Epigenetic adaptation drives monocyte differentiation into microglia-like cells upon engraftment into the retina.

The identification of specific markers for microglia has been a long-standing challenge. Recently, markers such as P2ry12, TMEM119, and Fcrls have been proposed as microglia-specific and widely used to explore microglial functions within various central nervous system (CNS) contexts. The specificity of these markers was based on the assumption that circulating monocytes retain their distinct signatures even after infiltrating the CNS. However, recent findings reveal that infiltrating monocytes can adopt microglia-like characteristics while maintaining a pro-inflammatory profile upon permanent engraftment in the CNS.In this study, we utilize bone marrow chimeras, single-cell RNA sequencing, ATAC-seq, flow cytometry, and immunohistochemistry to demonstrate that engrafted monocytes acquire expression of established microglia markers-P2ry12, TMEM119, Fcrls-and the pan-myeloid marker Iba1, which has been commonly mischaracterized as microglia-specific. These changes are accompanied by alterations in chromatin accessibility and shifts in chromatin binding motifs that are indicative of microglial identity. Moreover, we show that engrafted monocytes dynamically regulate the expression of CX3CR1, CCR2, Ly6C, and transcription factors PU.1, CTCF, RUNX, AP-1, CEBP, and IRF2, all of which are crucial for shaping microglial identity. This study is the first to illustrate that engrafted monocytes in the retina undergo both epigenetic and transcriptional changes, enabling them to express microglia-like signatures. These findings highlight the need for future research to account for these changes when assessing the roles of monocytes and microglia in CNS pathology.

A child with systemic onset juvenile idiopathic arthritis and nephrotic syndrome.

INTRODUCTION: Amyloidosis of the kidney is a rare complication in children with juvenile idiopathic arthritis (JIA), more commonly seen with systemic onset juvenile idiopathic arthritis (SOJIA). It usually presents with asymptomatic proteinuria. CASE REPORT: An 11.5-year-old boy with onset of SOJIA at 6 years of age came to our clinic with anasarca. Urinalysis and serum albumin suggested a diagnosis of nephrotic syndrome (NS) and kidney biopsy confirmed amyloidosis deposits. Treatment with injection tocilizumab was initiated. The proteinuria has decreased, and kidney functions are normal. CONCLUSION: Children with SOJIA should be monitored for proteinuria so that they can be offered timely appropriate therapy.

Clinical spectrum and outcome of children with deficiency of adenosine deaminase 2 (DADA2): multicentric experience from India.

OBJECTIVES: Deficiency of adenosine deaminase-2 (DADA2) is a monogenic disorder closely resembling polyarteritis nodosa (PAN) and can present to physicians across various specialties. Through this case series, we aim to describe the clinical spectrum and outcome of Indian children with DADA2. We aimed to study the clinical spectrum and outcome of Indian children with DADA-2. METHODS: The deidentified data from all participating centres were entered in an excel sheet, and the coordinating centre (All India Institute of Medical Sciences, New Delhi) screened the data for accuracy and completeness. RESULTS: We enrolled 16 children (11 females) in the study; the mean (SD) age at the time of onset of symptoms for males and females was 46.2 (47) and 73.6 (50.4) months, respectively. The most common clinical feature in this cohort was fever and rash in 80% of patients. More than half of children n, (%) [8, (53%)] had a CNS stroke. The other clinical features were hypertension [5(33%)], anaemia [3 (20%)] and arthralgia/arthritis in 4 (26%). These children were managed with various immunomodulators: steroids [13, (86%)], anti-TNF agents [(12, (80%)], cyclophosphamide [2 (13%)] and mycophenolate mofetil [3 (20%)]. The median (IQR) duration of follow-up for this cohort was 17 (10, 29) months. Fourteen children achieved remission and none had recurrent strokes after the initiation of anti-TNF drugs. CONCLUSION: DADA-2 closely resembles PAN; early age of onset and CNS stroke are striking differentiating features from classic PAN. Most children respond well to anti-TNF agents without serious adverse events during short-term follow-up.

