Dental Metal Matrix Composites: The Effects of the Addition of Titanium Nanoparticle Particles on Dental Amalgam.

Dental amalgams have been used by dentists for the restoration of posterior human teeth. However, there have been concerns about the release of mercury from amalgams into the oral cavity. The objective of the present research is to study the effect of titanium (Ti) nanoparticles on the microstructural mechanism of the release of mercury vapor in two commonly used brands of dental amalgam (the Dispersalloy: 11.8% Cu; the Sybralloy: 33% Cu). Ti powder was added to both the Dispersalloy and the Sybralloy in increments of 10 mg up to 80 mg. The addition of Ti powder to both brands of dental amalgam has been found to result in a considerable decrease in Hg vapor release. The decrease in the Hg vapor release due to Ti addition has been explained by the formation of strong Hg-Ti covalent bonds, which reduce the availability of Hg atoms for evaporation. The Ti atoms in excess of the solubility limit of Ti in Hg reside in the grain boundaries, which also reduces the evaporation of Hg from the amalgam. The binding of Hg with Ti via a strong covalent bond also results in a significant improvement in mechanical properties such as Vickers hardness.

DNMT3B PWWP mutations cause hypermethylation of heterochromatin.

The correct establishment of DNA methylation patterns is vital for mammalian development and is achieved by the de novo DNA methyltransferases DNMT3A and DNMT3B. DNMT3B localises to H3K36me3 at actively transcribing gene bodies via its PWWP domain. It also functions at heterochromatin through an unknown recruitment mechanism. Here, we find that knockout of DNMT3B causes loss of methylation predominantly at H3K9me3-marked heterochromatin and that DNMT3B PWWP domain mutations or deletion result in striking increases of methylation in H3K9me3-marked heterochromatin. Removal of the N-terminal region of DNMT3B affects its ability to methylate H3K9me3-marked regions. This region of DNMT3B directly interacts with HP1α and facilitates the bridging of DNMT3B with H3K9me3-marked nucleosomes in vitro. Our results suggest that DNMT3B is recruited to H3K9me3-marked heterochromatin in a PWWP-independent manner that is facilitated by the protein's N-terminal region through an interaction with a key heterochromatin protein. More generally, we suggest that DNMT3B plays a role in DNA methylation homeostasis at heterochromatin, a process which is disrupted in cancer, aging and Immunodeficiency, Centromeric Instability and Facial Anomalies (ICF) syndrome.

Strain imaging as an early predictor in acute myocardial infarction - An augmented cross-sectional study.

INTRODUCTION: Cardiac fibres are affected invariably in myocardial infarction, with longitudinal strain being the earliest to be detected in the ischaemic cascade. The present study aims to assess strain imaging in acute myocardial infarction (AMI) patients admitted to the cardiology department at our institute and correlate GLS and DESL findings with other markers for myocardial function. METHODS AND MATERIAL: This augmented cross-sectional study was conducted amongst the patients admitted with diagnosis of AMI. During the study period, 157 subjects were sampled through convenience sampling, and examined as well as tested with routine investigations at baseline. The subjects were then followed through at first, third and six months, and findings noted. Chi-square was used to assess the crude association between sample characteristics. Pearson correlation and student t-test were used to find association between continuous variables. RESULTS: After screening 564 patients, 157 patients were included in the study after fulfilment of inclusion and exclusion criteria. A significant difference was found in baseline GLS scores and NTproBNP levels at 6 months in alive patients with STEMI, t (21.728) = -5.717, p < .001. Out of the 50 NSTEMI patients, 35 (70 %) were positive for ESL, similarly out of 43 STEMI patients without any RWMA, ESL was positive in 39 (90.02 %) patients. CONCLUSIONS: GLS by STE has good correlation with LVEF, WMSI and NT pro-BNP and it is an independent predictor of mortality and heart failure among patients with AMI.

Echocardiography for Volume Assessment in Acute Myocardial Infarction.

