Streptococcus thermophiles, extensively utilized for producing dairy foods, has recently been regarded as a new promising probiotic candidate. In this study, the efficacy of Streptococcus thermophiles DMST-H2 (DMST-H2) for AAD treatment in mice was investigated. DMST-H2 was isolated from Chinese traditional yogurt, proved to be non-toxic, and presented tolerance against simulated gastrointestinal conditions in vitro. Additionally, genomic analysis revealed that it possessed genes related to acid tolerance, bile salt tolerance, adhesion, oxidative stress and bacteriocin production. The animal experiment results showed that both DMST-H2 treatment and natural recovery could reduce fecal water content. Compared with spontaneous recovery, DMST-H2 accelerated the recovery of the enlarged caecum and intestinal barrier injury from AAD, and further decreased endotoxin (ET), D-lactate (D-LA) and diamine oxidase (DAO) content in serum. Moreover, pro-inflammatory cytokines (TNF-α) were reduced, while interferon-γ (IFN-γ) and anti-inflammatory cytokines (IL-10) increased after treating with DMST-H2. Furthermore, DMST-H2 better restored the structure of intestinal flora. At the phylum level, Firmicutes increased and Proteobacteria decreased. These findings indicate that DMST-H2 could promote recovery in mice with antibiotic-associated diarrhea.Antibiotic-associated diarrhea (AAD) is the most common side effect of antibiotics and is routinely treated with probiotics in clinical.  Antibiotic treatment often causes diarrhea, which is called antibiotic-associated diarrhea (AAD). Various antibiotics result in AAD, especially aminopenicillins, cephalosporins, and clindamycin . AccordiLactobacillus rhamnosus GG and S. boulardii showed similar results [Lactobacillus rhamnosus GG had the best prevention effectiveness and tolerance on AAD, and Lactobacillus casei had better efficacy and medium tolerance in reducing the Clostridium difficile infection. Furthermore, the effects on AAD of both a single strain and a combination of strains were equal [Probiotics are non-pathogenic living microorganisms intended to colonize the intestinal tract and confer benefits on the host if given adequate amounts . Probiot results . A networe equal .Streptococcus thermophiles, one of the most considerable traditional fermented starters, is currently considered to have positive health effects [Streptococcus thermophiles were useful for the treatment and prevention of AAD. A commercial probiotic formula consisting of Bifidobacterium lactis and Streptococcus thermophilus reduced the frequency of AAD in infants significantly [Streptococcus thermophiles was efficacious in preventing AAD in average-risk patients [Streptococcus thermophiles improved the eradication rate of antibiotics to Helicobacter pylori and significantly minimized side-effect prevalence such as diarrhea and nausea/vomiting [Streptococcus thermophiles impacted the progression of C. difficile infection, which was present in 20% of AAD patients [Streptococcus thermophiles itself has an impact on AAD.  effects . Severalficantly . Probiotpatients . Probiotvomiting . Additiopatients . TherefoStreptococcus thermophiles DMST-H2 (DMST-H2) was isolated from a Chinese traditional yogurt, and showed potential probiotics. This study aims to evaluate the effects of DMST-H2 in AAD mice models from three aspects: (1) whether it can help to ameliorate the status of diarrhea; (2) whether it can help to repair tissue injury and regulate the inflammatory response; (3) whether it can help to regulate intestinal flora. This study enriches scientific understanding about DMST-H2 and facilitates its further application in antibiotic-associated diarrhea.−5 to 10−7) with sterile saline, and 100 μL of the dilution was plated on MRS ager medium . After anaerobic was cultured at 37 °C for 48 h, bacterial colonies were purified by re-streaking. The pure strains were cultured overnight in MRS medium  at 37 °C for 16S rDNA identification according to Yi et al. [The Chinese traditional yogurt was purchased from an Inner Mongolia family who makes traditional yogurt with no commercial starter. It was diluted  with a dose of 10.0 g/kg bw. Experimental observation of toxicity signs and death lasted for 14 days, and the mice were weighed every 7 days [An acute oral toxicity test of DMST-H2 was carried out by Guangzhou Quality Supervision and Testing Institute . Twenty healthy specific pathogen free Kunming (SPF KM) mice  were supplied by Ji’nan Peng Yue Laboratory Animal Breeding Co. Ltd. (License number: SCXK 2014-0007). The room temperature was 24 ± 2 °C, and relative humidity was 40–70%. All mice were fasted overnight and then gavaged with DMST-H2  according to manufacturer protocol. DNA concentration and purity were determined via Qubit fluorometer and Nanodrop 2000 spectrophotometer . Whole genome sequencing was performed on the MGISEQ-2000 platform and PacBio Sequel system at BGI . The obtained reads were assembled using Falcon v0.3.0 , proovread 2.12 , Celera Assembler 8.3 , SMRT Analysis v2.3.0  and GATK v1.6-13 . Gene prediction was performed using Glimmer v3.02 , RNAmmer 1.2 , tRNAscan-SE 1.3.1  and Rfam 9.1 . Functional annotation of genes were searched against the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Clusters of Orthologous Groups (COG). Antibiotic resistance genes were predicted using the Comprehensive Antibiotic Resistance Database (CARD). The Virulence Factors of Pathogenic Bacteria Database (VFDB) was used to identify the virulence factors. The genome sequence is available in the GenBank under the accession number CP063275.The resistance of DMST-H2 to simulated gastrointestinal circumstance was tested as previously described . DMST-H29 CFU/mL with sterile saline.The freeze-dried DMST-H2 and yogurt fermented with it were used in this study. The freeze-dried DMST-H2 powder was inoculated into 10% sterilized skim milk  and fermented at 42 °C until solid. Before administration, the viable cell count of DMST-H2 in powder and yogurt were adjusted to 2 × 10All animal procedures were performed following the Guidelines for Care and Use of Laboratory Animals of South China Agricultural University  and experiments were approved by the Animal Ethics Committee of Southern Medical University . SPF Male Balb/c mice  were provided by Southern Medical University . After one week of adapting , the mice were randomly divided into 4 groups with 8 mice per group as shown in Evaluating parameters of diarrhea symptoms were fecal consistency and fecal water content. Fecal consistency was measured on a 3 grade scale: formed, shaped and brown, score = 1; soft, does not pour, yellow, score = 2; liquid, yellow, score = 3 ,23. Feca−4–10−6 with sterile saline and then spread onto selective agar plates. TPY medium , LBS medium , bile aesculin azide medium  and EMB (eosin and methylene blue) medium  were used to detect Bifidobacterium, Lactobacillus, Enterococcus, and Enterobacteriaceae, respectively.Fresh feces on days 0 and 3 were diluted into 10The ileum fixed in formalin was stained by hematoxylin and eosin (HE) , and theThe levels of tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10) and interferon-γ (IFN-γ) were determined using mouse ELISA kits following the manufacturer’s instructions . Endotoxin (ET), D-lactate (D-LA) and Diamine oxidase (DAO) concentrations were also measured using mouse ELISA kits .Microbial DNA was extracted according to HiPure Stool DNA Kits instructions . The primers used for amplifying the 16S rDNA V3-V4 region were 341F: CCTACGGGNGGCWGCAG, 806R: GGACTACHVGGGTATCTAAT . After bp < 0.05. Figures were plotted in R project ggplot2 package (version 3.3.2) [A naive Bayesian model using the  RDP classifier 2.2  was used to classify the representative sequences  based onn 3.3.2) .treptococcus thermophilus ATCC 19258. The phylogenetic tree was showed in treptococcus thermophiles strains, thus DMST-H2 belonged to Streptococcus thermophiles.The 16S rDNA of the strain named DMST-H2 showed 99.93% similarity with SDMST-H2 contained a single circular chromosome of 1879014 base pairs (bp), with a G + C% content of 83.24%. A total of 2016 genes were identified with an average length of 775.81 bp. The chromosome harbored 18 rRNAs, 67 tRNAs, and 16 sRNAs, 24 minisatellite DNAs, and 2 microsatellite DNAs. 1497 genes were assigned to COGs, functioning in metabolism , information , cellular process , and poorly characterized  A. Furthe50) of DMST-H2 in KM mice was more than 10.0 g/kg bw. Therefore, DMST-H2 can be classified as actual non-toxic grade  genes were identified as putative virulence factor genes, they carried out the functions like amino acid transport and metabolism, nucleotide transport and metabolism, carbohydrate transport and metabolism, transcription, lipid transport and metabolism, translation, ribosomal structure and biogenesis, cell wall/membrane/envelope biogenesis, cell motility, posttranslational modification/protein turnover/chaperones, inorganic ion transport and metabolism, and signal transduction mechanisms according to COG. In fact, they could not be considered really harmful, because they could also represent essential probiotic traits for adhesion and protection ,37.The resistance of the gastrointestinal tract environment is a key factor for bacterial strains to be considered as probiotics . DMST-H2Genomic analysis is shown in For protection against reactive oxygen, the strain carried thioredoxin reductase (NADPH), glutathione reductase, peptide methionine sulfoxide reductase msrA/msrB , thiol pp < 0.05, Bifidobacteria, Lactobacilli, Enterococcus and Enterobacteriaceae were performed on days zero and three to examine AAD-related intestinal flora imbalance. Bifidobacteria and Lactobacilli were considered vital components of the gut microbiota and possessed many benefits [Bifidobacteria and Lactobacilli declined significantly (p < 0.05) with survival rates of only 6.50% and 8.93%, respectively. However, the survival rate of Enterococcus and Enterobacteriaceae still maintained 57.09% and 67.49%. In addition, the bacteria in the NC group remained relatively stable during the modeling period  reached 70.30 ± 2.80%, which was significantly higher than the NC group , but normal conditions were not restored  or DMST-H2 (ST and YT groups), fecal water content obviously decreased (p < 0.05), which might be related to the systemic inflammation of mice. ST and YT groups increased the content of IL-10 and IFN-γ and decreased the level of TNF-α significantly (p < 0.05). Nevertheless, only TNF-α was completely restored to the normal level . ST and YT groups completely cured it with the cecum index dropping to 2.25 ± 0.12% and 2.16 ± 0.37%, respectively  and decreased slightly in ST and YT groups . Remarkably, the 3D principal component analysis suggested differences among groups . The MC group formed a distinctive cluster from the NC, ST, and YT groups. Clusters from ST and YT groups were much closer to the NC group, thereby indicating that the bacterial community structures were more similar between the NC, ST, and YT groups  and Blautia (15.96%) were the dominant genera in the NC group. However, exposure to lincomycin hydrochloride caused higher levels of Bacteroides  and lower levels of Blautia  in the