Knowledge and Skill Retention in First-Year MBBS Students After Basic Life Support Training: A One-Year Longitudinal Study.

BACKGROUND: The Indian National Medical Council has incorporated the Basic Life Support (BLS) course in the foundation course of the undergraduate (MBBS) medical curriculum. However, medical teachers raise concerns about how training would affect the retention of Basic Life Support (BLS) abilities in the longer run. So, the current study assesses the knowledge and retention of BLS skills among first-year MBBS students over one year. METHODS: We included one hundred first-year MBBS students in our study who were trained for BLS, including theory, demonstrations and hands-on training using mannequins. Theoretical knowledge was assessed using pre-test and post-test questionnaires. At the same time, the skills were evaluated using Directly Observed Procedural Skills (DOPS) scores before, just after the training session, and again after one month, six months, and one year. Course feedback was also taken from the students after completing the sessions. RESULTS: There was a statistically significant difference between pre-and post-test knowledge scores, indicating that training improved their knowledge. (p < 0.001) There was also a statistically significant difference between pre-and post-test skills using DOPS (p < 0.001). There was no significant difference in the score when DOPS was conducted at one month, but a significant decrease in their skills was seen at six months and one year when compared with the Post Skill Score. (P < 0.001) CONCLUSIONS: The first-year medical students' knowledge and skills were enhanced by BLS training coupled with practical sessions. Such waning skills necessitate repeating the training at periodic intervals to reinform retention of skills acquired during BLS training.

Culture-Free Whole Genome Sequencing of Mycobacterium tuberculosis Using Ligand-Mediated Bead Enrichment Method.

Background: Direct whole genome sequencing (WGS) of Mycobacterium tuberculosis (Mtb) can be used as a tool to study drug resistance, mixed infections, and within-host diversity. However, WGS is challenging to obtain from clinical samples due to low number of bacilli against a high background. Methods: We prospectively collected 34 samples (sputum, n = 17; bronchoalveolar lavage, n = 13; and pus, n = 4) from patients with active tuberculosis (TB). Prior to DNA extraction, we used a ligand-mediated magnetic bead method to enrich Mtb from clinical samples and performed WGS on Illumina platform. Results: Mtb was definitively identified based on WGS from 88.2% (30/34) of the samples, of which 35.3% (12/34) were smear negative. The overall median genome coverage was 15.2% (interquartile range [IQR], 7.7%-28.2%). There was a positive correlation between load of bacilli on smears and genome coverage (P < .001). We detected 58 genes listed in the World Health Organization mutation catalogue in each positive sample (median coverage, 85% [IQR, 61%-94%]), enabling the identification of mutations missed by routine diagnostics. Mutations causing resistance to rifampicin, isoniazid, streptomycin, and ethambutol were detected in 5 of 34 (14.7%) samples, including the rpoB S441A mutation that confers resistance to rifampicin, which is not covered by Xpert MTB/RIF. Conclusions: We demonstrate the feasibility of magnetic bead-based enrichment for culture-free WGS of Mtb from clinical specimens, including smear-negative samples. This approach can also be integrated with low-cost sequencing workflows such as targeted sequencing for rapid detection of Mtb and drug resistance.

TET3 downregulation and low 5-hydroxymethylcytosine are epigenetic signatures of head and neck carcinoma.

