Accelerating attribute-focused process and product development through the development and deployment of autonomous process analytical technology platform system.

Enabling real-time monitoring and control of the biomanufacturing processes through product quality insights continues to be an area of focus in the biopharmaceutical industry. The goal is to manufacture products with the desired quality attributes. To realize this rigorous attribute-focused Quality by Design approach, it is critical to support the development of processes that consistently deliver high-quality products and facilitate product commercialization.

The prevalence of gastroesophageal reflux disease and laryngopharyngeal reflux in patients with dysphagia after radiotherapy for nasopharyngeal carcinoma.

Background: The prevalence of gastroesophageal reflux disease (GERD) and laryngopharyngeal reflux (LPR) in post-irradiated patients with nasopharyngeal carcinoma (NPC) is unknown.

Materials And Methods: In a cross-sectional study, 31 NPC and 12 control patients completed questionnaires for GERD/LPR before esophageal manometry and 24-h pH monitoring. The DeMeester score and reflux finding score (RFS) were used to define GERD and LPR, respectively.

Machine learning-coupled combinatorial mutagenesis enables resource-efficient engineering of CRISPR-Cas9 genome editor activities.

The genome-editing Cas9 protein uses multiple amino-acid residues to bind the target DNA. Considering only the residues in proximity to the target DNA as potential sites to optimise Cas9's activity, the number of combinatorial variants to screen through is too massive for a wet-lab experiment. Here we generate and cross-validate ten in silico and experimental datasets of multi-domain combinatorial mutagenesis libraries for Cas9 engineering, and demonstrate that a machine learning-coupled engineering approach reduces the experimental screening burden by as high as 95% while enriching top-performing variants by ∼7.

Oro-pharyngo-esophageal radionuclide scintigraphy predicts aspiration pneumonia risk and associated survival in post-irradiated nasopharyngeal carcinoma patients.

Objective: To demonstrate that oro-pharyngo-esophageal radionuclide scintigraphy (OPERS) not only detects tracheobronchial aspiration after swallowing, but also quantifies the amount of aspiration and subsequent clearance.

Methods: Data collected between 2014 and 2019 were reviewed for aspiration pneumonia at 12 and 24-months after OPERS. The predictive value for aspiration pneumonia on flexible endoscopic evaluation of swallowing (FEES), videofluoroscopic swallowing study (VFSS), and OPERS, and the overall survival of patients with or without aspiration were determined.

High-fidelity KKH variant of Staphylococcus aureus Cas9 nucleases with improved base mismatch discrimination.

The Cas9 nuclease from Staphylococcus aureus (SaCas9) holds great potential for use in gene therapy, and variants with increased fidelity have been engineered. However, we find that existing variants have not reached the greatest accuracy to discriminate base mismatches and exhibited much reduced activity when their mutations were grafted onto the KKH mutant of SaCas9 for editing an expanded set of DNA targets. We performed structure-guided combinatorial mutagenesis to re-engineer KKH-SaCas9 with enhanced accuracy.

A Combinatorial CRISPR-Cas9 Screen Identifies Ifenprodil as an Adjunct to Sorafenib for Liver Cancer Treatment.

Systematic testing of existing drugs and their combinations is an attractive strategy to exploit approved drugs for repurposing and identifying the best actionable treatment options. To expedite the search among many possible drug combinations, we designed a combinatorial CRISPR-Cas9 screen to inhibit druggable targets. Coblockade of the N-methyl-d-aspartate receptor (NMDAR) with targets of first-line kinase inhibitors reduced hepatocellular carcinoma (HCC) cell growth.

Assessment of pharyngeal motor function using a novel velopharyngeal squeeze maneuver and a novel endoscopic pharyngeal contraction grade scale in patients with dysphagia after radiotherapy for nasopharyngeal carcinoma.

Background: To investigate a novel velopharyngeal squeeze maneuver (VPSM) and novel endoscopic pharyngeal contraction grade (EPCG) scale for the evaluation of pharyngeal motor function.

