Arachnia rubra strain DSM 100122T. The genome is 3.32 Mb, with a GC content of 64.2%. The genome contains 3,005 predicted genes, including 2,923 predicted protein-coding genes.We report the complete genome of  Propionibacterium were recently divided into four genera, i.e., Propionibacterium, Acidipropionibacterium, Cutibacterium, and Pseudopropionibacterium , ultrasmall parasitic epibionts  systems are a major barrier to genetic transformation, and RM systems can be identified from the methylome obtained during single-molecule real-time (SMRT) genome sequencing . For DNA isolation, the strain was grown in a 50:50 mixture of Trypticase soy broth and brain heart infusion broth with 1% yeast extract. Genomic DNA was extracted using a MasterPure DNA isolation kit (Lucigen) with a modified protocol that included bead beating for cell lysis. SMRT sequencing was carried out on a Sequel instrument  with v3 chemistry, following standard SMRTbell template preparation protocols for base modification detection. Genomic DNA samples (5 to 10 μg) were sheared to an average size of 15 kbp via g-TUBE , end repaired, and ligated to hairpin-barcoded adapters prior to sequencing. Finally, prior to sequencing, the SMRTbell library was purified and size selected using AMPure PB beads to remove <3-kbp templates. Sequencing reads were processed using the Pacific Biosciences SMRT Link pipeline v8 (https://www.pacb.com/support/software-downloads) with Microbial Assembly under default parameters. A total of 145,877 subreads were obtained, covering 632,860,315 subread bases, with a mean read length of 4,329 bp and a read N50 value of 4,635 bp. The mean depth of coverage across the genome was 185×. A single circular contig of 3,316,958-bp length was assembled. The genomic GC content was 64.2%. The genome was annotated with the NCBI Prokaryotic Genome Annotation Pipeline (PGAP). A total of 3,005 genes were identified, including 2,923 predicted protein-coding genes, 56 predicted RNAs, and 26 predicted pseudogenes. Three motifs were identified as methylated throughout the genome, i.e., m6GCTGCA , m6BCTACGA , and m6TGGAAA (712 modified motifs). REBASE analysis assigned the type II methyltransferase M.Aru100122I as being responsible for the m6GCTGCA motif modification, while the remaining modifications could not be assigned unambiguously to the remaining methyltransferase identified within the genome. Additionally, the genome harbors open reading frames (Aru100122McrBC) consistent with an active type IV restriction system, which should be taken into consideration during genetic engineering.Strain DSM 100122CP072384 and SRA accession number SRR15979320. Base modification files were submitted with the GenBank submission, and the methylome analysis is available at REBASE with organism number 46978 for strain DSM 100122T. The BioProject accession number for this genome, as well as those of many other oral bacteria, is PRJNA282954.The genome sequence was deposited in GenBank under the accession number


Environmental enteric dysfunction (EED) is thought to occur from persistent intestinal inflammation. Studies also revealed the association of lipocalin-2 (LCN2) and low-density lipoprotein receptor-related protein-1 (LRP1) with intestinal inflammation. Therefore, we intended to explore the relationship of LCN2 and LRP1 with gut inflammation and biomarkers of EED in Bangladeshi malnourished children.A total of 222 children (length-for-age z-score (LAZ) <-1) aged 12–18 months were enrolled in this study in a cross-sectional manner. Among the participants, 115 were stunted (LAZ <-2) and 107 were at risk of being stunted (LAZ −1 to −2) children. Plasma and faecal biomarkers were measured using ELISA. Spearman’s rank correlation was done to see the correlation among LCN2, LRP1 and biological biomarkers.LCN2 correlates positively with myeloperoxidase , neopterin , calprotectin , Reg1B  and EED score . Whereas, LRP1 correlates negatively with myeloperoxidase , neopterin , alpha-1-antitrypsin , Reg1B  and EED score .Our findings imply that LCN2 might be a promising biomarker to predict gut inflammation and EED. Whereas, increased level of LRP1 may contribute to alleviating intestinal inflammation. Environmental enteric dysfunction (EED) is highly prevalent in low-income and middle-income countries, which affects approximately 40% of all children with age less than 5 years.EED causes linear growth faltering or stunting which has the manifestations including reduced neurodevelopmental and cognitive function and increased morbidity and mortality in childhood.The main challenge to tackle the EED is lack of non-invasive biomarker signature to diagnose the EED and take proper treatment initiative.There are significant positive correlation of lipocalin-2 and significant negative correlation of low-density lipoprotein receptor-related protein-1 (LRP1) with biomarkers of EED.Study findings strongly infer that lipocalin-2 might be a promising biomarker to predict gut inflammation and EED.Study findings also imply that LRP1 could be a therapeutic target to alleviate intestinal inflammation.Environmental enteric dysfunction (EED) is a subacute intestinal inflammation highly prevalent in low-income and middle-income countries, which is defined by crypt hyperplasia, villous atrophy, reduction of mucus layer and infiltration of lymphocytes in lamina propria.Escherichia coli than wild type.Salmonella typhimurium, Klebsiella pneumoniae and E. coli in different studies.LCN2 is a bacteriostatic peptide, a member of lipocalin super family, known as neutrophil gelatinase B-associated lipocalin.LRP1 is an endocytic receptor that facilitates the binding and endocytic transportation of a broad range of biological ligands as well as various molecules function in immune system.To perform the analysis for this study, data were taken from ‘Bangladesh environmental enteric dysfunction’ study. The study protocol was published earlier.Anthropometric and socioeconomic data were collected from study participants by trained staff. LAZ, weight-for length z-score (WLZ), weight-for age z-score (WAZ), mid-upper arm circumference (MUAC) and head circumference of the study participants were calculated as anthropometric indices. A total of 4 mL of blood was collected by medical doctor following all the aseptic conditions and transported to laboratory maintaining cold condition (−4°C). Plasma was collected by centrifugation of blood at 3000 ×g for 10 min. Aliquots of biological samples were stored at −80°C until biomarker analyses. Plasma biomarkers including LRP1 , sCD14 , C-reactive protein (CRP) , alpha-1 acid glycoprotein  and ferritin  were measured by using available ELISA kits. Atomic absorption spectrometry was used to measure the level of zinc in plasma. Stool sample were used to detect the level of biomarkers including LCN2 , neopterin (NEO) , myeloperoxidase (MPO) , alpha-1 antitrypsin (A1AT) , Reg1B , calprotectin  by using available ELISA kits. All the laboratory analyses were performed at the parasitology laboratory of icddr, b.2 test was performed for comparing categorical variables of the participants. Student’s t-test was performed to see the differences in length, weight, MUAC, head circumference, LAZ, WLZ, WAZ, maternal height and WAMI index .Data analyses were performed to present the characteristics of the study participants. Data were presented as median with IQRs when variables show skewed distributions, while categorical variables were showed as frequencies with percentages. The variables showing normal distribution were presented as mean with SD. Mann-Whitney U test was used to detect the differences in the age, family income, plasma biomarkers including CRP, alpha-1-acid glycoprotein (AGP), sCD14, LRP1, ferritin and zinc as well as faecal biomarkers including MPO, NEO, A1AT, calprotectin, Reg1B and LCN2; whereas χNo patient or public were involved with the research design, protocol development and participant’s enrolment. None of them will be involved during dissemination of findings.The levels of plasma biomarkers including LRP1, AGP, CRP, sCD14, ferritin and zinc and faecal biomarkers including MPO, NEO, A1AT, calprotectin, Reg1B and LCN2 were presented in A significant positive correlation was found among LCN2 and different biomarkers of intestinal inflammation like MPO, NEO, calprotectin and Reg1B. LCN2 also positively correlated with EED score. However, LRP1 was negatively and significantly correlated with MPO, NEO, A1AT, Reg1B and EED score. Correlations among different variables are illustrated in 10.1136/bmjpo-2021-001138.supp1Supplementary dataOur results reveal that LCN2 positively correlates with biomarkers of intestinal inflammation including MPO, NEO, calprotectin and Reg1B. These results are in accordance with the findings of another study, where LCN2 has been found to be positively correlated with MPO and calprotectin.In our analysis, we also measured the level of plasma LRP1. Correlation among the plasma LRP1 and faecal biomarkers showed that LRP1 is negatively correlated with MPO, NEO, A1AT and Reg1B. Moreover, LRP1 also negatively correlates with EED score. MPO, NEO, A1AT have been reported as biomarkers of intestinal inflammation and EEDThere were several limitations in our study. First, we could not measure the genetic expression of LCN2 and LRP1 of participants due to invasive collection process of intestinal cell. Second, we did not examine the consequence of suppression of LRP1 expression of human to investigate whether decreased expression may or may not contribute to gut inflammation because of ethical consideration. Lack of age-matched and sex-matched healthy children was also a limitation in this study. However, this was the first study that investigated the relationship of LCN2 and LRP1 with biomarkers of EED.Our findings suggest that LCN2 might be a potential biomarker to predict the gut inflammation and EED. However, increased level of LRP1 may contribute in alleviating intestinal inflammation that could be a potential therapeutic target to reduce the inflammation.