BACKGROUND: Ten-eleven translocases (TETs) are enzymes responsible for demethylation processes, playing a crucial role in maintaining the body's methylation balance. Dysregulation of TET expression can lead to abnormal methylation levels. Isocitrate dehydrogenases (IDH) are upstream genes involved in Kreb cycle responsible for production of α-ketoglutarate (α-KG). α-KG and vitamin C are cofactors of TET3 enzyme. There is limited data on the relationship between TET3 and its cofactor Vitamin C in head and neck carcinoma (H&NC). METHODS AND RESULTS: In this study, we have investigated the expression of the TET3 gene along with IDH1/2 genes involved in the Krebs cycle in the peripheral blood of 32 H&NC patients compared to 32 healthy controls. We estimated serum levels of TET3 protein and vitamin C and 5-hydroxymethylcytosine (5-hmC) percentage in DNA isolated from EDTA blood samples. Our findings revealed that TET3 and IDH1/2 were downregulated in H&NC patients compared to healthy controls. Serum levels of TET3 and Vitamin C were low in H&NC patients compared to healthy controls. Diminished levels of percentage 5-hmC were detected in EDTA blood samples of H&NC patients compared to controls. Spearman correlation analysis revealed a significant positive correlation between TET3 levels, vitamin C levels and 5-hmC percentage. CONCLUSION: The low levels of Vitamin C are believed to contribute to decreased activity of the TET3 gene and less conversion of 5-methylcytosine (5-mC) to 5-hmC. Dietary supplementation of Vitamin C may increase TET3 activity.

Animated Videos as a Tool for Teaching Uncommon ENT Diagnoses.

OBJECTIVE: The purpose of this study was to evaluate the knowledge of healthcare professionals and learners regarding the diagnosis and management of nasal septal hematomas (NSH). The secondary objective was to evaluate the effectiveness of a short-form animated video as an educational tool.  Methods: A cross-sectional survey study of healthcare professionals and medical students in the United States was undertaken from October 2022 to June 2023. A pre-test survey was distributed to assess participants' baseline knowledge of NSH management. An educational video on nasal septal hematoma management was presented, followed by a post-test survey to measure the effectiveness of the video.  Results: A total of 142 participant results were collected, 62 (43.7%) of which were attending physicians. There was a significant improvement in knowledge scores across the sample, with a median pre-test score of 83.0% (interquartile range (IQR) 33) and a median post-test score of 100.0% (IQR 17, p<0.001). Additionally, on a visual analog scale (VAS), comfort levels in managing NSH improved from 3.20 to 4.82 (p<0.001) for the entire sample.  Conclusion: NSH is a rare yet potentially devastating otolaryngologic emergency that requires prompt diagnosis and management. A short-form animated video can be an effective tool for educating emergency professionals on diagnosing and managing NSH.

The Role of Psychosocial Stress on Cardiovascular Disease in Women: JACC State-of-the-Art Review.

Psychosocial stress can affect cardiovascular health through multiple pathways. Certain stressors, such as socioeconomic disadvantage, childhood adversity, intimate partner violence, and caregiving stress, are especially common among women. The consequences of stress begin at a young age and persist throughout the life course. This is especially true for women, among whom the burden of negative psychosocial experiences tends to be larger in young age and midlife. Menarche, pregnancy, and menopause can further exacerbate stress in vulnerable women. Not only is psychosocial adversity prevalent in women, but it could have more pronounced consequences for cardiovascular risk among women than among men. These differential effects could reside in sex differences in responses to stress, combined with women's propensity toward vasomotor reactivity, microvascular dysfunction, and inflammation. The bulk of evidence suggests that targeting stress could be an important strategy for cardiovascular risk reduction in women.

Assessment of ultrasound guided erector spinae plane block for early post-operative analgesia for modified radical mastectomy: a prospective, randomized, controlled study.

