Insights into the mechanism of oligodendrocyte protection and remyelination enhancement by the integrated stress response.

central nervous system (CNS) inflammation triggers activation of the integrated stress response (ISR). We previously reported that prolonging the ISR protects remyelinating oligodendrocytes and promotes remyelination in the presence of inflammation. However, the exact mechanisms through which this occurs remain unknown. Here, we investigated whether the ISR modulator Sephin1 in combination with the oligodendrocyte differentiation enhancing reagent bazedoxifene (BZA) is able to accelerate remyelination under inflammation, and the underlying mechanisms mediating this pathway. We find that the combined treatment of Sephin1 and BZA is sufficient to accelerate early-stage remyelination in mice with ectopic IFN-γ expression in the CNS. IFN-γ, which is a critical inflammatory cytokine in multiple sclerosis (MS), inhibits oligodendrocyte precursor cell (OPC) differentiation in culture and triggers a mild ISR. Mechanistically, we further show that BZA promotes OPC differentiation in the presence of IFN-γ, while Sephin1 enhances the IFN-γ-induced ISR by reducing protein synthesis and increasing RNA stress granule formation in differentiating oligodendrocytes. Finally, pharmacological suppression of the ISR blocks stress granule formation in vitro and partially lessens the beneficial effect of Sephin1 on disease progression in a mouse model of MS, experimental autoimmune encephalitis (EAE). Overall, our findings uncover distinct mechanisms of action of BZA and Sephin1 on oligodendrocyte lineage cells under inflammatory stress, suggesting that a combination therapy may effectively promote restoring neuronal function in MS patients.

Potential of cinnamaldehyde essential oil as a possible antimicrobial against fowl typhoid in layers.

Fowl typhoid (FT) is an economically significant bacterial disease of layers leading to a drastic drop in egg production. Due to increased public health concerns about antibiotics in poultry feed, a search for new safe antimicrobials for treating fowl typhoid is crucial. The antimicrobial effect of cinnamaldehyde essential oil (CnEO) against fowl typhoid in layers was investigated in this experiment. The 60-week-old BV300-layer birds (n = 100) were divided into five groups: the non-challenged control group A, only cinnamaldehyde-treated group B (CnEO @ 1:8000 dilutions through drinking water for 60 days), the challenged group C, challenged plus cinnamaldehyde therapy group D (CnEO @ 1:8000 dilutions through drinking water from 16 to 30 dpi), and challenged plus antibiotic therapy group E (chloramphenicol @ 1 gm/5lit through drinking water from 16 to 30 dpi). Hens from all challenged groups were challenged with Salmonella Gallinarum (VTCCBAA588) @ 1 × 108 CFU/ml orally. Various parameters such as clinical signs, mortality, egg production and egg weight, colony-forming unit (CFU) count of cecal content, eggshell surface, and egg yolk were evaluated all through 60 days of an experimental trial. Results indicated that, in the case of the cinnamaldehyde therapeutic group, there was a significant improvement in egg production, mild clinical signs, lower feed conversion ratio (FCR), and a significantly lower bacterial count in ceca and on the eggshell surface compared to the control challenge group. Thus, CnEO @ 1:8000 dilutions through drinking water can be a potential antimicrobial for controlling fowl typhoid.

Comparative study between dynamic susceptibility contrast magnetic resonance imaging and arterial spin labelling perfusion in differentiating low-grade from high-grade brain tumours.