MC, ST and YT groups. Compared to the NC group, the genera Lachnoclostridium, Staphylococcus, Acinetobacter, Pseudomonas and Curvibacter increased in the MC group, which included species involved in pathogenesis. Additionally, genera of beneficial microorganisms Erysipelatoclostridium, Parasutterella and Parabacteroides were limited in the MC group. As for the ST and YT groups, Lachnoclostridium, Ruminiclostridium_5, and Streptococcus  increased . Though the number of different OTUs in the DMST-H2-treated groups (ST and YT) and MC group were similar, the species were distinguished. The significant highly abundant OTUs in the MC group were mainly within the phylum of Proteobacteria , and Bacteroidetes (genus Muribaculaceae and Bacteroides). Moreover, the highly decreased OTUs were within the phylum of Firmicutes , and Actinobacteria  (p < 0.05). In addition, OTUs markedly enriched in the ST group belonged to the phylum of Firmicutes , and Bacteroidetes , and decreased OTUs were mainly within the phylum of Actinobacteria  and Proteobacteria  (p < 0.05). In YT groups, the significant highly abundant OTUs were mainly within the phylum of Firmicutes , decreased OTUs were mainly within the phylum of Actinobacteria , Bacteroidetes , and Proteobacteria  (p < 0.05). After the statistics, a total of five genera were found to be significantly different between the ST and FY groups. Specifically, the ST group showed drastically higher levels of Parasutterella, Parabacteroides, Lachnospiraceae_NK4A136_group, and Sphingomonas, while a higher content of Coprobacillus was measured in the YT group. Furthermore, four-component analyses were also performed by the Kruskal-Wallis test, reveling that marked reductions in relative abundances of genera Blautia, Robinsoniella, Proteus and Parasutterella were noted in MC, ST and YT group compared to NC (p < 0.05). To further investigate the differences in bacteria species of these four groups, the relative abundances of all OTUs among the groups were compared in The above results showed that lincomycin hydrochloride treatment changed the composition of gut microbiota in various taxons. Natural recovery (MC group) resulting in the limited growth of Firmicutes, and the overgrowth of Proteobacteria which was generally regarded as a characteristic of dysbiosis. However, both ST and YT groups exhibited the opposite and performed a more similar population structure with the NC group. Therefore, DMST-H2 contributes to the recovery of ADD-induced intestinal dysbacteriosis.PICRUSt 2 predicted the metabolic processes of gut microbiota. The low value (0.08–0.22) of the nearest sequenced taxon index value (NSTI) indicates the accurate prediction . Figure Streptococcus thermophiles DMST-H2 from Chinese traditional yogurt. Genomic analysis and in vitro experimentation suggested that DMST-H2 had the potential ability to survive and adhere in the gastrointestinal tract, indicating the possibility of use as a probiotic. We successfully established the AAD mice model based on the results of increased fecal water content and fecal consistency score. Also, an even larger decrease in beneficial bacteria, such as Bifidobacteria and Lactobacilli, than the decrease in pathogenic bacteria like Enterococcus and Enterobacteriaceae was other evidence. Both natural recovery (MC group) and DMST-H2 treatment (ST and YT) groups decreased fecal water content significantly. However, DMST-H2 supplementation performed better at decreasing systemic inflammation, recovering intestinal injury and regulating the changes of intestinal flora.In this study, we isolated Clostridium butyricum and Bifidobacterium infantis could relieve systemic inflammation in the AAD mice by returning IL-10, IFN-γ, and TNF-α to normal levels [S. thermophilus strains were able to modulate the immune response of various human cell lines [S. thermophilus improved intestinal barrier function [S. thermophilus both prevented occludin degradation, rupture of tight cell junctions induced by E. coli in vitro, and decreased epithelial cell death [A growing body of evidence suggests that probiotics are prospective to prevent and treat AAD . Previoul levels . Besidesll lines ,55,56. Oll lines . Work byfunction . In addill death . Additioll death . ConsistBlautia and Parasutterella. Blautia could produce butyric acid, and butyrate will benefit the intestinal mucosa repair, increase the expression of ZO-1, and decrease the gut endotoxin levels in serum [Parasutterella is a core component of the human gut microbiota [In the case of dysbacteriosis, probiotics supplements could have a significant impact on reshaping the microbiota. Plenty of research on the AAD mice/rat model presented the same change of gut flora after antibiotic treatment: lower abundances of Firmicutes and over-representation of Proteobacteria ,62,63. Oin serum . Parasutcrobiota  and playcrobiota . Since tcrobiota , the chacrobiota  and our Streptococcus thermophiles DMST-H2, which is equipped with putative genes for adapting to gut transit stresses and showed tolerance to simulated gastrointestinal fluid in vivo. The animal experiment indicated that DMST-H2 had a potent effect on promoting recovery in AAD mice compared to natural recovery, and demonstrated it from three aspects: (1) DMST-H2 relieved diarrhea symptoms effectively, manifested in the reduction of fecal water content and fecal consistency score; (2) DMST-H2 positively recovered the inflammation and intestinal injury induced by AAD. TNF-α decreased while IL-10 and IFN-γ increased. Also, DMST-H2 lowered the cecal index, improved the intestinal barrier injury, and reduced ET, D-LA and DAO content in serum; (3) DMST-H2 better restored the microbial environment in the guts of mice with AAD, and the bacterial structures were much closer to the natural control group. Therefore, DMST-H2 deserves further research on AAD treatment. The joint analysis of microbiome and metabolomics will be carried out in further research for the purpose of analyzing the functionary mechanism of DMST-H2.Taken collectively, this study acquired a non-oral toxicity probiotic strain


The challenges in the diagnosis, prognosis, and monitoring of Gaucher disease (GD), an autosomal recessive inborn error of glycosphingolipid metabolism, can negatively impact clinical outcomes. This systematic literature review evaluated the value of glucosylsphingosine (lyso-Gb1), as the most reliable biomarker currently available for the diagnosis, prognosis, and disease/treatment monitoring of patients with GD. Literature searches were conducted using MEDLINE, Embase, PubMed, ScienceOpen, Science.gov, Biological Abstracts, and Sci-Hub to identify original research articles relevant to lyso-Gb1 and GD published before March 2019. Seventy-four articles met the inclusion criteria, encompassing 56 related to pathology and 21 related to clinical biomarkers. Evidence for lyso-Gb1 as a pathogenic mediator of GD was unequivocal, although its precise role requires further elucidation. Lyso-Gb1 was deemed a statistically reliable diagnostic and pharmacodynamic biomarker in GD. Evidence supports lyso-Gb1 as a disease-monitoring biomarker for GD, and some evidence supports lyso-Gb1 as a prognostic biomarker, but further study is required. Lyso-Gb1 meets the criteria for a biomarker as it is easily accessible and reliably quantifiable in plasma and dried blood spots, enables the elucidation of GD molecular pathogenesis, is diagnostically valuable, and reflects therapeutic responses. Evidentiary standards appropriate for verifying inter-laboratory lyso-Gb1 concentrations in plasma and in other anatomical sites are needed. GBA1 gene [Gaucher disease (GD) is an autosomal recessive disease of glycosphingolipid metabolism caused by a functional deficiency of the lysosomal enzyme β-glucocerebrosidase (glucosylceramidase (GBA); EC 3.2.1.45), resulting from variants in the GBA deficiency results in the progressive accumulation of the substrate glucosylceramide Gb1), along with a build-up of related glycosphingolipids almost exclusively in cells of the mononuclear phagocyte  system ,7. These, along wOne of the challenges in the care of patients with GD is the development of biomarkers as validated tools that aid diagnosis, prognosis, follow-up, treatment decisions, and pathophysiologic understanding ,10,11. AGBA genotype and residual GBA activity as a diagnostic and prognostic tool [Selecting appropriate candidate biomarkers has been hampered by the complexity of this monogenetic disorder, in which modifier genes, epigenetics, and external factors give rise to vast clinical and biochemical heterogeneity ,15,16,17tic tool ,21,22,23CHIT1 gene [The utility of more widely used plasma biomarkers is also limited. The hydrolase chitotriosidase and the chemokine (C-C motif) ligand 18 (CCL18) are secreted by activated macrophages, including Gaucher cells ,24, and IT1 gene .Over the last half-century, there has been ongoing focus on the occurrence of the sphingoid base in the primary lysosomal storage lipids as mediators of disease ,11,31. GGBA genotype, lipid storage deposition, and GD phenotype [There are at least three adaptations in Gb1 metabolism that occur during GBA deficiency—namely, the increased anabolism of Gb1 to gangliosides, excessive transglycosylation by cytosol-faced retaining β-glucosidase GBA2, and the active deacylation of Gb1 by acid ceramidase to lyso-Gb1  ten clinical (autopsy) studies ,62,63,93Evidence for lyso-Gb1 accumulation consistent with the pathology of GD was found in all ten autopsy studies  54,55,5,62,63,93GBA knockout mice models and in newborn neural cells from a type 3 GD mouse model in the presence of CBE [Seven of eight in vitro studies provided evidence for a role of lyso-Gb1 accumulation in GD . In culte of CBE ,101,102.In all five pharmacologically induced GD murine models, lyso-Gb1 was significantly elevated in the brain after exposure to the GBA inhibitor CBE ,76,81,85+ Gbaflox/flox) treated with therapeutic vectors containing the Gba gene, relative to control mice [In type 1 GD murine models, lyso-Gb1 was elevated in the spleen and liver ,77,80,92rol mice .Lyso-Gb1 accumulation was observed in the brain ,91,111, in vivo study [GD pathology is marked by the presence of Gaucher cells, neuropathology, and chronic low-grade inflammation . Evidencvo study , the effGBA−/− neurons had marked accumulation of Gb1 and lyso-Gb1, enlarged lysosomes, and an impaired ATP-dependent calcium-influx response [From a mechanistic viewpoint at the cellular level, lyso-Gb1 evoked HEK293 and RH7777 cells to form globoid cells (multinucleated macrophages) , a commoresponse . A wide response ,100,107.response .Over the past two decades, it has become apparent that there is an association between GD and Parkinson disease, with both diseases sharing neuropathological features . Lyso-GbResults from 14 of 17 experimental animal models of GD have shown that lyso-Gb1 is associated with: pro-inflammatory effects ; hepatosIn the CBE-induced murine GD model, high levels of lyso-Gb1 alone or in combination with Gb1 were accompanied by brain α-synuclein aggregation, neurodegeneration, microglia and complement C1q activation, and premature death ,85, withIn a lyso-Gb1-induced GD model reflecting lyso-Gb1 levels