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Acute myeloid leukemia (AML) is a complex clonal disorder characterized by clinical, genetic, metabolomic, and epigenetic heterogeneity resulting in the uncontrolled proliferation of aberrant blood-forming precursor cells. Despite advancements in the understanding of the genetic, metabolic, and epigenetic landscape of AML, it remains a significant therapeutic challenge. Functional profiling techniques, such as BH3 profiling (BP), gene expression profiling (GEP), proteomics, metabolomics, drug sensitivity/resistance testing (DSRT), CRISPR/Cas9, and RNAi screens offer valuable insights into the functional behavior of leukemia cells. BP evaluates the mitochondrial response to pro-apoptotic BH3 peptides, determining a cell’s apoptotic threshold and its reliance on specific anti-apoptotic proteins. This knowledge can pinpoint vulnerabilities in the mitochondria-mediated apoptotic pathway in leukemia cells, potentially informing treatment strategies and predicting therapeutic responses. GEP, particularly RNA sequencing, evaluates the transcriptomic landscape and identifies gene expression alterations specific to AML subtypes. Proteomics and metabolomics, utilizing mass spectrometry and nuclear magnetic resonance (NMR), provide a detailed view of the active proteins and metabolic pathways in leukemia cells. DSRT involves exposing leukemia cells to a panel of chemotherapeutic and targeted agents to assess their sensitivity or resistance profiles and potentially guide personalized treatment strategies. CRISPR/Cas9 and RNAi screens enable systematic disruption of genes to ascertain their roles in leukemia cell survival and proliferation. These techniques facilitate precise disease subtyping, uncover novel biomarkers and therapeutic targets, and provide a deeper understanding of drug-resistance mechanisms. Recent studies utilizing functional profiling have identified specific mutations and gene signatures associated with aggressive AML subtypes, aberrant signaling pathways, and potential opportunities for drug repurposing. The integration of multi-omics approaches, advances in single-cell sequencing, and artificial intelligence is expected to refine the precision of functional profiling and ultimately improve patient outcomes in AML. This review highlights the diverse landscape of functional profiling methods and emphasizes their respective advantages and limitations. It highlights select successes in how these methods have further advanced our understanding of AML biology, identifies druggable targets that have improved outcomes, delineates challenges associated with these techniques, and provides a prospective view of the future where these techniques are likely to be increasingly incorporated into the routine care of patients with AML.

Acute myeloid leukemia (AML) is clinically and genetically a heterogeneous disease characterized by clonal expansion of abnormal hematopoietic progenitors. Genomic approaches to precision medicine have been implemented to direct targeted therapy for subgroups of AML patients, for instance, IDH inhibitors for IDH1/2 mutated patients, and FLT3 inhibitors with FLT3 mutated patients. While next generation sequencing for genetic mutations has improved treatment outcomes, only a fraction of AML patients benefit due to the low prevalence of actionable targets. In recent years, the adoption of newer functional technologies for quantitative phenotypic analysis and patient-derived avatar models has strengthened the potential for generalized functional precision medicine approach. However, functional approach requires robust standardization for multiple variables such as functional parameters, time of drug exposure and drug concentration for making in vitro predictions. In this review, we first summarize genomic and functional therapeutic biomarkers adopted for AML therapy, followed by challenges associated with these approaches, and finally, the future strategies to enhance the implementation of precision medicine.

This review aims to explore the various antinutritional factors found in grains and oilseeds, their sources, and the different methods employed to mitigate their effects. Both traditional and innovative processing techniques have been utilized to reduce antinutrients. Traditional methods such as dehulling, soaking, boiling, pressure cooking, sprouting, and fermentation have proven effective in reducing specific antinutrients like α-galactosides, phytic acid, enzyme inhibitors, lathyrogens, lectins, saponins, and tannins. However, optimization of these techniques is necessary to maximize their effectiveness and minimize antinutrient content. The review also provides an overview of various analytical methods used to quantify antinutritional factors in grains and oilseeds. These methods encompass classical gravimetric and titrimetric analysis, potentiometric titrations, chromatographic and electrophoretic separation techniques, as well as spectroscopic methods, fluorescence, and biosensors. A systematic and chronological review of these analytical approaches and detection systems is presented. Furthermore, the review identifies research gaps and highlights potential areas for future exploration in the field of antinutrients in grains and oilseeds. By addressing these gaps, we can further enhance our understanding and develop more effective strategies to mitigate antinutrients, thus promoting the utilization of grains and oilseeds for their optimal health benefits.

