Effect of drill quality on biological damage in bone drilling.

Bone drilling is a universal procedure in orthopaedics for fracture fixation, installing implants, or reconstructive surgery. Surgical drills are subjected to wear caused by their repeated use, thermal fatigue, irrigation with saline solution, and sterilization process. Wear of the cutting edges of a drill bit (worn drill) is detrimental for bone tissues and can seriously affect its performance. The aim of this study is to move closer to minimally invasive surgical procedures in bones by investigating the effect of wear of surgical drill bits on their performance. The surface quality of the drill was found to influence the bone temperature, the axial force, the torque and the extent of biological damage around the drilling region. Worn drill produced heat above the threshold level related to thermal necrosis at a depth equal to the wall thickness of an adult human bone. Statistical analysis showed that a sharp drill bit, in combination with a medium drilling speed and drilling at shallow depth, was favourable for safe drilling in bone. This study also suggests the further research on establishing a relationship between surface integrity of a surgical drill bit and irreversible damage that it can induce in delicate tissues of bone using different drill sizes as well as drilling parameters and conditions.

Targeting Unc51-like Autophagy Activating Kinase 1 (ULK1) Overcomes Adaptive Drug Resistance in Acute Myelogenous Leukemia.

Despite effective new therapies, adaptive resistance remains the main obstacle in acute myelogenous leukemia (AML) therapy. Autophagy induction is a key mechanism for adaptive resistance. Leukemic blasts at diagnosis express higher levels of the apical autophagy kinase ULK1 compared with normal hematopoietic cells. Exposure to chemotherapy and targeted agents upregulate ULK1, hence we hypothesize that developing ULK1 inhibitors may present the unique opportunity for clinical translation of autophagy inhibition. Accordingly, we demonstrate that ULK1 inhibition, by genetic and pharmacologic means, suppresses treatment-induced autophagy, overcomes adaptive drug-resistance, and synergizes with chemotherapy and emerging antileukemia agents like venetoclax (ABT-199). The study next aims at exploring the underlying mechanisms. Mechanistically, ULK1 inhibition downregulates MCL1 antiapoptotic gene, impairs mitochondrial function and downregulates components of the CD44-xCT system, resulting in impaired reactive oxygen species (ROS) mitigation, DNA damage, and apoptosis. For further validation, several mouse models of AML were generated. In these mouse models, ULK1 deficiency impaired leukemic cell homing and engraftment, delayed disease progression, and improved survival. Therefore, in the study, we validated our hypothesis and identified ULK1 as an important mediator of adaptive resistance to therapy and an ideal candidate for combination therapy in AML. Therefore, we propose ULK1 inhibition as a therapeutically relevant treatment option to overcome adaptive drug-resistance in AML. IMPLICATIONS: ULK1 drives a cell-intrinsic adaptive resistance in AML and targeting ULK1-mediated autophagy can synergize with existing and emerging AML therapies to overcome drug-resistance and induce apoptosis.

Concomitant targeting of FLT3 and BTK overcomes FLT3 inhibitor resistance in acute myeloid leukemia through the inhibition of autophagy.

Strategies to overcome resistance to FMS-like tyrosine kinase 3 (FLT3)-targeted therapy in acute myeloid leukemia (AML) are urgently needed. We identified autophagy as one of the resistance mechanisms, induced by hypoxia and the bone marrow microenvironment via activation of Bruton tyrosine kinase (BTK). Suppressing autophagy/BTK sensitized FLT3- mutated AML to FLT3 inhibitor-induced apoptosis. Furthermore, co-targeting FLT3/BTK/aurora kinases with a novel multikinase inhibitor CG-806 (luxeptinib) induced profound apoptosis in FLT3-mutated AML by co-suppressing FLT3/BTK, antagonizing autophagy, and causing leukemia cell death in FLT3-wildtype AML by aurora kinase-mediated G2/M arrest and polyploidy, in addition to FLT3 inhibition. Thus, CG-806 exerted profound anti-leukemia activity against AML regardless of FLT3 mutation status. CG-806 also significantly reduced AML burden and extended survival in an in vivo patient-derived xenograft leukemia murine model of FLT3 inhibitor-resistant FLT3-ITD/TKD double-mutant primary AML. Taken together, these findings indicate that CG-806 has a unique mechanistic action and pre-clinical activity, which is presently undergoing clinical evaluation in both FLT3 wildtype and mutant AML.