observed in moderate to severely affected patients with untreated GD, decreased hemoglobin (Hb) and hematocrit was observed along with hepatosplenomegaly . In addiGBA1 knockout mouse model (Mx1-Cre+) detected a correlation between splenic lyso-Gb1 levels and splenomegaly [GBA2 knockout despite elevated Gb1 and lyso-Gb1 levels [Most pathologic associations of lyso-Gb1 in the conditional type 1 GD model were incidental, although two studies using the nomegaly ,77. Inte1 levels . These dflox/floxGBA; nestin-Cre model [+lnl/lnlGBA mice and improved survival [Similar to the type 1 GD models, most associations of lyso-Gb1 with pathology in the neuronopathic variants were incidental . In the re model , whereassurvival .Preclinical data and evidence from 20 patients with GD showed that the frequency of lyso-Gb1-specific T cells correlates with disease activity and therapeutic response .Excessive lyso-Gb1 has a role in dysregulating humoral immunity by promoting chronic B-cell activation and gammopathy, which can evolve into multiple myeloma, a relatively common blood cancer in patients with GD .Of the 74 articles included in the analysis, 21 reported data that pertained to plasma lyso-Gb1 as a biomarker in GD.Eleven studies provided information relevant to the diagnosis of GD ,47,49,50GBA mutation, with the exception of five patients homozygous for N370S. Fourteen homozygotes for the N370S mutation were also studied, two patients with saposin C deficiency, and three patients with type 3 GD [Two groups, Dekker et al.  and Rolfype 3 GD . Promineype 3 GD .Rolfs et al. retrospectively analyzed non-Jewish Caucasian patients from a single center in Germany . The marIn a US study, Murugesan et al. prospectively compared plasma lyso-Gb1 in patients with type 1 GD and healthy controls. In this cohort, the investigators found that a lyso-Gb1 cut-off of 4 ng/mL had 100% sensitivity and specificity as a diagnostic tool . The difFuller et al. described an accurate, reproducible, robust, and easy-to-perform assay for the determination of plasma lyso-Gb1 concentrations in a routine laboratory setting using N-palmitoyl-d3-lactosyl ceramide as an internal standard . This asChipeaux et al. prospectively recruited 64 patients with GD, 34 obligate carriers of GD, and 28 healthy controls from three centers in France . Plasma The findings of one additional study showed that it is possible for patients other than those with GD to have high lyso-Gb1 . In patiFour studies provided information on lyso-Gb1 as a prognostic biomarker of GD  26,33,3,50.GBA homozygous patients only), liver volume, and bone marrow fat fraction but not correlated with macrophage inflammatory protein 1-β, the presence of a spleen, skeletal complications, osteocalcin, and procollagen type 1 N propeptide in this study [In a prospective, case-control study, modest increases in plasma lyso-Gb1 were evident in mildly affected patients . Plasma is study .In a retrospective, case-control study, plasma lyso-Gb1 appeared to reflect the severity of the individual genetic variant . PatientLyso-Gb1 concentrations were also excessive in the cultured fibroblasts of a collodion patient with GD  with virtually no residual GBA activity versus more modest elevations among patients with type 1 GD and neuronopathic variants .GBA1 gene who received low-dose ERT (15 units/kg/month) revealed an exponential decay (Pearson product moment determination coefficient (r2), 0.84) in lyso-Gb1 plasma over 72 months [In 17 studies, cross-sectional and longitudinal data on lyso-Gb1 in treated and untreated patients with GD were compared ,51,52,532 months . ERT als2 months . A combi2 months . Howeverr), –0.83) and RBC membranes (−0.65) correlated inversely with hematocrit [Ten of the 17 studies provided different evidence levels of information on lyso-Gb1 as a response biomarker in GD  36,37,3,47,49,51matocrit . Nine stmatocrit ,47,49,51Regarding incidental findings in the periphery, as plasma lyso-Gb1 decreased upon treatment with either ERT or SRT, platelet counts, Hb, and the bone marrow fat fraction increased, whereas spleen and liver volumes decreased ,37,43,51r, −0.530), 25 , and 53 , and between decreasing plasma lyso-Gb1 concentrations and decreasing spleen volumes at weeks 25  and 101  [9/L at baseline (n = 22) to 146.5 × 109/L at week 209 (n = 10), whereas the median spleen volume decreased from 16.6 MN at baseline (n = 22) to 4.2 multiples of normal at week 209 (n = 12) [With respect to correlations, an exploratory pooled analysis of phase III clinical trials of treatment-naïve patients with type 1 GD receiving ERT revealed that there was moderate pairing between decreasing plasma lyso-Gb1 concentrations and increasing platelet counts at weeks 13  . In the (n = 12) . The ERT(n = 12) .r, 0.27), liver , and age  [In a prospective, single-center, longitudinal, case-control study of 169 patients with type 1 GD receiving ERT or SRT, plasma lyso-Gb1 concentration correlated significantly with indicators of severity of visceral disease: splenic volume  .In a prospective, multicenter, cross-sectional, case-control study, lyso-Gb1 content in RBCs was higher in untreated patients with type 1 and neuronopathic GD (0.69 p/mol per kg protein) than in healthy controls (0.15 p/mol per kg protein) and in patients who had been receiving ERT for at least 1 year (0.34 p/mol per kg protein) . Higher Correlations between lyso-Gb1 concentrations and other established biomarkers of GD were evaluated in six studies ,44,47,52By systematically evaluating preclinical and clinical publications relevant to lyso-Gb1 in GD up to March 2019, we have identified strong evidence for the use of lyso-Gb1 as a pathogenic mediator of disease and a diagnostic and pharmacodynamic biomarker, and some evidence for the use of lyso-Gb1 as a prognostic and disease-monitoring biomarker in GD. Although lyso-Gb1, chitotriosidase, and CCL18 correlate well with each other in patients with GD, lyso-Gb1 is more sensitive and specific than the other two biomarkers, irrespective of the chitotriosidase genotype ,50. LysoSince the literature search for this systematic review was conducted, additional data on the value of lyso-Gb1 as a biomarker in GD have been published. Lyso-Gb1 measured in dried blood spots was found to be highly sensitive and specific for GD in a case-control study conducted in the Russian Federation . FurtherGBA genotypes, a plasma lyso-Gb1 level spanning more than one order of magnitude correlated well with the presumed pathogenicity of the genotypes and with hepatosplenomegaly, thrombocytopenia, and bone pain [Data from these studies additionally suggest that lyso-Gb1 concentrations measured in dried blood spots have prognostic power ,119. Plaone pain . Secondlone pain . Thirdlyone pain .Four recent studies found that lyso-Gb1 measurement in dried blood spots is useful for monitoring patients with GD ,127,128.The value of plasma lyso-Gb1 as a pharmacodynamic and response biomarker was reported in three recent studies ,125,127.It is interesting to speculate as to why not all measures of disease severity correlated with plasma lyso-Gb1 . The genLyso-Gb1 measurement techniques continued to advance during the conduct of our review. Elevated concentrations of lyso-Gb1 could be detected in only 20 µL of plasma collected from patients with GD using reverse-phase LC-Differential Mobility Spectrometry-MS/MS, which enables the resolution of the corresponding stereoisomer . CompareOne important limitation of lyso-Gb1 analyses is the variable methodology employed by different laboratories or within the same laboratory over time, preventing the ability to directly compare findings. In addition, the storage conditions of the pathology samples from which lyso-Gb1 concentrations were measured cannot be assured in some early studies. High variability of longitudinal lyso-Gb1 measurements has been reported in multiple studies, with potential explanations including the impact of the circadian rhythm, effects of nutrition and/or physical activity, or effects of coexisting pathological conditions . Thus, tThe systematic literature review was conducted and reported in line with criteria stipulated by the Preferred Items for Systematic Review and Meta-Analyses Protocols (PRISMA-P) recommendations .Search terms were developed on the basis of Medical Subject Headings (MeSH), and free-text words and abbreviations related to lyso-Gb1. The specific search terms and Boolean strategy were as follows: (glucosylsphingosine OR GlcSph OR lyso-GL1 OR Lyso-Gb1 OR psychosine) AND Gaucher. The databases searched were the US National Library of Medicine (includes PubMed and MEDLINE), Embase, ScienceOpen, Science.gov, Biological Abstracts, and Sci-Hub. The searches were conducted between February and March 2019, with no restrictions with respect to the search period. Additional references identified from the reference lists of published articles were identified.Eligible articles included original research articles  reporting information on lyso-Gb1 in GD published at any time in the medical literature. Meta-analyses were included but systematic  or narrative reviews and single-patient case studies were excluded.® form developed for the systematic review. Extracted data included those related to study design, sample , lyso-Gb1 assay method, interventions, and type of information on lyso-Gb1 as a pathogenic mediator and biomarker in GD.Citations and abstracts retrieved from the searches were compiled to create a single list of references. Abstracts were reviewed by three researchers to determine the articles to be included. If information in the abstract was not sufficient to make a decision, the full text publication was obtained and reviewed. Data were extracted from each eligible publication using a standardized Microsoft ExcelA two-tiered approach to data analyses was applied on the basis of the US Food and Drug Administration/National Institutes of Health Biomarker Working Group definition of a biomarker as follows: “a defined characteristic that is measured as an indicator of normal biological processes, pathogenic processes, or responses to an exposure or intervention, including therapeutic interventions” . The twoA validated tool matched to the study type was used to assess the strength and validity of the empirical data for each individual study. The assessment of the quality of randomized controlled trials was carried out using recommendations from the National Institute for Health and Care Excellence (NICE) single technology appraisal (STA) manufacturer’s template . The NewAfter systematically searching for and evaluating an extensive list of manuscripts published since 1974 according to PRISMA-P guidelines, we conclude that circulating lyso-Gb1 is a selective and sensitive biomarker of GD throughout the course of the disease, as a companion diagnostic and pharmacodynamic biomarker. There is a high degree of confidence that lyso-Gb1 mediates GD pathophysiology but further study is required on its specific roles in this process. Although there is some evidence supporting a role of lyso-Gb1 as a prognostic and disease-monitoring biomarker in GD, the associations of lyso-Gb1 level with prognosis, secondary clinical events, and bone disease requires further research.