Rhizophora apiculata was characterized. The cp genome length was 164,343 bp in length, containing a typical structure of a large single copy (LSC) of 93,155 bp, a small single copy (SSC) of 19,376 bp, and two inverted repeats (IRs) of 25,906 bp, with a GC content of 34.9%. There were 131 genes were annotated in the cp genome, including 85 protein-coding genes, 38 tRNA genes, and 8 rRNA genes. A phylogenetic analysis using cp genomes of mangroves and ecologically associated species resolved R. apiculata in Rhizophora with R. stylosa and R. x lamarckii. This complete chloroplast sequence offers a promising tool for further species identification and evolutionary studies of Rhizophora, as well as for mangroves.The chloroplast (cp) genome sequence of  Mangroves are a diverse group of woody trees and shrubs that thrive in saline and daily inundation conditions in the inter-tidal zone of tropical and subtropical regions Ball . MangrovRhizophora consists of five species and at least 6 hybrid taxa, which are largely separated into Old and New World groups  are considered the most important of all mangroves across the Pacific tropical region  could provide useful and abundant information on genetic diversity and evolution based on our previous studies  is stored in Guangxi University, plant ecophysiology and evolution research group herbarium. A 350-bp paired-end library was then constructed and sequenced by Novogene  using an Illumina HiSeqX-ten system . After sequence filtering for read quality, about 1 Gb raw data were obtained, with a paired-end read length of 2 × 150 bp. A de novo assembly was performed using NOVOPlasty3.6 . The assembled sequence was then imported into Geneious R9 , to check manually as described previously . The final annotations were confirmed and integrated from results of CPGAVAS2 , following previously described methods . The cp genome exhibited a typical structure with a LSC of 93,155, a SSC 19,376 and two inverted repeats (IRa and IRb) of 25,906 bp, respectively. The overall GC content of the plastome was 34.9%, while the GC content in LSC, SSC, IRa, and IRb regions were 32.2%, 28.5%, 42.2%, respectively. In total, 131 genes were annotated, including 85 protein-coding genes, 38 tRNA genes, and 8 ribosomal RNA genes. Seventeen genes  contain 1–2 introns, respectively. While 17 genes were duplicated in the IR regions, including 6 protein-coding genes , 7 tRNA genes , and 4 rRNA genes . Notably, sequence of the translation initiation factor gene (infA) was found located within rps8, with the existence of several internal termination codons indicating a functional loss of infA in the cp genome of R. apiculata.The assembled cp genome of https://db.cngb.org/cnsa/) according to the reference , and CNSA (/11/17, aut-group . All theut-group , and a mut-group  with a G


NEGR1, NTNG1, XPNPEP2, NTNG2, CD109, OPCML, and PRND are critical genes. The miR-222-3p was highly expressed, probably leading to low NEGR1 and high PRND expression in THCA tissues. Low NEGR1 expression indicated favorable prognosis in THCA patients, and high PRND expression indicated poor prognosis. Seven critical genes were significantly related to gender, age, race, tumor stage, and lymph node metastasis. In addition, the seven-gene biomarker exhibited a certain diagnostic value. Finally, CD109 expression was closely correlated with immune cells, especially B cells and CD4+ T cells. The miR-222-3p and its critical target genes could be promising biomarkers for the prognosis of THCA and may emerge as key regulators of immune infiltration in THCA.Thyroid cancer (THCA) is a common endocrine malignancy. With increasing incidence and low mortality, balancing the therapeutic approach is an inevitable issue. This study aimed to confirm the role of miR-222-3p and its target genes in THCA survival and immune infiltration. From different expression analyses based on the GEO and TCGA databases, we predicted and subsequently identified the key target genes of miR-222-3p. We then explored the expression, enrichment, pairwise correlation, protein expression, survival analysis, principal component analysis, and immune significance of the critical genes using bioinformatics analysis. The present study demonstrated that  Thyroid cancer (THCA) is one of the most common endocrine malignancies and is a complex disease resulting from radiation exposure, abnormal hormone levels, abnormal iodine uptake, and genetic susceptibility. GloballPPP2R2A(MicroRNAs (miRNAs) are a class of endogenous non-coding RNA molecules, 19–25 nucleotides in length. Mature microRNAs can target the exact complementary sequence of the 3′-untranslated region (3′-UTR), resulting in the inhibition of translation and degradation of messenger ribonucleic acid (mRNA) . Many miPPP2R2A. Howeverhttps://www.gsea-msigdb.org/gsea/index.jsp), and protein-protein interaction (PPI) network analyses were conducted to analyze the consensus genes. We then conducted expressed validation of critical genes and analyses of related clinical data using UALCAN . The pairwise correlation and prognostic role of the critical target genes were validated based on the Gene Expression Profiling Interactive Analysis 2  and their protein expression were ascertained via the Human Protein Atlas . In addition, the diagnostic value of critical target genes was estimated using multiple-gene comparison and principal component analysis. Finally, because the tumor immune microenvironment in THCA has attracted increasing attention, we mined the Cell Type Identification By Estimating Relative Subsets Of RNA Transcripts  and Tumor Immune Estimation Resource  to demonstrate the relationship between miR-222-3p in THCA and immune infiltrate cells.This study investigated the differential miR-222-3p expression in THCA tissues and non-cancerous thyroid tissues, acquired the differentially expressed genes (DEGs) in the THCA dataset of the Cancer Genome Atlas (TCGA), predicted the possible target genes of miR-222-3p using miRWalk3.0, and identified the common genes of DEGs and the possible target genes as the consensus genes. Subsequently, Gene Ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) functional interpretation, Gene Set Enrichment Analysis  samples from Homo sapiens, and 2) sample size of each group ≥3. All data was normalized using the min-max normalization method. An unpaired To predict the possible target genes of miR-222-3p, we utilized the online analysis website miRWalk3.0, including four prediction algorithms, TargetScan, miRDB, mirtarbase, and TarPmiR. To imprp < 0.05.To elucidate the biological functions and signal transduction pathways of miR-222-3p and its target genes, GO annotation and KEGG functional interpretation were performed for the consensus genes using database for Annotation, Visualization and Integrated Discovery (DAVID) . The GSEThe PPI network and module analysis helped identify the critical genes of miR-222-3p and verified their interaction. We input the consensus genes into Cytoscape 3.7.2, and impBased on TCGA gene expression and clinical data, UALCAN was used to verify the expression of critical genes and perform the analyses of related clinical data in samples of THCA and normal thyroid . The 505p value and hazard ratio (HR) with 95% confidence intervals (95% CI). Meanwhile, we conducted a multiple gene comparison and principal component analysis for the critical genes to explore their diagnostic value.GEPIA2 could provide survival and integrated analysis, allowing researchers to select their genes and cancers of interest for evaluation of prognosis. We input the critical genes into the Survival Plots module, and the Kaplan-Meier curves were plotted with the log rank p < 0.05.To illustrate the association of miR-222-3p with the critical genes, we performed a Spearman’s correlation analysis based on 500 thyroid carcinoma samples utilizing the LinkedOmics database. The paihttp://www.proteinatlas.org), and the staining intensity of the antibody revealed the protein expression of critical genes in THCA in THCA. The corp < 0.001). The mean ± standard deviation were 0.25250 ± 0.18090 and 0.04267 ± 0.02269, respectively (p < 0.05), whereas no statistical significance was found in the two remaining datasets (gse62054 and gse97070).From the TCGA and GEO database, a total of 603 THCA and 120 normal thyroid tissue samples were compared in the present study. The miR-222-3p expression in THCA samples was significantly upregulated compared to that in normal thyroid tissue  were identified by edgeR. In addition, 2,105 possible target genes were predicted using the miRWalk3.0. Consequently, we regarded 303 overlapping genes as consensus target genes of miR-222-3p .p-value, indicating the closest connection with the tumor , cellular component (CC), and molecular function (MF), respectively, and a total of six significant pathways were verified by KEGG analysis. These noticeably enriched GO terms and KEGG pathways may play an important role in the mechanism of thyroid carcinoma, which might contribute to appropriate tailoring of treatment for patients. The BP GO term “nervous system development,” CC GO term “integral component of plasma membrane” and the MF GO term “calcium ion binding” had the smallest he tumor . For thes genes . The resgenes  were identified as critical genes, indicating that they may be important in the pathogenesis of THCA.The PPI network consisted of 302 nodes and 369 edges generated by entering 303 consensus genes into the Cytoscape 3.7.2 . The modNEGR1, NTNG1, and XPNPEP2) showed low expression, and the remaining critical genes  were highly expressed in THCA samples, comparing to normal thyroid samples. In terms of gender , whereas DFS was not significantly different , whereas the OS results were not significantly different  curve. Interestingly, high ifferent . On the ifferent . For theCD109 exhibited the highest expression level in THCA samples via multiple-gene comparison, followed by NTNG2, NEGR1, OPCML, XPNPEP2, PRND, and NTNG1, NTNG2 , and OPCML  were positively correlated with miR-222-3p, whereas NEGR1  and PRND  were negatively correlated. However, the correlation of NTNG1 (p = 0.29) and XPNPEP2 (p = 0.15) with miR-222-3p was not statistically significant. Interestingly, a negative correlation (cor = -0.19) only existed between NEGR1 and CD109, while the other groups exhibited positive correlations , gamma delta T cells, resting NK cells, activated NK cells, M0 macrophages, M1 macrophages, M2 macrophages, resting dendritic cells, activated dendritic cells, resting mast cells, activated mast cells, eosinophils, neutrophils) , CD4+ T cells , macrophages , neutrophils , and dendritic cells , and the highest negative correlation with CD8+ T cells   . Contrib< 0.001) . Moreovete cells . In addiand PRND .Kip1 in human thyroid papillary carcinomas, while abnormal cell cycle leads to malignant transformation. Moreover, Huang et al. indicated that miR-222 promotes tumor invasion and metastasis in papillary thyroid cancer by targeting PPP2R2A were identified as critical genes through module analysis using the Cytoscape software.In the present study, we aimed to elucidate the biological functions of miR-222-3p and its target genes in THCA. Differential expression analysis of miR-222-3p based on the GEO and TCGA database indicated the upregulation of miR-222-3p expression in THCA; 303 consensus genes were obtained by overlapping the target genes predicted by miRWalk3.0 and DEGs from the TCGA database. To search for enriched pathways of miR-222-3p and its target genes, we acquired three possible signature pathways using the GSEA analysis. Moreover, using the STRING public platform, the PPI network was constructed, and seven genes  of THCA, while PRND was associated with disease-free survival (DFS) and THCA. Neuronal growth regulator 1 (NEGR1) is a member of the immunoglobulin LON (IgLON) family, which pertains to GPI-anchored cell adhesion molecules (CAMs), consistent with the pathways of KEGG enrichment analysis, a paralog of the prion (PrP) protein, was upregulated in many malignant diseases, including astrocytomas, osteosarcoma, acute myeloid leukemia, and myelodysplastic syndromes(PRND