Background Errors caused by improper volume estimation increase acute mortality rates in acute myocardial infarction (AMI). We aimed to determine volume status in AMI patients using echocardiography and to correlate the findings with clinical outcomes. Methods This cross-sectional, single-center study was performed at a tertiary care center in India between August 2017 and September 2020 involving AMI patients. We performed echocardiography for all patients. Parameters such as left ventricle (LV) and atrium size, LV end-diastolic pressure, inferior vena cava (IVC) size and size variation, velocity stroke volume, and velocity time integral variation were measured. B-lines were recorded by scanning 32 regions on the anterior chest in the supine position using cardiac probes of echocardiography. Results A total of 184 patients were enrolled in the study with male predominance (82.1%). The mean age of patients was 58.2 ± 10.7 years. Dilated (>2.1 cm) and collapsible (<50%) IVC, and B-lines were significantly associated with heart failure (HF) (p<0.001; r=0.87 and p<0.001; r=0.74, respectively). The area under receiver operating characteristics (AUROC) curve to diagnose HF at a cut-off value of >10 for B-lines was 0.897 (0.842-0.951). AUROC curve for IVC size in diagnosing hypovolemia was 0.063 (0.000-0.130). Conclusions Volume status based on IVC size and B-lines detected by echocardiography has a strong prognostic value in AMI patients and should be included in the routine assessment of these patients.

BRCA1-BARD1 combines multiple chromatin recognition modules to bridge nascent nucleosomes.

Chromatin association of the BRCA1-BARD1 heterodimer is critical to promote homologous recombination repair of DNA double-strand breaks (DSBs) in S/G2. How the BRCA1-BARD1 complex interacts with chromatin that contains both damage induced histone H2A ubiquitin and inhibitory H4K20 methylation is not fully understood. We characterised BRCA1-BARD1 binding and enzymatic activity to an array of mono- and di-nucleosome substrates using biochemical, structural and single molecule imaging approaches. We found that the BRCA1-BARD1 complex preferentially interacts and modifies di-nucleosomes over mono-nucleosomes, allowing integration of H2A Lys-15 ubiquitylation signals with other chromatin modifications and features. Using high speed- atomic force microscopy (HS-AFM) to monitor how the BRCA1-BARD1 complex recognises chromatin in real time, we saw a highly dynamic complex that bridges two nucleosomes and associates with the DNA linker region. Bridging is aided by multivalent cross-nucleosome interactions that enhance BRCA1-BARD1 E3 ubiquitin ligase catalytic activity. Multivalent interactions across nucleosomes explain how BRCA1-BARD1 can recognise chromatin that retains partial di-methylation at H4 Lys-20 (H4K20me2), a parental histone mark that blocks BRCA1-BARD1 interaction with nucleosomes, to promote its enzymatic and DNA repair activities.

Myval: A Novel Transcatheter Heart Valve for the Treatment of Severe Aortic Stenosis.

Transcatheter aortic valve implantation (TAVI) performed using femoral arterial access is now a guideline recommended treatment for severe calcific aortic stenosis (AS) in elderly patients. Technological advancements and procedural refinements have focused on making TAVI simpler, safer, more effective and durable. Myval (Meril Lifesciences) is a new generation balloon-expandable transcatheter heart valve (THV) developed in India that possesses novel features to improve deliverability and aid precise deployment. Following the first-in-human study, Myval was approved in India for commercial implantation in October 2018 and was subsequently given a CE mark in April 2019. This article reviews the science, technology and up-to-date clinical evidence for the Myval THV.

Spatially Resolved Metabolites in Stable and Unstable Human Atherosclerotic Plaques Identified by Mass Spectrometry Imaging.