RPS14, RPS15A, RPS24, FAU, RPL27, RPL31, RPL34, RPL35A, RSL24D1, and EEF1B2. Sex difference of ribosome in stroke-induced peripheral immunosuppression may be the potential mechanism of sex disparities in outcome after IS, and women are more likely to have stroke-induced immunosuppression. RPS14, RPS15A, RPS24, FAU, RPL27, RPL31, RPL34, RPL35A, RSL24D1, and EEF1B2 may be novel prognostic biomarkers and potential therapeutic targets for IS.Ischemic stroke (IS) greatly threatens human health resulting in high mortality and substantial loss of function. Recent studies have shown that the outcome of IS has sex specific, but its mechanism is still unclear. This study is aimed at identifying the sexually dimorphic to peripheral immune response in IS progression, predicting potential prognostic biomarkers that can lead to sex-specific outcome, and revealing potential treatment targets. Gene expression dataset GSE37587, including 68 peripheral whole blood samples which were collected within 24 hours from known onset of symptom and again at 24-48 hours after onset (20 women and 14 men), was downloaded from the Gene Expression Omnibus (GEO) datasets. First, using Bioconductor R package, two kinds of differentially expressed genes (DEGs) (nonsex-specific- and sex-specific-DEGs) were screened by follow-up (24-48 hours) vs. baseline (24 hours). 30 nonsex-specific DEGs (1 upregulated and 29 downregulated), 79 female-specific DEGs (25 upregulated and 54 downregulated), and none of male-specific DEGs were obtained finally. Second, bioinformatics analysis of female-specific DEGs was performed. Gene Ontology (GO) functional annotation analysis shows that DEGs were mainly enriched in translational initiation, cytosolic ribosome, and structural constituent of ribosome. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis shows that the top 6 enrichment pathways are ribosome, nuclear factor-kappa B (NF-kappa B) signaling pathway, apoptosis, mineral absorption, nonalcoholic fatty liver disease, and pertussis. Three functional modules were clustered in the protein–protein interaction (PPI) network of DEGs. The top 10 key genes of the PPI network constructed were selected, including  Stroke was the second largest cause of death worldwide after ischemic heart disease and the second most common cause of worldwide disability-adjusted life years (DALYs). There were 80.1 million prevalent cases of stroke, which 84.4% contributed ischemic stroke (IS) and 13.7 million new stroke cases globally in 2016 , there iIn recent years, more and more attention has been paid to the pathogenesis of stroke-induced immunodeficiency syndrome (SIDS)  and stroBecause of the dynamic nature of transcriptional regulation, RNA levels represent not only features encoded in the genome but also the influence of the environment . Thus, sIn this study, we have downloaded one original microarray dataset GSE37587 from the Gene Expression Omnibus (GEO) datasets, including 68 peripheral whole blood samples which were collected within 24 hours from known onset of symptom and again at 24-48 hours after onset (20 women and 14 men). Using Bioconductor R package, nonsex-specific and sex-specific DEGs were got by follow-up (24-48 hours) vs. baseline (24 hours) with 30 nonsex-specific DEGs, 79 female-specific DEGs, and 0 male-specific DEGs. Then, Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment on female-specific DEGs were performed by the R software. Finally, the STRING online database protein–protein interaction (PPI) network was used to analyze the association of female-specific DEGs and discover the molecular interactions involved in IS progression. Meanwhile, according to the PPI network result, molecular complex detection (MCODE) modules was established and key gene analysis was performed by Cytoscape for identifying key genes of optimal significance. In conclusion, sex-specific DEGs associated with the IS progression were screened and an integrated analysis was conducted. Our study is aimed at identifying the gender dimorphic to peripheral immune response in IS progression, predicting potential prognostic biomarkers that can lead to sex-specific prognosis, and revealing potential treatment targets.https://www.ncbi.nlm.nih.gov/geo/), which platform is the GPL6883 Illumina HumanRef-8 v3.0 expression beadchip. Robust Multichip Average (RMA) normalization collation had been done with the series matrix file(s), including background correction, quantile normalization, and summarization. A total of 68 peripheral whole blood samples were included in the dataset, which were collected from n = 34, and magnetic resonance imaging (MRI) diagnosed IS patients ≥ 18 years of age within 24 hours from known onset of symptom and again at 24-48 hours after onset. IS patients, whose ethnicity are Caucasian, included 20 women with mean age of 72.15 ± 15.951 years and 14 men with mean age of 71.64 ± 13.054 years.The GSE37587 gene expression profile series matrix file(s) and SOFT formatted family file(s) were downloaded from GEO datasets . If multiple probes correspond to the same gene, only the probe with the highest mean expression value is retained, and 18631 genes were finally obtained. Third, the DEG analysis was performed using the Linear Models for Microarray Data (limma) package (http://www.bioconductor.org/). DEGs were selected with thresholds of ∣log2 (fold change) (FC) | >0.6 and adjusted p value <0.05. Two kinds of DEGs were screened: (1) nonsex-specific DEGs by follow-up (24-48 hours) vs. baseline (24 hours) and (2) sex-specific DEGs by female follow-up (24-48 hours) vs. female baseline (24 hours) and male follow-up (24-48 hours) vs. male baseline (24 hours).The analysis was carried out with the R language software (version 3.6.2). First, the dataset was normalized by logpAdjustMethod = “BH,”, pvalueCutoff = 0.05, and qvalueCutoff = 0.2 [For the sets of female-specific DEGs, GO analysis was performed using clusterProfiler in R package with ff = 0.2 . Then, wpAdjustMethod = “BH,”, and p value <0.05 was considered statistically significant.KEGG pathway enrichment analysis was performed using clusterProfiler with http://string-db.org/) (version 11.0), and only the interaction score > 0.4 (medium confidence) was considered significant. Interactions in STRING are derived from five main sources: genomic context predictions, high-throughput lab experiments, (conserved) coexpression, automated text mining, and previous knowledge in databases. The PPI network was visualized by the Cytoscape software (version 3.7.2).To evaluate the interactive relationships among the female-specific DEGs, we mapped them to the STRING database  from known onset of IS symptom and again at 24-48 hours (follow-up) after onset. 30 nonsex-specific DEGs (1 upregulated and 29 downregulated) were obtained by comparison of 34 follow-up and 34 baseline samples. 79 female-specific DEGs (25 upregulated and 54 downregulated) were obtained by comparison of 20 female follow-up and 20 female baseline samples. And none of male-specific DEGs were obtained by comparison of 14 male follow-up and 14 male baseline samples. The details of all DEGs are shown in ano plot , and thep value <0.05 and q value <0.2. The redundancy of enriched GO terms was removed using the function simplify. The results are shown in GO function annotation analysis was performed using clusterProfiler in R package, which was divided into three parts: biological process (BP), cell component (CC), and molecular function (MF). Those results were considered statistically significant if HSPB1 and DENR, nine downregulated genes RPS14, RPL35A, RPS24, RPL34, RPS15A, RPL27, RPL17, RPL31, and RSL24D1. The main BP enriched by the 11 genes are as follows: translational initiation, SRP-dependent cotranslational protein targeting to membrane, cotranslational protein targeting to membrane, nuclear-transcribed mRNA catabolic process, nonsense-mediated decay, and establishment of protein localization to endoplasmic reticulum. This indicates that in addition to the degradation of abnormal transcripts, these genes are mainly involved in protein biosynthesis: protein translation, postsynthesis secretion, and targeted transport in the cytoplasm. The most important CC enriched by all the nine downregulated genes is cytosolic ribosome. Meanwhile, the nine downregulated genes are enriched the only one MF result: structural constituent of ribosome; in other words, they encode the ribosomal structural protein.A total of 11 genes were enriched: two upregulated genes CTSW that is upregulated. Six signaling pathways with higher rich factor than others are shown in KEGG pathway enrichment analysis was performed using clusterProfiler in R package. A total of 18 genes are enriched in key signaling pathways, and all are downregulated genes except the only one The STRING online database was used to analyze the 79 female-specific DEGs, and the PPI network was visualized using the Cytoscape software, as shown in FAU, RPL31, TMA7, RPS24, PFDN5, RPS14, RPL34, RPL35A, EEF1B2, RPL27, RSL24D1, RPL17, and RPS15A, related to ribosome function, involving translation, elongation, and folding in the process of protein synthesis. The cluster 2 includes three genes, CD3E, IL2RB, and DDIT3, related to T cell adaptive immune and apoptosis. The cluster 3 includes three genes, LY96, CXCL8 , and BIRC2, related to NF kappa B signaling pathway.The cluster 1 included 13 genes, RPS15A, RPS14, RPL27, RPL35A, FAU, RPS24, RPL31, RPL34, EEF1B2 and RSL24D1, are supposed to be more important than others because of their key position on the PPI network. Key genes are shown in The top key genes of the PPI network constructed were selected using CytoHubba in the Cytoscape software. The top key genes, which have higher MCC degree, including Sex influence IS epidemiology, pathophysiology, treatment efficacy, and outcome. A recent clinical study found that minocycline, which is an inhibitor of cell death, had significantly better neurological outcomes in IS male patients only, with no significant clinical improvement to female patients . The stuOur study is aimed at identifying the sex-specific DEGs in peripheral blood in IS progression, predicting potential biomarkers that can lead to sex-specific peripheral immune response and prognosis, and revealing potential sex-specific treatment targets. The GSE37587 datasets included 68 peripheral whole blood samples which were collected 20 IS women and 14 IS men within 24 hours (baseline) from known onset of symptom and again at 24-48 hours (follow-up) after onset. In order to have further information whether the gene expression of sex-specific DEGs and nonsex-specific was consistent, we screened two kinds of DEGs: nonsex specific and sex specific. Through the follow-up of 34 patients compared with the baseline of 34 patients, we got the nonsex-specific DEGs of the progression of IS. Adopted method of Tian et al., which follow-up of 20 females were compared with baseline of 20 females and follow-up of 14 males compared with baseline of 14 males, respectively, we got the sex-specific DEGs of the progression post-IS. Interestingly, the results show that there are 30 nonsex-specific DEGs, 79 female-specific DEGs, and none of male-specific DEGs. There are 23 overlaps between female-specific and nonsex-specific DEGs . Only 7 GO analysis showed that the 11 genes enriched were mainly related to ribosomal and protein synthesis. Changes in the transcription level of ribosome-related genes have a broader impact on the downstream translation level. An in vitro experiment used ribosome profiling to explore the immediate transcription and translation change of neural cell gene expression to oxygen and glucose deprivation (OGD). It made a conclusion that the effect of OGD was widespread on translation than transcription . It can DDIT3 encoding a proapoptotic