in thyroid cancer. In our study, PRND had a high level expression in THCA, indicating poor DFS. Nevertheless, more research is needed to explore the target signature pathway.We discovered close relationships between critical genes and prognosis of THCA. Results confirmed that  analysis. Previouy cancers. By invecarcinoma. This stsyndromes. To dateNEGR1 and PRND exhibited prognostic significance in THCA, their pairwise correlations indicated that the remaining critical genes, CD109, NTNG1, NTNG2, XPNPEP2, and OPCML, also play important roles in the regulation of biological processes. The analyses of related clinical data revealed that critical genes were all significantly related to tumor stage and lymph node metastasis of THCA. In addition, multiple-gene comparison and principal component analysis of critical genes revealed that they had potential diagnostic value in THCA, which indicated that their conjoint analysis offered a method for effectively distinguishing THCA from normal thyroid tissue. Therefore, seven critical genes were suggested as biomarkers for the precise diagnosis of THCA. These results provide insights into the role of multiple genes and interactive networks in the multilevel regulation of the molecular mechanism of THCA.Even though CD109 had the highest positive correlation with B cells, CD4+ T cells, macrophages, neutrophils, and dendritic cells, and the highest negative correlation with CD8+ T cells. NTNG1 and XPNPEP2 also exhibited a high correlation with a variety of immune cells. Overall, the results of immune infiltrates illustrated that miR-222-3p was pertinent to the regulation of the immune microenvironment, mainly through targeting CD109, NTNG1, and XPNPEP2 in THCA.Previous studies revealed that immune cells in the tumor microenvironment play an essential role in the occurrence and development of THCA. As an iin vitro and in vivo experiments. Nevertheless, the results of this study are significant as they highlight on promising biomarkers of prognosis, diagnosis, and immune infiltrates.Inevitably, there were certain limitations to our study. As the datasets were obtained from different databases and platforms, an uncertain systematic bias might exist. Furthermore, although enriched analysis was performed to preliminarily investigate the regulatory function of miR-222-3p and its target genes in THCA, the particular mechanism between miR-222-3p and critical genes requires further NEGR1 expression and elevated PRND expression. Downregulated NEGR1 expression indicating a positive prognosis of THCA, whereas elevated PRND expression indicated poor prognosis. CD109 may be capable of regulating B cells, CD4+T cells, macrophages, neutrophils, and dendritic cells in immune microenvironment of THCA. Hence, the critical target genes of miR-222-3p, which indicate potential prognostic value, play an essential role in the infiltration of immune cells and could act as potential targets for the treatment of THCA patients.In summary, using integrated bioinformatics analysis, we demonstrated that miR-222-3p was upregulated in THCA. The elevated miR-222-3p expression may result in downregulated


Background: Derangement of liver blood tests (LBT) is frequent in patients with Coronavirus disease 2019 (COVID-19). We aimed to evaluate (a) the prevalence of deranged LBT as well as their association with (b) clinical severity at admission and (c) 30-day outcomes among the hospitalized patients with COVID-19. Methods: Consecutive patients with COVID-19 hospitalized in the regional referral center over the 12-month period were included. Clinical severity of COVID-19 at hospital admission and 30-day outcomes  were analyzed. Results: Derangement of LBT occurred in 2854/3812 (74.9%) of patients, most frequently due to elevation of AST (61.6%), GGT (46.1%) and ALT (33.4%). Elevated AST, ALT, GGT and low albumin were associated with more severe disease at admission. However, in multivariate Cox regression analysis, when adjusted for age, sex, obesity and presence of chronic liver disease, only AST remained associated with the risk of dying  independently of comorbidity burden and COVID-19 severity at admission. Patients with more severe liver injury more frequently experienced defined adverse outcomes. Conclusions: Deranged LBTs are common among patients hospitalized with COVID-19 and might be used as predictors of adverse clinical outcomes. Coronavirus disease 2019 (COVID-19) is a multisystemic disease, with pandemic features . Along wWhereas elevation of aminotransferases has been commonly seen in hospitalized patients with COVID-19, liver failure represents a rare development usually encountered among patients with already known liver cirrhosis, or as the part of multiorgan failure caused by severe inflammatory response syndrome and septic shock ,6,7,8,9.Therefore, we aimed to evaluate (a) prevalence of deranged LBT at admission to hospital, as well as their association with (b) clinical severity, and (c) 30-day outcomes among the hospitalized patients with COVID-19, reflecting the real-life experience from the largest regional COVID-19 hospital in Croatia. This study included 3.812 consecutive patients with COVID-19 who were hospitalized in Dubrava University Hospital over the period from 19 March 2020 to 19 March 2021. Dubrava University Hospital was completely re-purposed to serve exclusively as the regional tertiary COVID-19 center during COVID-19 pandemic. All patients had a positive nasopharyngeal swab on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by polymerase chain reaction (PCR) or antigen test and were admitted through the hospital’s emergency department. Standardized clinical work-up was performed for each patient, including medical history taking, clinical examination, obtaining blood biochemistry, peripheral oxygen saturation, chest X-ray, and electrocardiogram. Other examinations were performed as indicated based on the clinical picture and decision of the attending physician. Included were the patients with available laboratory and clinical parameters collected within the 24 h from admission, sufficient to assess the severity of COVID-19, presence of comorbidity, along with LBTs (aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), total bilirubin (Bil), serum albumin , prothrombin time (PT)), complete blood count (CBC), who were followed for 30-day from the admission to hospital. Patients were admitted to the intensive care unit (ICU), or to the regular ward based on the severity of clinical picture at presentation, as assessed in emergency department according to the national guidelines that incorporated severity of pneumonia and modified early warning score (MEWS) ,21. Upon2)/FiO2) ≤ 300 mmHg), (b) presence of sepsis or septic shock, with/without organs’ failure, or (c) MEWS ≥ 5, and these patients required ICU admission. Comorbidities were assessed as individual entities and were summarized using the Charlson comorbidity index [2. All LBTs were recorded at hospital admission. The following were considered as normal ranges: Bilirubin ≤ 20 µmol/L, AST ≤ 30 IU/L in females and ≤ 38 IU/L for males, ALT ≤ 36 for females and ≤48 for males, GGT ≤ 35 IU/L for females and ≤ 55 IU/L for males, ALP ≤ 153 IU/L for females and ≤ 142 IU/L for males, albumin ≥ 35 g/L, and PT (quick) ≥ 70%. In accordance to the proposed nomenclature, the term LBT referred to all these tests, whereas the term “liver enzymes” referred to AST, ALT, GGT and AP [Severity of COVID-19 at admission was graded using the classification from the national guidelines for treatment of COVID-19, version 2, issued on 19 November 2020 by the Ministry of Health [ty index . Easternty index . ObesityT and AP . “Liver 2 test and the Χ2 test for trend. Survival analyses were based on the Kaplan–Meier method. Survival curves were compared using the Cox–Mantel version of the log-rank test for univariate and the Cox regression analysis for multivariate analyses. The log-rank test for trend was used for assessing the trend of gradual increase in mortality with higher degree of derangement of liver specific parameters. p-values < 0.05 were considered statistically significant. All analyses were performed using the MedCalc statistical software version 20 .The presented analyses are retrospective in nature. Normality of distribution of numerical variables was tested using the Kolmogorov–Smirnov test. All numerical variables were non-normally distributed and were presented as median and interquartile range (IQR) and were compared between groups using the Mann–Whitney U or the Kruskal–Wallis ANOVA test where appropriate. The Jonckheere–Terpstra test for trend was used to assess rising or degrading trends of specific parameters over disease severity categories. Correlation between numerical variables was assessed using the Spearman rank correlation, relationships with Rho ≥ 0.2 considered as meaningful. Categorical variables are presented as frequency and percentage and were compared between groups using the ΧThis study was conducted in accordance with the World Medical Association Declaration of Helsinki and the study protocol was approved by the Institutional Ethics Committee, No. 2020/1012-10. Due to retrospective design signed informed consent was waived by the Ethics Committee.A total of 3812 COVID-19 patients were analyzed. Median age was 74 years, IQR (64–82). There were 2148/3812 (56.3%) males and 1664/3812 (43.7%) females, and 1023 (28.6%) of patients were obese. There was a significant burden of co-morbidities as 2658 (69.7%), 1154 (30.3%), 617 (16.2%) and 474 (12.4%) patients had arterial hypertension, diabetes, congestive heart failure and chronic kidney disease, respectively. One hundred and six (2.8%) of the patients had history of chronic liver disease, and of them 49 (1.3%) had cirrhosis. Four hundred and thirty-four (11.4%) patients were current smokers. A total of 3390 (88.9%) patients had pneumonia at admission, and of them 3136 (82.3%) required oxygen supplementation. Median length of hospitalization was 10 days IQR (6–16). Rates of ICU admission, need for HFNC oxygenation and MV were 23.1%, 19.9% and 17.3%, respectively. A total of 1315 (34.5%) patients died during the 30-day period. Patients’ characteristics at admission to hospital and their outcomes are shown in p < 0.05 for all analyses). Median AST levels were 41 IQR (28–64). A total of 1432 (38.4%), 1922 (51.5%) and 377 (10.1%) patients presented with normal, 1–3 × elevated and >3 × elevated AST levels on admission, respectively. Higher AST was significantly associated with male sex, obesity, presence of chronic liver disease and liver cirrhosis, higher CRP, and higher ferritin . Median ALT levels were 31 IQR (19–52). A total of 2521 (67.4%), 1049 (28%) and 172 (4.6%) patients presented with normal, 1–3 × elevated and >3 × elevated ALT levels on admission, respectively. Higher ALT was significantly associated with male sex, younger age, obesity, with active or previous smoking, higher hemoglobin, and higher ferritin . Median GGT levels were 42 IQR (24–81). A total of 1934 (53.9%), 1212 (33.8%) and 441 (12.3%) patients presented with normal, 1–3 × elevated and >3 × elevated GGT levels on admission, respectively. Higher GGT was significantly associated with younger age, male sex, obesity, presence of chronic liver disease and liver cirrhosis, alcohol use, active or previous smoking, use of pre-admission antibiotic therapy, and with higher ferritin . Median ALP levels were 72 IQR (56–97). A total of 2862 (90.3%), 270 (8.5%) and 39 (1.2%) patients presented with normal, 1–3 × elevated and >3 × elevated ALP levels on admission, respectively. Higher ALP was significantly associated with non-obesity, presence of chronic kidney disease, presence of chronic liver disease and liver cirrhosis, use of oral anticoagulant therapy, use of pre-admission antibiotic therapy, and with higher D-dimers . Median total bilirubin levels were 11.4 IQR (8.6–15.9). A total of 2586 (85.3%), 375 (12.4%) and 69 (2.3%) patients presented with normal, 1–3 × elevated and >3 × elevated total bilirubin levels on admission, respectively. Higher total bilirubin was significantly associated with