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Synthetic nanosized hydroxyapatite (HA) particles (

Antimicrobial resistance is a major challenge in the field and threat to human life. Many patients are suffering from cancer, infection and other diseases simultaneously. Therefore, early detection of infection can lead to treatment of these patients with an appropriate antibiotic. Hence, the development of a specific imaging molecule can increase the speed of infection analysis and thereby application of proper antibiotic. The present work involves the optimization of labelling conditions for an antibiotic of cephalosporin family, cephradine with technetium-99m (99mTc) and establishment of quality control tests. Labelling of cephradine was also determined by applying MALDI-TOF mass spectrometry. Evaluation of in vitro binding with S. aureus bacteria was carried out. Animal model was used to conduct in vivo binding studies. For this, infected animals were injected with the radiolabelled ligand and images were captured by Gamma camera, to observe target to non-target uptake of radiolabelled complex. Furthermore, we optimized various parameters to achieve best labelling efficacy and stability of cephradine. Our results show that cephradine can be used as potential infection imaging agent for advanced clinical care. [ABSTRACT FROM AUTHOR]

DFT calculations were performed to predict the structures of two copper(II) azide complexes, [Cu(en)2(N3)2] (1) and [Cu(Tmen)(N3)2]2 (2), and to calculate their atomic charges and spin densities using natural bond orbital (NBO) analysis (where, en = 1,2-diaminoethane and Tmen = N,N,N',N'-tetramethylethylenediamine). A reliable assignment of the IR spectrum of complex 2 was made by DFT computations. The SEM–EDX measurements of the two complexes are also reported. The DFT optimization of 1 and 2 reveals that the theoretical models of water solvent are more effective in reproduction of the experimental structures. The results of SEM–EDX analysis described the surface morphologies and confirmed the elemental stoichiometry of the complexes. [ABSTRACT FROM AUTHOR]

Inorganic metal ions and the proteins play an essential role in living systems. Different hormones and cellular messengers regulate the hemostasis of organisms. Calcium, magnesium, iron and zinc are abundantly distributed in human bodies and play vital role in many biochemical processes. Albumin and ferritin are also the most abundant proteins present in our bodies and carrying out important physiological processes. A disease state or a medical condition can perturb the normal levels of metal ions and proteins in living systems. In the present study, we have analyzed the serum concentration of Ca, Mg, Fe, Zn, albumin and ferritin of hyperthyroid patients. This study includes 184 collected samples (52 normal subjects and 132 hyperthyroid patients), with age group ranging between 11 to 75 years. During sample collection, only those patients were included for whom hyperthyroidism was initially diagnosed but not taking any medicines yet. The hyperthyroid state was determined by the correlation of FT3, FT4 and TSH. Serum Fe, Ca and albumin levels were determined by spectrophotometric method, while Zu and Mg by atomic absorption spectroscopy (AAS) and ferritin by radioimmunology assay (RIA). Obtained results demonstrated the significant differences in the studied parameters as compared to the normal subjects. [ABSTRACT FROM AUTHOR]

Ubiquitin protein was identified and characterized from the seeds of Sisymbrium (S.) irio. Results of LC-MS/MS spectrometry revealed that S. irio ubiquitin (Siu) shares around 77% sequence identity with already reported ubiquitin proteins from Triticum aestivum, Avena sativa, Arabidopsis thaliana and others. Siu revealed a molecular weight of 17 kDa on non-reduced SDS-PAGE while a reduced band of approximately 9 kDa in the presence of ß-mercaptoethanol. A pI of 9.2 was calculated by isoelectric focusing measurement. Circular Dichroism (CD) spectroscopic studies showed that the secondary structure of Siu comprised of about 6.5% a-helix, 39.3% ß-sheet, 0.7% turn, 53.5% random coil which indicates that it is mostly ß-sheeted. Similarly, a hydrodynamic radius (RH) of 2.44 nm was calculated by Dynamic Light Scattering (DLS) for monodispersive Siu which further infers the presence of a dimeric status of 9 kDa native protein in solution. Multiple sequence alignment of 20 different ubiquitin sequences revealed that ubiquitin sequence is well conserved in nature with very few amino acid differences, e.g. R42K, Q49D, and Q62D. Homology modeling of Siu (11-28, and 42-76) residues suggested overall conserved secondary and tertiary conformation and other structural motifs. This report describes the purification, characterization and structural analysis of an ubiquitin protein from seeds of S. irio. [ABSTRACT FROM AUTHOR]