transcription factor DNA damage-inducible transcript 3  and the BIRC2 encoding cIAP1, which is a member of the inhibitor of apoptosis family that inhibits apoptosis [CTSW, encoding a cysteine proteinase associated with cytotoxic T and natural killer cell [The results of the KEGG enrichment analysis of female-specific DEGs are highly consistent with the GO analysis results. The most significant result is the ribosome followed by NF-kappa B signaling pathway and apoptosis and finally, the mineral absorption, nonalcoholic fatty liver disease, and pertussis. Ribosome is the place where biological protein synthesis takes place. It consists of two major components which the small ribosomal subunits in charge of reading the mRNA, and the large subunits in charge of joining amino acids to form a polypeptide chain. Each subunit consists of varieties of ribosomal proteins and one or more ribosomal RNA (rRNA) molecules. Ribosome-related genes downregulated lead to protein synthesis and secretion disorders. The NF-kappa B signaling pathway has a broad role in the regulation of numerous biological processes. NF-kappa B-dependent gene expression is required for proper development and formation of the immune system, innate immune responses, initiation of the adaptive immune response, pathogen recognition, inflammation, lymphoid organogenesis, transcription of cytokines, and immediate antimicrobial responses –22. Downpoptosis , are botler cell , 27, is CD3E, IL2RB, and DDIT3. CD3e molecule, epsilon , encoded by CD3E, together with CD3-gamma, -delta, and -zeta and the T cell receptor alpha/beta and gamma/delta heterodimers, forms the T cell receptor-CD3 complex, which couples antigen recognition to several intracellular signal-transduction pathways. CD3E plays an essential role in T cell development and interleukin-2 production in T cells [IL2RB, is involved in T cell-mediated immune responses. Activation of the receptor increases proliferation of CD8+ effector T cells [CD3E and IL2RB suggests the downregulation of T cell adaptive immune response. DDIT3 encodes DNA damage-inducible transcript 3 (DDIT3) that belongs to the family of CCAAT/enhancer-binding proteins (C/EBPs) [BCL-2 and a transcriptional activator to upregulate expression of proapoptotic genes such as BAX [LY96, CXCL8 , and BIRC2. The MD-2 protein , encoded by LY96, associates with toll-like receptor 4 (TLR-4) on the cell surface and confers responsiveness to lipopolysaccharide (LPS) [α, IL-1β, IL-6, IL-18, and IL-12 [CXCL8 (IL8), is a chemokine produced by macrophages mainly. Its primary function is to induce chemotaxis in target cells, primarily neutrophils, causing them to migrate toward the site of infection. cIAP1, encoded by BIRC2, is a member of the inhibitor of apoptosis family that inhibits apoptosis by interfering with the activation of caspases [Three modules were obtained from the functional module analysis of the PPI network. The clusters 1 and 3 are basically consistent with the results of KEGG enrichment analysis. The cluster 1 is closely related to ribosome function, and the important genes involved will be discussed in the following selection of key genes. The cluster 2 includes three genes,  T cells . Interle T cells . The dow(C/EBPs) . It is a(C/EBPs) . DDIT3 p(C/EBPs) . Under Eh as BAX , 32. Thede (LPS) . LPS is nd IL-12  and chemcaspases . Meanwhicaspases . In addicaspases . It can https://www.genecards.org/). RPS14, RPS15A, and RPS24 encode 40S ribosomal proteins. FAU encodes a fusion protein consisting of the ubiquitin-like protein fubi at the N terminus and ribosomal protein S30 at the C terminus, which is posttranslationally processed to generate free fubi and free ribosomal protein S30. Whereas the function of fubi is currently unknown, ribosomal protein S30 is a component of the 40S subunit of the cytoplasmic ribosome and displays antimicrobial activity [RPL27, RPL31, RPL34, and RPL35A encode 60S ribosomal proteins. The above key genes participate in the structure of ribosome and protein synthesis. RSL24D1 encodes probable ribosome biogenesis protein (RLP24), which localizes to the nucleolus and is thought to play a role in the biogenesis of the 60S ribosomal subunit. EEF1B2 encodes a translation elongation factor, named eukaryotic translation elongation factor 1 beta 2, which involved in the transfer of aminoacylated tRNAs to the ribosome. We infer that the downregulation of key genes will affect the protein synthesis of peripheral blood immune cells and thus cause immunosuppression.The results of key gene analysis further confirm the results of GO, KEGG, and functional module analysis. 10 top key genes screened, which all of them are downregulated, related to ribosome or translation. We obtained the basic information of these genes through the Human Gene Database GeneCards  family, which are molecular scaffolding proteins binding to a regulatory subunit (RII) of cAMP-dependent protein kinase A (PKA), directing the kinase to discrete subcellular locations [AKAP7 (AKAP15/18) are essential components of neuronal and cardiac phosphatase complexes, ion channels, cardiac Ca2+ handling, and renal water transport [AKAP7 were significantly associated with poststroke blood-brain barrier (BBB) disruption 24 hours posthospital admission, which suggested that AKAP7 may be a prognostic biomarker for poststroke BBB complications [After a comprehensive exploring of female-specific DEGs, we studied main nonsex-specific DEGs. According to the introduction of GeneCards, ocations , 45. Altransport . A new sications . In thisThe innovation of this study is to take the lead in analyzing the difference of gene expression between men and women in the early stage of stroke recovery through high-throughput biochip transcriptome datasets and to find that women are more responsive in stroke-induced immunosuppression, and ribosome-related genes play the most important roles in stroke-induced immunosuppression. This finding provides a new insight to evaluate the prognosis and potential accurate treatment targets. Ribosome-related genes may be a novel prognostic biomarker and a potential therapeutic target for IS. However, the specific mechanism of ribosome in SIDS needs to be further studied in vitro and in vivo.RPS14, RPS15A, RPS24, FAU, RPL27, RPL31, RPL34, RPL35A, RSL24D1, and EEF1B2 may be novel prognostic biomarkers and potential therapeutic targets for IS.Sex difference of ribosome in stroke-induced peripheral immunosuppression may be the potential mechanism of sex disparities in outcome after IS, and women are more likely to have SIDS after IS.


This error was, regrettably, not caught during peer or editorial review. As a result, the decimal point has been removed from "94.4 mg/m2" as seen in the following sentence located in the results section: "The optimum cut-off value for age was 60 years, sensitivity 60% and specificity 56%; the same values for cumulative oxaliplatin dosage were, respectively, 944 mg/m2, 72%, and 68%."The corrected sentence now reads: "The optimum cut-off value for age was 60 years, sensitivity 60% and specificity 56%; the same values for cumulative oxaliplatin dosage were, respectively, Cureus sincerely regrets this error.


The role of the gut microbiome in human health is becoming apparent. The major functional impact of the gut microbiome is transmitted through the microbial metabolites that are produced in the gut and interact with host cells either in the local gut environment or are absorbed into circulation to impact distant cells/organs. Short-chain fatty acids (SCFAs) are the major microbial metabolites that are produced in the gut through the fermentation of non-digestible fibers. SCFAs are known to function through various mechanisms, however, their signaling through free fatty acid receptors 2 and 3  is a new therapeutic approach. FFAR2/3 are widely expressed in diverse cell types in human and mice, and function as sensors of SCFAs to change several physiological and cellular functions. FFAR2/3 modulate neurological signaling, energy metabolism, intestinal cellular homeostasis, immune response, and hormone synthesis. FFAR2/3 function through Gi and/or Gq signaling, that is mediated through specific structural features of SCFAs-FFAR2/3 bindings and modulating specific signaling pathway. In this review, we discuss the wide-spread expression and structural homologies between human and mice FFAR2/3, and their role in different human health conditions. This information can unlock opportunities to weigh the potential of FFAR2/3 as a drug target to prevent human diseases. The gut microbiome and its contribution to human health is an emerging area and much remains to be learned about the interactions between microbial cells with a host. The major communication between the microbiome and host cells takes place through the metabolites produced by the gut microbiome . These mi/o coupled signaling [q/11 coupled signaling [FFAR2/3 are cell surface receptors that can play a very significant role in intracellular cell signaling ,47. Bothignaling ,48. Onlyignaling ,49. TherHere we performed an extensive search of the literature and compiled detailed information about the expression (gene and protein) patterns and comparative structural analysis developed by using our own in-silico designed model setup of the FFAR2/3 receptors. We described the expression information from literature as well as from our cell line repository. In addition, we have performed a literature search using PubMed, Google Scholar, and Web of Knowledge using different combinations of keywords FFAR2, FFAR3, GPR43, GPR41, microbiota, short-chain fatty acids, diet, fibers, human health, diabetes, obesity, colon, intestine, adipose tissue, liver, lungs, disease, metabolic disease, stem cell, monocyte, Colitis, expression, structure, modeling, mice, knockout, human, agonist, antagonist, activator, inhibitor, software, cell line, enteroendocrine, ligand, docking, tight junction, inflammation, mucus, brain, neuron, and organoid with diverse combinations.Protein sequences and structural information were obtained from the protein database site and modeled by using modeler software and I-TASSER to develop the models for further docking with ligands, and also comparing and verifying with specific ligands in human and mice. More comprehensive information about FFAR2/3 in-silico-interactive analysis have also been published elsewhere ,51,52. TFFAR2 and FFAR3 are present in close proximity on chromosome 1 (19q13.12) in humans and chromosome 7  in mice. The phylogenetic tree and detail information on FFAR2/3 chromosomal location, gene length, and position, number of exons, and protein length are shown in During the initial days of FFAR2 discovery, it was found to be intensively expressed in human immune cells such as peripheral blood mononuclear cells (PBMCs) and polymorphonuclear cells (PMNs) with maximum expression in neutrophils ,55,56,57FFAR2 expresses in hypothalamus , bone maFFAR2 expresses in the hypothalamus, cerebral cortex , pituitaFFAR3 expression is detected in the central nervous system (CNS) , brain eIn rodents, FFAR3 expression is detected in superior cervical ganglia (SCG) and celiac sympathetic-mesenteric ganglia (CSMG) of autonomic nervous systems selected randomly from different tissue regions ,101,102,FFAR3 expression is detected in bovine cerebral cortex , hypothaComprehensive expression analysis of FFAR2 and FFAR3 in different tissues and cell lines of mice and humans are presented in Multiple emerging sources indicate that FFAR2/3 can be novel targets to prevent and/or treat several human diseases ,115,116.2-adrenergic receptor as template. [90, Ile145, Arg180, Arg255, and Glu166 [90, Ile145, and Glu166 [180 and Arg255 are positively charged orthosteric sites which interact with negatively charged glutamine residue (Glu171) to stabilize