Purpose: Our aim was to distinguish between low-grade and high-grade brain tumours on the basis of dynamic susceptibility contrast (DSC) magnetic resonance imaging (MRI) perfusion and arterial spin labelling (ASL) perfusion and to compare DSC and ASL techniques. Material and methods: Forty-one patients with brain tumours were evaluated by 3-Tesla MRI. Conventional and perfusion MRI imaging with a 3D pseudo-continuous ASL (PCASL) and DSC perfusion maps were evaluated. Three ROIs were placed to obtain cerebral blood value (CBV) and cerebral blood flow (CBF) in areas of maximum perfusion in brain tumour and normal grey matter. Histopathological diagnosis was considered as the reference. ROC analysis was performed to compare the diagnostic performance and to obtain a feasible cut-off value of perfusion parameters to differentiate low-grade and high-grade brain tumours. Results: Normalised perfusion parameters with grey matter (rCBF or rCBV lesion/NGM) of malignant lesions were significantly higher than those of benign lesions in both DSC (normalised rCBF of 2.16 and normalised rCBV of 2.63) and ASL (normalised rCBF of 2.22) perfusion imaging. The normalised cut-off values of DSC (rCBF of 1.1 and rCBV of 1.4) and ASL (rCBF of 1.3) showed similar specificity and near similar sensitivity in distinguishing low-grade and high-grade brain tumours. Conclusions: Quantitative analysis of perfusion parameters obtained by both DSC and ASL perfusion techniques can be reliably used to distinguish low-grade and high-grade brain tumours. Normalisation of these values by grey matter gives us more reliable parameters, eliminating the different technical parameters involved in both the techniques.

Additive-free Aqueous Dispersions of Two-Dimensional Materials with Glial Cell Compatibility and Enzymatic Degradability.

Water-dispersible two-dimensional (2D) materials are desirable for diverse applications. Aqueous dispersions make processing safer and greener and enable evaluation of these materials on biological and environmental fronts. To evaluate the effects of 2D materials with biological systems, obtaining dispersions without additives is critical and has been a challenge. Herein, a method was developed for obtaining additive-free aqueous dispersions of 2D materials like transition metal dichalcogenides and hexagonal boron nitride (h-BN). The nanosheet dispersions were investigated through spectroscopic and microscopic methods, along with the role of size on stability. The aqueous media enabled investigations on cytocompatibility and enzymatic degradation of molybdenum disulphide (MoS2 ) and h-BN. Cytocompatibility with mixed glial cells was observed up to concentrations of 100 µg mL-1 , suggesting their plausible usage in bioelectronics. Besides, biodegradation using human myeloperoxidase (hMPO) mediated catalysis was investigated through Raman spectroscopy and electron microscopy. The findings suggested that additive-free 2H-MoS2 and h-BN were degradable by hMPO, with 2H-phase exhibiting better resistance to degradation than the 1T-phase, while h-BN exhibited slower degradation. The findings pave a path for incorporating 2D materials in the burgeoning field of transient bioelectronics.

In Vitro Model to Investigate Communication between Dorsal Root Ganglion and Spinal Cord Glia.

Chronic discogenic back pain is associated with increased inflammatory cytokine levels that can influence the proximal peripheral nervous system, namely the dorsal root ganglion (DRG). However, transition to chronic pain is widely thought to involve glial activation in the spinal cord. In this study, an in vitro model was used to evaluate the communication between DRG and spinal cord glia. Primary neonatal rat DRG cells were treated with/without inflammatory cytokines (TNF-α, IL-1ß, and IL-6). The conditioned media were collected at two time points (12 and 24 h) and applied to spinal cord mixed glial culture (MGC) for 24 h. Adult bovine DRG and spinal cord cell cultures were also tested, as an alternative large animal model, and results were compared with the neonatal rat findings. Compared with untreated DRG-conditioned medium, the second cytokine-treated DRG-conditioned medium (following medium change, thus containing solely DRG-derived molecules) elevated CD11b expression and calcium signal in neonatal rat microglia and enhanced Iba1 expression in adult bovine microglia. Cytokine treatment induced a DRG-mediated microgliosis. The described in vitro model allows the use of cells from large species and may represent an alternative to animal pain models (3R principles).

Internalized stigma among patients with substance use disorders at a tertiary care center in India.

Internalized stigma among individuals with substance use disorders is a major barrier for accessing mental health services. This study aimed to assess internalized stigma among individuals with substance use disorders and to assess the relationship of internalized stigma with the quality of life. This cross-sectional study recruited 201 patients with a clinical diagnosis of at least opioid or alcohol use disorder according to Diagnostic and Statistical Manual 5 at a public-funded tertiary care center in India. The study participants were interviewed using a sociodemographic questionnaire, the Internalized Stigma of Mental Illness Scale (ISMIS), and the World Health Organization's Quality of Life (WHOQOL-Bref) questionnaire. Seven participants (3.5% of the sample) had mild stigma according to ISMI scores, 62 (30.8%) had moderate stigma, and 132 (65.7%) had severe stigma. The various quality-of-life domains generally had a negative correlation with the internalized stigma scores. Participants using opioids as the primary substance of use were more likely to have severe internalized stigma. The experience of internalized stigma and dissatisfaction with quality of life is quite high among people suffering with substance use disorders in India. These results emphasize the need for interventions to reduce internal perception of stigma and improve the quality of life of individuals with substance use disorders.

