Ion Mobility-Resolved Collision-Induced Dissociation and Electron Transfer Dissociation of N-Glycopeptides: Gathering Orthogonal Connectivity Information from a Single Mass- Selected Precursor Ion Population

Glycopeptide-level mass spectrometry (MS) and tandem mass spectrometry (MS/MS) analyses are commonly performed to establish site-specific protein glycosylation profiles that are of central importance to gaining structure-function insights on glycoproteins. Confoundingly, the complete characterization of glycopeptide connectivity usually requires the acquisition of multiple MS/MS fragmentation spectra. Complementary ion fragmentation techniques such as collision-induced dissociation (CID) and electron transfer dissociation (ETD) are often applied in concert to address this need. While structurally informative, the requirement for acquisition of two MS/MS spectra per analyte places considerable limitations upon the breadth and depth of large-scale glycoproteomic inquiry. Here, a previously developed method of multiplexing CID and ETD is applied to the study of glycopeptides for the first time. Integration of the two dissociation methods was accomplished through addition of an ion mobility (IM) dimension that disperses the two stages of MS/MS in time. This allows the two MS/MS spectra to be acquired within a few milliseconds of one another, and to be deconvoluted in post-processing. Furthermore, the method allows both fragmentation readouts to be obtained from the same precursor ion packet, thus reducing the inefficiencies imposed by separate CID and ETD acquisitions and the relatively poor precursor ion to fragment ion conversion typical of ETD. N-linked glycopeptide ions ranging in molecular weight from 1800 to 6500 u were generated from four model glycoproteins that collectively encompassed paucimannosidic, high mannose, and complex types of N-glycosylation. In each case, IM-resolved CID and ETD events provided complete coverage of the glycan topology and peptide sequence coverages ranging from 48.4% (over 32 amino acid residues) to 85.7% (over eight amino acid residues). The potential of this method for large-scale glycoproteomic analysis is discussed

CHARACTERIZATION AND ANTIBACTERIAL ACTIVITY OF ZnO NANOPARTICLES SYNTHESIZED BY CO PRECIPITATION METHOD

Objective: In the present study the antibacterial activity of zinc oxide (ZnO) nanoparticles was investigated against gram negative (Escherichia coli and Proteus vulgaris) and gram positive (Staphylococcus aureus and Streptococcus mutans) organisms.Methods: The synthesis of ZnO nanoparticles was carried out by co-precipitation method using zinc sulfate and sodium hydroxide as precursors. These nanoparticles were characterized by XRD (X-Ray Diffraction), FTIR (Fourier Transform Infrared Radiation), UV-Visible spectroscopy and SEM (Scanning Electron Microscope) with EDX (Energy Dispersive X-ray analysis). As well as antibacterial activity and minimum inhibitory concentration of the nanoparticles were carried out by agar well diffusion method and broth dilution method respectively against gram negative (Escherichia coli and Proteus vulgaris) and gram positive (Staphylococcus aureus and Streptococcus mutans) bacteria.Results: The average crystallite size of ZnO nanoparticles was found to be 35 nm by X-ray diffraction. The vibration bands at 450 and 603 cm-1 which were assigned for ZnO stretching vibration were observed in FTIR spectrum. The optical absorption band at 383 nm was obtained from UV-Visible spectrum. Spherical shape morphology was observed in SEM studies. The antibacterial assay clearly expressed that E. coli showed a maximum zone of inhibition (32±0.20 mm) followed by Proteus vulgaris (30±0.45 nm) at 50 mg/ml concentration of ZnO nanoparticles.Conclusion: Zinc oxide nanoparticles have exhibited good antibacterial activity with gram negative bacteria when compared to gram positive bacteria

Influence of Mortar Rheology on Aggregate Settlement

The influence of the rheology of fresh concrete on the settlement of aggregate is examined. Fresh concrete exhibits a yield stress that, under certain conditions, prevents the settlement of coarse aggregate, although its density is larger than that of the suspending mortar. Calculations, based on estimates of the yield stress obtained from slump tests, predict that aggregate normally used in concrete should not sink. To test this prediction, the settlement of a stone in fresh mortar is monitored. The stone does not sink in the undisturbed mortar (which has a high yield stress), but sinks when the mortar is vibrated, presumably due to a large reduction in its yield stress. This implies that during placement of concrete, the aggregate settles only while the concrete is being vibrated. A unique experimental method for measuring aggregate settlement is also introduced and demonstrated

Optimizing Sequence Coverage for a Moderate Mass Protein in Nano-Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry

Sample pretreatment was optimized to obtain high sequence coverage for human serum albumin (HSA, 66.5 kDa) when using nano-electrospray ionization quadrupole time-of-flight mass spectrometry (nESI-Q-TOF-MS). Use of the final method with trypsin, Lys-C and Glu-C digests gave a combined coverage of 98.8%. The addition of peptide fractionation resulted in 99.7% coverage. These results were comparable to those obtained previously with matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The sample pretreatment/nESI-Q-TOF-MS method was also used with collision-induced dissociation to analyze HSA digests and to identify peptides that could be employed as internal mass calibrants in future studies of modifications to HSA