Erector spinae plane block is a recently introduced block with a wide range of indications. The aim of the present study was to assess the efficacy and safety of ultrasound-guided erector spinae plane block on early post-operative pain relief in patients undergoing modified radical mastectomy. We conducted a prospective, randomized, controlled study in a tertiary care institute. Sixty-five patients were enrolled. Final analysis was performed on 58 patients randomized into two groups. Ultrasound-guided erector spinae plane block with 25 mL of 0.25% bupivacaine was given using a 18 gauge needle. No block was given in the other group. All patients received general anesthesia. Primary outcome measure was 24-hour analgesic consumption. Secondary outcome measures included intra-operative fentanyl consumption, time to first analgesic request, Visual Analog Scale score, nausea score, sedation score, wound quality and patient satisfaction score. Post-operative additional analgesics and intra-operative fentanyl were significantly reduced in patients receiving erector spinae plane block. Time to rescue analgesia was significantly delayed. Visual Analog Scale score was significantly lower at all time intervals. Post-operative nausea at various time intervals was also less. Patient satisfaction score was also noted on a 5-point scale and there was a statistical significant difference. We concluded that erector spinae plane block is a safe technique and provides good analgesia in breast surgery.

Mechanisms of the Gewald Synthesis of 2-Aminothiophenes from Elemental Sulfur.

The Gewald reaction is a well-established one-pot method to access 2-aminothiophenes from carbonyl compounds, activated acetonitriles, and elemental sulfur. To elucidate the reaction's poorly understood mechanism, with regard to the decomposition of sulfur and polysulfide intermediates, we have performed a comprehensive computational study using density functional theory (DFT) calculations at the M06-2X (or ωB97X-D)/aug-cc-pV(T + d)Z/SMD(C2H5OH) level of theory. The results show that the reaction is initiated by a Knoevenagel-Cope condensation, followed by opening of the elemental sulfur, leading to polysulfide formation. The polysulfide intermediates can interconvert and decompose using various mechanisms including unimolecular cyclization, nucleophilic degradation, and scrambling. Protonation of the polysulfides changes their electrophilic behavior and provides a kinetically favorable pathway for their decomposition. This protonation-induced intermolecular degradation is feasible for polysulfides of all lengths, but unimolecular decomposition is kinetically favored for long polysulfides (≥6 sulfur atoms). None of the pathways provide any thermodynamic benefit due to the lack of resonance-stabilized leaving group, and a complex equilibrium of polysulfides of all lengths is expected in solution. Cyclization of the monosulfide with aromatization to the thiophene product is the only driving force behind the reaction, funneling all of the various intermediates into the observed product in a thermodynamically controlled process.

Transpersulfidation or H2S Release? Understanding the Landscape of Persulfide Chemical Biology.

Persulfides (RSSH) are biologically important reactive sulfur species that are endogenously produced, protect key cysteine residues from irreversible oxidation, and are important intermediates during different enzymatic processes. Although persulfides are stronger nucleophiles than their thiol counterparts, persulfides can also act as electrophiles in their neutral, protonated form in specific environments. Moreover, persulfides are electrophilic at both sulfur atoms, and the reaction with a thiolate can lead to either H2S release with disulfide formation or alternatively result in transpersulfidation. Despite the broad acceptance of these reaction pathways, the specific properties that control whether persulfides react through the H2S-releasing or transpersulfidation pathway remain elusive. Herein, we use a combined computational and experimental approach to directly investigate the reactivity between persulfides and thiols to answer these questions. Using density functional theory (DFT) calculations, we demonstrate that increasing steric bulk or electron withdrawal near the persulfide can shunt persulfide reactivity through the transpersulfidation pathway. Building from these insights, we use a synthetic persulfide donor and an N-iodoacetyl l-tyrosine methyl ester (TME-IAM) trapping agent to experimentally monitor and measure transpersulfidation from a bulky penicillamine-based persulfide to a cysteine-based thiol, which, to the best of our knowledge, is the first direct observation of transpersulfidation between low-molecular-weight species. Taken together, these combined approaches highlight how the properties of persulfides are directly impacted by local environments, which has significant impacts in understanding the complex chemical biology of these reactive species.

No Association Between AGT Gene Polymorphisms with Hypertension in a South African Population.