An Observational Study of Treatment Seeking Users of Natural Opiates from India.

BACKGROUND: The type of opioid used influences the severity and complications experienced. Natural opiates (opium and poppy husk) use is a socio-culturally accepted phenomenon reported in India. However, studies on their profile, quality of life, and addiction severity are limited. OBJECTIVES: The objective of this cross-sectional, observational study was to assess the socio-demographic profile, clinical profile, addiction severity, and quality of life of treatment-seeking natural opiate users. METHOD: Hundred subjects aged 18-65 years using opium or poppy husk seeking outpatient treatment at a tertiary addiction treatment center in India were interviewed to collect information on their socio-demography, natural opiate, and other substance use. Additionally, their addiction severity and quality of life were assessed using Addiction Severity Index-Lite and WHO Quality of Life-Bref instrument, respectively. RESULTS: All subjects were male with a mean age of 44.6 (±11.0) years. Majority (97%) used poppy husk daily orally. Curiosity/experimentation (63%) was the most common reason for starting opiate use. The past month rates of tobacco, alcohol, cannabis, and sedative-hypnotics use was 58%, 33%, 3%, and 12%, respectively. Only 4% injected any opioid. Inability to afford opiates (72%) was the most common reason for seeking treatment. Rates of medical, familial, social, psychological, and legal complications were low, while the WHOQOL-BREF scores fell between 40 and 50 across various domains. Conclusions/Importance: Natural opiate users may constitute distinct subgroup of opioid users with fewer/no complications despite long duration of uninterrupted use. These findings would be important in planning management strategies for people dependent on natural opiates.

Smart stylet: the development and use of a bedside external ventricular drain image-guidance system.

BACKGROUND: Placement accuracy of ventriculostomy catheters is reported in a wide and variable range. Development of an efficient image-guidance system may improve physician performance and patient safety. OBJECTIVE: We evaluate the prototype of Smart Stylet, a new electromagnetic image-guidance system for use during bedside ventriculostomy. METHODS: Accuracy of the Smart Stylet system was assessed. System operators were evaluated for their ability to successfully target the ipsilateral frontal horn in a phantom model. RESULTS: Target registration error across 15 intracranial targets ranged from 1.3 to 4.6 mm (mean 3.1 mm). Using Smart Stylet guidance, a test operator successfully passed a ventriculostomy catheter to a shifted ipsilateral frontal horn 20/20 (100%) times from the frontal approach in a skull phantom. Without Smart Stylet guidance, the operator was successful 4/10 (40%) times from the right frontal approach and 6/10 (60%) times from the left frontal approach. In a separate experiment, resident operators were successful 2/4 (50%) times when targeting the shifted ipsilateral frontal horn with Smart Stylet guidance and 0/4 (0%) times without image guidance using a skull phantom. CONCLUSIONS: Smart Stylet may improve the ability to successfully target the ventricles during frontal ventriculostomy.

Combination of non-endoscopic parameters as predictors of large esophageal varices.