the protein structure for proper binding with ligands [The crystal structure of FFAR2 is not yet determined and the structure was predicted using human βemplate. . Human Femplate. ,129. Stremplate. ,118, andemplate. . The actd Glu166 ,52. The  ligands .75, Gln148, Tyr238, Arg65, Arg180, Tyr90, and Arg255 according to our in-silico analysis. Both mouse and human FFAR2 show the protein sequential similarity from the 95-111 AAs position except at the 105th position where mouse FFAR2 consists of methionine whereas human FFAR2 is of isoleucine. The significant change in the secondary structure of human and mice FFAR2 is observed at the C-terminal end. The C-terminal of the receptors consists of most conserved minimotifs and short peptides that regulate receptor binding efficiency, posttranslational modification, and trafficking with unique biochemical and physiological properties [Similar to humans, mouse FFAR2 is also a 7-TM protein made of 330 AAs  and showoperties . The det140, Arg180, His242, and Arg255 [201, Glu113, and His115, and associated with protein-ligand interactions [Mesocricetus auratus, Cavia porcellus, and Oryctolagus cuniculus) are not yet known and studies are needed to comprehend their location, structure, and functionalities.The FFAR2-like protein in chicken contains 367 AAs, encoded with the gene of 1105 bp nucleotides as per National Center for Biotechnology Information (NCBI)’s latest update. The chicken FFAR2 paralog homology model shows four active AA residues at the positions Hisd Arg255 . These rractions . The hom2-adrenergic receptor [96, Tyr151, and Leu171 are involved in specific ligand binding [171 residues provides stabilization to negatively charged arginine residues at the second extracellular loop (EL2) [Human FFAR3 structure was also predicted based on the crystal structure of the human βreceptor . Human Freceptor ,129. Simreceptor ,118 whicreceptor . AA resi binding  with SCAop (EL2) . The olyop (EL2) ,131. Mou3) is twice more than the volume of FFAR2 (41 Å3) [2 and higher aromatic SASA value by 63 Å2 as compared to human FFAR3 [3) and surface areas (510 Å2) as compared to FFAR3 binding pockets [3) is larger than FFAR2 (332 Å3) [90, Ile145, and Glu166 in FFAR2 and Phe96, Tyr151, and Leu171. Thus, SAR information shows that FFAR2 and FFAR3 are lying in very close proximity to each other and can interact with the same chemical compound to compensate for each other’s biological response [The structure-activity relationship (SAR) study showed that the endogenous binding site volume of human homology FFAR3 (105 Å (41 Å3) . SAR hel (41 Å3) . Along w (41 Å3) . SASA an (41 Å3) . Human Fan FFAR3 . Based o pockets . However(332 Å3)  whereas (332 Å3) . Therefo(332 Å3) , predictresponse . Thus, m2- or sp-hybridized α-carbon [140 and His242 residues to alanine in the fourth and sixth TM domains decrease the binding potential of SCFAs to FFAR2 [180, Arg255) mutation at the top of either five and/or seven TM helix is important for facilitating the interactions of SCFAs with human and mice FFAR2 [SCFAs are the orthosteric ligand of FFAR2/3 as they bind to endogenous binding sites . SCFAs’ α-carbon . That meα-carbon  by showito FFAR2 ,134. Usice FFAR2   of mouse FFAR2 through homology modeling (unpublished data). The mouse FFAR2 interacts with SCFAs by forming H-bonds, such as acetate-made H-bonds with Tyr90, Gln148, Trp75, and Arg65; propionate-made H-bonds with Tyr90, Ile145, Arg255, Tyr238, Arg180, and His242; and butyrate-made H-bonds with Trp75, Gln148, Tyr238, Arg65, Arg180, Tyr90, and Arg255. Similar to humans, the mouse FFAR2 activated by acetate and butyrate by making H-bonds with Arg180, Tyr190, and His242 residual sites [So far, no in-silico study demonstrated the SCFAs’ binding site with FFAR2 receptor in mice. Recently, we determined the common active binding site residues  and 6-TM (His242) is important in deciding the binding efficacy of SCFAs in human FFAR3, as indicated by mutagenesis studies replacing these AAs with alanine [SAR data showed that human FFAR3 receptors prefer saturated or ali-cyclic moieties of SCAs for ideal binding . Histidi alanine ,134. SCF alanine ,123,126.Although the interactions of SCFAs with FFAR2/3 are similar, they still show a degree of selectiveness in these interactions ,50. In a2+ and the phosphorylated extracellular signal-regulated kinase (ERK)1/2 pathway [242 site to alanine, mutation within the binding pockets at His140, Val179, Tyr90, Tyr165, and Tyr238 residual sites to alanine significantly reduced agonist property of Cmp1 for mice FFAR2 [Synthetic ligands such as CATPB and GLPG0974 act as allosteric antagonists to the FFAR2 receptor by reducing Ca pathway ,138,139. pathway ,140. Cmp pathway ,137. Aloce FFAR2 , as showDEFINITIONS:Orthosteric ligands. The ligand which binds to a receptor at an endogenous active site.Allosteric ligands. The ligand which binds to a receptor other than an endogenous site.Allosteric agonist ligands. Allosteric ligand that activate the receptors in the absence of orthosteric ligands by binding other than on an active site.Ago-allosteric ligands. The ligand binds allosterically to activate a receptor in the absence of an orthosteric ligand equal to an allosteric agonist and also activates the receptor in the presence of an orthosteric ligand as a positive allosteric modulator (potentiate agonist-mediated receptor response).Inverse agonist. An inverse agonist is a ligand that binds to the same receptor as an agonist but induces a pharmacological inhibitory response.Inverse agonist. The ligand binds to the receptor as an agonist but develops pharmacological function opposite to that of agonist.66, Phe89, Leu173, Tyr238, and Val259 residues [66, Phe89, Leu173, Val259, Tyr90, Ile145, Asn239, and His242 [242) in human FFAR2 serves as a key residual site to classify whether a ligand will show allosteric or orthosteric activity [2+ level via Gαq signaling [The CFMB (previously known as phenylacetamide 1) , AMG-770residues , while Ad His242  with humactivity . 2CTAP, ignaling ,144. How246, Met80, and His182 provide supports to these ligand-binding grooves [4-CMTB is an ago-allosteric modulator ligand for human FFAR2 as it increases the binding efficacy of SCFAs  and also activates the human FFAR2 receptor of its own  ,145,146. grooves .3-MQC are antagonists for mouse FFAR3, however detailed in-silico analysis has yet to be done [Well-known human FFAR3 agonist 1-MCPC forms H-bonds at different binding residues to activate the FFAR3 receptor shown in  be done ,147. To 166, Leu183, and Cys184 with corresponding FFAR3 residues such as Leu, Met, and Ala using site-directed mutagenesis favored the binding of FFAR3 agonists 1-MCPC and 3-pentenoic acid, while these compounds were not able to bind wild type FFAR2 [Interestingly, in many instances, FFAR2 and FFAR3 activities are interchangeable and/or compensatory, this is because of similar chemical and structural characteristics. For example, substation of FFAR2 amino acid residues such as Glupe FFAR2 . Howeverspp., Prevotella spp., Ruminococcus spp., Blautia hydrogenotrophica, ClostridiumBacteroides (B.) thetaiotaomicron, Akkermansia (A.) muciniphila, Bifidobacterium  spp., and Streptococcus spp. produce acetate from pyruvate via acetyl-CoA and/or the reductive acetyl-CoA pathway [spp., Firmicutes, Roseburia inulinivorans, Ruminococus spp., Clostridiales (C.) Lacterium, Eubacterium (Eu.) spp., Coprococcus spp., Dialister succinatiphilus, Phascolarctobaterium succinatutens, A. muciniphila, Clostridium spp., Coproccus catus, Clostridium sp., Roseburia insulinivorans, RuminococusB. thetaiotaomicron, Roseburia  spp., and Eu. halli from succinate, acrylate and/or propanediol pathways [spp., C. symbiosum, Faecalibacterium prasnitzii, BacteroidetesC. tyrobutyricum, Roseburia intestinalis, Eu. rectale, Roseburia insulinivorans, Clostridiales bacterium, Anaerostripes hadrus, Coprococcus  spp., and Coprococcus spp. by butyrate kinase and/or the butyryl-CoA:acetate CoA-transferase biosynthesis route [The gut microbiota is an important source of SCFAs that exhibit several health beneficial effects such as immune ,44,56, m pathway ,151. Thepathways ,151. Simis route ,151. Theis route , but alsis route , white ais route ,24, cardis route , hepatocis route ,18, skelis route ,20, alveis route ,22,152, is route , bone mais route , and splis route ,22. One is route ,49,121, is route ,29,153, is route ,44 whichThe biological functions regulated by FFAR2/3 signaling are immunity ,114,154,In humans, FFAR2 exhibits an anti-inflammatory response against metabolic diseases ,56,155. FFAR2 signaling activation led to increase immunoglobulin (Ig)A  production to protect intestinal epithelium against foreign pathogenic microbe invasion ,158. SunCitrobacter rodentium infection with delayed in expression of interferon gamma (INFγ)  through rapidly accelerated fibrosarcoma which activates the MAPK/ERK pathway [FFAR3 signaling activated by acetate and propionate reduces the production of pro-inflammatory cytokine  secretion , and enh pathway . However pathway ,174. TheCapra hircus fed with a high concentrate diet (60%) increases LPS and SCFAs production that activate FFAR2/3 signaling to produce cytokines and chemokines which in turn lead to cecal inflammation [In mice macrophages (Raw 264.7), activation of FFAR3 signaling by SCFAs reduces the proinflammatory cytokines and increases nitric oxide synthase (iNOS) secretion . Moreoveammation . These rOverall, these observations indicate that both FFAR2/3 are closely associated with complex mechanism of immune response ,178,179,FFAR2/3 signaling significantly contribute in gut hormone homeostasis through gut-hepatic ,169 and 2+ levels. In addition, FFAR2 KO mice show a decrease in GLP-1 and insulin secretion leads to impair glucose tolerance even under SCFA treatment [FFAR2 activation increases GLP-1 and PYY synthesis in human, rodents and guinea pig L-cells ,181,182.reatment . Howeverreatment , suggestreatment . In addireatment ,172. Thereatment . These rreatment , indicat3- secretion via activating the 5-HT4 receptor, and muscarinic M1 and M3 receptors [In addition, FFAR2 signaling also involved in the GI tract buffering, especially on the conjunction of stomach and duodenum where acid of stomach poured down in the duodenum. A study showing that FFAR2 agonist phenylacetamide 1 increases the duodenal HCOeceptors , which t3- and GLP-2 hormonal synthesis whereas FFAR3 antagonist (CF3-MQC) counteract the AR420626 effect by reducing the enteropathy condition [Tolhurst et al.  also repondition , indicatEven so, increased synthesis of PYY and GLP-1 hormone by Roux-en-Y gastric bypass (RYGB) surgery leads to overexpression of both FFAR2 and FFAR3 in the intestine . This inEmerging evidence indicates that FFAR2/3 signaling significantly contributes to nutrient absorption ,76 and hFFAR2 signaling contributes to (i) maintaining intestinal integrity which in turn reduces leaky gut, and (ii) regulating the colonic motility through intestinal 5-hydroxytryptamine (5-HT) release ,76 and g- T-cells to Fox3+ Treg cells through overexpression of FoxP3 protein, IL-10 and Helios transcription factor to provide protection against T1D (or autoimmune activity) by increasing the number of autoreactive T cells and Treg cells [C. rodentium infection induced intestinal inflammation in FFAR2 KO mice [Min/+) model [Also, FFAR2 KO-NOD mice have a higher rate of T1D development as compared to FFAR2 WT-NOD mice . Howevereg cells . In humaeg cells ,189. SCF KO mice ,190. How KO mice . In addi+) model ,192. How+) model  showed t-/- mice, the inflammatory response was significantly reduced as compare to their WT [Lactobacillaceae and clostridium) in young swine models [FFAR3 maintains intestinal integrity by activating the cytokines and chemokines through the MEK-ERK pathway . In FFARtheir WT . Grape se models . SCFAs ae models . The SCFe models ,44,189. After the deorphanization, many research groups have reported that neither FFAR2 nor FFAR3 are expressed in CNS ,101. HowClostridium tyrobutyricum (responsible for production of butyrate) and Bacteroides thetaiotaomicron (mainly produce acetate and propionate) in germ-free mice decreases BBB permeability through boosting Ocln mRNA expression in the frontal cortex and hippocampus [FFAR2 regulates the blood brain barrier (BBB) permeability . Butyratpocampus . Even FFpocampus . Howeverpocampus . This ispocampus . Howeverpocampus . As SCFAβγ-phospholipase C (PLC)-β3-ERK1/2-synapsin 2-β at serine 426 pathway to enhance norepinephrine release from sympathetic nerve endings [FFAR3 controls sympathetic neurons which in turn regulate whole body metabolic homeostasis . FFAR3 i endings . FFAR3-d endings . Further endings . In addi endings ,196.