Acid hydrolysis conditions for quantification of meningococcal X polysaccharide in a pentavalent vaccine using HPAEC-PAD/ESI-MS.

A selective and sensitive method was evaluated for quantitation of meningococcal X (Men X) polysaccharide in pentavalent meningococcal A, C, W, Y and X conjugate vaccine using different acid hydrolysis conditions like HCl, TFA, HF, HF-TFA, and HF-HCl. High-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) using CarboPac PA10 column was used to identify the hydrolyzed products based on retention time and its comparison with monosaccharide standards. Complete release of glucosamine (GlcN) from Men X in monovalent bulk and pentavalent vaccine samples was achieved using HF hydrolysis at 80 °C for 2 h. The Men X HF-hydrolyzed polysaccharide to glucosamine along with the reference standard was identified using collision-induced dissociation (CID) electrospray mass spectroscopy and the MS/MS fragments of m/z 162, m/z 144 and m/z 84. Meningococcal polysaccharide concentration was determined with a correlation coefficient r2 >0.99 using polysaccharide reference standard. The serogroups A, W, and Y were converted to their monosaccharides units and quantified using this method however, milder acid hydrolysis 0.1 M HCl 80 °C 2 h for release of sialic acid for Men C polysaccharide was found to be more suitable. These methods will provide necessary tools and prove to be beneficial to laboratories developing new saccharide-based vaccine combinations.

Quantitation of free cyanide using ion exchange chromatography in Neisseria meningitidis serogroups A, C, W, Y and X conjugates used in vaccine manufacture.

Polysaccharide vaccines essentially used in the prevention of bacterial infections are known to be good immunogens when conjugated to an immunogenic protein using various cyanylating agents. Analysis of residual cyanide in polysaccharide conjugate vaccines is an ardent task due to the complexity of the sample matrices and the lack of suitable methods. We report a selective ion chromatography method with electrochemical detection using IonPac AS7 column for estimation of residual cyanide in meningococcal serogroups A, C, W, Y and X bulk conjugates in presence of other interfering ions. Gold electrode and Ag/AgCl reference electrode ensures sensitivity and reproducibility of cyanide quantitation. The calibration curve of the method is linear having r2 ≥0.990 over the concentration range 1.45 ng/mL to 93.10 ng/mL. The recovery of cyanide in bulk conjugates ranged between 96.0% and 108.9%. The limits of detection and quantitation were 0.50 ng/mL and 1.45 ng/mL which corresponds to 0.31 ng/µg and 0.91 ng/µg of polysaccharide respectively. The method validation and feasibility study were performed using Men W and Men X bulk conjugates respectively with in house residual cyanide specification due to unavailability of pharmacopeia guidelines. The method is reproducible and can accurately quantify residual cyanide in purified meningococcal bulk conjugates.

Use of artificial intelligence in determination of bone age of the healthy individuals: A scoping review.

BACKGROUND: Bone age assessment, as an indicator of biological age, is widely used in orthodontics and pediatric endocrinology. Owing to significant subject variations in the manual method of assessment, artificial intelligence (AI), machine learning (ML), and deep learning (DL) play a significant role in this aspect. A scoping review was conducted to search the existing literature on the role of AI, ML, and DL in skeletal age or bone age assessment in healthy individuals. METHODS: A literature search was conducted in PubMed, Scopus, and Web of Science from January 2012 to December 2022 using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses-Extension for Scoping Reviews (PRISMA-ScR) and Joanna Briggs Institute guidelines. Grey literature was searched using Google Scholar and OpenGrey. Hand-searching of the articles in all the reputed orthodontic journals and the references of the included articles were also searched for relevant articles for the present scoping review. RESULTS: Nineteen articles that fulfilled the inclusion criteria were included. Ten studies used skeletal maturity indicators based on hand and wrist radiographs, two studies used magnetic resonance imaging and seven studies used cervical vertebrae maturity indicators based on lateral cephalograms to assess the skeletal age of the individuals. Most of these studies were published in non-orthodontic medical journals. BoneXpert automated software was the most commonly used software, followed by DL models, and ML models in the studies for assessment of bone age. The automated method was found to be as reliable as the manual method for assessment. CONCLUSIONS: This scoping review validated the use of AI, ML, or DL in bone age assessment of individuals. A more uniform distribution of sufficient samples in different stages of maturation, use of three-dimensional inputs such as magnetic resonance imaging, and cone beam computed tomography is required for better training of the models to generalize the outputs for use in the target population.

Nonextraction Correction of Class II Malocclusion by Pendulum Appliance.

A 13-year-old female patient, presented with the chief complaint of forwardly placed upper front teeth. On examination and analysis of relevant records, she was diagnosed as an Angle's Class II malocclusion on a skeletal Class I base. It was decided to treat the patient with a nonextraction treatment approach with the help of maxillary molar distalization followed by fixed mechanotherapy.