BACKGROUND AND AIM: Bleeding from gastroesophageal varices is a life-threatening complication of cirrhosis. Prophylactic therapy for large varices considerably reduces such bleeding. Current guidelines recommend upper GI endoscopy as the gold standard for detecting high-grade varices. However, in peripheral centers, this is often unfeasible or inaccessible thus requiring non-endoscopic predictors of large oesophageal varices. METHODS: This prospective study involved 100 newly diagnosed cirrhosis patients without any history of variceal bleeding. Demographic, clinical, bio-chemical and ultrasonographic parameters were recorded. Esophageal varices were classified endoscopically as small and large. Multivariate logistic regression was performed to identify predictors of variceal size and presence. Cut-offs of significant indicators were determined by ROC analysis. RESULTS: 69% patients had small and 31% had large varices. Univariate analysis revealed significant differences between the grade of spleen, blood parameters, platelet count, international normalized ratio, serum albumin, spleen size, portal vein diameter (PVD) and platelet count to spleen diameter ratio in the two groups. Multivariate analysis revealed that INR [OR: 2.432(95% CI: 1.192 - 4.958)], splenomegaly at USG [OR: 2.138 (95% CI: 0.662 - 6.911)] and PVD [OR: 1.318 (95% CI: 0.937 - 1.853)] were the most significant predictors for large varices. CONCLUSIONS: Multivariate prediction of large varices based on a combination of non-endoscopic parameters can be utilized in place of single parameter based predictions.

When a Sibling Has Autism: Narrative Review of Interventions for Typically Developing Siblings.

Being a typically developing sibling to a child with autism spectrum disorder (ASD-Sib) can be associated with several behavioral and mental health problems. With this understanding, researchers are beginning to focus on sibling-oriented interventions. This review seeks to evaluate the current literature on such interventions. We searched digital databases and identified eight relevant studies. The sample (n = 247) had an age range of 4-16 years. The outcomes were generally positive, with improvements in different facets of ASD-Sibs. Results differed due to varied intervention styles, outcome variables, methods, and samples. Our findings highlight the potential for improvement in ASD-Sib's knowledge of autism and their psychological functioning and hence call for further research with robust methods.

Cascade Oxypalladation/1,3-Palladium Shift to Access Cyclopentene-Fused Isocoumarins.

Fused isocoumarins are frequently found in several natural products and pharmaceuticals. Herein, a cascade annulation of 2-alkynylbenzoate-tethered cyclic 1,3-diones via sequential trans-oxypalladation, carbonyl insertion, 1,3-Pd shift, and ß-hydride elimination is reported. This method provides efficient access to highly diastereoselective tetracyclic cyclopentene-fused isocoumarins containing two contiguous quaternary stereocenters. A plausible reaction mechanism is proposed on the basis of mechanistic studies, including deuterium labeling experiments. Studies toward enantioselective synthesis using a chiral Bpy ligand gave encouraging initial results.

Pathways to care and barriers in treatment among patients with Dissociative disorders.

INTRODUCTION: Dissociative disorder patients often present with sudden and embarrassing symptoms, and it is difficult for the patient and care giver to understand initially, recognize the need for help and reach for appropriate treatment timely. This can result in high risk of engaging in dangerous behaviors such as self-harm and suicidal acts, impaired global functioning, and poor quality of life. Knowledge about the types of barriers which are there in treatment seeking, can help in planning strategies for their removal and to facilitate the treatment process. METHODS: Cross-sectional study among patients (n=133) with Dissociative disorders which were recruited from January 2023 to June 2023 in a tertiary care hospital. Pathways to care and barriers in treatment for Dissociative disorders were assessed by interviewing patients using semi-structured proforma. The Dissociative Experience Scale and World Health Organization Disability Assessment Schedule 2.0 (WHODAS 2.0)) were used to assess disease severity and impact of illness on various domains of life respectively. Group comparison was made to assess differences in social- clinical profile of patients choosing different modalities of treatment. RESULTS: 133 patients of Dissociative disorders with mean age 29.6±9.2, showed their first-choice of help seeking from general practitioner/ neurologist (40.6%), traditional faith healers (35.3%), psychiatrist (18.1%) and 5.2% preferred alternative treatments. This trend changed with 2nd and 3rd contact of help seeking with greater preference for psychiatrist in their 2nd (n=45, 33.8%) and 3rd (n=69, 51.8%) contact. The median duration of untreated illness was 56 weeks (IQR 24-182 weeks). Social-clinical profile of patients varied with their choice of treatment, having lower education level (P = 0.013), longer duration of untreated illness (p=0.003), more severity of symptoms (p=0.032) and greater disability scores(p=0.002) in patients whose first treatment choice was traditional faith healers. More than 70% patients faced availability barriers, stigma, unawareness about mental illness and influence of others in treatment of choice as barriers in initiating and continuing treatment. CONCLUSION: Patients with Dissociative disorders seek treatment from a multitude of healthcare providers including traditional faith healers, general physicians, and alternative medicine practitioners before reaching psychiatrist and undergoes various barriers in treatment. There is need to implement necessary measures for sensitization and awareness about Dissociative disorders to prevent prolonged and undue delays in initiation of appropriate management.