BACKGROUND: Impairments in carbohydrate, lipid, and amino acid metabolism drive features of plaque instability. However, where these impairments occur within the atheroma remains largely unknown. Therefore, we sought to characterize the spatial distribution of metabolites within stable and unstable atherosclerosis in both the fibrous cap and necrotic core. METHODS: Atherosclerotic tissue specimens from 9 unmatched individuals were scored based on the Stary classification scale and subdivided into stable and unstable atheromas. After performing mass spectrometry imaging on these samples, we identified over 850 metabolite-related peaks. Using MetaboScape, METASPACE, and Human Metabolome Database, we confidently annotated 170 of these metabolites and found over 60 of these were different between stable and unstable atheromas. We then integrated these results with an RNA-sequencing data set comparing stable and unstable human atherosclerosis. RESULTS: Upon integrating our mass spectrometry imaging results with the RNA-sequencing data set, we discovered that pathways related to lipid metabolism and long-chain fatty acids were enriched in stable plaques, whereas reactive oxygen species, aromatic amino acid, and tryptophan metabolism were increased in unstable plaques. In addition, acylcarnitines and acylglycines were increased in stable plaques whereas tryptophan metabolites were enriched in unstable plaques. Evaluating spatial differences in stable plaques revealed lactic acid in the necrotic core, whereas pyruvic acid was elevated in the fibrous cap. In unstable plaques, 5-hydroxyindoleacetic acid was enriched in the fibrous cap. CONCLUSIONS: Our work here represents the first step to defining an atlas of metabolic pathways involved in plaque destabilization in human atherosclerosis. We anticipate this will be a valuable resource and open new avenues of research in cardiovascular disease.

Determination and Calculations of Mercury Vapor Concentration and Energy Released from Freshly Condensed Dental Amalgams Having Various Copper Percentages within the Alloy.

Dental amalgam is an alloy consisting of a mixture of fine metallic powder of silver, tin, zinc, copper, and a trace amount of palladium in combination with about fifty percent elemental mercury that forms a matrix phase. Dental amalgams consisting of a high-copper content are the most common types of alloys currently utilized for the restoration of decayed, broken, and fractured posterior human teeth. The present research objective was primarily to improve the material properties by determining and analyzing the amount of mercury vapor released from dental amalgam received from eight different commercial brands. The mechanical hardness of the alloys was found to increase with an increase in copper content in the amalgam. The effect of copper addition on material aging was also studied. During the release of mercury vapor, the corresponding energies associated with the release of mercury vapor from each sample were determined for each successive measurement. The results indicated that increasing the copper content of the amalgam counters the release of mercury vapor from posterior teeth and improves the hardness properties.

The interplay between nonalcoholic fatty liver disease and atherosclerotic cardiovascular disease.

Therapeutic approaches that lower circulating low-density lipoprotein (LDL)-cholesterol significantly reduced the burden of cardiovascular disease over the last decades. However, the persistent rise in the obesity epidemic is beginning to reverse this decline. Alongside obesity, the incidence of nonalcoholic fatty liver disease (NAFLD) has substantially increased in the last three decades. Currently, approximately one third of world population is affected by NAFLD. Notably, the presence of NAFLD and particularly its more severe form, nonalcoholic steatohepatitis (NASH), serves as an independent risk factor for atherosclerotic cardiovascular disease (ASCVD), thus, raising interest in the relationship between these two diseases. Importantly, ASCVD is the major cause of death in patients with NASH independent of traditional risk factors. Nevertheless, the pathophysiology linking NAFLD/NASH with ASCVD remains poorly understood. While dyslipidemia is a common risk factor underlying both diseases, therapies that lower circulating LDL-cholesterol are largely ineffective against NASH. While there are no approved pharmacological therapies for NASH, some of the most advanced drug candidates exacerbate atherogenic dyslipidemia, raising concerns regarding their adverse cardiovascular consequences. In this review, we address current gaps in our understanding of the mechanisms linking NAFLD/NASH and ASCVD, explore strategies to simultaneously model these diseases, evaluate emerging biomarkers that may be useful to diagnose the presence of both diseases, and discuss investigational approaches and ongoing clinical trials that potentially target both diseases.

