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Antibodies are critical tools for research into extracellular vesicles (EVs) and other extracellular nanoparticles (ENPs), where they can be used for their identification, characterization, and isolation. However, the lack of a centralized antibody platform where researchers can share validation results thus minimizing wasted personnel time and reagents, has been a significant obstacle. Moreover, because the performance of antibodies varies among assay types and conditions, detailed information on assay variables and protocols is also of value. To facilitate sharing of results on antibodies that are relevant to EV/ENP research, the EV Antibody Database has been developed by the investigators of the Extracellular RNA Communication Consortium (ERCC). Hosted by the ExRNA Portal (https://exrna.org/resources/evabdb/), this interactive database aggregates and shares results from antibodies that have been tested by research groups in the EV/ENP field. Currently, the EV Antibody Database includes modules for antibodies tested for western Blot, EV Flow Cytometry, and EV Sandwich Assays, and holds 110 records contributed by 6 laboratories from the ERCC. Detailed information on antibody sources, assay conditions, and results is provided, including negative results. We encourage ongoing expert input and community feedback to enhance the databases utility, making it a valuable resource for comprehensive validation data on antibodies and protocols in EV biology.

Inherited genetic disorders of the liver pose a significant public health burden. Liver transplantation is often limited by the availability of donor livers and the exorbitant costs of immunosuppressive therapy. To overcome these limitations, nucleic acid therapy provides a hopeful alternative that enables gene repair, gene supplementation, and gene silencing with suitable vectors. Though viral vectors are the most efficient and preferred for gene therapy, pre-existing immunity debilitating immune responses limit their use. As a potential alternative, lipid nanoparticle-mediated vectors are being explored to deliver multiple nucleic acid forms, including pDNA, mRNA, siRNA, and proteins. Herein, we discuss the broader applications of lipid nanoparticles, from protein replacement therapy to restoring the disease mechanism through nucleic acid delivery and gene editing, as well as multiple preclinical and clinical studies as a potential alternative to liver transplantation.

Abstract Goals of high efficiency, morphological analysis, and the ability to produce organic solar cell (OSC) sub‐modules using halogen‐free solvents are demanding. In this study, a robust conjugated polymer with thienothiophene π‐spacer with pendant alkyl side chain (NapBDT‐C12) was synthesized and used to fabricate sub‐modules. Excellent efficiencies were demonstrated by a NapBDT‐C12 integrated ternary blend, which was used to produce stable small‐area‐to‐sub‐module devices using O‐xylene. The efficiency of the NapBDT‐C12 added small‐area ternary devices (PM6:NapBDT‐C12:L8‐BO) was 18.71%. Owing to the controlled homogeneity of the blend with favorable nanoscale film morphology, enhanced carrier mobilities, and exciton dissociation/splitting properties, contributed to the efficiencies of small‐area‐to‐sub‐module OSCs. Moreover, a 55 cm2 sub‐module with an efficiency of 14.69% was accomplished by bar coating using O‐xylene under ambient conditions. This study displays the potential of a ternary blend based OSC device to produce high efficiency scalable sub‐modules at ambient conditions.

RNA N6-methyladenosine (m6A) methylation is the most abundant and conserved RNA modification in eukaryotes. It participates in the regulation of RNA metabolism and various pathophysiological processes. Non-coding RNAs (ncRNAs) are defined as small or long transcripts which do not encode proteins and display numerous biological regulatory functions. Similar to mRNAs, m6A deposition is observed in ncRNAs. Studying RNA m6A modifications on ncRNAs is of great importance specifically to deepen our understanding of their biological roles and clinical implications. In this review, we summarized the recent research findings regarding the mutual regulation between RNA m6A modification and ncRNAs (with a specific focus on microRNAs, long non-coding RNAs, and circular RNAs) and their functions. We also discussed the challenges of m6A-containing ncRNAs and RNA m6A as therapeutic targets in human diseases and their future perspective in translational roles.