Qualitative study to explore the perspectives and mental health experiences of first episode psychosis patients and their caregivers in North India.

BACKGROUND: The onset of psychosis brings unfamiliar experiences that can be disturbing for patients and their caregivers. Few studies from India (only one from North India) have examined these experiences from the perspective of the patient and caregiver. We explored experiences of first episode psychosis (FEP) patients and their caregivers within a North Indian context. METHOD: Semi-structured interviews were conducted in 2019 with ten FEP patients and their caregivers (total n=20) receiving out-patient care in a tertiary care centre. Topic guides focused on concerns/complaints, symptoms, help-seeking, and barriers and facilitators to treatment. Interviews were audio recorded, transcribed, and analysed using qualitative content analysis. RESULTS: Main categories of responses from patients and caregivers included: initial complaints for seeking help, initial emotional response, barriers to seeking treatment, perceived dysfunction and improvement, experienced stigma, understanding about illness, early follow-up, preventive measures and awareness programs. Caregivers undergo myriad of emotional reactions including anger, anxiety, guilt, and confusion. Symptoms other than psychotic symptoms were the primary complaint upon seeking help, and there was lack of understanding about the psychosocial model of care (role of medications acknowledged with little awareness regarding psychosocial interventions in recovery). Persisting occupational dysfunction despite perceived symptomatic improvement was described by both patients and caregivers. CONCLUSION: North Indian patients with FEP lack awareness of symptoms. Therefore, onus for seeking help often falls on their caregivers. Psychoeducation from first contact with services and increasing awareness about psychotic illness within the community might help address lack of awareness about symptoms, mental health services, early signs of relapse, and importance of psychosocial interventions in achieving functional recovery.

Pilot study to test the feasibility and clinical efficacy of a psychosocial care programme for patients with psychosis in low-resource settings.

BACKGROUND: Home-based psychosocial care has the potential to improving outcomes in patients with schizophrenia and related disorders (SCZ). There is lack of India data for such care in early psychosis. We developed the "Saksham" programme, a bespoke self-managed home-based psychosocial care model, available in two formats: manual-based and mobile-application based. With the anticipated success of recruitment of early psychosis cases in our setting, we plan to test the such intervention in this population in future trials. AIM: To assess the feasibility of the Saksham programme intervention in people with SCZ and its clinical efficacy as an adjunct to treatment as usual. METHODS: Seventy-five patient-caregiver pairs (total n=150) were recruited. Patients received either: treatment-as-usual (TAU) (n=25), manual-based Saksham intervention+TAU (n=25), or app-based Saksham intervention+TAU (n=25). Feasibility (i.e. acceptability, practicality, demand, implementation and integration) was assessed at three-months. Participants were assessed for psychopathology, illness-severity, cognition, functioning, disability, and caregiver-coping at baseline, one-month, and three-month. The percentage changes over time were compared across three groups. RESULTS: More found the mobile application-based intervention acceptable and easy-to-use than the manual-based intervention (92 % vs 68 %, and 76 % vs 68 %, respectively). Psychopathology and caregiver-burden improved significantly in all three groups (p<0.05). Cognition, disability, functioning, and caregiver burden improved significantly in the two Saksham intervention groups, with greater improvement in the Saksham app group (p<0.05). CONCLUSION: Home-based intervention is feasible and acceptable in a low-resource setting, with preliminary evidence for effectiveness. These findings need corroboration with randomised controlled trials in early psychosis to ameliorate course of illness.

Healthcare in the Modern Era: Launching a Telemedicine-Based OPD Consultation in Rural Pune (Process, Results, and Challenges).