Coverage of mass drug administration (MDA) and operational issues in elimination of lymphatic filariasis in selected districts of Jharkhand, India.

Background: Despite annual rounds of mass drug administration (MDA) being carried out every year since 2004 to break the transmission of lymphatic filariasis, India has not achieved elimination status in many areas of country. This study was conducted to determine the operational issues in the implementation of MDA for elimination of lymphatic filariasis in selected districts of Jharkhand. Methods: Two districts of Jharkhand state were selected for present cross-sectional study. Multi-staged cluster sampling was adopted to select study participants. Data were collected with use of prescribed questionnaire of National Vector Borne Disease Control Programme (NVBDCP) from the study participants. Data related to socio-demographic details, coverage and compliance of MDA and operational issues were collected. Results: Findings of independent assessment show that percentage of MDA coverage and compliance for Hazaribag district was 96.02% and 88.90%, respectively. On the other hand, percentage of MDA coverage and compliance was found to be only 67.06% and 48.44% in Chatra district. Overall drug consumption was significantly associated with age group (p-value = 0.045), educational status (p-value = <0.0001) and socio-economic status (p-value <0.0001) of eligible population. Most common reason for not swallowing the drugs was found to be absence of family members at the time of MDA rounds followed by no visit of house by drug distributors. Conclusions: Coverage and compliance of MDA was better in Hazaribag district as compared to Chatra district. In order to increase compliance, socio-demographic factors must also be addressed apart from other operational issues.

Modulation of Structural, Electronic, and Optical Properties of Titanium Nitride Thin Films by Regulated In Situ Oxidation.

Epitaxial titanium nitride (TiN) and titanium oxynitride (TiON) thin films have been grown on sapphire substrates using a pulsed laser deposition (PLD) method in high-vacuum conditions (base pressure <3 × 10-6 T). This vacuum contains enough residual oxygen to allow a time-independent gas phase oxidation of the ablated species as well as a time-dependent regulated surface oxidation of TiN to TiON films. The time-dependent surface oxidation is controlled by means of film deposition time that, in turn, is controlled by changing the number of laser pulses impinging on the polycrystalline TiN target at a constant repetition rate. By changing the number of laser pulses from 150 to 5000, unoxidized (or negligibly oxidized) and oxidized TiN films have been obtained with the thickness in the range of four unit cells to 70 unit cells of TiN/TiON. X-ray photoelectron spectroscopy (XPS) investigations reveal higher oxygen content in TiON films prepared with a larger number of laser pulses. The oxidation of TiN films is achieved by precisely controlling the time of deposition, which affects the surface diffusion of oxygen to the TiN film lattice. The lattice constants of the TiON films obtained by x-ray diffraction (XRD) increase with the oxygen content in the film, as predicted by molecular dynamics (MD) simulations. The lattice constant increase is explained based on a larger electrostatic repulsive force due to unbalanced local charges in the vicinity of Ti vacancies and substitutional O. The bandgap of TiN and TiON films, measured using UV-visible spectroscopy, has an asymmetric V-shaped variation as a function of the number of pulses. The bandgap variation following the lower number of laser pulses (150-750) of the V-shaped curve is explained using the quantum confinement effect, while the bandgap variation following the higher number of laser pulses (1000-5000) is associated with the modification in the band structure due to hybridization of O2p and N2p energy levels.

Mechanisms of continual efferocytosis by macrophages and its role in mitigating atherosclerosis.