Purpose: Hypertension is a leading cause of cardiovascular-related morbidity and mortality worldwide, with a prevalence increasing at an alarming rate in both middle- and low-income countries. Various environmental and genetic factors have been attributed to play a significant role in the increasing prevalence of hypertension. Single nucleotide polymorphisms (SNPs) in the angiotensinogen (AGT) gene are reported to have a significant association with hypertension; however, there are limited studies done on South African populations. Therefore, this case-control study aimed to investigate the association between AGT SNPs (rs2004776, rs3789678, rs5051 and rs7079) with hypertension in a study population of isiXhosa-speaking participants from the Eastern Cape Province in South Africa. Materials and Methods: The SNPs were genotyped in 250 hypertensive cases and 237 normotensive controls, using TaqMan genotyping assays. Results: For the SNP rs2004776, the frequency of CC genotype (18.4%) and C allele (44%) in hypertensive cases showed no significant differences (p = 0.52, χ2 = 1.32), when compared to the normotensive control group (CC: 19.8% and C allele: 43%). Similar results were obtained for the genotypic and allelic frequencies between hypertensive cases and normotensive controls for rs3789678 (p = 0.88, χ2=0.26) and rs5051 (p = 0.57, χ2=1.12), and rs7079 (p = 0.33, χ2=2.23). These findings demonstrate that there were no significant associations between the SNPs rs2004776, rs3789678, rs7079, rs5051 with hypertension in our study population. Conclusion: These findings suggest that AGT gene polymorphisms are not associated with the development of hypertension in the studied population. The present study represents the first genetic report to investigate the AGT gene polymorphisms with hypertension in an isiXhosa-speaking South African population.

SpatialPrompt: spatially aware scalable and accurate tool for spot deconvolution and domain identification in spatial transcriptomics.

Efficiently mapping of cell types in situ remains a major challenge in spatial transcriptomics. Most spot deconvolution tools ignore spatial coordinate information and perform extremely slow on large datasets. Here, we introduce SpatialPrompt, a spatially aware and scalable tool for spot deconvolution and domain identification. SpatialPrompt integrates gene expression, spatial location, and single-cell RNA sequencing (scRNA-seq) dataset as reference to accurately infer cell-type proportions of spatial spots. SpatialPrompt uses non-negative ridge regression and graph neural network to efficiently capture local microenvironment information. Our extensive benchmarking analysis on Visium, Slide-seq, and MERFISH datasets demonstrated superior performance of SpatialPrompt over 15 existing tools. On mouse hippocampus dataset, SpatialPrompt achieves spot deconvolution and domain identification within 2 minutes for 50,000 spots. Overall, domain identification using SpatialPrompt was 44 to 150 times faster than existing methods. We build a database housing 40 plus curated scRNA-seq datasets for seamless integration with SpatialPrompt for spot deconvolution.

Comparative evaluation of the antibacterial activity of curcumin-coated gutta-percha versus nanocurcumin-coated gutta-percha against Escherichia coli: An in vitro study.

Introduction: Due to its biological and antibacterial qualities, many plants, including curcumin, are used as phytomedicines in dentistry. They are primarily used as intracanal medication in endodontics to prevent probable chemical side effects and also to address antimicrobial resistance. Curcumin nanoformulations have improved antibacterial activity and improved dispersion, making them the superior form of curcumin. The purpose of this study was to assess curcumin and nanocurcumin's antibacterial properties. As a gutta-percha coating, they are to be tested against Escherichia coli. Materials and Methods: The study employs the standard strain of E. coli, ATCC 25922. The antibacterial activity of gutta-percha cones against E. coli is assessed after coating them with suspensions of curcumin and nanocurcumin. Scanning electron microscopy is utilized to evaluate the coatings' continuity. Results: The gutta-percha cones that are untreated, coated with curcumin, and coated with nanocurcumin exhibit significantly different levels of antibacterial activity. There is statistically significant variation in their antibacterial activity. Conclusion: (1) Compared to curcumin-coated and untreated gutta-percha cones, those coated with nanocurcumin exhibit a stronger antibacterial activity. (2) Compared to uncoated gutta-percha cones, gutta-percha cones coated with curcumin exhibit more antibacterial action.