male sex, presence of congestive heart failure, presence of chronic liver disease and liver cirrhosis, alcohol use, oral anticoagulant therapy use . Median albumin levels were 32 g/L IQR (28–35). A total of 114 (4.9%), 515 (22.1%) and 1698 (73%) patients presented with albumin levels of ≥40 g/L, 35–39 g/L and <35 g/L, respectively. Lower albumin was significantly associated with older age, female sex, arterial hypertension, congestive heart failure, chronic kidney disease, chronic liver disease and liver cirrhosis, higher Charlson comorbidity index, active or previous smoking, use of oral anticoagulant therapy, use of pre-admission antibiotic therapy, higher WBC, lower hemoglobin, higher CRP, higher ferritin, and higher D-dimers . Median PT values were 100% IQR (89–109%). Considering that 27.5% of patients were receiving oral anticoagulant therapy, further analysis was narrowed to the subgroup without exposure to these drugs. Accordingly, lower PT values were significantly associated with male sex, presence of congestive heart failure, presence of chronic liver disease and liver cirrhosis, use of pre-admission antibiotic therapy, lower hemoglobin, and higher D-dimers  to be considered meaningful. p < 0.05 for all analyses). However, total bilirubin showed no association with the COVID-19 severity at admission. Median time from the first symptoms of COVID-19 to admission was 5 days IQR (1–9). Only higher ALT was associated with longer disease duration prior to admission, whereas lower albumin was associated with worse ECOG functional status. Higher AST, ALT, GGT and lower ALP, albumin and PT were associated with severe clinical presentation of COVID-19 , whereas ALP and PT show no associations with these outcomes (p > 0.05). When analyzed as continuous variables, higher AST, ALP, bilirubin and lower albumin and PT were associated with inferior 30-day survival, while ALT and GGT were not associated with inferior 30-day survival .Associations of clinical outcomes with LBTs are shown in p for trend <0.05 for all analyses except for ALT). Hazard ratios and confidence intervals for the associations of each LBT with 30-day mortality stratified categorically by the degree of derangement are presented in We further investigated associations of LBTs with 30-day mortality using the time to event survival analyses stratified by the degree of derangement from normal values . As depicted in Considering the relationship of LBTs with 30-day mortality, a Cox regression analysis model controlling for age, sex, obesity, Charlson comorbidity index, MEWS severity, chronic liver disease, liver cirrhosis, AST, ALT, GGT, ALP, total bilirubin, albumin and PT was created to assess independent associations. Model is shown in p < 0.001 for overall difference and p < 0.001 for trend for all outcomes, A total of 2650 patients had available synchronous data on all liver enzymes  and bilirubin among whom a total of 314 (11.8%) had combined elevation of both bilirubin and any of the enzymes corresponding to the liver injury, 1732 (65.4%) had enzyme elevation without elevated bilirubin, and in 604 (22.8%) neither enzymes nor bilirubin were elevated. There was a significant trend of increased frequency of adverse outcomes  over higher degree of liver lesion , followed by GGT in 46.1% and ALT in 33.4% of patients. This is much higher prevalence of deranged LBTs than initially reported in studies coming from China (14.9%), but comparable to data from United States ranging from 40–67% ,3,25,26.1.6%, folDeranged LBTs are associated with more severe clinical presentation of COVID-19, which is in keeping with the reports from other authors ,10,25. IThe most important result of our study is clear association between the serum levels of LBTs and clinical outcomes, indicating strongly for their prognostic significance. Indeed, significant gradual increase in mortality was observed with higher degree of derangement of all investigated LBTs, except for ALT . HoweverConflicting data have been published regarding the prognostic impact of elevated LBTs. Whereas in some studies, elevated LBTs were associated with more severe clinical presentation but they were lacking follow-up data to analyze mortality, and others reported various associations to ICU admission, need for mechanical ventilation and death ,13,14,25The pathophysiological role of the liver in COVID-19 remains still not fully elucidated, as histological data are scarce, due to very infrequent liver biopsy taking. Whereas ACE2 receptors, as the gate for SARS-CoV-2 entry in the cell, are most abundantly expressed on the cholangiocytes, hepatocellular profile of elevated LBTs has been commonly reported, and histological changes (in the limited series of patients) were more in favor of deranged liver circulation (microthrombosis), mitochondrial dysfunction and mild hepatitis, rather than biliary injury ,6,28,29.Liver involvement in pathogenesis of COVID-19 and prognostic importance of LBTs is furtherly supported by our results showing incremental increase in frequency of adverse outcomes  when patients were assessed according to the severity of liver lesion. Indeed, patients with liver injury (defined here by elevated bilirubin in addition to elevated liver enzymes) had worse prognosis when compared to patients with the isolated elevation of liver enzymes (not accompanied by the elevated bilirubin), and to patients with normal both liver enzymes and bilirubin. Based on our results and available data from other studies we believe it is still not possible to claim if liver contributes to the severity of inflammatory response to SARS CoV-2 infection, and hence to the severity of COVID-19, or is it only indicator and the part of generalized severe inflammatory response to the virus. In any case, patients with biochemical indicators of more severe liver injury appear to be under increased risk of severe clinical course including the need for ICU treatment, MV and death.In contrast to the high prevalence of biochemical abnormalities suggestive of liver involvement in COVID-19, the prevalence of chronic liver disease (2.8%) and cirrhosis 1.3%) observed in our cohort was low [% observen = 49), as compared to the entire cohort of 3812 patients analyzed here.Interestingly, the association between the presence of cirrhosis and mortality risk was not independent when adjusted to other covariables in multivariate regression analysis as shown in High mortality in our cohort might result from the selection criteria for the admission, as our hospital was the major tertiary referral center taking care of patients with most severe forms of COVID-19, as well as for those with other urgent conditions complicated with COVID-19. Indeed, almost 90% of patients had pneumonia, 82% required oxygen supplementation, and at peak pandemic almost 25% needed ICU admission or were receiving HFNC oxygenation outside ICU. In addition, the analyzed population was old, with median age of 74 and burdened by multiple co-morbidities. The limitations of our study are single center experience, selection of most severe/critical COVID-19 cases, or COVID-19 cases with comorbidities that required hospital level of care, which are representative of the tertiary COVID-19 hospital, and inability to longitudinally assess dynamics of particular measurements over time. In addition to this, we did not regularly perform liver imaging in patients with deranged LBT to furtherly explore liver status, given almost universal prevalence and self-limited course of deranged liver biochemistry in typical cases. Nevertheless, this study was performed in the one of the largest cohort of patients reported so far, representative for the Caucasian population with almost universal prevalence of pneumonia, all of whom underwent standardized diagnostic procedures and therapy. Therefore, the obtained results might be considered robust, with high statistical power.In conclusion, patients who are hospitalized due to COVID-19 usually have elevated LBTs at admission. Only elevated AST is the independent predictor of death. There is an incremental trend for higher rates of ICU admission, MV and 30-day mortality among the patients with more severe liver injury when compared to those with normal liver biochemistry.


Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a successful intracellular pathogen that is responsible for the highest mortality rate among diseases caused by bacterial infections. During early interaction with the host innate cells, M. tuberculosis cell surface antigens interact with Toll like receptor 4 (TLR4) to activate the nucleotide-binding domain, leucine-rich-repeat containing family, pyrin domain-containing 3 (NLRP3) canonical, and non-canonical inflammasome pathways. NLRP3 inflammasome activation in the alveoli has been reported to contribute to the early inflammatory response that is needed for an effective anti-TB response through production of pro-inflammatory cytokines, including those of the Interleukin 1 (IL1) family. However, overstimulation of the alveolar NLRP3 inflammasomes can induce excessive inflammation that is pathological to the host. Several studies have explored the use of medicinal plants and/or their active derivatives to inhibit excessive stimulation of the inflammasomes and its associated factors, thus reducing immunopathological response in the host. This review describes the molecular mechanism of the NLRP3 inflammasome activation in the alveoli during M. tuberculosis infection. Furthermore, the mechanisms of inflammasome inhibition using medicinal plant and their derivatives will also be explored, thus offering a novel perspective on the alternative control strategies of M. tuberculosis-induced immunopathology. Mycobacterium tuberculosis has proven to be a successful pathogen over the years, responsible for an estimated 1.2 million deaths, with 10 new million cases annually. This opportunistic pathogen is also accountable for another 0.2 million estimated deaths among HIV-TB co-infected individuals [M. tuberculosis as a human pathogen is due to the emergence, persistence and transmission of genetically diverse drug resistant strains in different geographic locations [ocations ,4, and focations .M. tuberculosis bacillus and the alveolar lining present an opportunity for containment of infection by innate cells, such as type I and type II pulmonary epithelial cells [M. tuberculosis is a successful pathogen due to its ability to bypass the host innate defence mechanisms, which eventually leads to the granuloma formation [In the initial stages of pulmonary TB infection, physical barriers and innate immune response are present to prevent and control the infection. Initial interaction between the al cells , residenal cells ,8. The hal cells ,10 that al cells . Residenal cells . Surrounal cells ,14. M. tormation .M. tuberculosis [M. tuberculosis cell surface lipid antigens with the host [M. tuberculosis, thus, offering an effective control strategy [Innate cells express a range of host pathogen recognition receptors (PRRs) that interact with pathogen associated molecular patterns (PAMPs) in rculosis ,17,18. Ithe host ,20. Actithe host ,22. Sevestrategy ,24. Parastrategy ,26. ThusM. tuberculosis infection models. The current review reveals the molecular mechanism behind inflammasome activation on resident alveolar macrophages and pulmonary epithelial cells that results in IL1 cytokine production, which have been shown to contribute to the alveolar inflammatory response during M. tuberculosis infection. Furthermore, medicinal plants and their active derivatives that can be explored to prevent overstimulation of the alveolar NLRP3 inflammasome and their mechanisms of action will be reported as a novel host-directed therapy against M. tuberculosis.Several studies have shown the effectiveness of medicinal plants and their bioactive derivatives as a promising approach to TB treatment since these are naturally sourced compounds with minimal side effects ,28,29,30M. tuberculosis PAMPs and mammalian PRRs is crucial for the bacilli to gain entry into the cells and initiate the pathogenesis process. Several studies have shown that macrophages