Inhibition of mitochondrial complex I reverses NOTCH1-driven metabolic reprogramming in T-cell acute lymphoblastic leukemia.

T-cell acute lymphoblastic leukemia (T-ALL) is commonly driven by activating mutations in NOTCH1 that facilitate glutamine oxidation. Here we identify oxidative phosphorylation (OxPhos) as a critical pathway for leukemia cell survival and demonstrate a direct relationship between NOTCH1, elevated OxPhos gene expression, and acquired chemoresistance in pre-leukemic and leukemic models. Disrupting OxPhos with IACS-010759, an inhibitor of mitochondrial complex I, causes potent growth inhibition through induction of metabolic shut-down and redox imbalance in NOTCH1-mutated and less so in NOTCH1-wt T-ALL cells. Mechanistically, inhibition of OxPhos induces a metabolic reprogramming into glutaminolysis. We show that pharmacological blockade of OxPhos combined with inducible knock-down of glutaminase, the key glutamine enzyme, confers synthetic lethality in mice harboring NOTCH1-mutated T-ALL. We leverage on this synthetic lethal interaction to demonstrate that IACS-010759 in combination with chemotherapy containing L-asparaginase, an enzyme that uncovers the glutamine dependency of leukemic cells, causes reduced glutaminolysis and profound tumor reduction in pre-clinical models of human T-ALL. In summary, this metabolic dependency of T-ALL on OxPhos provides a rational therapeutic target.

Analysis of supply chain resilience drivers in oil and gas industries during the COVID-19 pandemic using an integrated approach.

The COVID-19 pandemic has significantly affected the supply chains (SCs) of many industries, including the oil and gas (O&G) industry. This study aims to identify and analyze the drivers that affect the resilience level of the O&G SC under the COVID-19 pandemic. The analysis helps to understand the driving intensity of one driver over those of others as well as drivers with the highest driving power to achieve resilience. Through an extensive literature review and an overview of experts' opinions, the study identified fourteen supply chain resilience (SCR) drivers of the O&G industry. These drivers were analyzed using the integrated fuzzy interpretive structural modeling (ISM) and decision-making trial and evaluation laboratory (DEMATEL) approaches. The analysis shows that the major drivers of SCR are government support and security. These two drivers help to achieve other drivers of SCR, such as collaboration and information sharing, which, in turn, influence innovation, trust, and visibility among SC partners. Two more drivers, robustness and agility, are also essential drivers of SCR. However, rather than influencing other drivers for their achievement, robustness and agility are influenced by others. The results show that collaboration has the highest overall driving intensity and agility has the highest intensity of being influenced by other drivers.

Targeting the NOTCH1-MYC-CD44 axis in leukemia-initiating cells in T-ALL.

The NOTCH1-MYC-CD44 axis integrates cell-intrinsic and extrinsic signaling to ensure the persistence of leukemia-initiating cells (LICs) in T-cell acute lymphoblastic leukemia (T-ALL) but a common pathway to target this circuit is poorly defined. Bromodomain-containing protein 4 (BRD4) is implicated to have a role in the transcriptional regulation of oncogenes MYC and targets downstream of NOTCH1, and here we demonstrate its role in transcriptional regulation of CD44. Hence, targeting BRD4 will dismantle the NOTCH1-MYC-CD44 axis. As a proof of concept, degrading BRD4 with proteolysis targeting chimera (PROTAC) ARV-825, prolonged the survival of mice in Notch1 mutated patient-derived xenograft (PDX) and genetic models (ΔPTEN) of T-ALL. Single-cell proteomics analysis from the PDX model, demonstrated quantitative reduction of LICs (CD34+ CD7+ CD19-) and downregulation of the NOTCH1-MYC-CD44 axis, along with cell cycle, apoptosis and PI3K/Akt pathways. Moreover, secondary transplantation from PDX and ΔPTEN models of T-ALL, confirmed delayed leukemia development and extended survival of mice engrafted with T-ALL from ARV-825 treated mice, providing functional confirmation of depletion of LICs. Hence, BRD4 degradation is a promising LIC-targeting therapy for T-ALL.

Pan-RAF Inhibition Shows Anti-Leukemic Activity in RAS-Mutant Acute Myeloid Leukemia Cells and Potentiates the Effect of Sorafenib in Cells with FLT3 Mutation.