Atherosclerotic cardiovascular disease is the leading cause of death worldwide. Rupture-prone atheromas that give rise to myocardial infarction and stroke are characterized by the presence of a necrotic core and a thin fibrous cap. During homeostasis, cellular debris and apoptotic cells are cleared quickly through a process termed "efferocytosis". However, clearance of apoptotic cells is significantly compromised in many chronic inflammatory diseases, including atherosclerosis. Emerging evidence suggests that impairments in efferocytosis drive necrotic core formation and contribute significantly to plaque vulnerability. Recently, it has been appreciated that successive rounds of efferocytosis, termed "continual efferocytosis", is mechanistically distinct from single efferocytosis and relies heavily on the metabolism and handling of apoptotic cell-derived cargo. In vivo, selective defects in continual efferocytosis drive secondary necrosis, impair inflammation resolution, and worsen atherosclerosis. This Mini Review focuses on our current understanding of the cellular and molecular mechanisms of continual efferocytosis and how dysregulations in this process mediate nonresolving inflammation. We will also discuss possible strategies to enhance efferocytosis when it fails.

Genetic manipulation of photosynthesis to enhance crop productivity under changing environmental conditions.

Current global agricultural production needs to be increased to feed the unconstrained growing population. The changing climatic condition due to anthropogenic activities also makes the conditions more challenging to meet the required crop productivity in the future. The increase in crop productivity in the post green revolution era most likely became stagnant, or no major enhancement in crop productivity observed. In this review article, we discuss the emerging approaches for the enhancement of crop production along with dealing to the future climate changes like rise in temperature, increase in precipitation and decrease in snow and ice level, etc. At first, we discuss the efforts made for the genetic manipulation of chlorophyll metabolism, antenna engineering, electron transport chain, carbon fixation, and photorespiratory processes to enhance the photosynthesis of plants and to develop tolerance in plants to cope with changing environmental conditions. The application of CRISPR to enhance the crop productivity and develop abiotic stress-tolerant plants to face the current changing climatic conditions is also discussed.

Community-based assessment of risk factors for cardiovascular diseases in a tribal population of Tharus in Bihar, India.

Background: The problems of cardiovascular diseases are on the rise and the tribal population are also not an exception to it. The present study aims to estimate the prevalence of risk factors of cardiovascular diseases and to find association of hypertension and diabetes with various risk factors among the Tharu tribe of Bihar, India. Methodology: The present study was an observational, cross-sectional study conducted on 252 individuals of 20-60 years age group. Risk factors related to cardiovascular diseases were assessed. Results: Among 252 participants, the prevalence of hypertension and diabetes was 32.5% and 4.8%, respectively. Around 30% of study participants were smokers, 39% were habitual of chewing tobacco, and about half of the population were not involved in active physical activities (49.2%). Raised blood glucose level was significantly associated with gender, age, smoking status, and tobacco chewing while hypertension was significantly associated with age, smoking status, and body mass index. Conclusions: Increased number of various risk factors among the tribal population makes them vulnerable to cardiovascular diseases and other complications.

Click Chemistry Inspired Synthesis of Hydroxyanthracene Triazolyl Glycoconjugates.

Novel hydroxyanthracene-based terminal alkynes 3 and 5a/b were synthesized by the acetylide addition reaction at the 9,10-position of anthraquinone 1 under mild conditions. The developed alkynes 3, 5a, and 5b on Huisgen azide-alkyne cycloaddition reaction with azido-sugars 6 in the presence of Cu(I) catalyst provided a series of triazole fasten hydroxyanthracene glycoconjugates 7, 8, and 9, respectively, in good yields. The representative compounds 9 and 7h were successfully deprotected under room-temperature conditions to liberate the corresponding free glycoconjugates 10 and 11, respectively. Further, structures of a few compounds were unmaliciously evidenced by their single-crystal X-ray.

Recurrent respiratory epithelial adenomatoid hamartoma of the nasal cavity.

Respiratory epithelial adenomatoid hamartoma (REAH) refers to a rare entity characterized by excessive proliferation of the normal glandular elements of the respiratory epithelium present as mass lesions in various body sites. The pathophysiology of the disorder is still debated. The condition can closely mimic inverted papilloma, adenocarcinoma, and nasal polyposis clinically, radiologically, and pathologically. However, distinction from the above disorders is important in view of the excellent prognosis associated with complete excision of REAH. Recurrence is uncommon with complete excision, and a high-risk pathologic transformation is not expected with this lesion. We report a case of recurrent REAH managed with repeat surgical endoscopic excision. The patient is disease free 4 years after re-excision of the lesion.