Methods: During endoscopic examination of 77 post-irradiated nasopharyngeal carcinoma patients and control subjects, VPSM was rated and lateral pharyngeal wall movement graded with EPCG scale during swallowing. Pharyngeal constriction ratio (PCR) measured by videofluoroscopy was used for correlation.

Extensible combinatorial CRISPR screening in mammalian cells.

The CRISPR-Cas system coupled with Combinatorial Genetics (CombiGEM) enables systematic analysis of high-order genetic perturbations that are important for understanding biological processes and discovering therapeutic target combinations. Here, we present detailed steps and technical considerations for building multiplexed guide RNA libraries and carrying out a combinatorial CRISPR screen in mammalian cells. We also present an analytical pipeline, CombiPIPE, for mapping two- and three-way genetic interactions.

Development and pilot evaluation of the Cognition domain of the Hong Kong Comprehensive Assessment Scales for Toddlers.

: To describe the development of the Cognition domain of the Hong Kong Comprehensive Assessment Scales for Toddlers (HKCAS-T).: Participants included 345 toddlers aged 18-41 months, with 258 recruited from Maternal and Child Health Centers (MCHCs) and 87 with cognitive delay recruited from Child Assessment Centers (CACs). They were individually administered the 83-item pilot version by medical practitioners or educational psychologists between 2017 and 2019 in MCHCs and CACs in Hong Kong.

A Three-Way Combinatorial CRISPR Screen for Analyzing Interactions among Druggable Targets.

We present a CRISPR-based multi-gene knockout screening system and toolkits for extensible assembly of barcoded high-order combinatorial guide RNA libraries en masse. We apply this system for systematically identifying not only pairwise but also three-way synergistic therapeutic target combinations and successfully validate double- and triple-combination regimens for suppression of cancer cell growth and protection against Parkinson's disease-associated toxicity. This system overcomes the practical challenges of experimenting on a large number of high-order genetic and drug combinations and can be applied to uncover the rare synergistic interactions between druggable targets.

Pearls of experience for safe and efficient hospital practices in otorhinolaryngology-head and neck surgery in Hong Kong during the 2019 novel coronavirus disease (COVID-19) pandemic.

The 2019 novel coronavirus disease (COVID-19) epidemic originated in Wuhan, China and spread rapidly worldwide, leading the World Health Organization to declare an official global COVID-19 pandemic in March 2020. In Hong Kong, clinicians and other healthcare personnel collaborated closely to combat the outbreak of COVID-19 and minimize the cross-transmission of disease among hospital staff members. In the field of otorhinolaryngology-head and neck surgery (OHNS) and its various subspecialties, contingency plans were required for patient bookings in outpatient clinics, surgeries in operating rooms, protocols in wards and other services.

Management of dysphagia in the patient with head and neck cancer during COVID-19 pandemic: Practical strategy.

The global pandemic of 2019 novel coronavirus disease (COVID-19) has tremendously altered routine medical service provision and imposed unprecedented challenges to the health care system. This impacts patients with dysphagia complications caused by head and neck cancers. As this pandemic of COVID-19 may last longer than severe acute respiratory syndrome (SARS) in 2003, a practical workflow for managing dysphagia is crucial to ensure a safe and efficient practice to patients and health care personnel.

Publisher Correction: Combinatorial mutagenesis en masse optimizes the genome editing activities of SpCas9.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Combinatorial mutagenesis en masse optimizes the genome editing activities of SpCas9.

The combined effect of multiple mutations on protein function is hard to predict; thus, the ability to functionally assess a vast number of protein sequence variants would be practically useful for protein engineering. Here we present a high-throughput platform that enables scalable assembly and parallel characterization of barcoded protein variants with combinatorial modifications. We demonstrate this platform, which we name CombiSEAL, by systematically characterizing a library of 948 combination mutants of the widely used Streptococcus pyogenes Cas9 (SpCas9) nuclease to optimize its genome-editing activity in human cells.