express complement (CR) [M. tuberculosis infection. Reduced M. tuberculosis bacterial uptake was observed in macrophages isolated from CR3-knockout mice compared to the wild type [M. tuberculosis in human macrophages. Initially, the Fcγ receptors were thought to contribute to M. tuberculosis killing because they mediate uptake of Immunoglobulin G (IgG)-opsonised mycobacteria [M. tuberculosis uptake in macrophages through opsonic and non-opsonic phagocytosis.Initial interaction between ent (CR) ,37 and Fent (CR)  during Mild type . Moreoveild type ,41,42 habacteria , howeverbacteria . CollectM. tuberculosis surface [M. tuberculosis mannosylated LAM promotes phagocytosis in macrophages [M. tuberculosis in alveolar macrophages include Surfactant Protein A [M. tuberculosis surface as excellently reviewed by Faridgohar and Nikoueinejad [M. tuberculosis remains to be investigated as this can contribute to better understanding of the inflammatory response induced by genetically diverse clinical strains.Mannose receptors (MR) are transmembrane C-type  lectins that recognise and bind to the expressed mannose sugars in the lipoarabinomannan (LAM) of  surface ,45,46,47rophages , which mrotein A ,49, Dectrotein A ,51, NOD2rotein A , and CD1rotein A . TLRs [5rotein A ,55 are oueinejad . DespiteM. tuberculosis infection. Bermudez and Goodman [M. tuberculosis use microtubules and microfilament pathways to gain entry into pulmonary epithelial cells. This was shown by significant reduction of intracellular bacilli when these pathways were blocked [M. tuberculosis infection. Through the production of a Dectin-1 receptor protein, they showed that these non-phagocytic cells might use this receptor for recognition of M. tuberculosis. The M. tuberculosis strains of East-African Indian and Euro-American lineages both induced down-regulation of the NOD1 receptor of the A549 pulmonary epithelial cells at 72 h post-infection [M. tuberculosis. Mannose and DC-SIGN receptors have not been linked to M. tuberculosis invasion of pulmonary epithelial cells and this area remains to be investigated.Previously, not much was known about specific receptors expressed by pulmonary epithelial cells during  Goodman  initiall blocked . Lee et  blocked  revealednfection . HoweverM. tuberculosis cell wall-associated structures, such as acylated lipoproteins , ESAT-6, mycolic acid (TLR2), acylated lipomannan (TLR4), and trehalose dimycolates , respectively [M. tuberculosis. Inhibition of TLR2-mediated response by mycolic acid and mce1 operon mutant significantly reduce the production of both IL8 and MCP-1, which suggests that TLR2 is essential to effective inflammatory response of pulmonary epithelial cells to M. tuberculosis. Previously, we showed an increased expression of TLR2, TLR3, TLR5, and TLR8 in pulmonary epithelial cells at 48 h post-infection by genetically diverse M. tuberculosis clinical strains of East-Asian and Euro-American lineages, leading to production of a diverse range of inflammatory cytokines. Upregulation of TLR4 was only induced by the Beijing (East-Asian) and F11 (Euro-American) strains, while TLR1, TLR6, TLR7, and TLR9 were downregulated by all strains [TLRs are involved in the recognition of different ectively . Sequeirectively  showed t strains .M. tuberculosis at 72 h post-infection of A549 pulmonary epithelial cells. Furthermore, the East-African Indian strain induced increased expression of TLR2, TLR5, and TLR9; while the Euro-American strain induced down-regulation of TLR4, TLR5, and TLR9. Both transcriptomic studies [M. tuberculosis in pulmonary epithelial cells at 48 and 72 h post infection and these PRRs may be the main receptors used by this pathogen to gain entry into these cells.A transcriptomic study performed by Hadifa et al.  revealed studies ,60 indicM. tuberculosis PAMPs, the MyD88, TIR-domain-containing adapter-inducing interferon-β (TRIF), TRIF-related adapter molecule (TRAM), Toll-interleukin 1 receptor domain containing adaptor protein (TIRAP), B-cell adaptor for PI3K (BCAP), and sterile alpha and TIR motif containing (SARM) are adaptor molecules that are central components of the TLR signalling pathway [M. tuberculosis induced high enrichment of the TLR signalling pathways in pulmonary epithelial cells with increased expression of the MyD88 adaptor molecule [M. tuberculosis by alveolar macrophages to the MyD88-signalling pathway [M. tuberculosis surface antigens with TLR4 triggers the NF-kB signalling pathway to induce expression of IL1β and IL18, whose maturation and production is dependent on the inflammasome assembly [Upon stimulation of the alveolar macrophages and pulmonary epithelial cells TLRs by  pathway . Previoumolecule . Several pathway ,64 with  pathway ,66. Thus pathway . Moreoveassembly .M. tuberculosis [Inflammasome is a multi-protein complex intracellular structure, which induces maturation of inflammatory cytokines, IL1β, and IL18 through the activation of caspase-1 and caspase-4/11 . The actrculosis ,72. The rculosis ,67.M. tuberculosis activate TLR4 that initiate the canonical inflammasome in alveolar macrophages and pulmonary epithelial cells. Infection by genetically diverse clinical strains of M. tuberculosis revealed an upregulation of NFKB1 and NFKB2 transcriptional factors that lead to the transcription of IL1β, while IL18 was slightly down-regulated at 48 h infection by East-Asian and Euro-American lineages [M. tuberculosis Beijing strains of the East-Asian lineage were differentially recognised by TLR2 and TLR4 in macrophages resulting in distinct cytokine profiles, including IL1β [M. tuberculosis interact with the TLR4, resulting in the activation of the NF-κB transcriptional factor, which is responsible for the transcription of pro-inflammatory cytokines, including IL1 by the alveolar macrophages and pulmonary epithelial cells. Furthermore, M. tuberculosis lineage-specific differences of the canonical inflammasome activation suggest that genetically diverse strains may induced diverse inflammasome patterns in the alveolar lining.The canonical inflammasome pathway is initiated through the activation of caspase 1 by NLRP3, which responds to stimulation by IL1R, Tumor necrosis factor (TNF) or a TLR ligand that binds its cognate receptor, which results in the translocation of NF-κB into the nucleus. Thereafter, the expression of NLRP3 and pro-IL1β are induced, while pro-IL18 is constitutively expressed within cells. Several studies ,55,58,60ing IL1β , activating IL1β . CollectM. tuberculosis [M. tuberculosis-stimulated pulmonary epithelial cells, which indicated the upregulation of IL18 expression at both transcriptional and post-transcriptional levels, suggesting the involvement of CASP1 enzymatic activity [Mycobacterium tuberculosis curli pili (MTP) mutant and complemented strains of M. tuberculosis [Mycobacterium and it associated factors is well described and characterised in alveolar macrophages [The second signal of the canonical inflammasome pathway involves the assembly of the NLRP3 inflammasome complex, which results in the recruitment of adaptor molecule, CARD and pro-caspase-1 in order to induce the processing and secretion of IL1β and IL18 cytokines. Pulmonary epithelial cells exhibit increased regulation of NLRP3 (by only F11 and H37Rv strains) PYCARD (ASC) and CASP1 during early infection by clinical strains of rculosis . This uprculosis  in pulmoactivity . There wrculosis , suggestrculosis ,60. Stimrophages ,77,78, arophages .Citrobacter rodentium, Escherichia coli, Legionella pneumophila, Salmonella typhimurium, and Vibrio cholera [Leishmania amazonensis [M. tuberculosis in pulmonary epithelial cells from transcriptomics and proteomics studies; this area remains to be investigated. However, the NLRP3 CASP4/11-dependant inflammasome is activated in macrophages during M. tuberculosis infection [M. tuberculosis stimulate alveolar macrophages canonical and non-canonical NLRP3 inflammasome, leading to production of inflammatory cytokines, IL-1β, and IL18 in a TLR4-NF-kB dependent manner. Furthermore, despite increased expression of the NLRP3 inflammasome transcripts and subsequence IL1 cytokine production in pulmonary epithelial cells, the presence of this complex and other types of inflammasomes such as NLRP6, NLRC4, and AIM2 remains to be investigated and confirmed in future studies. The activation of a canonical and non-canonical alveolar NLRP3 molecular mechanism during M. tuberculosis infection is depicted in Non-canonical NLRP3 inflammasome mediated by CASP4/11  has been cholera  and othezonensis . The patnfection . CollectM. tuberculosis infections. However, this mechanism remains to be confirmed for pulmonary epithelial cells. IL1 are among the group of pro-inflammatory cytokines that are essential in host defence against M. tuberculosis [M. tuberculosis and BCG strains [M. tuberculosis, while blocking IL1β production relieved pulmonary inflammation [M. tuberculosis that is pathological to the host, thus novel strategies must be exploited to regulate this response.It is apparent that alveolar macrophages produce IL1 cytokines ,76,82,83rculosis . Mice wirculosis . IL18 isrculosis ,85 and hrculosis ,87. More strains . Despite strains  that canammation . Signifiammation , while hammation . TherefoM. tuberculosis infection [M. tuberculosis infections. Medicinal plants and/or their bioactive derivatives that are used to inhibit NLRP3 inflammasome gene transcription, formation of the inflammasome complex and production of NLRP3 inflammasome proteins (including IL1 cytokines) are reviewed below.The high prevalence of TB worldwide is partly due to development of drug resistant strains, rendering the current treatment regimen ineffective ,94,95. Snfection ,101,102.nfection . TherefoMichelia compressa and Michelia champaca plants from the Magnolia family [M. tuberculosis infection. The NF-κB transcriptional factor and NLRP3 inflammasome that are essential for transcription of IL1β, IL18, and pro-caspase 11 transcripts were downregulated, accompanied by reduction of inflammatory cytokines (IL1β and TNF-α) in a dose-dependent manner. This study concluded that the micheliolide downregulates the activation of NLRP3 inflammasome by modulating the inflammatory response induced by M. tuberculosis through the PI3K/Akt/NF-κB pathway.a family  possess a family ,107,108.a family  used an Garcinia yunnanensis Hu plant [M. tuberculosis H37Ra (avirulent) infected macrophages. This inhibition resulted in significantly reduced TLR-NF-κB inflammatory mediators, IL1β, TNF-α, IL6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) [M. tuberculosis.NLRP3 inflammasome-specific transcript can be inhibited through the adaptor molecules that signal to activate transcriptional factor, NF-κB. Guttiferone K is the active compound isolated from Hu plant  and has Hu plant . The ant (COX-2) . The GutM. tuberculosis models. These include Arctigenin [M. tuberculosis in vitro and in vivo models to identify their potential use as host-directed immunomodulatory plant derivatives that can be used to prevent lung pathology during infection.There are many medicinal plant bioactive derivatives that act as NLRP3 inflammasome transcript inhibitors identified in non-ctigenin , Silymarctigenin , Rutin [ctigenin , Genistectigenin , Aloe emctigenin , Anemosictigenin , epigallctigenin , Resveractigenin , Silibinctigenin , Artemisctigenin , Icariinctigenin , Polydatctigenin , Cinnamactigenin , Huangkuctigenin  whose adAndrographis paniculate was observed in macrophages co-cultured with pulmonary epithelial cells that had significantly reduced IL1β cytokine production during M. tuberculosis infection. This IL1β reduction in macrophages resulted in declining chemokine (IL8 and MCP-1) expression in neighbouring pulmonary epithelial cells. The proposed molecular mechanism behind IL1β inhibition is the activation of macrophage autophagy to degrade NLRP3, thus inhibiting the inflammasome activation and IL1β production. Moreover, andrographolide downregulated the phosphorylation of Akt/mTOR and NF-𝜅B p65 subunit, which will interfere with the NLRP3 inflammasome transcript expression that is essential for the maturation of the