Abstract The fabrication of environmentally benign, solvent‐processed, efficient, organic photovoltaic sub‐modules remains challenging due to the rapid aggregation of the current high performance non‐fullerene acceptors (NFAs). In this regard, design of new NFAs capable of achieving optimal aggregation in large‐area organic photovoltaic modules has not been realized. Here, an NFA named BTA‐HD‐Rh is synthesized with longer (hexyl‐decyl) side chains that exhibit good solubility and optimal aggregation. Interestingly, integrating a minute amount of new NFA (BTA‐HD‐Rh) into the PM6:L8‐BO system enables the improved solubility in halogen‐free solvents (o‐xylene:carbon disulfide (O‐XY:CS2)) with controlled aggregation is found. Then solar sub‐modules are fabricated at ambient condition (temperature at 25 ± 3 °C and humidity: 30–45%). Ultimately, the champion 55 cm2 sub‐modules achieve exciting efficiency of >16% in O‐XY:CS2 solvents, which is the highest PCE reported for sub‐modules. Notably, the highest efficiency of BTA‐HD‐Rh doped PM6:L8‐BO is very well correlated with high miscibility with low Flory‐Huggins parameter (0.372), well‐defined nanoscale morphology, and high charge transport. This study demonstrates that a careful choice of side chain engineering for an NFA offers fascinating features that control the overall aggregation of active layer, which results in superior sub‐module performance with environmental‐friendly solvents.

Modern computational environments have been getting more and more complex and dynamic; hence, mechanisms of task scheduling are increasingly sophisticated in order to guarantee optimal resource usage and reduced latency. Traditional heuristic-based approaches, although very fast, often fail to meet the intricacies of the dependencies and real-time variability that exist in large-scale systems and hence result in far from optimal task completion timestamp and resource inefficiencies. Existing machine learning methods have improved but still remain unstable, lacking domain-specific adjustability for robust scheduling across a very broad workload spectrum. We provide an integrated scheduling framework that integrates Proximal Policy Optimization, Graph Neural Networks, hybrid rule-based and machine learning techniques, and synthetic data generation with Generative Adversarial Networks in this paper. PPO has been chosen to strike a balance between exploration and exploitation while considering efficient task scheduling in a dynamic environment. The GNNs are used to model complicated relations of tasks with resources, describing dependencies usually ignored by traditional models. This hybrid approach exploits domain knowledge via rule-based filtering and refines scheduling decisions with machine learning to bring flexibility and accuracy into the same platform. They are also applied to generate realistic synthetic workload traces. These augment the real data and, therefore, improve the robustness and generalization of the scheduling models. Our framework improves important performance metrics of the job completion timestamp by 20%-30%, resource utilization by 15%-25%, the latency of scheduling by 25%, and enhancing the accuracy of task prioritization by 30%. These results underline the potential of hybrid AI approaches to advance task scheduling mechanisms, offering much more adaptability, efficiency, and resiliency for computational systems today.

Background/Objectives: Hemophilia B is a hereditary bleeding disorder due to the production of liver malfunctional factor IX (FIX). Gene therapy with viral vectors offers a cure. However, applications are limited due to pre-existing antibodies, eligibility for children under 12 years of age, hepatotoxicity, and excessive costs. Lipid nanoparticles are a potential alternative owing to their biocompatibility, scalability, and non-immunogenicity. However, their therapeutic applications are still elusive due to the poor transfection efficiencies in delivering plasmid DNA into primary cells and target organs in vivo. To develop efficient liver-targeted lipid nanoparticles, we explored galactosylated lipids to target asialoglycoprotein receptors (ASGPRs) abundantly expressed on hepatocytes. Methods: We developed 12 novel liposomal formulations varying the galactose lipid Gal-LNC 5, cationic lipid MeOH16, DOPE, and cholesterol. We evaluated their physicochemical properties, toxicity profiles, and transfection efficiencies in hepatic cell lines. Among the formulations, Gal-LNC 5 could efficiently transfect the reporter plasmid eGFP in hepatic cell lines and specifically distribute into the liver in vivo. Toward developing functional factor IX, we cloned Padua mutant FIX-L in a CpG-free backbone to enhance the expression and duration. Results: We demonstrated superior expression of FIX with our galactosylated lipid nanoparticle system. Conclusions: The current research presents a specialized lipid nanoparticle system viz. Gal-LNC which is a specialized lipid nanoparticle system for liver-targeted gene therapy in hemophilia B patients that has potential for clinical use. The Gal-LNC successfully delivers a CpG-free Padua FIX gene to liver cells, producing therapeutically relevant levels of FIX protein. Among its benefits are the ideal qualities of stability, targeting the liver specifically, and maximizing efficiency of transfection. Optimization of liver-targeting lipid nanoparticle systems and function FIX plasmids will pave the way for novel lipid nanoparticle-based gene therapy products for hemophilia B and other monogenic liver disorders.