βγ signaling pathway and inhibits N-type Ca2+ channels, which in turn reduces neuronal catecholamine release in rat sympathetic nervous systems. Moreover, in proximal colonic mucosa of rats, FFAR3 is associated with cholinergic-mediated secretory response in enteric nervous system [Won et al.  suggestss system . Thus, Fs system . Moreoves system . FFAR3 es system . These eSeveral cellular and molecular pathways involved in adipogenesis, lipolysis, glucose homeostasis, insulin sensitivity, and energy metabolism are regulated by FFAR2/3 signaling  3,11,28,121,148.Firmicutes: Bacteroidetes ratio along with lower plasma FFA level [FFAR2 is responsible for energy accumulation in adipose tissues, adipogenesis, and metabolic syndrome disease pathogenesis . In-vitrFA level ,199. MorFA level . Apart fFA level . During FA level .Additionally, in the differentiated 3T3-L1 cells, FFAR2 activation by propionate enhances adipogenesis via peroxisome proliferator-activated receptor gamma 2 (PPAR-γ2) pathway and the presence of FFAR2 siRNA  which inhibits the adipogenesis process . In addiA human multipotent adipose tissue-derived stem (hMADS) model reveals that activated FFAR3 by acetate significantly reduces lipolysis through decreasing hormone-sensitive lipase phosphorylation . In micePancreatic beta-cells are crucial to regulate blood glucose homeostasis by producing insulin. Therefore, maintaining and preserving beta-cell mass and functions remain critical. Beta-cell proliferation and differentiation is important for maintaining beta-cell population, while beta-cell functions are important for efficiently releasing insulin in response to glucose. In T2D, beta-cell proliferation, differentiation, and functions are deteriorated, which ultimately causes a decrease in insulin secretion and hyperglycemia in long-term diabetics. The coupling effect of FFAR2/3 receptors plays a fundamental role in the regulation of glucose-stimulated insulin secretion (GSIS) ,73,203 aStarting from an early embryonic stage, maternal gut microbiota-SCFA-FFAR2 signaling plays a crucial role in regulating metabolic syndrome, as FFAR2 KO mice embryos have lower insulin and higher glucose level, and are more susceptible to obesity and diabetes in adulthood . Additioi/o pathway [Gut microbiota changes in obese humans are associated with increased FFAR3/ Gi signaling to inhibit insulin synthesis . These c pathway ,210. Opp pathway . Similar pathway .Overall, these findings indicate that FFAR2/3 signaling is critical to regulate pancreatic beta-cells either by changing their proliferation, differentiation, insulin synthesis, and regulating their functions in terms of GSIS, which in turn maintains better glucose homeostasis, however, the their precise role in regulating proliferation and differentiation are poorly understood.Dysbiotic gut microbiota with reduced SCFAs are related with suppression of FFAR2/3 signaling—that are known to regulate an array of biological pathways participation energy metabolism, adipogenesis, appetite control, intestinal cellular homeostasis, gut motility, glucose metabolism, and inflammatory response. Alterations in these biological pathways are hallmarks of several human diseases such diabetes, obesity, IBS/IBD, Crohn’s disease, atherosclerosis, gout, asthma, cardiovascular diseases, arthritis, hypertension, and colitis, therefore, targeting FFAR2/3 signaling can provide promising therapeutic strategies for these human diseases. The immune cell during metabolic diseases such as obesity and T2D causes chronic inflammatory response which provides an insight crucial mechanism for further disease progression. However, the role of FFAR2/3 signaling in these diseases remain controversial and needs to be further studied for better understanding of their role to devise the therapeutic importance of FFAR2/3 agonist/antagonists. For example, the majority of studies show that activation of FFAR2/3 signaling ameliorates obesity/T2D pathology, however, some studies show the opposite. For example, HFD feeding to FFAR2 KO mice shows improved oral glucose tolerance test (OGTT) and insulin sensitivity along with lower fat mass and increased lean mass compared to wild type (WT) mice ,30,33, iThe pharmacological modification of these SCFAs receptors by endogenous or synthetic ligands provides an opportunity to counteract these gastrointestinal disorders in humans. However, overlapping expression of FFAR2 and FFAR3 in the same tissues/cells, and their similar affinity to specific endogenous ligands develop puzzled outcomes to understand the role of FFAR2/3 in particular biological functioning. Thus, the future studies must aim to develop highly specific and efficacious small molecules to modulate pharmacological actions of FFAR2/3 signaling, and can display a promising strategy to prevent, manage and/or treat human diseases including diabetes, obesity, Crohn’s disease, atherosclerosis, gout, asthma, cardiovascular diseases, arthritis, hypertension, and colitis.


Psoriasis is a systemic inflammatory disease caused by crosstalk between various cells such as T cells, neutrophils, dendritic cells, and keratinocytes. Antimicrobial peptides (AMPs) such as β-defensin, S100, and cathelicidin are secreted from these cells and activate the innate immune system through various mechanisms to induce inflammation, thus participating in the pathogenesis of psoriasis. In particular, these antimicrobial peptides enhance the binding of damage-associated molecular patterns such as self-DNA and self-RNA to their receptors and promote the secretion of interferon from activated plasmacytoid dendritic cells and keratinocytes to promote inflammation in psoriasis. Neutrophil extracellular traps (NETs), complexes of self-DNA and proteins including LL-37 released from neutrophils in psoriatic skin, induce Th17. Activated myeloid dendritic cells secrete a mass of inflammatory cytokines such as IL-12 and IL-23 in psoriasis, which is indispensable for the proliferation and survival of T cells that produce IL-17. AMPs enhance the production of some of Th17 and Th1 cytokines and modulate receptors and cellular signaling in psoriasis. Inflammation induced by DAMPs, including self-DNA and RNA released due to microinjuries or scratches, and the enhanced recognition of DAMPs by AMPs, may be involved in the mechanism underlying the Köbner phenomenon in psoriasis. Psoriasis is a representative autoimmune or inflammatory skin disorder characterized by well-delineated, raised areas of red or salmon-pink papulosquamous plaques covered by white or silvery scales . PsoriasPsoriasis affects not only the skin but also nails and joints. Up to 30% of psoriasis patients develop psoriatic arthritis (PsA) . In addiIn cellular and molecular pathology, psoriasis is regarded as a T cell-mediated skin disease, involving both T-helper (Th)1 and Th17 cells to promote inflammation by producing cytokines, including TNF-α interferon (IFN)-γ, interleukin (IL)-12, IL-17A, IL-22, and IL-23 ,10. AntiAMPs have essential roles in skin immunity, enabling epithelial surfaces to cope with many microbial challenges . AMPs arPsoriatic lesions highly express AMPs such as cathelicidin, β-defensins, S100 proteins, RNase 7, lysozyme, elafin, and neutrophil gelatinase-associated lipocalin ,20. ContCathelicidin was the first AMP identified in mammalian skin . A singlAs natural antibiotics, AMPs target the essential cell wall or cell membrane structures of microorganisms through enzymatic or nonenzymatic disruption. Simultaneously, AMPs can also function as potent immune regulators by signaling through chemokine receptors and inhibiting or enhancing Toll-like receptor (TLR) signaling . FurtherThe expression, secretion, and activity of most AMPs are tightly controlled. Cathelicidins are synthesized as propeptides and are activated into various CAMPs including NL-8, LR-10, KR-10, IK-14, LL-17, LL-23, KR-20, KS-27, KS-30, and LL-37 by serine proteases . HoweverDefensins are a type of cationic microbial peptide and contain six conserved cysteine residues that form three pairs of intramolecular disulfide bonds ,19. In c2+ signaling, cytoskeletal membrane interaction, enzyme activities, cell cycle progression, differentiation, and inflammatory responses. Twenty-one S100 proteins are known, of which S100A7 (psoriasin), S100A8 , S100A9 , S100A12 , and S100A15 have antimicrobial effects and their expression levels are increased in the lesional skin and serum of psoriasis patients [+ T lymphocytes [S100 proteins are another group of AMPs important in psoriasis and comprise a family of low molecular weight (9–13 kDa) proteins characterized by the presence of two calcium-binding helix-loop-helix motif . S100 prpatients . In partpatients . S100A7 phocytes . Hegyi ephocytes . The rolShao et al. reported that lipocalin-2 (Lcn2), an AMP derived from keratinocytes and neutrophils, was highly expressed in the lesional skin of psoriatic patients. In vitro, Lcn2 stimulated human neutrophils to produce proinflammatory mediators such as IL-6, IL-8, TNF-α, and IL-1α via a specific receptor, 24p3R, on neutrophils . ElghariLactococcus, Streptococcus, and Streptomyces produce factors, known as bacteriocins, which are another type of AMP and inhibit the growth of other bacterial strains and species that could compete for nutrients and other resources. Staphylococcus epidermidis, the dominant bacterium cultured from skin microflora, produces the AMP phenol-soluble modulin (PSM)-γ. PSMγ causes membrane leakage in target bacteria, indicating that it functions in a manner similar to that of host-derived AMPs [S. epidermidis from which the peptide is derived. In addition, PSMs enhance the bactericidal activity of human neutrophils by inducing their neutrophil extracellular traps (NETs) [AMPs provide defense and resistance to infection by killing pathogenic bacteria. AMPs also determine the microbiota composition and limit access of the microbiota to host tissues. Surprisingly, an important component of the surface antimicrobial shield of the skin is produced by the resident microorganisms themselves. Gram-positive bacteria such as ved AMPs . Interess (NETs) , suggestS. epidermidis in vivo was observed on the surface of the nasal cavity. Nasal colonization by S. aureus was inhibited in individuals whose nasal passages were colonized with specific strains of S. epidermidis that produce a serine protease capable of destroying biofilms formed by S. aureus [S. epidermidis blocks the S. aureus quorum-sensing system that controls the production of various virulence factors [Another important example of the protective action of . aureus . For exa factors . The selIn psoriasis skin lesions, keratinocytes, neutrophils, dendritic cells, and T cells have their own roles in generating unique skin symptoms by expressing AMPs, with cell-specific AMP production modulating intracellular and intercellular reactions in