Bioenergetic Failure Drives Functional Exhaustion of Monocytes in Acute-on-Chronic Liver Failure.

Objective: The monocyte-macrophage system is central to the host's innate immune defense and in resolving injury. It is reported to be dysfunctional in acute-on-chronic liver failure (ACLF). The disease-associated alterations in ACLF monocytes are not fully understood. We investigated the mechanism of monocytes' functional exhaustion and the role of umbilical cord mesenchymal stem cells (ucMSCs) in re-energizing monocytes in ACLF. Design: Monocytes were isolated from the peripheral blood of ACLF patients (n = 34) and matched healthy controls (n = 7) and patients with compensated cirrhosis (n = 7); phagocytic function, oxidative burst, and bioenergetics were analyzed. In the ACLF mouse model, ucMSCs were infused intravenously, and animals were sacrificed at 24 h and day 11 to assess changes in monocyte function, liver injury, and regeneration. Results: Patients with ACLF (alcohol 64%) compared with healthy controls and those with compensated cirrhosis had an increased number of peripheral blood monocytes (p < 0.0001) which displayed significant defects in phagocytic (p < 0.0001) and oxidative burst capacity (p < 0.0001). ACLF patients also showed a significant increase in the number of liver macrophages as compared with healthy controls (p < 0.001). Bioenergetic analysis showed markedly reduced oxidative phosphorylation (p < 0.0001) and glycolysis (p < 0.001) in ACLF monocytes. Patients with monocytes having maximum mitochondrial respiration of <37.9 pmol/min [AUC = 0.822, hazard ratio (HR) = 4.5] and baseline glycolysis of ≤42.7 mpH/min (AUC = 0.901, HR = 9.1) showed increased 28-day mortality (p < 0.001). Co-culturing ACLF monocytes with ucMSC showed improved mitochondrial respiration (p < 0.01) and phagocytosis (p < 0.0001). Furthermore, ucMSC therapy increased monocyte energy (p < 0.01) and phagocytosis (p < 0.001), reduced hepatic injury, and enhanced hepatocyte regeneration in ACLF animals. Conclusion: Bioenergetic failure drives the functional exhaustion of monocytes in ACLF. ucMSCs resuscitate monocyte energy and prevent its exhaustion. Restoring monocyte function can ameliorate hepatic injury and promote liver regeneration in the animal model of ACLF.

Defects in energy metabolism are associated with functional exhaustion of bone marrow mesenchymal stem cells in cirrhosis.

OBJECTIVES: Cellular and functional exhaustion of bone marrow mesenchymal stem cells (BM-MSC) is significantly associated with the loss of HSCs and hepatic osteodystrophy in cirrhosis. The molecular mechanisms underlying the dysfunction of BM-MSCs are not well understood. We investigated the underlying mechanisms of cellular and functional exhaustion of BM-MSCs in cirrhosis. METHODS: The MSCs were isolated retrospectively from bone marrow of decompensated alcoholic cirrhosis patients {(Trial registration: ClinicalTrials.gov NCT01902511) (n=10; MELD=16.2±2.3; CTP=8.7±2.3)} and age and gender-matched healthy controls (n=8). Global gene expression profile of healthy bone marrow MSCs (hBM-MSCs) and cirrhosis patients BM-MSCs (cBM-MSCs) were done by mRNA sequencing. XFe24-bioanalyzer analyzed the bioenergetic potential of cells. Level of different cytokines and growth factors in BM-plasma and MSCs secretome were analyzed by Luminex-based bead array. RESULTS: Analysis of differentially expressed genes showed significant (P<0.01) up-regulation of genes associated with ubiquitination and catabolism of proteins; TNF signaling, insulin resistance, and down-regulation of genes associated with DNA repair, protein processing, cell cycle, and mitochondrial respiration in cBM-MSCs in comparison to hBM-MSCs. Compared to hBM-MSCs, cBM-MSCs showed a significant defect in glycolysis due to insulin resistance and poor glucose uptake (P=0.002). This led to compromised self-renewal capacity and cellular loss of MSCs in cirrhosis. cBM-MSCs also showed a significant impairment in Oxidative phosphorylation (OXPHOS) due to mitochondrial dysfunction leading to defects in the osteogenic differentiation with early aging and senescence. CONCLUSION: Compromised energy metabolism due to inflammatory and metabolic stress-induced insulin resistance underlies the cellular and functional exhaustion of BM-MSCs in cirrhosis.