Summary: The clinical effectiveness of BH3 mimetics therapy is limited by the inevitable emergence of acquired resistance. We present a protocol to model in vivo acquired resistance to BH3 mimetics in patient-derived xenograft (PDX) mouse models of acute myeloid leukemia. Using resistant PDXs as a valuable model, we next introduce a protocol for dynamic BH3 profiling (DBP) method. DBP allows functional identification of effective drug therapies based on measurements of drug-induced apoptosis signaling to overcome in vivo BH3 mimetics resistance.For complete details on the use and execution of this protocol, please refer to Bhatt et al. (2020).

Objectives: The existing biomarkers used to promptly identify graft dysfunction after kidney transplantation lack consistency. Neutrophil gelatinase-associated lipocalin (NGAL) appears to be a promising biomarker but its levels measured from serum and urine have demonstrated varying predictive values. Our study aimed to explore the potential of NGAL as a biomarker in predicting graft dysfunction in kidney transplant patients, including live and deceased donor recipients. Methods: A single-centered observational cohort study with live and deceased kidney recipients as participants was conducted between 2018 and 2022 at a tertiary care hospital in Southern India. Serum creatinine levels were monitored daily; creatinine reduction on day two and day seven were calculated. The recipients were categorized based on graft recovery into three groups: delayed graft function (DGF), slow graft function (SGF), or immediate graft function (IGF). Analysis of serum and urine NGAL was conducted two hours after the transplant and their predictive values were evaluated by the area under the curves (AUC) method. Results: Of the 40 participants, 34 (85.0%) received their transplant from live-related donors, while six (15.0%) received kidneys from deceased donors. DGF occurred in four (10.0%) patients, SGF in 12 (30.0%), and 24 (60.0%) patients achieved IGF. Serum NGAL demonstrated higher sensitivity compared to urine NGAL. At a cut-off value of 678 ng/mL (AUC = 0.77), serum NGAL showed 90.0% sensitivity and 53.0% specificity. Urine NGAL had 70.0% sensitivity and 74.0% specificity at a cutoff value of 489 ng/mL (AUC = 0.72). Conclusions: Kidney recipients in SGF and DGF categories had elevated levels of serum and urine NGAL compared to those without IGF. Although serum NGAL showed higher sensitivity than urine NGAL in predicting graft dysfunction, both markers lacked the specificity needed for accurate predictions.

BACKGROUND: Pterygium is very common in India and is usually removed by limbal conjunctival autograft transplantation (LCAT), which, despite being the first-line therapy, is still associated with recurrences of up to 18%. OBJECTIVES: To compare the safety and efficacy of topical cyclosporine A (CsA) and interferon (IFN) alpha-2b in the prevention of postoperative recurrence of pterygium. METHODS: A total of 40 patients with primary pterygium were randomized into two equal groups, Group C and Group I. Both the groups underwent LCAT, with Group C kept on topical cyclosporine 0.05% (CsA) 4 times daily and Group I on topical IFN alpha 2b 0.2 million IU 4 times daily postoperatively for 3 months. Pre- and posttreatment best-corrected visual acuity (BCVA), recurrence, and complications were assessed at day 1, week 1, 1 month, and 3 months. RESULTS: The mean preoperative BCVA of 0.51 ± 0.18 and 0.51 ± 0.23 improved to 0.13 ± 0.13 and 0.13 ± 0.13 in Group C and Group I, respectively, after 3 months of treatment (P < 0.0001). Recurrence was seen in 2 cases in Group C and in 1 case in Group I at 3 months. No significant complications occurred in either of the groups. CONCLUSION: Topical CsA and IFN Alpha-2b are newer efficacious adjuvants with LCAT for prevention of postoperative pterygium recurrence.

Abstract Purpose: Osteomyelitis of the orbital bones presenting as an orbital cellulitis is a rare form of extrapulmonary tuberculosis (TB). We report a rare case of tubercular osteomyelitis of the orbital bones presenting as a periorbital cellulitis. Case Report: A seven-year-old female child presented to our tertiary eye care center with swelling involving the right eyelids and the right cheek for two months. She had been provisionally diagnosed elsewhere as pre-septal cellulitis and had been given oral antibiotics. We clinically diagnosed her as orbital cellulitis, but her non-responsiveness to intravenous antibiotics prompted us to get a contrast enhanced computed tomography (CECT) of the orbit and paranasal sinuses, which was suggestive of tubercular etiology. However, the patient had no foci for TB elsewhere. We used a relatively new, but rapid test, called Cartridge-based Nucleic Acid Amplification Test (CBNAAT) on the pus aspirate which was positive for TB. Thereafter, the patient was started on anti-tubercular treatment to which she responded wonderfully. Conclusion: A high index of suspicion should be kept for TB infection in cases of orbital cellulitis with unusual clinical behavior in an endemic region such as India.