inflammasome complex [Andrographis paniculate plant [Medicinal plants and their derivatives can exert their anti-inflammatory activity in the NLRP3 inflammasome by directly inhibiting production of proteins that regulate this inflammatory pathway, including production of IL1 cytokines. The anti-inflammatory activity of andrographolide, the labdane diterpenoid isolated in the stems, and leaves of  complex . Androgrte plant  with mante plant .Panax ginseng, Curcuma longa, Gardenia jasminoides, Mangifera indica, Salvia miltiorrhiza medicinal plants, respectively.Several in vitro and in vivo models have identified other medicinal plant derivatives that have been found to inhibit NLRP3 inflammasome proteins and subsequent IL1 production; and these include Ginsenosides (Retinoblastoma and Compound K) ,128, CurScutellaria baicalensis possess an anti-NLRP3 inflammasome activity during M. tuberculosis through decreased phosphorylated protein kinase B and mammalian target of rapamycin, decreasing NLRP3 inflammasome and subsequently IL1β production [Abrus precatorius subsp. Africanus, Ficus sur, Pentanisia prunelloides, and Terminalia phanerophlebia were shown to inhibit the COX-2 enzyme, thus may contribute to the anti-NLRP3 inflammasome response during M. tuberculosis infection [Several medicinal plants and/or their bioactive derivatives can interfere with the formation of the inflammasome complex or the components that are needed for a complete NLRP3 inflammasome to facilitate maturation of the IL1 cytokines. The flavonoid Baicalin, isolated from oduction . The COXoduction . The crunfection . The bioQuamoclit angulate plant [M. tuberculosis models exploring these bioactive plant derivatives remain to be investigated for their potential use as regulators of the alveolar NLRP3 inflammasome.NLRP3 inflammasome complex inhibitors in other models include Sweroside , Oridonite plant , 4- Sulfte plant , Obovatote plant , Berberite plant  (Figure M. tuberculosis infection models. There is a growing understanding that M. tuberculosis infection is a multifaceted condition that is also driven by the host inflammatory response. Thus, controlling the alveolar NLRP3 inflammasome seem to be one of the novel strategies for host directed control measures of pathological response induced by M. tuberculosis.There is growing appreciation of medicinal plants and their bioactive molecules in management of inflammatory response regulated by the NLRP3 inflammasome as excellently reviewed by several authors ,162,163.M. tuberculosis that results in the production of IL1 inflammatory cytokines. Overstimulation of the alveolar NLRP3 inflammasome may contribute to the exaggerated inflammatory response that is pathological to the host. Recently, several studies have shown that medicinal plants and/or their derivatives, Andrographolide, Baicalin, Micheliolide, Guttiferone K, Abrus precatorius subsp. Africanus, Ficus sur, Pentanisia prunelloides, and Terminalia phanerophlebia can inhibit the NLRP3 inflammasome at transcriptional and post-transcriptional levels in M. tuberculosis models. Therefore, these plants and bioactive derivatives can be explored for anti-TB host directed adjuvants to control the M. tuberculosis-induced inflammatory response. Furthermore, many studies have identified many regulators of the NLRP3 inflammasome that remains to be tested on both in vitro and in vivo M. tuberculosis models. Future studies should investigate the ideal route of administration of these medicinal plants and their bioactive derivatives due to challenges associated with oral availability of medicinal drugs. Exploring alternative strategies such as using medicinal plants and their bioactive derivatives to regulate the host inflammatory response in TB may bring us closer to winning the battle against M. tuberculosis.The activation of the NLRP3 inflammasome in alveolar macrophages and potentially in pulmonary epithelial cells is an essential response of innate cells to


Lifestyle improvement, including through exercise, has been recognized as an important mode of therapy for the suppression of diabetic kidney disease (DKD). However, the detailed molecular mechanisms by which exercise exerts beneficial effects in the suppression of DKD have not yet been fully elucidated. In this study, we investigate the effects of treadmill exercise training (TET) for 8 weeks  on kidney injuries of type 2 diabetic male rats with obesity (Wistar fatty (fa/fa) rats: WFRs) at 36 weeks of age. TET significantly suppressed the levels of albuminuria and urinary liver-type fatty-acid-binding protein (L-FABP), tubulointerstitial fibrosis, inflammation, and oxidative stress in the kidneys of WFRs. In addition, TET mitigated excessive apoptosis and restored autophagy in the renal cortex, as well as suppressed the development of morphological abnormalities in the mitochondria of proximal tubular cells, which were also accompanied by the restoration of AMP-activated kinase (AMPK) activity and suppression of the mechanistic target of rapamycin complex 1 (mTORC1). In conclusion, TET ameliorates diabetes-induced kidney injury in type 2 diabetic fatty rats. The number of patients with type 2 diabetes mellitus (T2DM) has been increasing worldwide. Vascular complications due to diabetes include microvascular disorders, including retinopathy, neuropathy, and kidney disease, and macrovascular disorders caused by atherosclerosis . Among tLifestyle interventions serve as important basic treatments for patients with T2DM and DKD. Among the various lifestyle interventions, increasing physical activity or regular exercise is generally a widely accepted method for the improvement of health outcomes, including overall well-being, quality of life, and cardiometabolism . In CKD For this study, we investigated whether regular physical exercise using a treadmill may exert renoprotective effects, focusing on the alteration of anti-inflammatory/oxidative stress, apoptosis, and autophagy in Wistar fatty (fa/fa) rats.Kidney injury molecule 1 (KIM-1) antibody was purchased from R&D Systems, Inc. . Cluster of differentiation 68 (CD68) antibody was purchased from Serotec . Nitrotyrosine antibody was purchased from Trans Genic Inc. . Nuclear factor-erythroid 2-related factor 2 (Nrf2) antibody was purchased from Abcam . Heme oxygenase 1 (HO-1), cleaved caspase 3, phospho-S6 ribosomal protein (Ser 235/236) (p-S6RP), S6RP, phospho-AMP-activated protein kinase (Thr172) (p-AMPK), and AMPK antibodies were obtained from Cell Signaling Technology . p62/SQSTM1 (p62) was obtained from Medical & Biological Laboratories . β-actin was purchased from Sigma-Aldrich .The study was approved by the Research Center for Animal Life Science of Kanazawa Medical University. All experiments were performed in accordance with the principles of laboratory animal care. Male and female Wistar lean (fa/+) rats  were maintained in temperature-controlled (23 ± 1 °C) rooms on a 12-h light/dark cycle with free access to water and chow, as previously described .n = 8), non-diabetic and non-exercise control; (2) sedentary WFRs ; and (3) WFRs with treadmill exercise training . Rats in the WFRs-T group were allowed to run on a 0% gradient treadmill  [2max), as previously reported [At 34 weeks of age, male non-diabetic WLRs and male diabetic Wistar fatty (fa/fa) rats (WFRs) were randomly divided into three groups: (1) sedentary WLRs  . The trereported ,20. Bodyreported . Rats wereported . At the HbA1c levels were measured by a DCA 2000 Analyzer  at the end of the experiment ,21. UrinParaffin sections of the kidney were stained with Masson’s trichrome (MT) reagent, and immunohistochemical staining was performed by using KIM-1 antibodies (1:100), CD68 antibodies (1:50), and nitrotyrosine antibodies (1:100) . For theImmunoblotting was performed by using nitrotyrosine antibodies (1:1000), Nrf2 antibodies (1:1000), HO-1 antibodies (1:1000), cleaved caspase 3 antibodies (1:1000), p62/SQSTM1 antibodies (1:1000), p-S6RP antibodies (1:2000), S6RP antibodies (1:1000), p-AMPK antibodies (1:1000), and AMPK antibodies (1:1000). Isolation of total RNA from the renal cortex, cDNA synthesis, and quantitative real-time PCR were performed [p < 0.05 was considered to indicate significant differences.Data are expressed as means ± standard deviation (SD). ANOVA followed by Tukey’s multiple comparison test was used to determine the significance of pairwise differences among the three groups. The characteristics of the considered rats at 44 weeks of age are shown in Renal fibrosis, evaluated by MT staining, and collagen III expression in the renal cortex were significantly increased in the kidneys of the WFRs compared with the WLRs A–C. In aThe intensity of CD68 immunohistochemical staining in tubulointerstitial areas A,B and mThe intensity of nitrotyrosine immunohistochemical staining in the kidney of the WFRs was higher than that of the WLRs. In addition, the nitrotyrosine, Nrf2, and HO-1 protein levels were significantly increased in the renal cortex of the WFRs compared with the WLRs A–G. TET Mitochondrial morphological changes, including fragmentation or swelling, were observed in the proximal tubular cells of WFRs. TET suppressed the development of all these mitochondrial alterations A. The clIn this study, TET ameliorated urinary albumin and L-FABP excretion as well as kidney injuries, including tubular cell damage and tubulointerstitial fibrosis, inflammation, and oxidative stress, in WFRs. In addition, TET resulted in improvements by mitigating excessive apoptosis and restoring impaired autophagy in the renal cortex as well as mitochondrial morphology with respect to abnormalities in proximal tubular cells, which were also accompanied by AMPK activation and mTORC1 suppression. The beneficial effects on the kidney of TET were exerted independent of glucose levels, because there was no difference in the levels of HbA1c and blood glucose under ad libitum feeding between sedentary WFRs and WFRs with TET.2−). Moreover, HO-1 is regulated by Nrf2, a key transcription factor in the regulation of cellular redox balance [yA  mice [Chronic inflammation and oxidative stress play crucial roles in DKD pathogenesis and are closely linked to tubular cell damage and fibrosis ,4,23. In balance  that fun balance . In this balance ,21,26—a A y mice .Accumulating evidence demonstrates that the reduction in AMPK activity and mTORC1 activation in animals and humans, under caloric excess or diabetic conditions, is involved in the impairment of autophagy ,28,29,30In our study, TET reduced fat weights and increased gastrocnemius muscle weights, as observed in the WFRs-T, compared with the WFRs. However, HbA1c levels did not significantly differ between the WFRs with TET and sedentary WFRs. Thus, TET might provide a greater renoprotective effect independent of glucose levels. Previously, Gosh et al. also demonstrated that exercise using a motorized exercise wheel system for 1 h every day in db/db mice improved diabetes-induced kidney injuries including renal cell excess apoptosis, without changes in plasma glucose or insulin status . In our This study had several limitations, as follows: We performed this study using a level of exercise intensity that did not result in the exhaustion of the rats . The intensity of exercise was considered to be moderate and approximately 50–60% of the maximal oxygen consumption . HoweverTET ameliorated functional and histological kidney injuries in WFRs, including albuminuria, increased urinary L-FABP excretion, tubular cell damage, and tubulointerstitial fibrosis, inflammation, and oxidative stress. In addition, TET mitigated excessive apoptosis and restored autophagy in the renal cortex as well as mitochondrial morphology in proximal tubular cells, which was accompanied by AMPK activation and mTORC1 suppression. TET might also provide a greater renoprotective effect independent of glucose levels. However, further studies are necessary to clarify a causality among the TET-induced improvement of inflammation/oxidative stress, decrease in the excess of apoptosis, and the restoration of autophagy through AMPK activation and mTORC1 suppression, in the kidney of WFRs.