Development of loop-mediated isothermal method and comparison with conventional PCR assay for rapid on spot identification of tissue of cattle origin.

Loop-mediated isothermal amplification (LAMP) is a diagnostic method for meat speciation with rapid and minimal equipment requirements. In this study, we developed cattle-specific tube-based LAMP assays targeting mitochondrial Cyt b gene sequence, compared with conventional PCR assay for specificity, sensitivity, and validation of the assay was made. The LAMP reaction was carried at 64 °C for 45 min, and results were confirmed by SYBR Green I dye and agarose gel-electrophoresis. The specificity of the assays was cross-tested with DNA of buffalo, goat, sheep, and pork. The amplification was observed with samples from cattle only without cross-reactivity with other meat species. The analytical sensitivity of LAMP and PCR method for cattle DNA detection was 0.0001 ng and 1 ng, respectively. Repeatability of the assay was achieved on samples from known/blind and admixture meat with other than cattle at the relative percentage of 20%, 10%, 5%, and 1%. The study concluded that the developed assay can be easily employed for the rapid identification of tissue of cattle origin in meat and meat products in low resource areas.

Establishment of a murine model of acute-on-chronic liver failure with multi-organ dysfunction.

BACKGROUND AND AIMS: Acute-on-chronic liver failure (ACLF) is a distinct clinical entity with high probability of organ failure and mortality. Since patients generally present late, experimental models are needed to understand the pathophysiology and natural course of the disease. METHODOLOGY: To reproduce the syndrome of ACLF, chronic liver disease was induced in C57BL6 mice (6-8 weeks; approximately 20-24 g weight) by intraperitoneal administration of carbon tetrachloride (CCl4) for 10 weeks followed by an acute injury with acetaminophen (APAP) and lipopolysaccharide (LPS). Blood, ascitic fluid, and organs were collected to study cell death, regeneration, and fibrosis. RESULTS: At 24 h post-APAP/LPS infusion, the liver tissue showed increased hepatocyte ballooning and endothelial cell TUNEL positivity. This was followed by progressive hepatocyte necrosis from perivascular region at day 7 to lobular region by day 11. ACLF (day 7 and day 11) animals showed increase in bilirubin (p < 0.05), prothrombin time (p < 0.0001), blood ammonia (p < 0.001), and portal pressure post-acute hepatocellular injury similar to human ACLF. Ascites was noticed by day 11 with median serum-ascites albumin gradient of 1.2 (1.1-1.3) g/dL. In comparison to cirrhosis, ACLF group (day 7 and day 11) showed significant decrease in Sirius red (p ≤ 0.0001), collagen1 (p < 0.0001), and a-SMA proportionate area (p < 0.0001) with loss of hepatocytes regeneration (p < 0.005). At day 11, ACLF animals also showed significant increase in serum creatinine (p < 0.05) and acute tubular necrosis suggestive of organ failure, compared to cirrhotic animals. CONCLUSION: The CCL4/APAP/LPS (CALPS) model of ACLF mimics the clinical, biochemical, and histological features of ACLF with demonstrable progressive hepatocellular necrosis, liver failure, impaired regeneration, development of portal hypertension, and organ dysfunction in an animal with chronic liver disease.