Clinical utility of the risperidone formulations in the management of schizophrenia

Risperidone is one of the early second-generation antipsychotics that came into the limelight in the early 1990s. Both the oral and long-acting injectable formulations have been subject to numerous studies to assess their safety, efficacy, and tolerability. Risperidone is currently one of the most widely prescribed antipsychotic medications, used for both acute and long-term maintenance in schizophrenia. Risperidone has better efficacy in the treatment of psychotic symptoms than placebo and possibly many first-generation antipsychotics. Risperidone fares better than placebo and first-generation antipsychotics in the treatment of negative symptoms. Risperidone’s long acting injectable preparation has been well tolerated and is often useful in patients with medication nonadherence. Risperidone has a higher risk of hyperprolactinemia comparable to first-generation antipsychotics (FGAs) but fares better than many second-generation antipsychotics with regards to metabolic side effects. In this article, we briefly review the recent literature exploring the role of risperidone formulations in schizophrenia, discuss clinical usage, and highlight the controversies and challenges associated with its use

Optimizing Sequence Coverage for a Moderate Mass Protein in Nano-Electrospray Ionization Quadrupole Time-of-Flight Mass Spectrometry

Sample pretreatment was optimized to obtain high sequence coverage for human serum albumin (HSA, 66.5 kDa) when using nano-electrospray ionization quadrupole time-of-flight mass spectrometry (nESI-Q-TOF-MS). Use of the final method with trypsin, Lys-C and Glu-C digests gave a combined coverage of 98.8%. The addition of peptide fractionation resulted in 99.7% coverage. These results were comparable to those obtained previously with matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The sample pretreatment/nESI-Q-TOF-MS method was also used with collision-induced dissociation to analyze HSA digests and to identify peptides that could be employed as internal mass calibrants in future studies of modifications to HSA

The potential of urinary metabolites for diagnosing multiple sclerosis

A definitive diagnostic test for multiple sclerosis (MS) does not exist; instead physicians use a combination of medical history, magnetic resonance imaging, and cerebrospinal fluid analysis (CSF). Significant effort has been employed to identify biomarkers from CSF to facilitate MS diagnosis; however none of the proposed biomarkers have been successful to date. Urine is a proven source of metabolite biomarkers and has the potential to be a rapid, non-invasive, inexpensive, and efficient diagnostic tool for various human diseases. Nevertheless, urinary metabolites have not been extensively explored as a source of biomarkers for MS. Instead, we demonstrate that urinary metabolites have significant promise for monitoring disease-progression, and response to treatment in MS patients. NMR analysis of urine permitted the identification of metabolites that differentiate experimental autoimmune encephalomyelitis (EAE)-mice (prototypic disease model for MS) from healthy and MS drug-treated EAE mice

Design and Synthesis of Different Aryl Substituted 1,3,4-Oxadiazole-imidazo[1,5-a]pyridine Derivatives as Anticancer Agents

A new series of 1,3,4-oxadiazole incorporated imidazo[1,5-a]pyridine derivatives was prepared. Anticancer activity of all the obtained compounds was investigated by employing MTT assay. Among them, six compounds showed most prominent anticancer activity than etoposide

A Mixed-Method Study to Determine the Benefits of Periconceptional Folic Acid Supplementation and Effects of Folic Acid Deficiency in Mothers on Birth Outcomes.

BACKGROUND: Evidence from high income countries shows mothers who are supplemented with folic acid in their periconceptional period and early pregnancy have significantly reduced adverse outcomes like birth defects. However, in India there is a paucity of data on association of birth defects and folic acid supplementation. We identified a few important questions to be answered using separate scientific methods and then planned to triangulate the information. OBJECTIVE: In this paper, we describe the protocol of our study that aims to determine the association of folic acid and pregnancy outcomes like neural tube defects (NTDs) and orofacial clefts (OFCs). We decided to fill the gaps in knowledge from India to determine public health consequences of folic acid deficiency and factors influencing dietary and periconceptional consumption of folic acid. METHODS: The proposed study will be carried out in five stages and will examine the questions related to folic acid deficiency across selected locations in South and North India. The study will be carried out over a period of 4 years through the hierarchical evidence-based approach. At first a systematic review was conducted to pool the current birth prevalence of NTDs and orofacial clefts OFCs in India. To investigate the population prevalence, we plan to use the key informant method to determine prevalence of NTDs and OFCs. To determine the normal serum estimates of folic acid, iron, and vitamin B12 among Indian women (15-35 years), we will conduct a population-based, cross-sectional study. We will further strengthen the evidence of association between OFCs and folic acid by conducting a hospital-based, case-control study across three locations of India. Lastly, using qualitative methods we will understand community and health workers perspective on factors that decide the intake of folic acid supplements. RESULTS: This study will provide evidence on the community prevalence of birth defects and prevalence folic acid and vitamin B12 deficiency in the community. The case-control study will help understand the association of folic acid deficiency with OFCs. CONCLUSIONS: The results from this study are intended to strengthen the evidence base in childhood disability for planning and policy initiatives