This study describes the purification and spectroscopic analysis of a trimeric (~ 180 kDa) 11S globulin protein from the seeds of Cucumis (C.) sativus L. The predominant seeds storage protein of C. sativus L. is a salt soluble globulin (~ 50 kDa), which is composed of large (28-31 kDa) and small (19-22 kDa) subunits linked together through disulfide linkage. The N-terminal amino acid sequence of small subunit showed high homology (83%) to that of many monocotyledonous and dicotyledonous plant 11S globulins (also commonly known as Cruciferins). Further random amino acids sequences were obtained from MALDI-Q-TOF MS/MS and multiple sequence alignment exhibited 72% sequence similarity with cruciferin of Cucurbita maxima. C. sativus Cruciferin (CsCr) from seed was first partially purified by ammonium sulfate precipitation (40% saturation constant) and further by gel filtration chromatography. Purified CsCr exhibited a typical single polypeptide of approximately 50 kDa monomeric protein under non-reduced condition of SDSPAGE while produces two bands of major molecular weights of 28 kDa (also called a-polypeptides) and 19 kDa (ß-polypeptides) under non-reduced conditions which is very much typical of 11S globulins and confirming the presence of disulfide linkages between two sub-units. Highly purified CsCr was produced in 25 mM Phosphate buffer of pH 7.0 and subjected to Dynamic Light Scattering (DLS) measurement which showed the monodispersive nature of the 11S globulins with hydrodynamic radius of approx. 5.7 nm confirming the trimeric nature (~ 180 kDa) of the protein. CsCr was further subjected to Circular Dichroism (CD) spectra which indicated the presence of a-helices and ß-sheets in the native conformation of approximately 180 kDa proteins. This is a first report describing the purification and spectroscopic study of an 11S Globulin/Cruciferin protein from seeds of C. sativus. [ABSTRACT FROM AUTHOR]

Snake venom is a myriad of biologically active proteins and peptides. Three finger toxins are highly conserved in their molecular structure, but interestingly possess diverse biological functions. During the course of evolution the introduction of subtle mutations in loop regions and slight variations in the three dimensional structure, has resulted in their functional versatility. Cytotoxin-1 (UniProtID: P01467), isolated from Naja mossambica mossambica, showed the potential to inhibit chymotrypsin and the chymotryptic activity of the 20S proteasome. In the present work we describe a molecular model of cytotoxin-1 in complex with chymotrypsin, prepared by the online server ClusPro. Analysis of the molecular model shows that Cytotoxin-1 (P01467) binds to chymotrypsin through its loop I located near the N-terminus. The concave side of loop I of the toxin fits well in the substrate binding pocket of the protease. We propose Phe10 as the dedicated Pi site of the ligand. Being a potent inhibitor of the 20S proteasome, cytotoxin-1 (P01467) can serve as a potential antitumor agent. Already snake venom cytotoxins have been investigated for their ability as an anticancer agent. The molecular model of cytotoxin-1 in complex with chymotrypsin provides important information towards understanding the complex formation. [ABSTRACT FROM AUTHOR]