Antibodies are critical tools for research into extracellular vesicles (EVs) and other extracellular nanoparticles (ENPs), where they can be used for their identification, characterization, and isolation. However, the lack of a centralized antibody platform where researchers can share validation results thus minimizing wasted personnel time and reagents, has been a significant obstacle. Moreover, because the performance of antibodies varies among assay types and conditions, detailed information on assay variables and protocols is also of value. To facilitate sharing of results on antibodies that are relevant to EV/ENP research, the EV Antibody Database has been developed by the investigators of the Extracellular RNA Communication Consortium (ERCC). Hosted by the ExRNA Portal (https://exrna.org/resources/evabdb/), this interactive database aggregates and shares results from antibodies that have been tested by research groups in the EV/ENP field. Currently, the EV Antibody Database includes modules for antibodies tested for western Blot, EV Flow Cytometry, and EV Sandwich Assays, and holds 110 records contributed by 6 laboratories from the ERCC. Detailed information on antibody sources, assay conditions, and results is provided, including negative results. We encourage ongoing expert input and community feedback to enhance the database's utility, making it a valuable resource for comprehensive validation data on antibodies and protocols in EV biology.

β-thalassemia/HbE results from mutations in the β-globin locus that impede the production of functional adult hemoglobin. Base editors (BEs) could facilitate the correction of the point mutations with minimal or no indel creation, but its efficiency and bystander editing for the correction of β-thalassemia mutations in coding and non-coding regions remains unexplored. Here, we screened BE variants in HUDEP-2 cells for their ability to correct a spectrum of β-thalassemia mutations that were integrated into the genome as fragments of HBB. The identified targets were introduced into their endogenous genomic location using BEs and Cas9/homology-directed repair (HDR) to create cellular models with β-thalassemia/HbE. These β-thalassemia/HbE models were then used to assess the efficiency of correction in the native locus and functional β-globin restoration. Most bystander edits produced near target sites did not interfere with adult hemoglobin expression and are not predicted to be pathogenic. Further, the effectiveness of BE was validated for the correction of the pathogenic HbE variant in severe β0/βE-thalassaemia patient cells. Overall, our study establishes a novel platform to screen and select optimal BE tools for therapeutic genome editing by demonstrating the precise, efficient, and scarless correction of pathogenic point mutations spanning multiple regions of HBB including the promoter, intron, and exons.

The cellular response to stress is an important determinant of disease pathogenesis. Uncovering the molecular fingerprints of distinct stress responses may identify novel biomarkers and key signaling pathways for different diseases. Emerging evidence shows that transfer RNA-derived small RNAs (tDRs) play pivotal roles in stress responses. However, RNA modifications present on tDRs are barriers to accurately quantifying tDRs using traditional small RNA sequencing. Here, AlkB-facilitated methylation sequencing is used to generate a comprehensive landscape of cellular and extracellular tDR abundances in various cell types during different stress responses. Extracellular tDRs are found to have distinct fragmentation signatures from intracellular tDRs and these tDR signatures are better indicators of different stress responses than miRNAs. These distinct extracellular tDR fragmentation patterns and signatures are also observed in plasma from patients on cardiopulmonary bypass. It is additionally demonstrated that angiogenin and RNASE1 are themselves regulated by stressors and contribute to the stress-modulated abundance of sub-populations of cellular and extracellular tDRs. Finally, a sub-population of extracellular tDRs is identified for which AGO2 appears to be required for their expression. Together, these findings provide a detailed profile of stress-responsive tDRs and provide insight about tDR biogenesis and stability in response to cellular stressors.

Naturally occurring point mutations in the HBG promoter switch hemoglobin synthesis from defective adult beta-globin to fetal gamma-globin in sickle cell patients with hereditary persistence of fetal hemoglobin (HPFH) and ameliorate the clinical severity. Inspired by this natural phenomenon, we tiled the highly homologous HBG proximal promoters using adenine and cytosine base editors that avoid the generation of large deletions and identified novel regulatory regions including a cluster at the -123 region. Base editing at -123 and -124 bp of HBG promoter induced fetal hemoglobin (HbF) to a higher level than disruption of well-known BCL11A binding site in erythroblasts derived from human CD34+ hematopoietic stem and progenitor cells (HSPC). We further demonstrated in vitro that the introduction of -123T > C and -124T > C HPFH-like mutations drives gamma-globin expression by creating a de novo binding site for KLF1. Overall, our findings shed light on so far unknown regulatory elements within the HBG promoter and identified additional targets for therapeutic upregulation of fetal hemoglobin.