Coronavirus disease 2019 (COVID-19) vaccinations have been associated with a higher risk of thromboembolic events. There have been no reports of central retinal artery occlusion (CRAO) after vaccination with the indigenously developed Covaxin, and worldwide, there has been only one such isolated case after administration of the AstraZeneca vaccine. We report a case of a 44-year-old healthy man who presented with sudden painless vision loss in his left eye 10 days after receiving Covaxin. His best-corrected visual acuity was minimal perception of light, with a relative afferent pupillary defect. Fundus examination revealed arterial attenuation and macular cherry red spot, suggesting an acute CRAO. Optical coherence tomography showed macular swelling and disorganization of the inner layers due to ischemic sequelae. Blood work was normal and cardiovascular examination was unremarkable. The patient was kept on follow-up. To our knowledge, this is the first case of an isolated CRAO after Covaxin administration, but further studies are needed to evaluate this potential association.

Abstract Background Axial SpA and Enthesitis related arthritis (ERA) patients show strong HLA-B27 association, gut dysbiosis, high toll like receptor (TLR)2 and 4 expression on monocytes, pro-inflammatory cytokine production and elevated levels of TLR4 endogenous ligands [tenascin-c (TNC) and myeloid related protein (MRP)8/14] in serum. Hence, we aimed to understand if these diseases have similar or different monocyte response. Methods Fifty adult axial SpA, 52 ERA patients and 25 healthy controls (HC) were enrolled. Cytokine-producing monocyte frequency before and after stimulation with lipopolysaccharide (LPS), peptidoglycan (PG), TNC or MRP8 were measured in whole blood (WB) and synovial fluid mononuclear cells (SFMC) by flow cytometry. Also, IL-6, TNF, MMP3, TNC and MRP8/14 levels were measured in unstimulated and TLR ligand stimulated WB cultures supernatant by ELISA. Finally, the mRNA expression levels of TNF and IL-6 were measured post stimulation with LPS, TNC and MRP8. Results At baseline, ERA and axial SpA patients showed similar TNF-α producing monocyte frequency which was higher than HC. MRP8 simulation led to increased TNF-α producing monocyte frequency in ERA than axial SpA. TNC and MRP8 stimulation led to similar IL-6 producing monocyte frequency in axial SpA and ERA patients. Baseline TNF and IL-6 producing monocyte frequency also modestly correlated with disease activity scores. TNF and IL-6 producing monocyte frequency increased in response to TLR stimulation in SFMC from both patients. In culture supernatants, axial SpA and ERA patients showed similar TNF production at baseline. MRP8 and TNC stimulation led to higher TNF production from ERA. Baseline IL-6 and MMP3 production was higher in ERA while TLR stimulation led to similar IL-6 and MMP3 production from axial SpA and ERA. TNC stimulation led to higher MMP3 production in ERA. mRNA expression in response to TLR stimulation was observed to be similar in axial SpA and ERA. TNC production was higher in ERA at baseline, while MRP8/14 production was higher in axial SpA than ERA post stimulation. Conclusion ERA patients have similar monocyte response to exogenous and endogenous TLR ligands as patients with axial SpA. This suggests that differences between pediatric and adult-onset SpA are minimal and they may have a common pathogenesis.

Distinct subgroups of diffuse large B-cell lymphoma (DLBCL) genetically resemble specific mature B-cell populations that are blocked at different stages of the immune response in germinal centers (GCs). The activated B-cell (ABC)-like subgroup resembles post-GC plasmablasts undergoing constitutive survival signaling, yet knowledge of the mechanisms that negatively regulate this oncogenic signaling remains incomplete. In this study, we report that microRNA (miR)-181a is a negative regulator of nuclear factor k-light-chain enhancer of activated B-cells (NF-κB) signaling. miR-181a overexpression significantly decreases the expression and activity of key NF-κB signaling components.Moreover, miR-181a decreases DLBCL tumor cell proliferation and survival, and anti-miR-181a abrogates these effects. Remarkably, these effects are augmented in the NF-κB dependent ABC-like subgroup compared with the GCB-cell (GCB)-likeDLBCLsubgroup.Concordantly, in vivo analyses of miR-181a induction in xenografts results in slower tumor growth rate and prolonged survival in the ABC-like DLBCL xenografts compared with the GCB-like DLBCL. We link these outcomes to relatively lower endogenous miR-181a expression and to NF-κB signaling dependency in the ABC-like DLBCL subgroup. Our findings indicate that miR-181a inhibits NF-κB activity, and that manipulation of miR-181a expression in the ABC-like DLBCL genetic backgroundmay result in a significant change in the proliferation and survival phenotype of this malignancy. [ABSTRACT FROM AUTHOR]