RAF molecules play a critical role in cell signaling through their integral impact on the RAS/RAF/MEK/ERK signaling pathway, which is constitutively activated in a sizeable subset of acute myeloid leukemia (AML) patients. We evaluated the impact of pan-RAF inhibition using LY3009120 in AML cells harboring mutations upstream and downstream of RAF. LY3009120 had anti-proliferative and pro-apoptotic effects and suppressed pERK1/2 levels in leukemic cells with RAS and FLT3 mutations. Using reverse protein phase array analysis, we identified reductions in the expression/activation of cell signaling components downstream of RAF (activated p38) and cell cycle regulators (Wee1/cyclin B1, Cdc2/Cdk1, activated Rb, etc.). Notably, LY3009120 potentiated the effect of Ara-C on AML cells and overcame bone marrow mesenchymal stromal cell-mediated chemoresistance, with RAS-mutated cells showing a notable reduction in pAKT (Ser473). Furthermore, the combination of LY3009120 and sorafenib resulted in significantly higher levels of apoptosis in AML cells with heterozygous and hemizygous FLT3 mutations. In conclusion, pan-RAF inhibition in AML using LY3009120 results in anti-leukemic activity, and combination with Ara-C or sorafenib potentiates its effect.

Experimental study on the mechanical properties of biological hydrogels of different concentrations.

BACKGROUND: Biological hydrogels provide a conducive three-dimensional extracellular matrix environment for encapsulating and cultivating living cells. Microenvironmental modulus of hydrogels dictates several characteristics of cell functions such as proliferation, adhesion, self-renewal, differentiation, migration, cell morphology and fate. Precise measurement of the mechanical properties of gels is necessary for investigating cellular mechanobiology in a variety of applications in tissue engineering. Elastic properties of gels are strongly influenced by the amount of crosslinking density. OBJECTIVE: The main purpose of the present study was to determine the elastic modulus of two types of well-known biological hydrogels: Agarose and Gelatin Methacryloyl. METHODS: Mechanical properties such as Young's modulus, fracture stress and failure strain of the prescribed gels with a wide range of concentrations were determined using tension and compression tests. RESULTS: The elastic modulus, failure stress and strain were found to be strongly influenced when the amount of concentration in the hydrogels was changed. The elastic modulus for a lower level of concentration, not considered in this study, was also predicted using statistical analysis. CONCLUSIONS: Closed matching of the mechanical properties of the gels revealed that the bulk tension and compression tests could be confidently used for assessing mechanical properties of delicate biological hydrogels.

Experimental investigation on the effect of drill quality on the performance of bone drilling.

Bone drilling is a well-known process in operative fracture treatment and reconstructive surgery. The cutting ability of the drill is lost when used for multiple times. In this study, the effect of different levels of drill wear on bone temperature, drilling force, torque, delamination around the drilling region and surface roughness of the hole was investigated using a series of experiments. Experimental results demonstrated that the wear of the drill is strongly related to the drilling force, torque, temperature and surface roughness of the drilled hole. Statistical analysis was performed to find the effect of various factors on multiple response variables in the bone drilling process. The favorable conditions for bone drilling are obtained when feed rate, drill speed and the roughness of the cutting edge of the drill were fixed at 30 mm, 2000 rpm and up to 2 mm, respectively. Further, analysis of variance (ANOVA) was performed to determine the factor with a significant impact on the response variables. F-test and p-value indicated that the feed rate had the highest effect on grey relational grade followed by the roughness of the drill. This study suggests that the sharp drill along with controlled drilling speed and feed rate may be used for safe and efficient surgical drilling in bone.

In-vitro experimental study of histopathology of bone in vibrational drilling.

Drilling is a common surgical procedure for fracture treatment and reconstruction in multiple surgical fields, including orthopaedics, neurology, and dentistry. Drilling delicate tissue (such as bone) with a hard metallic tool is considered notorious for inducing mechanical and thermal damage, which can adversely affect osseointegration and may weaken the bond between the bone and implant, or other fixative devices anchoring the bone. The aim of this study is to explore the benefits of vibrational drilling (VD) in overcoming the complications associated with conventional drilling (CD). Drilling tests were performed on fresh cortical bone with the intention of investigating the effect of a range of frequencies, in combination with drilling speed and feed rate, on biological damage around the drilling region using histological sections of skeletally mature bone. The study examined the most influential factors and optimal combination of parameters for safe and efficient drilling in bone. Results from Taguchi grey relational analysis showed that a lower drilling speed and feed rate combined with a frequency of 20 kHz were favourable parameters for safe drilling in bone. Accordingly, VD using controlled parameters may be an alternative to CD in bone surgical procedures.