Extracellular vesicles (EVs) mediate intercellular signaling by transferring their cargo to recipient cells, but the functional consequences of signaling are not fully appreciated. RBC-derived EVs are abundant in circulation and have been implicated in regulating immune responses. Here, we use a transgenic mouse model for fluorescence-based mapping of RBC-EV recipient cells to assess the role of this intercellular signaling mechanism in heart disease. Using fluorescent-based mapping, we detected an increase in RBC-EV-targeted cardiomyocytes in a murine model of ischemic heart failure. Single cell nuclear RNA sequencing of the heart revealed a complex landscape of cardiac cells targeted by RBC-EVs, with enrichment of genes implicated in cell proliferation and stress signaling pathways compared with non-targeted cells. Correspondingly, cardiomyocytes targeted by RBC-EVs more frequently express cellular markers of DNA synthesis, suggesting the functional significance of EV-mediated signaling. In conclusion, our mouse model for mapping of EV-recipient cells reveals a complex cellular network of RBC-EV-mediated intercellular communication in ischemic heart failure and suggests a functional role for this mode of intercellular signaling.

Abstract Spike glycoprotein of SARS-CoV-2 variants plays a critical role in infection and transmission through its interaction with human angiotensin converting enzyme 2 (hACE2) receptors. Prior findings using molecular docking and biomolecular studies reported varied findings on the difference in the interactions among the spike variants with the hACE2 receptors. Hence, it is a prerequisite to understand these interactions in a more precise manner. To this end, firstly, we performed ELISA with trimeric spike glycoproteins of SARS-CoV-2 variants including Wuhan Hu-1(Wild), Delta, C.1.2 and Omicron. Further, to study the interactions in a more specific manner by mimicking the natural infection, we developed hACE2 receptors expressing HEK-293T cell line, evaluated their binding efficiencies and competitive binding of spike variants with D614G spike pseudotyped virus. In line with the existing findings, we observed that Omicron had higher binding efficiency compared to Delta in both ELISA and Cellular models. Intriguingly, we found that cellular models could differentiate the subtle differences between the closely related C.1.2 and Delta in their binding to hACE2 receptors. Our study using the cellular model provides a precise method to evaluate the binding interactions between spike sub-lineages to hACE2 receptors.

Abstract Exercise has well-recognized beneficial effects on the whole body. Previous studies suggest that exercise could promote tissue regeneration and repair in various organs. In this review, we have summarized the major effects of exercise on tissue regeneration primarily mediated by stem cells and progenitor cells in skeletal muscle, nervous system, and vascular system. The protective function of exercise-induced stem cell activation under pathological conditions and aging in different organs have also been discussed in detail. Moreover, we have described the primary molecular mechanisms involved in exercise-induced tissue regeneration, including the roles of growth factors, signaling pathways, oxidative stress, metabolic factors, and non-coding RNAs. We have also summarized therapeutic approaches that target crucial signaling pathways and molecules responsible for exercise-induced tissue regeneration, such as IGF1, PI3K, and microRNAs. Collectively, the comprehensive understanding of exercise-induced tissue regeneration will facilitate the discovery of novel drug targets and therapeutic strategies.

Extracellular vesicles (EVs) are membrane-bound nanoparticles with different types of cargo released by cells and postulated to mediate functions such as intercellular communications. Recent studies have shown that long non-coding RNAs (lncRNAs) or their fragments are present as cargo within EVs. LncRNAs are a heterogeneous group of RNA species with a length exceeding 200 nucleotides with diverse functions in cells based on their localization. While lncRNAs are known for their important functions in cellular regulation, their presence and role in EVs have only recently been explored. While certain studies have observed EV-lncRNAs to be tissue- and disease-specific, it remains to be determined whether or not this is a global observation. Nonetheless, these molecules have demonstrated promising potential to serve as new diagnostic and prognostic biomarkers. In this review, we critically evaluate the role of EV-derived lncRNAs in several prevalent diseases, including cancer, cardiovascular diseases, and neurodegenerative diseases, with a specific focus on their role as biomarkers.

