Human Mesenchymal Stem Cell Processing for Clinical Applications Using a Closed Semi-Automated Workflow.

Human mesenchymal stem cells (hMSCs) are currently being explored as a promising cell-based therapeutic modality for various diseases, with more market approvals for clinical use expected over the next few years. To facilitate this transition, addressing the bottlenecks of scale, lot-to-lot reproducibility, cost, regulatory compliance, and quality control is critical. These challenges can be addressed by closing the process and adopting automated manufacturing platforms. In this study, we developed a closed and semi-automated process for passaging and harvesting Wharton's jelly (WJ)-derived hMSCs (WJ-hMSCs) from multi-layered flasks using counterflow centrifugation. The WJ-hMSCs were expanded using regulatory compliant serum-free xeno-free (SFM XF) medium, and they showed comparable cell proliferation (population doubling) and morphology to WJ-hMSCs expanded in classic serum-containing media. Our closed semi-automated harvesting protocol demonstrated high cell recovery (~98%) and viability (~99%). The cells washed and concentrated using counterflow centrifugation maintained WJ-hMSC surface marker expression, colony-forming units (CFU-F), trilineage differentiation potential, and cytokine secretion profiles. The semi-automated cell harvesting protocol developed in the study can be easily applied for the small- to medium-scale processing of various adherent and suspension cells by directly connecting to different cell expansion platforms to perform volume reduction, washing, and harvesting with a low output volume.

Acceleration of Translational Mesenchymal Stromal Cell Therapy Through Consistent Quality GMP Manufacturing.

Human mesenchymal stromal cell (hMSC) therapy has been gaining immense interest in regenerative medicine and quite recently for its immunomodulatory properties in COVID-19 treatment. Currently, the use of hMSCs for various diseases is being investigated in >900 clinical trials. Despite the huge effort, setting up consistent and robust scalable manufacturing to meet regulatory compliance across various global regions remains a nagging challenge. This is in part due to a lack of definitive consensus for quality control checkpoint assays starting from cell isolation to expansion and final release criterion of clinical grade hMSCs. In this review, we highlight the bottlenecks associated with hMSC-based therapies and propose solutions for consistent GMP manufacturing of hMSCs starting from raw materials selection, closed and modular systems of manufacturing, characterization, functional testing, quality control, and safety testing for release criteria. We also discuss the standard regulatory compliances adopted by current clinical trials to broaden our view on the expectations across different jurisdictions worldwide.

Association between reduced visual-motor integration performance and socioeconomic factors among preschool children in Malaysia: A cross-sectional study.

Visual-motor integration (VMI) is related to children's academic performance and school readiness. VMI scores measured using the Beery-Bucktenicka Developmental Test of Visual-Motor Integration (Beery-VMI) can differ due to differences in cultural and socioeconomic backgrounds. This study compared the VMI scores of Malaysian preschoolers with the corresponding US norms and determined the association between their VMI scores and socioeconomic factors. A cross-sectional study was conducted among 435 preschoolers (mean age: 5.95±0.47 years; age range: 5.08-6.83 years) from randomly selected public and private preschools. VMI scores were measured using Beery-VMI in the preschools' classrooms. Information on the socioeconomic characteristics of the preschoolers was obtained using a parent-report questionnaire. One sample t-test was used to compare their VMI scores with the corresponding US norms. Multivariate logistic regression models were used to explore the influence of socioeconomic factors on the preschoolers' VMI scores. Overall, Malaysian preschoolers' VMI performance was similar to the US standardized norms (p>0.05). Children from low-income families were twice likely to obtain lower than average VMI scores than those from higher-income families (OR = 2.47, 95%CI 1.05, 5.86). Children enrolled at public preschools were more likely to obtain a lower than average VMI score than those who enrolled at private preschools (OR = 2.60, 95%CI 1.12, 6.06). Children who started preschool at the age of six were more likely to obtain lower than average VMI scores than those who started at an earlier age (OR = 4.66, 95%CI 1.97, 11.04). Low maternal education level was also associated with lower than average VMI score (OR = 2.60, 95%CI 1.12, 6.06). Malaysian preschoolers' Beery-VMI performance compared well to their US counterparts. Some socioeconomic factors were associated with reduced VMI scores. Those from disadvantaged socioeconomic backgrounds are more likely to have reduced VMI performance, potentially adversely affecting their school readiness, cognitive performance, and future academic achievements.

A systematic study of stereotypy in epileptic seizures versus psychogenic seizure-like events.

OBJECTIVE: The objective of this study was to quantify the features of stereotypy in epileptic seizures and compare it with that of stereotypy in psychogenic nonepileptic seizure-like events (PNES) confirmed by video-electroencephalography (VEEG) monitoring. METHODS: Video-electroencephalography monitoring records of 20 patients with temporal lobe seizures (TLS) and 20 with PNES were retrospectively reviewed (n = 138 seizures, 48 TLS and 90 PNES). We analyzed the semiology of 59 behaviors of interest for their presence, duration, sequence, and continuity using quantified measures that were entered into statistical analysis. RESULTS: We identified discontinuity as the parameter that was clearly distinct between PNES and epileptic TLS events: there were significantly more frequent pauses of behavior (i.e., "on-off" pattern) in PNES compared with TLS (P = 0.012). The frequency of pauses during an event was diagnostic of PNES events. For instance, the presence of 2 "pauses" during an episode determines a 69% probability of the seizure being nonepileptic. Moreover, PNES events had significantly greater duration (143 s) than TLS events (68 s) (excluding outliers, P = 0.002) and greater duration variability from one event to another in the same subject (P = 0.005). SIGNIFICANCE: Our work provides the first quantified measure of behavioral semiology during epileptic and nonepileptic seizures and offers novel behavioral measures to differentiate them from each other.