Introduction Telemedicine serves as a means of overcoming geographical barriers and increasing access to specialist care. This study focuses on the impact of telemedicine on the early diagnosis and treatment of patients, as well as its effect on patient satisfaction. In addition, the study examines the obstacles and facilitators that influence the implementation of telemedicine. Objectives The primary objectives of this study are to assess the effectiveness of telemedicine in facilitating early diagnosis and treatment for patients in need of specialist consultations, to evaluate patient satisfaction with specialist care delivered through telemedicine, and to identify the factors that influence the successful implementation of telemedicine in rural healthcare centers. Methodology An exploratory feasibility study was carried out at two rural health training centers (RHTCs) over a one-year period, enrolling 400 patients requiring specialist consultations. The study involved establishing a telemedicine center, implementing teleconsultations, and collecting data through patient interviews and self-administered questionnaires. Results A majority of teleconsultations, over 79%, were deemed valuable by medical officers, resulting in improved management, better counseling, and earlier diagnoses. More than 76% of patients found telemedicine to be acceptable due to the reduction in travel time and cost. The most common health concerns among patients were diabetes, hypertension, and skin disorders. The study also revealed several challenges, including limited specialist personnel, waiting times, prescription limitations, and connectivity issues. Discussion Telemedicine has proven to be a valuable tool for rural healthcare delivery, providing patients with access to specialist consultations and improving patient outcomes. Both patients and medical officers reported positive experiences with telemedicine. The findings of this study align with existing literature, which highlights the benefits of telemedicine in managing chronic diseases and increasing patient satisfaction. However, it is crucial to address challenges, such as personnel limitations and connectivity issues, to optimize telemedicine's effectiveness. Conclusion Telemedicine offers great potential for enhancing access to specialist care and achieving universal healthcare in rural areas. Despite its limitations, telemedicine demonstrates promising outcomes and warrants further development and optimization to ensure its successful implementation in rural healthcare centers.

Protocol-based assessment and management of first episode psychosis: Comparison of short and medium-term outcomes in psychopathology, quality of life, functioning and family burden across two sites in India.

BACKGROUND: Standard assessment and management protocols exist for first episode psychosis (FEP) in high income countries. Due to cultural and resource differences, these need to be modified for application in low-and middle-income countries. AIMS: To assess the applicability of standard assessment and management protocols across two cohorts of FEP patients in North and South India by examining trajectories of psychopathology, functioning, quality of life and family burden in both. METHOD: FEP patients at two sites (108 at AIIMS, North India, and 115 at SCARF, South India) were assessed using structured instruments at baseline, 3, 6 and 12 months. Standard management protocols consisted of treatment with antipsychotics and psychoeducation for patients and their families. Generalised estimating equation (GEE) modelling was carried out to test for changes in outcomes both across and between sites at follow-up. RESULTS: There was an overall significant improvement in both cohorts for psychopathology and other outcome measures. The trajectories of improvement differed between the two sites with steeper improvement in non-affective psychosis in the first three months at SCARF, and affective symptoms in the first three months at AIIMS. The reduction in family burden and improvement in quality of life were greater at AIIMS than at SCARF during the first three months. CONCLUSIONS: Despite variations in cultural contexts and norms, it is possible to implement FEP standard assessment and management protocols in North and South India. Preliminary findings indicate that FEP services lead to significant improvements in psychopathology, functioning, quality of life, and family burden within these contexts.

Factors associated with external ventricular drain placement accuracy: data from an electronic health record repository.

BACKGROUND: We evaluated external ventricular drain placement for factors associated with placement accuracy. Data were acquired using an electronic health record data requisition tool. METHOD: Medical records of all patients who underwent ventriculostomy from 2003 to 2010 were identified and evaluated. Patient demographics, diagnosis, type of guidance and number of catheter passes were searched for and recorded. Post-procedural hemorrhage and/or infection were identified. A grading scale was used to classify accuracy of catheter placements. A multiple logistic regression model was developed to assess features associated with accurate catheter placement. RESULTS: One hundred nine patients who underwent 111 ventriculostomies from 2003 to 2010 were identified. Patient diagnoses were classified into vascular (63 %), tumor (21 %), trauma (14 %), and cyst (2 %). Procedures were performed freehand in 90 (81 %), with the Ghajar guide in 17 (15 %), and with image guidance in 4 (4 %) patients. Eighty-eight (79 %) catheters were placed in the correct location. Trauma patients were more likely to have catheters misplaced (p = 0.007) whereas patients in other diagnostic categories were not significantly associated with misplaced catheters. Post-procedural hemorrhage was noted in 2 (1.8 %) patients on post-procedural imaging studies. Five (4.5 %) definite and 6 (5.4 %) suspected infections were identified. CONCLUSIONS: External ventricular drain placement can be performed accurately in most patients. Patients with trauma are more likely to have catheters misplaced. Further development is required to identify and evaluate procedure outcomes using an electronic health record repository.