Leber’s congenital amaurosis (LCA), one of the most severe inherited retinal dystrophies, is typically associated with extremely early onset of visual loss, nystagmus, and amaurotic pupils, and is responsible for 20% of childhood blindness. With advances in molecular diagnostic technology, the knowledge about the genetic background of LCA has expanded widely, while disease-causing variants have been identified in 38 genes. Different pathogenetic mechanisms have been found among these varieties of genetic mutations, all of which result in the dysfunction or absence of their encoded proteins participating in the visual cycle. Hence, the clinical phenotypes also exhibit extensive heterogenicity, including the course of visual impairment, involvement of the macular area, alteration in retinal structure, and residual function of the diseased photoreceptor. By reviewing the clinical course, fundoscopic images, optical coherent tomography examination, and electroretinogram, genotype-phenotype correlations could be established for common genetic mutations in LCA, which would benefit the timing of the diagnosis and thus promote early intervention. Gene therapy is promising in the management of LCA, while several clinical trials are ongoing and preliminary success has been announced, focusing on RPE65 and other common disease-causing genes. This review provides an update on the genetics, clinical examination findings, and genotype-phenotype correlations in the most well-established causative genetic mutations of LCA.  Inherited retinal dystrophies (IRDs) are a group of diseases caused by genetic mutations that are characteristic of photoceptor dysfunction and eventual death of retinal cells. Different extents of retinal degeneration result in widely varied presentations, from milder night blindness or color blindness to profound visual impairment . Among tGUCY2D) for studies published before 30 June. We included articles published in Science Citation Index (SCI) journals and focused on reporting the clinical presentation of LCA cases with specific genetic mutations. Thus, this review aims to summarize the clinical manifestations and genetic updates of LCA, as well as show the genotype-phenotype correlations in LCA to promote early diagnosis and facilitate bridging therapies in the near future. To establish the diagnosis of LCA in children with suspected IRDs, detailed ophthalmic history, imaging studies, electrophysical examinations, and, most importantly, molecular confirmation is needed. Compared to other retinal dystrophies, LCA is even more heterogeneous in its disease course and presentation. A milder phenotype of LCA with relative visual preservation has been considered as early-onset severe retinal dystrophy, although their genetic basis overlaps significantly . Owing tGUCY2D mutation. The patient may present with photophobia or nyctalopia, which could be gene specific, as described by Hanein et al. [Typically, patients with LCA present with poor vision and nystagmus or the absence of fixation at as early as 6 weeks of age. Manifestation of vision loss in patients with LCA ranges from functional visual acuity to light perception only, and approximately three-quarters of these are stationary. Transient fluctuation in vision with delayed visual maturation is occasionally observed in the first or second decade of life until progressive decline occurs . The mosn et al. . Similarn et al. .GUCY2D, and a residual rod response in RPE65 [CRB1, GUCY2D). The disrupted anatomical structure of the retina includes decreased thickness in different layers, especially in the outer nuclear layer (ONL), loss of integrity in the ellipsoid zone, and disorganized macular atrophy [GUCY2D and RPE65, which indicates a possible therapeutic window for genetic remodeling [Electrophysical studies are important in assessing patients with suspected LCA in their early life. Although a non-detectable electroretinogram (ERG) response is typical in LCA, a residual cone response could be detected with mutation in in RPE65 . Fundoscin RPE65 . Originain RPE65 . The ret atrophy . The cenmodeling . In summary, typical findings defining LCA includes (1) early and severe reduction of vision (from non-light perception to 20/400) associated with nystagmus in a large number of patients; (2) fundus appearance, ranging from normal, maculopathy, to typical retinitis pigmentosa (RP)-like abnormalities; and (3) severely reduced to non-detectable full field ERG responses . BecauseGUCY2D, in LCA cases, the Online Mendelian Inheritance (OMIM) database has identified 19 types of LCA with mutations in specific genes [Linkage analysis, identity-by-descent mapping, and the candidate gene approach are used to identify the mutation loci in LCA cases . Since tic genes . The incCRX, IMPDH1, and OTX2 [GUCY2D, AIPL1, RD3, KCNJ13), retinoid cycle , ciliary transportation , photoreceptor morphogenesis , guanine synthesis (IMPDH1), and photoreceptor differentiation (OTX2) [USP45 (LCA type 19) and other genes is still controversial and further studies are required [With the advances in molecular genetics, approximately three-quarters of the LCA cases could be identified with specific mutations ,18. Mostand OTX2 . These wn (OTX2) . The mecLCA type  and otheGUCY2D, RPE65, CRB1, CEP290, and RDH12 [CEP290, GUCY2D, and RPE65 are generally more frequent in Caucasian populations [CRB1, which accounts for 13.6% of LCA cases, has been reported as the leading causative genetic mutation in Chinese cohorts, followed by GUCY2D [CEP290 than in the European cohort, but more in RPGRIP1. The most common variant of the CEP290 mutation found in the European population, c.2991+1655A>G, was not detected in the Chinese cohort [CRB1, NMNAT1, and RPGRIP1 were reported as the most common mutations in a Japanese cohort [CEP290 and GUCY2D were the leading causative mutations in Australia, followed by NMNAT1 [Common mutations, usually occurring with a frequency of approximately 10% or higher, include nd RDH12 . The freulations ,21,22. Oy GUCY2D . Meanwhie cohort . CRB1, Ne cohort . CEP290 y NMNAT1 . Moreovey NMNAT1 . These fAlthough patients with LCA exhibit an extended range of intra- and interfamilial heterogenicity, genotype-phenotype correlations have been identified in several common mutations ,16. EstaGUCY2D, the first gene that was identified to be correlated with LCA, is localized on chromosome 17p13.1, and encodes retinal guanylate cyclase-1, which is involved in photoreceptor recovery phase in phototransduction [GUCY2D is expressed in the photoreceptor outer segment with a higher level in cone cells than in rod cells [GUCY2D mutation accounts for 6–21% of LCA cases. GUCY2D-associated LCA cases are known to have very poor but stationary vision early in life, with nystagmus, oculodigital signs, and prominent photophobia [GUCY2D result in complete deficiency of enzyme function [GUCY2D mutation on exon 2, which encodes the extracellular domain, had a milder phenotype with better visual acuity [GUCY2D mutation [sduction . GUCY2D od cells . The GUCtophobia ,29. Apprtophobia . While nl acuity . Althougl acuity . High hyl acuity ,31. Fundl acuity . Despitel acuity . The fovl acuity . The ONLl acuity . In patil acuity . This fumutation . RPE65 gene on chromosome 1p31.2 encodes retinoid isomerase, which is a 65 kDa enzyme expressed abundantly in RPE and is responsible for vitamin A metabolism in the retinoid cycle [RPE65 mutation accounts for 4–16% of LCA cases, which is relatively rare in the Chinese population but more prevalent in the Caucasian and Indian populations [RPE65-associated LCA includes profound nyctalopia at birth; however, some residual cone-mediated vision has been observed in these cases. Therefore, possibly an alternative supply of 11-cis-retinal, apart from RPE65, in cone photoreceptors has been proposed [RPE65-associated LCA cases present with vision worse than counting fingers initially, and transient improvement could be noted in early life. However, visual impairment gradually progresses from the second to the third decade of life [RPE65 mutation [The id cycle . This deulations ,37,38. Tproposed . About 1 of life ,39,40. P of life . Myopia mutation . Depigmemutation ,42,43. Pmutation . Preservmutation ,45. Thinmutation ,44. HypeCRB1 gene on chromosome 1q31.3 encodes Crumbs homologue 1, which is expressed in the inner segment of the photoreceptors and Muller cells to maintain adequate morphogenesis and polarity in retinal development [CRB1 gene have been associated with multiple retinal dystrophies, including RP, foveal retinoschisis, macular dystrophy, and LCA. Mutations within the Ca2+-binding EGF-like domain are often associated with more severe disease [CRB1 mutations, which are higher in the Chinese population [CRB1-associated LCA cases present relatively preserved visual function, nyctalopia, and high hyperopia [CRB1-associated RP cases, coats-like vasculopathy could also be present in LCA with CRB1 mutations [CRB1 mutation, which indicates retinal disorganization caused by developmental defects [CRB1 mutations from other types of LCA and facilitates genetic diagnosis. The ERG response is generally diminished in these cases [The elopment . Therefo disease ,47. Apprpulation ,23. CRB1yperopia ,49. Simiutations . Thicken defects ,49. ThisCEP290 gene on chromosome 12q21.32 encodes a 290 kD centrosomal protein that localizes to the basal bodies of the photoreceptors connecting the cilium and is responsible for its formation, stability, and transportation function. Mutations in CEP290 are associated with several systemic diseases, including Joubert syndrome, Senior-Løken syndrome, and Meckel syndrome [CEP290-associated LCA cases in which the majority (62–89%) of patients had only light perception or less [CEP290 mutation demonstrate an extinguished ERG response.The syndrome ,51. The syndrome . Vision  or