BETP degradation simultaneously targets acute myelogenous leukemia stem cells and the microenvironment.

Anti-leukemic effect of BET/BRD4 (BETP) protein inhibition has been largely attributed to transcriptional downregulation of cellular anabolic/anti-apoptotic processes but its effect on bone marrow microenvironment, a sanctuary favoring persistence of leukemia stem/progenitor cells, is unexplored. Sustained degradation of BETP with small-molecule BET proteolysis-targeting chimera (PROTAC), ARV-825, resulted in marked downregulation of surface CXCR4 and CD44, key proteins in leukemia-microenvironment interaction, in AML cells. Abrogation of surface CXCR4 expression impaired SDF-1α directed migration and was mediated through transcriptional down-regulation of PIM1 kinase that in turn phosphorylates CXCR4 and facilitates its surface localization. Down-regulation of CD44/CD44v8-10 impaired cystine uptake, lowered intracellular reduced glutathione and increased oxidative stress. More importantly, BETP degradation markedly decreased CD34+CD38-CD90-CD45RA+ leukemic stem cell population and alone or in combination with Cytarabine, prolonged survival in mouse model of human leukemia including AML-PDX. Gene expression profiling and single cell proteomics confirmed down regulation of the gene signatures associated with 'stemness' in AML and Wnt/ß-catenin, Myc pathways. Hence, BETP degradation by ARV-825 simultaneously targets cell intrinsic signaling, stromal interactions and metabolism in AML.

Bromodomain inhibitors: what does the future hold?

Cancer cells are addicted to mutations that cause gain of function in oncogenes and loss of function in tumor suppressors, so that these cells are reliant on aberrant signaling pathways and transcription. Protein-protein and DNA-protein interactions that cause chromatin remodeling are another source of the deregulation of critical signaling and transcriptional regulators, altering epigenetic signatures and creating additional vulnerabilities. Owing to mutations in multiple epigenetic regulators in hematologic malignancies, cancer cells are highly addicted to altered transcription. These vulnerabilities have been targeted by several epigenetic drugs, including hypomethylating agents, but the idea of targeting bromodomain proteins has emerged relatively recently. Because bromodomain proteins recognize acetylated lysine on histones and recruit transcription complexes on the chromatin, targeting these proteins may serve as a strategy to target transcription, irrespective of the presence of epigenetic mutations. Here, we review the existing literature to explain the rationale of using bromodomain inhibitors in hematologic malignancies. We discuss the evolution of bromodomain inhibitors, with an in-depth evaluation of bromodomain and extraterminal domain (BET) proteins, the most prominent bromodomain family, and also highlight the prospect of targeting non-BET proteins. In the later sections, we comment on the combinatorial targeting of BET proteins to overcome the effects of multiple signaling pathways. Finally, we emphasize the newer concepts, such as dual-kinase inhibition and selective bromodomain targeting, and technologies, such as protein degradation, that are expected to influence the future generation of bromodomain inhibitors.

The Effect of Lipopolysaccharide on Noxa Expression Is Mediated through IRF1, 3, and 7.

Lipopolysaccharide (LPS), a component of the cell wall of gram-negative bacteria, elicits the secretion of cytokines, such as interferons, that stimulate the host defense system. Previously, we demonstrated that interferons induce interferon regulatory factors (IRFs) 1, 3, and 7, which regulate the transcription of Noxa and alter the expression profiles of Bcl-2 family proteins in tumors. However, the immediate consequences of LPS stimulation on Noxa and BH3 expression in tumor cells remain uncharacterized. In this study, we determined that LPS induced Noxa expression in CT26 cells. Furthermore, studies in HCT116 parental and HCT116 p53-deficient cells revealed that LPS-mediated Noxa was independent of p53. Meanwhile, IRF1, 3, and 7 in CT26, HCT116 parental, and HT116 p53-deficient cells were upregulated by LPS stimulation, suggesting that LPS induces the expression of these IRFs in a p53-independent manner. The responsiveness of IRF1, 3, 4, and 7 binding to the Noxa promoter region to LPS indicated that IRF1, 3, and 7 activated Noxa expression, whereas IRF4 repressed Noxa expression. Together, these results suggest that LPS directly affects Noxa expression in tumor cells through IRFs, implicating that it may contribute to LPS-induced tumor regression.

