune system. The Histone-like DNA binding protein is one of the most abundant nucleoid-associated proteins in Helicobacter pylori (H. pylori). The protein -referred here as Hup- is also secreted in vitro by H. pylori, thus it may have its role in disease pathogenesis. This is possible only if Hup interact with some human proteins including Small-Ubiquitin-like-Modifier (SUMO) proteins. Studies have established that SUMO-proteins participate in various innate-immune pathways and thus promote an efficient immune response to combat pathogenic infections. Sequence analysis revealed the presence of two SUMO interacting motifs (SIMs) and several positively charged lysine residues on the protein surface of Hup. Additionally, SUMO-proteins epitomize negatively charged surface which confers them the ability to bind to DNA/RNA binding proteins. Based on the presence of SIMs as well as charge complementarity between the proteins, it is legitimate to consider that Hup protein would bind to SUMO-proteins. The present study has been undertaken to establish this interaction for the first time using NMR in combination with ITC and other biophysical techniques.


Subject(s)
Bacterial Proteins/chemistry , DNA-Binding Proteins/chemistry , Helicobacter pylori/chemistry , Nuclear Magnetic Resonance, Biomolecular , SUMO-1 Protein/chemistry , Amino Acid Motifs , Bacterial Proteins/metabolism , DNA-Binding Proteins/metabolism , Helicobacter pylori/metabolism , Humans , Protein Binding , SUMO-1 Protein/metabolism
18.
J Phys Chem B ; 122(26): 6763-6770, 2018 07 05.
Article in English | MEDLINE | ID: mdl-29878782

ABSTRACT

The interaction of amphiphilic ionic liquids containing an 1-alkyl-3-methylimidazolium cation ([C12MIM]+), which shows acute cytotoxicity toward marine and bacterial life, with zwitterionic 1-palmitoyl-2-oleoyl- sn-glycero-3-phospho-choline (POPC) and anionic 1-palmitoyl-2-oleoyl- sn-glycero-3-phospho- rac-glycerol (POPG) membranes was investigated. Biophysical parameters of this interaction were quantified by fluorescence spectroscopy, isothermal titration calorimeter, and solution-state NMR measurements. [C12MIM]+ inserts into the membrane and induces vesicle leakage at relatively low concentration (<1 mM). Zwitterionic POPC membranes are more leakage-prone, but the binding of [C12MIM]+ cations is stronger to anionic POPG membranes. A higher rate of exchange of membrane-bound [C12MIM]+ is suspected to play a key role in membrane leakage. Furthermore, solid-state NMR spectroscopy was employed to determine the location of [C12MIM]+ in lipid membranes and its impact on the structure and dynamics of the bilayer. The study provides a molecular understanding of the membrane permeabilizing effect of the [C12MIM]+ mediated by its detergent-like structure.

19.
Drug Res (Stuttg) ; 68(5): 296-300, 2018 May.
Article in English | MEDLINE | ID: mdl-29100264

ABSTRACT

The present work describes a rapid and green microwave mediated method for the synthesis and a simple and precise isocratic reverse phase HPLC method for the estimation of the biologically significant dihydropyridines. The conventional synthesis of these dihydropyridines has been previously reported from our lab. The analysis of a standard solution (1 mg/ml) was accomplished on a symmetry (4.6 mm I.D x 250 mm) C-18 column using mobile phase acetonitrile:water:triethylamine (TEA) (70:30:0.1 v/v/v) at a flow rate of 0.7 ml/min. Detection was monitored at 354 nm. The retention time for all the compounds was accomplished as less than 10 min. The compounds showed the linear response over the concentration range 10-100 µg/ml. The study is aimed to develop a rapid method for the quantification of these potent molecules. Various parameters like linearity (10-100 µg/ml), USP tailing and plate count were found to be satisfactory. The investigated parameters were studied with the freshly prepared solutions.


Subject(s)
Dihydropyridines/analysis , Dihydropyridines/chemical synthesis , Microwaves , Chromatography, High Pressure Liquid , Chromatography, Reverse-Phase
20.
J Clin Diagn Res ; 9(1): UC01-6, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25738062

ABSTRACT

CONTEXT: Brachial plexus block is a suitable alternative to general anaesthesia for patient undergoing upper extremity surgery. Ropivacaine the S-enantiomer emerged as a possible replacement of Bupivacaine without undesirable toxic effects. It provides similar duration of sensory analgesia with early recovery of motor block. AIMS: Comparision of onset, duration of sensory- motor block and any adverse effects between 0.5% Bupivacaine and 0.5% Ropivacaine in axillary brachial plexus block. SETTINGS AND DESIGN: Prospective randomized study. MATERIALS AND METHODS: This study was carried out in 50 patients between 18-55 y, comparable in demographic variables was randomly allocated to two groups of 25 each. Group I received 30ml 0.5% Bupivacaine, Group II received 30 ml 0.5% Ropivacaine in axillary brachial plexus block for forearm surgeries. Onset, Duration of sensory-motor block, Heart rate, Blood pressure, Oxygen saturation and Respiratory rate were recorded. STATISTICAL ANALYSIS: Statistical analysis used was Statistical Package for Social Sciences version 15.0, Chi-square test was used to evaluate the proportional data. Odds ratio/risk ratios have been calculated wherever necessary. Parametric data has been evaluated using Student t-test while non-parametric data has been evaluated using Mann-Whitney U-test. RESULTS: Onset of motor blockade was earlier in ropivacaine group (5 min) as compared to bupivacaine group (20 min), Higher levels of motor blockade, Mean onset time for motor block was significantly shorter in ropivacaine group (14.88±3.35 min) as compared to bupivacaine group (22.92±3.79 min), Mean duration of block was significantly longer in bupivacaine group (408.40±50.39 min) as compared to ropivacaine group (365.60±34.29 min) (p=0.001), Onset of sensory block was observed from 5 min itself in ropivacaine group as compared to bupivacaine group (10 min), Duration of sensory block was significantly longer in bupivacaine group (450.40±54.50 min) as compared to ropivacaine group (421.20±38.33 min) . CONCLUSION: On the basis of present study, conclusions were drawn that onset of action of sensory, motor block was early in Ropivacaine group with faster recovery of motor functions as compared to Bupivacaine group. No adverse effects were noted in either groups. This study suggests that Ropivacaine is a suitable alternative to Bupivacaine for forearm surgeries under Brachial Plexus Block.

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