Functional and Radiological Outcomes of Miniature Plate Osteosynthesis in Metacarpal and Phalangeal Fractures of Hand: A Prospective Study.

INTRODUCTION:  There are several operative modes to address hand fractures to gain better anatomical and functional results including external fixator, percutaneous K-wire fixation, lag screw fixation, tension band wiring, intra-medullary nails or wires, and plate-screw fixation. We evaluated the results of plate osteosynthesis in fractures of metacarpals and phalanges in a prospective manner. MATERIAL AND METHODS:  A total 50 adults (19-60 years) of either sex having 58 fractures managed by miniature plate osteosynthesis and followed for a minimum six months (6-19), including metacarpal and phalangeal fractures (unstable or serial fractures), intra-articular fractures, fracture-dislocation of proximal interphalangeal and distal interphalangeal joints with joint incongruity or subluxation were enrolled while contaminated compound fractures, pathological fractures and cases of reimplantation were excluded from study. Clinical assessment was done using the American Society for Surgery of the Hand (ASSH), total active flexion (TAF), total active range of motion (TAM) score, and the Disabilities of the Arm, Shoulder, and Hand (quick DASH) score while sequential radiographs were done at each follow-up. RESULT: All the fractures had perfect union clinically as well as radiologically with a mean duration of six weeks while functional outcomes in reference to clinical scores were observed excellent and fair in all cases. None of the cases had any loss of reduction, loosening of the implant, or other major complications. CONCLUSIONS: Miniature plate fixation provides enough stability in metacarpal and phalanges fractures, thereby allowing immediate active movements, which leads to excellent functional outcomes and early return to normal activities.

Correlation of Venous Blood Sugar Measured by Lab Method and Capillary Blood Sugar Measured by Glucometer in Neurosurgical Patients Receiving Dexamethasone.

Background Brain is vulnerable to extreme blood glucose levels that may occur due to multiple factors in neurosurgical patients; perioperative use of dexamethasone is the most common. Thus, frequent monitoring of blood sugar levels is advocated. This study aimed to assess correlation between venous blood sugar measured by lab method and capillary blood sugar by glucometer at various time intervals. Materials and Methods This prospective and observational study was conducted in 20 adult patients of either sex, American Society of Anesthesiologists grade I to III, scheduled to undergo brain tumor resection. The patients who were already on dexamethasone and received intraoperatively 8 mg dexamethasone were enrolled. Standard anesthesia technique and intraoperative monitoring were followed in all patients. Venous sample was withdrawn and blood sugar analyzed in laboratory, while at the same time capillary blood sugar was tested by glucometer. The sampling was done at baseline, 1 hourly after dexamethasone administration till 4 hours and then 8, 12, and 24 hours. The correlation between the two values was assessed. Results During the study, 160 venous and 160 capillary blood sugar levels were analyzed. Though capillary blood sugar levels were slightly higher than venous sugar levels, there was strong correlation between the two (Pearson correlation coefficient) with p -value less than 0.05 except at 24 hours when two values were not correlated. Conclusion Capillary blood sugar levels by glucometer have good correlation with venous sugar levels; therefore, this method may be adopted routinely for frequent blood sugar estimation as it is reliable, easy, and practical.

Effects of pain education on disability, pain, quality of life, and self-efficacy in chronic low back pain: A randomized controlled trial.