Targeting autophagy to overcome chemoresistance in acute myleogenous leukemia.

Therapeutic inhibition of macroautophagy/autophagy is expected to increase chemosensitivity of cancers and alter tumor-stroma interdependence. The hypoxic, metabolically challenged bone marrow microenvironment confers chemoresistance to leukemia cells. The impact of autophagy inhibition in the context of microenvironment-mediated resistance in leukemia is less explored. Our recent studies demonstrated that co-culture of acute myelogenous leukemia (AML) cells with marrow-derived mesenchymal stromal cells (MSC) induces autophagy in AML cells and increases resistance to genotoxic agents (cytarabine and idarubicin). Genetic silencing of ATG7 in AML enhances the sensitivity to these genotoxic agents, an effect that was more pronounced with concomitant silencing of ATG7 in AML and MSCs. Mechanistically, the increased sensitivity of AML cells to genotoxic agents is associated with alteration of BCL2 family proteins, particularly transcriptional upregulation of PMAIP1/NOXA. In a disseminated AML model in immunocompromised mice, ATG7 knockdown in AML cells results in better survival compared to control mice when treated with chemotherapy. Our studies support the therapeutic role of autophagy inhibition, specifically ATG7 inhibition, in AML.

Atg7 suppression enhances chemotherapeutic agent sensitivity and overcomes stroma-mediated chemoresistance in acute myeloid leukemia.

Autophagy is a cellular adaptive mechanism to stress, including that induced by chemotherapeutic agents. Reverse phase protein array suggested that high expression of the essential autophagy-related protein, Atg7, was associated with shorter remission in newly diagnosed acute myeloid leukemia (AML) patient samples, indicating a role in chemoresistance. Knockdown of Atg7 in AML cells using short hairpin RNA markedly increased apoptosis and DNA damage following treatment with cytarabine and idarubicin. Interestingly, coculture of AML cells with stromal cells increased autophagy and chemoresistance in the AML cells exposed to chemotherapeutic agents, and this was reversed following Atg7 knockdown. This effect was further enhanced by concomitant knockdown of Atg7 in both AML and stromal cells. These findings strongly suggest that Atg7, and likely microenvironment autophagy in general, plays an important role in AML chemoresistance. Mechanistic studies revealed that Atg7 knockdown induced a proapoptotic phenotype in AML cells, which was manifested by an increased NOXA expression at the transcriptional level. Finally, in a mouse model of human leukemia, Atg7 knockdown extended overall survival after chemotherapy. Thus, the inhibition of Atg7 appears to be a valid strategy to enhance chemosensitivity, and it could indeed improve outcomes in AML therapy.

MDM2 Inhibitor, Nutlin 3a, Induces p53 Dependent Autophagy in Acute Leukemia by AMP Kinase Activation.

MDM2 (mouse double minute 2) inhibitors that activate p53 and induce apoptosis in a non-genotoxic manner are in clinical development for treatment of leukemias. P53 can modulate other programmed cell death pathways including autophagy both transcriptionally and non-transcriptionally. We investigated autophagy induction in acute leukemia by Nutlin 3a, a first-in-class MDM2 inhibitor. Nutlin 3a induced autophagy in a p53 dependent manner and transcriptional activation of AMP kinase (AMPK) is critical, as this effect is abrogated in AMPK -/- mouse embryonic fibroblasts. Nutlin 3a induced autophagy appears to be pro-apoptotic as pharmacological (bafilomycin) or genetic inhibition (BECLIN1 knockdown) of autophagy impairs apoptosis induced by Nutlin 3a.

DUSP6 is a novel transcriptional target of p53 and regulates p53-mediated apoptosis by modulating expression levels of Bcl-2 family proteins.

p53 regulates various cellular responses through transcriptional regulation of distinct sets of target genes. Dual specificity phosphatase 6 (DUSP6) is a cytosolic phosphatase that inactivates the extracellular-signal-regulated kinase 1/2 (ERK1/2). This study demonstrates that p53 transactivates DUSP6 in human colorectal HCT116 cells to regulate ERK1/2 in p53-mediated cell death. DUSP6 is transactivated by p53 overexpression and genotoxic agents, and chromatin immunoprecipitation revealed two p53-binding sites in the DUSP6 promoter responsible for DUSP6 induction. Expression of shDUSP6 inhibited 5'-FU-induced cell death, whereas overexpression of DUSP6 increased susceptibility to 5'-FU. 5'-FU treatment dephosphorylated ERK in a DUSP6-dependent manner, resulting in destabilization of Bcl-2 and stabilization of Bad. These results provide insights on the modulatory role of p53 in the survival pathway by up-regulating DUSP6.