Does Low Dose Angiotensin Converting Enzyme Inhibitor Prevent Pneumonia in Older People With Neurologic Dysphagia--A Randomized Placebo-Controlled Trial.

Objective: To examine if angiotensin converting enzyme inhibitor reduces the risk of pneumonia in older patients on tube-feeding because of dysphagia from cerebrovascular diseases.

Design: Randomized placebo-controlled trial.

Setting: Acute and subacute geriatrics units, speech therapists' clinic, and nursing home.

The joint effects of risk status, gender, early literacy and cognitive skills on the presence of dyslexia among a group of high-risk Chinese children.

This study sought to examine factors that are predictive of future developmental dyslexia among a group of 5-year-old Chinese children at risk for dyslexia, including 62 children with a sibling who had been previously diagnosed with dyslexia and 52 children who manifested clinical at-risk factors in aspects of language according to testing by paediatricians. The age-5 performances on various literacy and cognitive tasks, gender and group status (familial risk or language delayed) were used to predict developmental dyslexia 2 years later using logistic regression analysis. Results showed that greater risk of dyslexia was related to slower rapid automatized naming, lower scores on morphological awareness, Chinese character recognition and English letter naming, and gender (boys had more risk).

Early predictors of dyslexia in Chinese children: familial history of dyslexia, language delay, and cognitive profiles.

Background: This work tested the rates at which Chinese children with either language delay or familial history of dyslexia at age 5 manifested dyslexia at age 7, identified which cognitive skills at age 5 best distinguished children with and without dyslexia at age 7, and examined how these early abilities predicted subsequent literacy skills.

Method: Forty-seven at-risk children (21 who were initially language delayed and 26 with familial risk) and 47 control children matched on age, IQ, and mothers' education were tested on syllable awareness, tone detection, rapid automatized naming, visual skill, morphological awareness, and word reading at age 5 and subsequently tested for dyslexia on a standard Hong Kong measure at age 7.

Results: Of those with an early language delay, 62% subsequently manifested dyslexia; for those with familial risk, the rate of dyslexia was 50%.

The cognitive profile of Chinese children with mathematics difficulties.

This study examined how four domain-specific skills (arithmetic procedural skills, number fact retrieval, place value concept, and number sense) and two domain-general processing skills (working memory and processing speed) may account for Chinese children's mathematics learning difficulties. Children with mathematics difficulties (MD) of two age groups (7-8 and 9-11 years) were compared with age-matched typically achieving children. For both age groups, children with MD performed significantly worse than their age-matched controls on all of the domain-specific and domain-general measures.

Differential expression of tissue repair genes in the pathogenesis of chronic obstructive pulmonary disease.

Rationale: The airflow limitation that defines severity of chronic obstructive pulmonary disease (COPD) is caused by a combination of small airway obstruction and emphysematous lung destruction.

Objectives: To examine the hypothesis that small airway obstructive and emphysematous destructive lesions are produced by differential expression of genes associated with tissue repair.

Methods: The expression of 54 genes associated with repair of repetitively damaged tissue was measured in 136 paired samples of small bronchioles and surrounding lung tissue separated by laser capture microdissection.

Early swallowing problems in a cohort of patients with nasopharyngeal carcinoma: Symptomatology and videofluoroscopic findings.

Objective: To study the incidence and the degree of swallowing dysfunction in patients with nasopharyngeal carcinoma (NPC) who underwent radiation therapy treatment.

Institution: The study was conducted in the Prince of Wales Hospital, a tertiary teaching hospital of the Chinese University of Hong Kong.

Materials And Methods: From October 1999 to July 2001, a cohort of 20 consecutive patients with newly diagnosed NPC was prospectively studied.

Influence of a portable audio-biofeedback device on structural properties of postural sway.

Background: Good balance depends on accurate and adequate information from the senses. One way to substitute missing sensory information for balance is with biofeedback technology. We previously reported that audio-biofeedback (ABF) has beneficial effects in subjects with profound vestibular loss, since it significantly reduces body sway in quiet standing tasks.