Muscle atrophy is debilitating and can be induced by several stressors. Unfortunately, there are no effective pharmacological treatment until now. MicroRNA (miR)-29b is an important target that we identified to be commonly involved in multiple types of muscle atrophy. Although sequence-specific inhibition of miR-29b has been developed, in this study, we report a novel small-molecule miR-29b inhibitor that targets miR-29b hairpin precursor (pre-miR-29b) (Targapremir-29b-066 [TGP-29b-066]) considering both its three-dimensional structure and the thermodynamics of interaction between pre-miR-29b and the small molecule. This novel small-molecule inhibitor has been demonstrated to attenuate muscle atrophy induced by angiotensin II (Ang II), dexamethasone (Dex), and tumor necrosis factor α (TNF-α) in C2C12 myotubes, as evidenced by increase in the diameter of myotube and decrease in the expression of Atrogin-1 and MuRF-1. Moreover, it can also attenuate Ang II-induced muscle atrophy in mice, as evidenced by a similar increase in the diameter of myotube, reduced Atrogin-1 and MuRF-1 expression, AKT-FOXO3A-mTOR signaling activation, and decreased apoptosis and autophagy. In summary, we experimentally identified and demonstrated a novel small-molecule inhibitor of miR-29b that could act as a potential therapeutic agent for muscle atrophy.

Purpose: To assess the knowledge of nursing staff on ocular care in medical intensive care unit (ICU) and to compare the incidence of ocular surface disorders in patients of medical ICU pre- and post-training. Methods: Two hundred patients admitted in medical ICU for more than 24 h underwent a detailed ocular examination along with documentation of ICU stay, ventilation status, and Glasgow coma scale (GCS) scoring. An assessment on ocular care knowledge among the nursing staff of medical ICU was done. They were further given training in the form of audio-visuals and demonstration, and an eye care protocol was given to them. The second phase of the study was carried out in the same manner. A comparison was made between pre-training and post-training incidence of ocular surface disorders among ICU patients. Results: Ventilated patients had more eye discharge. Incidence of eye discharge was more in patients with duration of stay of more than 7 days in ICU. Ocular surface disorders closely correlated with the degree of lagophthalmos. There was significant reduction in ocular morbidity following ocular care training of the nursing staff. Conclusion: Eye care is a very important part of nursing care in sedated and ventilated patients in the ICU. Ophthalmic consultations are routinely needed in ICU subjects who are hospitalized for more than 1 week or if the ICU staff suspects any eye problems.

Sarcoidosis is an inflammatory granulomatous disease of unknown etiology involving any organ or tissue along with any combination of active sites, even the most silent ones clinically. The unpredictable nature of the sites involved in sarcoidosis dictates the highly variable natural history of the disease and the necessity to cluster cases at diagnosis based on clinical and/or imaging common characteristics in an attempt to classify patients based on their more homogeneous phenotypes, possibly with similar clinical behavior, prognosis, outcome, and therefore with therapeutic requirements. In the course of the disease's history, this attempt relates to the availability of a means of detection of the sites involved, from the Karl Wurm and Guy Scadding's chest x-ray staging through the ACCESS, the WASOG Sarcoidosis Organ Assessment Instruments, and the GenPhenReSa study to the 18F-FDG PET/CT scan phenotyping and far beyond to new technologies and/or the current “omics.” The hybrid molecular imaging of the 18F-FDG PET/CT scan, by unveiling the glucose metabolism of inflammatory cells, can identify high sensitivity inflammatory active granulomas, the hallmark of sarcoidosis—even in clinically and physiologically silent sites—and, as recently shown, is successful in identifying an unexpected ordered stratification into four phenotypes: (I) hilar–mediastinal nodal, (II) lungs and hilar–mediastinal nodal, (III) an extended nodal supraclavicular, thoracic, abdominal, inguinal, and (IV) all the above in addition to systemic organs and tissues, which is therefore the ideal phenotyping instrument. During the “omics era,” studies could provide significant, distinct, and exclusive insights into sarcoidosis phenotypes linking clinical, laboratory, imaging, and histologic characteristics with molecular signatures. In this context, the personalization of treatment for sarcoidosis patients might have reached its goal.

Abstract Muscle atrophy is a frequently observed complication, characterized by the loss of muscle mass and strength, which diminishes the quality of life and survival. No effective therapy except exercise is currently available. In our previous study, repressing miR-29b has been shown to reduce muscle atrophy. In our current study, we have constructed artificially engineered extracellular vesicles for the delivery of CRISPR/Cas9 to target miR-29b (EVs-Cas9-29b). EVs-Cas9-29b has shown a favorable functional effect with respect to miR-29b repression in a specific and rapid manner by gene editing. In in vitro conditions, EVs-Cas9-29b could protect against muscle atrophy induced by dexamethasone (Dex), angiotensin II (AngII), and tumor necrosis factor-alpha (TNF-α). And EVs-Cas9-29b introduced in vivo preserved muscle function in the well-established immobilization and denervation-induced muscle atrophy mice model. Our work demonstrates an engineered extracellular vesicles delivery of the miR-29b editing system, which could be potentially used for muscle atrophy therapy.