An insight into new glycotherapeutics in glial inflammation: Understanding the role of glycosylation in mitochondrial function and acute to the chronic phases of inflammation.

INTRODUCTION: Glycosylation plays a critical role during inflammation and glial scar formation upon spinal cord injury (SCI) disease progression. Astrocytes and microglia are involved in this cascade to modulate the inflammation and tissue remodeling from acute to chronic phases. Therefore, understating the glycan changes in these glial cells is paramount. METHOD AND RESULTS: A lectin microarray was undertaken using a cytokine-driven inflammatory mixed glial culture model, revealing considerable differential glycosylation from the acute to the chronic phase in a cytokine-combination generated inflamed MGC model. It was found that several N- and O-linked glycans associated with glia during SCI were differentially regulated. Pearson's correlation hierarchical clustering showed that groups were separated into several clusters, illustrating the heterogenicity among the control, cytokine combination, and LPS treated groups and the day on which treatment was given. Control and LPS treatments were observed to be in dense clusters. This was further confirmed with lectin immunostaining in which GalNAc, GlcNAc, mannose, fucose and sialic acid-binding residues were detected in astrocytes and microglia. However, the sialyltransferase inhibitor inhibited this modification (upregulation of the sialic acid expression), which indeed modulates the mitochondrial functions. CONCLUSIONS: The present study is the first functional investigation of glycosylation modulation in a mixed glial culture model, which elucidates the role of the glycome in neuroinflammation in progression and identified potential therapeutic targets for future glyco therapeutics in neuroinflammation.

Models and approaches to comprehend and address glial inflammation following spinal cord injury.

Spinal cord injury (SCI) culminates in chronic inflammation and glial scar formation driven by the activation of microglia and astrocytes. Current anti-inflammatory strategies to treat glial activation associated with SCI have several limitations. Existing in vitro and ex vivo models studying molecular mechanisms associated with inflammation focus only on the acute phase. However, the progression of glial cell-derived inflammation over the acute-to-chronic phases has not been assessed. Understanding this progression will help establish a framework for evaluating therapeutic strategies. Additionally, new models could be useful as high-throughput screening (HTS) platforms. This review aims to highlight currently available models and future methods that could facilitate screening of novel therapeutics for SCI.

Statistically-aided development of protein A affinity chromatography for enhancing recovery and controlling quality of a monoclonal antibody.

Protein A chromatography is widely used for isolation of monoclonal antibodies (mAbs) from cell culture components. In this study, the effect of different process parameters of the Protein A purification namely, binding pH, elution pH, flow rate, neutralization pH and tween concentration, on the concentration and quality of the purified mAb were evaluated. Using design of experiments approach, the critical process parameters of protein A chromatography were identified and experimentally optimized. Their impact on quality attributes, such as size variants and charge variants, of the mAb was studied. Multivariate data analysis was subsequently performed using multiple linear regression and partial least squares regression methods. It was observed that the elution pH primarily governed the concentration of the purified mAb and the content of monomers and aggregates, while the tween concentration primarily influenced the main peak of the charge variants. This is the first study that evaluates the impact of tween concentration in buffers on the protein A chromatography purification step. These studies helped in identifying the design space and defining the target robust and optimal setpoints of the responses, which were subsequently verified experimentally. These setpoints not only passed the target criteria but also resulted in the highest recoveries during the investigation. Through this statistically-aided approach, an optimized and robust protein A chromatography process was rationally developed for purification of mAbs, while achieving the desired product quality. This study highlights the influence of multiple parameters of the protein A purification process on critical quality attributes of mAbs, such as the size and charge variants, which has been a very scarcely explored area.