An Intractable Case of Prototheca Keratitis and Chronic Endophthalmitis in Stevens-Johnson Syndrome With Boston Type 1 Keratoprosthesis.

PURPOSE: To report a case of recalcitrant Prototheca spp. keratitis and endophthalmitis in a patient with Stevens-Johnson syndrome (SJS) and Boston type 1 keratoprosthesis (B1 KPro). METHODS: Case report. RESULTS: A 67-year-old female with a history of SJS with chronic severe ocular surface disease in her monocular left eye underwent placement of a B1 KPro. Her course was subsequently complicated by persistent Prototheca spp. keratitis and endophthalmitis requiring ongoing treatment with triazole and polyene antifungal agents and multiple surgical revisions of her keratoprosthesis. CONCLUSIONS: Protothecosis is an algal infection that is rarely pathologic in humans, but when present can be difficult to manage. We present the second known case of persistent chronic Prototheca spp. keratitis and endophthalmitis in a patient with long-standing SJS and B1 KPro.

Spectrum of ocular manifestations in cobalamin C and cobalamin A types of methylmalonic acidemia.

BACKGROUND: Cobalamin C disease (cblC), which leads to methylmalonic acidemia with homocystinuria, is the most common inherited disorder of vitamin B12 metabolism. Reported ocular findings associated with cblC have been maculopathy, pigmentary retinopathy, and optic nerve atrophy. Cobalamin A disease (cblA) which causes an isolated methylmalonic acidemia without homocystinuria is rarer than cblC. This is the first detailed report of the ocular findings associated with cblA. We also describe the spectrum of ocular findings in our cblC patients. MATERIALS AND METHODS: A case series describing the ophthalmologic clinical course of six patients with a diagnosis of cobalamin C type and one patient with cobalamin A type of methylmalonic acidemia. Patients were diagnosed through biochemical laboratory testing and genetic analysis was conducted on most patients. Longitudinal fundus findings, optical coherence tomography (OCT), autofluorescence, and electrophysiology were followed in the patients. RESULTS: The cblA patient demonstrated a relatively mild ocular phenotype with late-onset and slowly progressing temporal disc pallor and peripapillary atrophy in the second decade of life. The patient maintained good visual acuity and central vision, without evidence of maculopathy. The six cblC patients demonstrated a range of ocular findings from unremarkable and mild phenotypes to significant retinopathy, including bull's eye maculopathy, severe maculopathy with punched out chorioretinal atrophy, peripheral bone spicules, and optic nerve atrophy. CONCLUSIONS: The spectrum of ocular manifestations seen with inherited disorders of cobalamin metabolism is wide, ranging from mild optic nerve atrophy to severe macular or retinal degeneration. This heterogeneity may in part reflect the associated biochemical phenotype, such as that observed between our cblA and cblC patients. We also observed heterogeneity within the cblC type in agreement with previous reports.

Charting Phelan's 'To Suffer a Sea Change'.

Physicians and healthcare workers usually perceive their medical record entries as documentation rather than construction. In the following article, we extract a medical record from a narrative, Peggy Phelan's pathography of glaucoma, 'To Suffer a Sea Change'. From information about encounters described by Phelan, an ophthalmologist reconstructs progress notes similar to those that would be key to a glaucoma patient's medical record. Rather than condemning the arcane pointilism of the medical record as a poverty of language, or isolating the pathography as an academic text, we hope to instead appreciate what their collaborative dialogue offers the study of disease. While the points of divergence between these texts will demonstrate failures in communication, they will also unearth an enriched dialogue.

Blepharophimosis syndrome with absent tear production.

The purpose of this report was to present a case of congenital alacrima in a patient with blepharophimosis-ptosis-epicanthus inversus syndrome (BPES). A 9-month-old boy presented with characteristic clinical findings of BPES confirmed by genetic testing. On further history taking and evaluation, the patient was noted to have no tear production, despite clinically present palpebral lobes of the lacrimal glands. BPES is an autosomal dominant condition characterized by narrowed horizontal palpebral fissures, severe bilateral symmetric ptosis, epicanthus inversus, and telecanthus. To the authors' knowledge, this represents the second reported case of congenital alacrima in this syndrome. The first case described in the literature was in a 9-month-old girl who had congenital absence of the lacrimal glands. BPES may present with alacrima requiring vigilant lifelong lubrication and careful consideration in decisions for eyelid surgery including ptosis repair.

Pseudoduplication of the optic disk.

PURPOSE: The purpose of this study was to report a case of a chorioretinal coloboma mimicking a second optic disk in an asymptomatic 12-year-old boy. METHODS: This is a case report. RESULTS: An asymptomatic 12-year-old boy presented with a chorioretinal coloboma of the right eye giving the impression of a second optic disk. The patient also had a superior visual field defect. CONCLUSION: True optic nerve duplication is a rare entity that can be mimicked by other etiologies, including choroidal colobomas and postinflammatory lesions, requiring careful examination by clinicians.