Background: Fungi and insect pests ruin stored crop grain, which results in millions of dollars of damage, presenting an ongoing challenge for farmers in addition to diminishing the safety of stored food. A wide-range defensive system against pathogens is needed to reduce or even eliminate the dependence of the crop yield upon the use of pesticides. Plant defensins (γ-thionins) are antimicrobial peptides (AMPs) that are a component of the host defense system. They are known to interact with cell membranes to exhibit antifungal and insecticidal activity. They exhibit a broad range of activities against fungi and insects and are effective at low concentrations. Thionins act on membranes, greatly reducing the development of pathogen resistance., Objective: The aim of this study is to investigate a bioactive molecule that acts against fungal pathogens and stored grain insect pests., Methods: γ-thionin protein was extracted from Brassica oleracea L. var. capitata f. alba (white cabbage) seed powder in phosphate buffer (100 mM, pH 7.0) and was identified by MALDI-TOF/TOF. The crude extract was subjected to 70% ammonium sulfate saturation followed by gel filtration chromatography. The disc diffusion assay along with a microtiter bioassay was used to determine the antifungal activity of the protein against phytopathogenic fungi. The insecticidal efficacy was evaluated by feeding insect pests with food contaminated with the purified protein. Additionally, an in silico molecular structure prediction study of the protein was performed using Auto Dock Vina for molecular docking of the protein with either fungal membrane moieties or α-amylase from Tenebrio molitor L. MD simulations of protein-ligand complexes were conducted using Schrodinger's Desmond module., Results: γ-Thionin (BoT) was purified from white cabbage seeds and showed 100% homology with thionin (Brassica oleracea L. var. viridis) and 80% homology with defensin-like protein 1 (Raphanus sativus L.), respectively. BoT significantly inhibited the mycelial growth of Aspergillus niger van Tieghem and Aspergillus flavus Link at a concentration of 2 μM. Similarly, 0.12 μM BoT treatment resulted in significant mortality of Tribolium castaneum Herbst and Sitophilus oryzae L. Molecular docking and MD simulation of BoT confirmed the strong binding affinity with fungal membrane moieties (phosphatidylinositol 4,5-bisphosphate and phosphatidic acid), which causes disruption of the cell membrane and leakage of the cellular contents, leading to cell death. BoT blocked the active site of α-amylase, and as a result of the inactivation of this gut enzyme, the digestive systems of insects were disturbed, resulting in their deaths., Conclusion: This study revealed that γ-thionin is a good antifungal and insecticidal agent that could be used as an alternate to fungicides and insecticides., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

A Kunitz-type trypsin inhibitor protein has been purified and characterized from seeds of Acacia nilotica L. LC-MS/MS analysis of Acacia nilotica trypsin inhibitor ( An TI) provided the N-terminal fragment of 11 amino acids which yielded 100% identity with already reported Kunitz-type trypsin inhibitor protein of Acacia confusa ( Ac TI) in UniProtKB database search. SDS-PAGE showed a single band of ~21 kDa under nonreduced condition and appearance of a daughter band (17 kDa) in the presence of β -mercaptoethanol indicating the presence of interchain disulfide linkage typical for Kunitz-type trypsin inhibitors. An TI was purified from seed extract by using a combination of anion exchange and gel filtration chromatography. Since An TI showed maximum homology with Ac TI, a molecular structure of Ac TI was predicted which showed highly β -sheeted molecular conformation similar to crystallographic structure of Enterolobium contortisiliquum trypsin inhibitor ( Ec TI). An TI (20 µg) produces significant population inhibition against different human pathogenic bacteria along strong antifungal activity (50 µg). Entomotoxin potential of An TI was evaluated against two stored grain insect pests Tribolium castaneum (Herbst) (Tenebrionidae: Coleoptera) and Sitophilus oryzae (Linnaeus) (Curculionidae: Coleoptera). Statistically significant mortality of T. castaneum adults was observed at 1.5 mg after 15 days in comparison to control. Additionally, number of total eggs, larvae, pupae, adults, and their male/female ratio were also severely reduced in comparison to control. Similarly, two generation progeny of S. oryzae was studied after mixing An TI with rice kernels. Mean percent mortality of adult population was significantly higher after 9 days of exposure in comparison to control group. An TI significantly reduced the F1 generation while little mortality was observed for F2 generation. Exploration of such potent molecules is the prerequisite of our time regarding the anticipation of postantibiotic era and the development of insect resistance against chemical pesticides., Competing Interests: CONFLICT OF INTEREST: none declared, (Copyright © 2020 The Author(s).)

The mol-ecular structure of the title compound, [HgCl2(C3H8N2S)2], has point group symmetry 2, with the twofold rotation axis passing through the Hg(II) atom. The latter is coordinated by two Cl atoms and two N,N'-di-methyl-thio-urea (Dmtu) ligands through their S atoms, defining a distorted tetra-hedral coordination sphere with bond angles in the range 102.47 (4)-118.32 (4)°. Intra- and inter-molecular hydrogen bonds of the type N-H⋯Cl with S(6) and R 2 (2)(12) ring motifs are present. The inter-molecular contacts make up polymeric chains extending parallel to [101].