less . While n or less . Photoph or less . Other a or less . Apart f or less ,14. Cide or less . Cyst-liRDH12 gene on chromosome 14q24.1, is expressed predominantly in the inner segment of the photoreceptor and participates in the retinoid cycle by converting 11-cis retinol to 11-cis retinal. Mutations in RDH12 account for approximately 3–10% of LCA cases [RDH12-associated LCA cases demonstrate poor but functional vision with transient improvement lasting less than 10 years. Progressive decline in vision occurs after the second decade of life [Retinol dehydrogenase 12, which is encoded by the CA cases ,15. RDH1 of life . Other r of life . Garg et of life . A thinn of life ,59. An e of life .SPATA7 gene on chromosome 14q31.3 encodes a protein with a single transmembrane protein, termed spermatogenesis-associated protein 7. SPATA7 is expressed in the cytoplasm of the inner segment of photoreceptors of mature retina in animal models, but its function in the visual cycle has not been fully elucidated [SPATA7 has been identified not only in LCA but also in juvenile RP cases, while the severity of visual impairment largely differs [SPATA7 has been reported to be low as 3% in LCA cases [SPATA7, which had a phenotype resembling RPE65-associated LCA. The cases presented with stationary poor vision, nystagmus, nyctalopia, and hyperopia. Fundus examination revealed peripheral chorioretinal atrophy, bone spicule pigmentation, and a central foveal hyperautofluorescent ring. ERG was extinguished in both the rod and cone responses [The ucidated . The mut differs . The preCA cases . Perraulesponses ,61.AIPL1 gene on chromosome 17p13.2 encodes Aryl-hydrocarbon-interacting-protein-like 1, which is a cochaperone specific to photoreceptors that stabilize retinal cGMP phosphodiesterase in the phototransduction process. Both the rod and cone photoreceptors express the AIPL1 gene during development, but only the rod photoreceptor continues its expression in the adult retina [AIPL1 produce an LCA phenotype, while a frameshift mutation (class III) results in the milder phenotype of juvenile RP [AIPL1 account for about 5% of LCA cases. The variant p.W278X (c.834G>A) has been commonly reported [AIPL1-associated LCA cases is considered to be severe, with rapidly progressive visual impairment to only light perception in most patients. Patients often present with nystagmus, photophobia, photoattraction, keratoconus, and PSC [AIPL1 mutation [The t retina . Missensenile RP . Mutatioreported ,62,63. T and PSC . Althoug and PSC ,66,67. T and PSC ,65,68. Emutation . LCA5 gene on chromosome 6q14.1 encodes lebercilin, which is expressed widely in the tissues composed of cilia, including photoreceptors. However, mutations in LCA5 have been identified to be confined to retinal dystrophy without systemic association, while most of the cases meet the diagnosis of LCA [LCA5 mutation accounts for only 1–2% of the LCA cases [LCA5-associated LCA cases present with impaired vision, nyctalopia, and severe hyperopia. Fundus examination typically reveals diffuse mottling of the RPE at the periphery when aging and a frankly normal macula [RPE65 phenotype, were common in the case series presented by Mackay et al. [LCA5 mutation, which exhibited coloboma-like macular atrophy [The s of LCA ,71. The CA cases . LCA5-asl macula . Jacobsol macula . Scattery et al. . An ERG  atrophy .RPGRIP1 gene on chromosome 14q11.2 regularly binds to the RP GTPase at the C-terminal interacting domain, to anchor on the connecting cilium in the retinal photoreceptor cells [RPGRIP1 account for approximately 5% of LCA cases when null mutations occur, while less severe cone-rod dystrophy results from variants causing missense mutations [RPGRIP1-associated LCA cases present with gradually declining impaired vision, photophobia, and nystagmus. Attenuated vessels, chorioretinal atrophy, and peripheral pigmentation may be observed on fundus examination in childhood [The protein product of the or cells ,75. Mutautations . RPGRIP1hildhood . Granulahildhood . OCT imahildhood ,79. Howehildhood . In prevhildhood ,78. CRX) gene on chromosome 19q13.3 encodes a homeobox transcription factor, which is essential for early ocular development by regulating the differentiation and maintenance of photoreceptor cells [CRX accounts for up to 1% of LCA cases, which is autosomal dominant [The cone-rod homeobox  [IMPDH1-associated LCA cases that presented with mottling or depigmented fundus. One of the patients also had systemic hypotonia and growth retardation [The protein product of the l tissue . Mutatioreported . IMPDH1-% (2/24) . They reardation . RD3 gene on chromosome 1q32.3 is only expressed in retinal photoreceptors and is essential for the transportation of guanylate cyclase between the inner and outer segments [RD3 gene [RD3-associated LCA cases present nyctalopia and poor vision at birth, with progressive decline to barely precepting light. The visual function is comparably reduced as in the GUCY2D mutation because of the sharing dysfunction in guanylate cyclase [The segments . Less thRD3 gene ,90. RD3- cyclase . Central cyclase . LRAT gene on chromosome 4q32.1, which converts all-trans retinol to all-trans retinyl esters in the RPE phase of the retinoid cycle. Mutations in the LRAT gene have been reported in approximately 1–2% of LCA cases, and their phenotype is similar to that of the RPE65 mutation [LRAT-associated LCA cases usually present with nyctalopia with poor vision but absence of photophobia. On fundus examination, a grossly normal but mild mottling appearance is observed in the posterior pole, and a demarcation margin with diffuse granular depigmentation can be seen outside the macula [Lecithin retinol acyltransferase is the product of the mutation ,93. LRATe macula . Borman e macula . TULP1 gene on chromosome 6q21.31, which is expressed specifically in the retina and is localized in the cilia of the photoreceptor’s inner segment, including both rods and cones. Mutations in TULP1 account for approximately 1% of LCA cases [The tubby-like protein 1 is encoded by the CA cases . TULP-asCA cases . A hyperCA cases . The ERGCA cases ,96. KCNJ13 gene on chromosome 2q37.1 encodes protein Kir7.1, which is localized on the apical process of RPE and functions as a potassium channel [KCNJ13 present with nyctalopia, nystagmus, and cataracts [The  channel . LCA casataracts . Funduscataracts ,99. PRPH2 gene on chromosome 6p21.1 encodes peripherin 2, which is present in the disc rims of the outer segment of the photoreceptors. Mutations in PRPH2 are related to RP, retinitis punctata albescens, and several macular dystrophies [PRPH2 has been reported in LCA in a limited manner. Poor vision, nyctalopia, and nystagmus have been observed in the reported case [The trophies . Mutatioted case ,102. Funted case . The famted case . The ERGIQCB1 gene on 3q13.33 encodes nephrocystin, which is localized in the cilia in the kidney and retinal photoreceptors and interacts with CEP290 in ciliary function [IQCB1 have onset of LCA in early childhood, and some of them develop renal dysfunction in adolescence [The function . Patientlescence . These plescence . Eccentrlescence . OTX2 and GDF6 are more known for severe ocular malformations, such as microphthalmia, because of their essential role in embryonic ocular development; however, milder cases with retinopathy have been diagnosed as LCA as well [IFT140 gene cause ciliopathy, which is associated with retinal, renal, and skeletal abnormalities  [Many LCA-associated mutations have a relatively lower prevalence, and limited cases have been reported in the literature. Mutations in  as well ,107. Mutyndrome) . HoweverGUCY2D, RPE65, AIPL1, RPGRIP1, LCA5, CEP290, and RDH12, mostly by using adeno-associated virus (AAV) vector-mediated or lentiviral-vector-mediated gene augmentation therapy [GUCY2D, RPE65, and CEP290 are ongoing [RPE65 mutations [RPE65 gene to restore the deficiency in the RPE65 protein in patients with LCA with a biallelic RPE65 mutation. Early intervention is essential for success because more retinal cells may be viable during early life [Gene augmentation therapy is the main source of potential treatment for LCA. Animal studies have been conducted on commonly involved genes, including  therapy . Moreove ongoing . To dateutations ,112,113.rly life ; howeverrly life ,111. CEP290 gene. A pioneer CRISPR -mediated gene editing therapy focusing on the c.2991+1655A>G variant has been launched for phase I clinical trials. Another approach with intravitreal injection of mRNA, which serves as an anti-sense oligonucleotide for genetic editing, has been introduced to restore the expression level of CEP290 (NCT03140969) [LRAT- and RPE65-associated LCA cases [Being one of the most identified variants in western countries, gene therapy trials have also focused on c.2991+1655A>G in the 3140969) ,116. PhaCA cases . Stem-ceCA cases . LCA is the most severe and early onset form of IRDs and is a major cause of childhood blindness. Hence, it is important to find an effective treatment soon. However, its highly heterogeneous phenotypic and genetic spectra have made it difficult to diagnose and plan individualized treatment. Owing to the modern advances in next-generation sequencing, the efficiency and accuracy of identifying causative gene mutations have been largely elevated. Therefore, the genotype-phenotype correlations can be explored more precisely when incorporating clinical assessments and image examinations. Nonetheless, the correlations between phenotypical features and intra-genetic variability or ethnical difference are even less established and further research is required in the future. Here, we provide a comprehensive review, anticipating to enhance the diagnosis and treatment of potential LCA cases in clinical practice, as well as the development of available therapeutic interventions.