Autophagy regulation in cancer development and therapy.

Autophagy is a cellular process to degrade long-lived or malfunctioning proteins and obsolete or damaged organelles. It maintains cellular homeostasis and helps cells survive stressful conditions. Tumor suppressors mostly positively regulate autophagy, whereas oncogene products usually inhibit autophagy. Alterations in key autophagy genes have also been shown to affect cancer development. However, the role of autophagy in cancer depends on the status of the cells and can either suppress or promote tumor growth. In the present review, we report on the current state of knowledge about the reciprocal regulation of autophagy and the potential role of autophagy played in cancer development and therapy.

Suppression of IRF4 by IRF1, 3, and 7 in Noxa expression is a necessary event for IFN-γ-mediated tumor elimination.

IFN-γ plays a critical role in tumor immunosurveillance by affecting either immune cells or tumor cells; however, IFN-mediated effects on tumor elimination are largely unknown. In this study, we showed that IFN regulatory factors (IRF) modulated by IFNs up- and downregulated Noxa expression, a prodeath BH3 protein, in various cancer cells. Inhibition of Noxa expression using short hairpin RNA in tumor cells leads to resistance against lipopolysaccharide (LPS)-induced tumor elimination, in which IFN-γ is known as a critical effecter in mice. Chromatin immunoprecipitation analysis in both CT26 cells and SP2/0 cells, sensitive and resistant to LPS-induced tumor elimination, respectively, revealed that the responsiveness of IRF1, 3, 4, and 7 in the Noxa promoter region in response to IFN-γ might be crucial in LPS-induced tumor elimination. IRF1, 3, and 7 were upregulated by IFN-γ and activated Noxa expression, leading to the death of Noxa wild-type baby mouse kidney (BMK) cells but not of Noxa-deficient BMK cells. In contrast, IRF4 acts as a repressor for Noxa expression and inhibits cell death induced by IRF1, 3, or 7. Therefore, although IFN-γ alone are not able to induce cell death in tumor cells in vitro, Noxa induction by IFN-γ, which is regulated by the balance between its activators (IRF1, 3, and 7) and its repressor (IRF4), is crucial to increasing the susceptibility of tumor cells to immune cell-mediated cytotoxicity.

The E1B19K oncoprotein complexes with Beclin 1 to regulate autophagy in adenovirus-infected cells.

The mechanisms underlying adenovirus-mediated autophagy are currently unknown. Recently, members of the Bcl-2 protein family have been associated with autophagy. It was also reported that the Bcl-2 homology-3 (BH3) domain encompassed by both Beclin 1 and Bcl-2-like proteins is essential for their pro-autophagy or anti-autophagy functions. Here, we report for the first time that E1B19K, the adenovirus BH3 domain protein, interacts with Beclin 1 to initiate autophagy. Using immunoprecipitation assays we showed that expression of E1B19K in the host cell disrupted the physical interactions between Beclin 1 and Bcl-2 proteins. The displacement of Bcl-2 was coincident with the recruitment of PI3KC3 to the Beclin 1/E1B19K complexes. As a result of the changes in the components of the Beclin 1 interactome, there was activation of PI3KC3, as showed by the identification of PI3K-mediated lipid phosphorylation, and subsequent formation of autophagosomes. Importantly, the BH3 functional domain of E1B19K protein was required for the heterodimerization with Beclin 1. We also showed that transfer of E1B19K was sufficient to trigger autophagy in cancer cells. Consistent with these data, mutant adenoviruses encompassing a deletion of the E1B19K gene produced a marked deficiency in the capability of the virus to induce autophagy as showed by examining the lipidation and cleavage of LC3-I as well as the subcellular localization of LC3-II, the decrease in the levels of p62, and the formation of autophagosomes. Our work offers new information on the mechanisms of action of the adenoviral E1B19K protein as partner of Beclin 1 and positive regulator of autophagy.