psoriasis.Various external stimuli result in rapid innate immune responses by keratinocytes, leading to the production of an array of pro-inflammatory cytokines or chemokines such as IFN-β, IL-1β, IL-36, TNF, IL-6, IL-8, IL-25, and CXCL10 ,66,67. THarder et al. isolated human β-defensin (hBD)-2  and hBD-Neutrophil extracellular traps (NETs) are of central importance in psoriasis. NETs are a complex of self-DNA and proteins, including LL-37 and proteases released from neutrophils after NET-specific cell death (NETosis). NETs were originally reported to be triggered by bacterial components and inflammatory mediators such as IL-8 and type I IFN. NETs are “webs” that capture bacteria and thus exhibit antibacterial action ,77. LambPlasmacytoid dendritic cells (pDCs) are a unique subpopulation which infiltrate psoriatic skin rapidly ,82. The mDCs are activated by diverse cytokines, including IL-6, TNF-α, and IFN-α, as well as LL-37–RNA complexes. Activated mDCs secrete a mass of inflammatory cytokines such as IL-12 and IL-23 in psoriasis ,87, whic+ or CD8+ T cells which responded to LL-37 [Many investigators have demonstrated that psoriasis lesions contain increased numbers of T cells . T cellsto LL-37 . These cto LL-37 . On the to LL-37 , implyinTh17 and Th1 cytokines play roles in chronic inflammation in psoriasis. AMPs enhance the production of some of these cytokines and modulate receptors and cellular signaling in psoriasis.Type 1 interferons (IFN-α and IFN-β) are key cytokines that activate autoimmunity, such as systemic lupus erythematosus, and are activated in response to viral infection . IFN-α aType 1 IFNs can be induced following the activation of endosomal TLR7 and TLR9, or cytosolic cGAS-STING (cyclic GMP-AMP synthase-stimulator of interferon genes), by host, viral, or bacterial DNA. Type 1 IFNs are also induced by the activation of endosomal TLR8 by ssRNA, by the activation of endosomal TLR3, mitochondrial RIG1  by host or viral dsRNA, and by the activation of plasma membrane TLR4 by bacterial lipopolysaccharide (LPS) . Cell reLande et al. showed that LL-37 enhances the recognition of self-DNA through TLR9 in pDCs and enhances inflammation . This ovMorizane et al. reported that LL-37 and extracellular DNA strongly induce type I IFN production from keratinocytes via TLR9 . There aIt remains unclear how extracellular self-RNA released from injured cells reaches intracellular TLR3. We found that the complex formed by LL-37 and dsRNA binds to scavenger receptors and causes uptake of the complex into the intracellular space . We alsoCurrent genetic analysis tools and methodologies are lending insights into the genetic background of psoriasis, and molecular biology techniques are providing effective biologics for psoriasis treatments. These technological developments have shown promising links between AMPs in clinical phonotypes and psoriasis treatments.Psoriasis is thought to be triggered by both genetic and environmental factors. Strong association between psoriasis and the human leukocyte antigen (HLA)-C*06:02 allele has been reported in various races. Mabuchi et al. predicted sequences of peptides which are included in LL-37 and binds strongly to HLA-*C06:02 using computer models . InteresYuan et al. found that the levels of anti-LL-37 and anti-ADAMTS-L5 autoantibodies were significantly elevated in patients with PsA compared to non-PsA controls, suggesting that these molecules may be involved in the pathogenesis of PsA . Frasca Narrowband-UVB treatment induces vitamin D production and subsequent secretion of cathelicidin . Cyclosp2D3. The reaction is stimulated locally by the activation of TLR2 or local cytokines, such as TNF or type I IFNs [2D3 to VDRE. These observations suggest that AMP expression might be influenced by serum vitamin D levels [A particularly surprising observation came with the recognition that the human cathelicidin gene is under transcriptional control of a vitamin D response element (VDRE) ,109. Fole I IFNs ,111. ThiD levels , dietaryD levels , or vitaD levels . This meD levels ,116.+CD25+ regulatory T cells [Vitamin D is important in the relationship between psoriasis and cathelicidin. Vitamin D derivatives are used externally to treat psoriasis and may promote cell differentiation and suppress proliferation. Recently, vitamin D was reported to suppress the expression of hBD2, hBD3, IL-17A/F and IL-8 in psoriasis plaques  and indu T cells . These r T cells , which i T cells . Whether T cells .Anti-TNF agents are highly effective in the treatment of psoriasis, but 2–5% of treated patients develop psoriasis-like skin lesions called “paradoxical psoriasis” . Conrad Psoriasis is not just an inflammation of the epidermis or an immune disease derived only from T cells, but a systemic inflammatory disease caused by crosstalk between various cells such as keratinocytes, neutrophils, dendritic cells, and T cells. Contrary to their name, “antimicrobial” peptides are often involved in the pathogenesis of psoriasis by activating the innate immune system and triggering inflammation by various mechanisms independent of infection. At present, there is no established treatment targeting antimicrobial peptides, and blocking or degradation of AMPs is expected to be a novel treatment for psoriasis.


SERPINE1) gene corepressor Rev-Erbα increased PAI-1, which then increased the tPA gene PLAT via a PAI-1/LRP1/PKA/p-CREB1 pathway. This pathway was partially counterbalanced by increased DACH1, a PLAT-negative regulator. We focused on the PAI-1/PLAT pathway, which mitigates the reduction in fibrinolysis in obesity. Thus, silencing hepatocyte PAI-1, CREB1, or tPA in obese mice lowered plasma tPA and further impaired fibrinolysis. The PAI-1/PLAT pathway was present in primary human hepatocytes, and associations among PAI-1, tPA, and PLAT in livers from obese and lean humans were consistent with these findings. Knowledge of PAI-1 and tPA regulation in hepatocytes in obesity may suggest therapeutic strategies for improving fibrinolysis and lowering the risk of thrombosis in this setting.Fibrinolysis is initiated by tissue-type plasminogen activator (tPA) and inhibited by plasminogen activator inhibitor 1 (PAI-1). In obese humans, plasma PAI-1 and tPA proteins are increased, but PAI-1 dominates, leading to reduced fibrinolysis and thrombosis. To understand tPA–PAI-1 regulation in obesity, we focused on hepatocytes, a functionally important source of tPA and PAI-1 that sense obesity-induced metabolic stress. We showed that obese mice, like humans, had reduced fibrinolysis and increased plasma PAI-1 and tPA, due largely to their increased hepatocyte expression. A decrease in the PAI-1 (  Obesity, which has now reached epidemic proportions, increases risks for arterial, venous, and microvascular thrombosis, including coronary thrombosis , 2, stroSERPINE1) gene corepressor Rev-Erbα. Moreover, reducing hepatocyte PAI-1 in obese mice blocked the increase in both liver tPA and functionally active plasma tPA, indicating the existence of a hepatocyte PAI-1/tPA pathway in obesity. Although this tPA response pathway is partially counteracted by a coexisting tPA-suppressive pathway mediated by the corepressor DACH1, we provide evidence that the PAI-1/tPA pathway limits the degree of impairment of fibrinolysis in obesity. Accordingly, the fibrinolysis defect in obesity becomes even worse when the PAI-1/tPA pathway is genetically targeted in mice. The mechanism of the PAI-1/tPA pathway involves induction of the tPA gene PLAT through a pathway in which PAI-1, through its receptor LRP1, activates the PLAT inducer cAMP-responsive element-binding protein 1 (CREB1). The PAI-1/PLAT pathway is present in primary human hepatocytes, and livers, but not adipose tissue, from obese and lean humans showed associations among DACH1, tPA, and PLAT that are consistent with these findings. These findings add insight into the regulation of fibrinolysis in obesity, thereby suggesting new strategies for lowering thrombotic risk in this condition.One source of PAI-1 is adipocytes, and elevated adipocyte PAI-1 has been proposed as the dominant source for increased PAI-1 and the consequent fibrinolysis defect in obesity , 14, 15.https://doi.org/10.1172/JCI135919DS1). These findings are consistent with data in obese humans  were lower in the livers of obese versus lean mice   . Consistlat mRNA . Importad injury , all witd injury .PLAT mRNA , the major cellular receptor for PAI-1, can increase intracellular cAMP levels and the activity of protein kinase A (PKA) , the kinLAT mRNA . FurtherT rPAI-1 . We thenT rPAI-1 . WT rPAI did not . With reitor H89 . TogethePLAT) gene expression in obesity is regulated negatively by DACH1 and positively by PAI-1, which may represent a “compensatory” feedback pathway; (d) the net result of these opposing modes of hepatocyte PLAT regulation in obesity is an increase in plasma tPA protein, mimicking the findings in humans; and (e) although the PAI-1–mediated impairment of fibrinolysis in obesity is dominant, this impairment would be even worse without the hepatocyte PAI-1/PLAT pathway.Obesity has reached epidemic proportions worldwide and is a major contributor to a number of widespread chronic diseases, notably type 2 diabetes, cardiovascular disease, and nonalcoholic steatohepatitis (NASH) , 43. AmoSERPINE1) and protein levels among various tissues . AAV8 vectors were delivered at 1 × 1011 genome copies/mouse, and experiments were commenced 3 to 6 weeks later. fl/flCreb1 mice were generated as previously described .The /fl mice , Dach1fl/fl mice , and Cre/fl mice  were houescribed . The micescribed –32, 74  and recorded by LabChart software (ADInstruments). The time to total occlusion was defined as the interval between the application of FeCl3 and stable occlusion of the artery, defined as 0 blood flow for 3 minutes  was centrifuged for 15 minutes at 2300 g at 4°C. After careful removal of the supernatant fraction, each pellet (euglobulin fraction) was resuspended in 55 μL sodium borate/NaCl (pH 9.0) and transferred to a single well on a flat‑bottom 96-well microtiter assay plate; 50 μL of 25 mM CaCl2 was added to each well, and then the absorbance was recorded at 405 nm every 10 minutes, with 3-second shakes before each reading, at room temperature for 16 hours. Clot-lysis time was calculated as the time to achieve 50% of clot lysis   .Human primary hepatocytes were obtained from the Liver Tissue Cell Distribution System at the University of Pittsburgh . Primary mouse hepatocytes were isolated from 10-week-old WT C57BL/6J mice, as described previously , 81. AllHuman liver specimens were acquired from patients undergoing bariatric surgery or clinically indicated laparoscopic procedures at the New York Presbyterian Hospital, Columbia University Irving Medical Center ref. . The livP values were calculated using 2-tailed Student’s t test for data that passed the normality test or the Mann-Whitney rank-sum U test for data that were not normally distributed. One-way ANOVA with post hoc Tukey’s test was used to evaluate differences among groups when 3 or more groups were analyzed.All results are presented as mean ± SEM. All mouse experiments were conducted with the approval of the IACUC of Columbia University Medical Center. The use of human cells and specimens in this study was approved by the IRB at the Columbia University Irving Medical Center. All participants provided written informed consent.ZZ and IT designed the research. ZZ, KN, SG, XW, and ST conducted the research. RML provided critical reagents related to PAI-1. ST, MB, BS, and AK organized patient recruitment and human liver sample collection. LO provided critical reagents and advice related to DACH1. ZZ, KN, SG, LO, JAL, and IT analyzed the data. ZZ, SG, JAL, and IT wrote the manuscript. All authors read and commented on the manuscript.