The b-chromatic number b(G) of a graph G is the maximum k for which G has a proper vertex coloring using k colors such that each color class contains at least one vertex adjacent to a vertex of every other color class. In this paper, we have mainly investigated on the b-chromatic number of the Mycielskian of regular graphs. In particular, we have obtained the exact value of the b-chromatic number of the Mycielskian of some classes of graphs. This includes a few families of regular graphs, graphs with b(G) = 2 and split graphs. In addition, we have found bounds for the b-chromatic number of the Mycielskian of some more families of regular graphs in terms of the bchromatic number of their original graphs. Also we have found b-chromatic number of the generalized Mycielskian of some regular graphs.

In order to accomplish machine visualization, object detection algorithms strive to recognise all target items in the image and derive the categories and object location information. Numerous ways to solve this problem have been presented where most of them are inspired by computer vision and deep learning methodologies. Existing techniques, on the other hand, consistently fail to recognise small, dense objects and even fail to detect objects with random geometric modifications. Object skeletons can help in object representation and detection. Skeleton is a natural object’s inherent visual description that contains comprehensive shape semantics. They supplement the object outline by providing additional information, such as how the object scale in thickness that varies between object components. However, extracting object skeletons from natural photos is difficult because the extractor must be able to capture both local and non-local visual contexts in order to calculate the scale of each skeleton pixel. To solve this issue, a Cusp Pixel Labelled Model with Precise Tuned Outline using Machine Learning (CPLM-PTOML) is proposed in this paper that accurately detects the cusp points of the object in the image by extracting the skeleton of the object to recognize the exact object resided in the image. A multi scale associated outputs to each stage of the image by monitoring the relationship between the receptive field sizes of the different layers in the image and the skeleton scales they can capture. The model is trained via multi-task learning, with one job being skeleton localization, which determines whether or not a pixel is a skeleton pixel, and the other being skeleton scale prediction, which predicts the scale of each skeleton pixel. At various phases, supervision is imposed by directing the scale-associated side outputs toward the ground-truth skeletons at the appropriate scales. The proposed model is compared with the traditional model in terms Data Training Samples, Object Feature Extraction Time Levels, Feature Extraction Accuracy, Cusp Pixel Labelling Time Levels, Cusp Pixel Identification Accuracy, Cusp Point Linking Accuracy, Image Considered and Cusp Point Recognition levels and the results exhibit that the proposed model exhibits better outcome.

In this work, we propose generic and novel adaptations to the binary Plaintext-Checking (PC) oracle based side-channel attacks for Kyber KEM. These attacks operate in a chosen-ciphertext setting, and are fairly generic and easy to mount on a given target, as the attacker requires very minimal information about the target device. However, these attacks have an inherent disadvantage of requiring a few thousand traces to perform full key recovery. This is due to the fact that these attacks typically work by recovering a single bit of information about the secret key per query/trace. In this respect, we propose novel parallel PC oracle based side-channel attacks, which are capable of recovering a generic P number of bits of information about the secret key in a single query/trace. We propose novel techniques to build chosen-ciphertexts so as to efficiently realize a parallel PC oracle for Kyber KEM. We also build a multi-class classifier, which is capable of realizing a practical side-channel based parallel PC oracle with very high success rate. We experimentally validated the proposed attacks (upto P = 10) on the fastest implementation of unprotected Kyber KEM in the pqm4 library. Our experiments yielded improvements in the range of 2.89× and 7.65× in the number of queries, compared to state-of-the-art binary PC oracle attacks, while arbitrarily higher improvements are possible for a motivated attacker, given the generic nature of the proposed attacks. We further conduct a thorough study on applicability to different scenarios, based on the presence/absence of a clone device, and also partial key recovery. Finally, we also show that the proposed attacks are able to achieve the lowest number of queries for key recovery, even for implementations protected with low-cost countermeasures such as shuffling. Our work therefore, concretely demonstrates the power of PC oracle attacks on Kyber KEM, thereby stressing the need for concrete countermeasures such as masking for Kyber and other lattice-based KEMs.