BACKGROUND: Low back pain stands as a prevalent contributor to pain-related disability on a global scale. In addressing chronic low back pain (CLBP), there is a growing emphasis on incorporating psychological strategies into the management process. Among these, pain education interventions strive to reshape pain beliefs and mitigate the perceived threat of pain. This randomized controlled trial sought to assess the effects of pain education on various aspects, including pain levels, disability, quality of life, self-efficacy, and prognostic characteristics in individuals grappling with CLBP. METHODS: The clinical trial, retrospectively registered with the Clinical Trials Registry of India (CTRI/2021/08/035963), employed a two-arm parallel randomized design. Ninety-two participants with CLBP were randomly assigned to either the standard physiotherapy care with a pain education program or the control group. Both groups underwent a 6-week intervention. Assessment of pain intensity (using NPRS), disability (using RMDQ), self-efficacy (using the general self-efficacy scale), and well-being (using WHO 5I) occurred both before and after the 6-week study intervention. FINDINGS: Post-intervention score comparisons between the groups revealed that the pain education intervention led to a significant reduction in disability compared to the usual standard care at 6 weeks (mean difference 8.2, p < 0.001, effect size Cohen d = 0.75), a decrease in pain intensity (mean difference 3.5, p < 0.001, effect size Cohen d = 0.82), and an improvement in the well-being index (mean difference 13.7, p < 0.001, effect size Cohen d = 0.58). CONCLUSION: The findings suggest that integrating a pain education program enhances the therapeutic benefits of standard physiotherapy care for individuals dealing with chronic LBP. In conclusion, the clinical benefits of pain education become apparent when delivered in conjunction with standard care physiotherapy during the management of chronic low back pain.

Defining albumin as a glycoprotein with multiple N-linked glycosylation sites.

BACKGROUND: Glycosylation is an enzyme-catalyzed post-translational modification that is distinct from glycation and is present on a majority of plasma proteins. N-glycosylation occurs on asparagine residues predominantly within canonical N-glycosylation motifs (Asn-X-Ser/Thr) although non-canonical N-glycosylation motifs Asn-X-Cys/Val have also been reported. Albumin is the most abundant protein in plasma whose glycation is well-studied in diabetes mellitus. However, albumin has long been considered a non-glycosylated protein due to absence of canonical motifs. Albumin contains two non-canonical N-glycosylation motifs, of which one was recently reported to be glycosylated. METHODS: We enriched abundant serum proteins to investigate their N-linked glycosylation followed by trypsin digestion and glycopeptide enrichment by size-exclusion or mixed-mode anion-exchange chromatography. Glycosylation at canonical as well as non-canonical sites was evaluated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) of enriched glycopeptides. Deglycosylation analysis was performed to confirm N-linked glycosylation at non-canonical sites. Albumin-derived glycopeptides were fragmented by MS3 to confirm attached glycans. Parallel reaction monitoring was carried out on twenty additional samples to validate these findings. Bovine and rabbit albumin-derived glycopeptides were similarly analyzed by LC-MS/MS. RESULTS: Human albumin is N-glycosylated at two non-canonical sites, Asn68 and Asn123. N-glycopeptides were detected at both sites bearing four complex sialylated glycans and validated by MS3-based fragmentation and deglycosylation studies. Targeted mass spectrometry confirmed glycosylation in twenty additional donor samples. Finally, the highly conserved Asn123 in bovine and rabbit serum albumin was also found to be glycosylated. CONCLUSIONS: Albumin is a glycoprotein with conserved N-linked glycosylation sites that could have potential clinical applications.

Management of Extraocular Retinoblastoma: ICMR Consensus Guidelines.

Retinoblastoma (RB) is the most common intraocular malignancy of childhood. Advanced stage presentation of RB is common in low middle-income countries (LMICs) due to lack of awareness, social taboos associated with enucleation, seeking alternative conservative treatment options, and poor accessibility to health care. Over the last few decades, there have been significant advancements in the management of extraocular RB (EORB) which have improved outcomes and helped in minimizing treatment-related toxicities. The incorporation of multimodality approaches including chemotherapy, surgery, and radiotherapy (RT) has shown promising results; however, prognosis remains poor especially in LMICs. In this article, authors have discussed the ICMR consensus guidelines on the management of EORB, including metastatic RB.