Transforming Growth Factor-beta1 Gene Research Articles

Nephroprotective Effect of Ferula assa-foetida Oleo Gum Resin on Type 2 Diabetic Rats.

Diabetic nephropathy is one of the main causes of kidney failure in the end stage of diabetes worldwide. On the other hand, asafoetida is a gum whose hypoglycemic effects have been proven. The present study was conducted with the aim of using asafoetida to prevent diabetic nephropathy. Diabetes was induced by a high-fat diet (60%) and streptozotocin injection (35 mg/kg) in rats. Diabetic rats were treated with an oral dose of 50 mg/kg of asafoetida for 8 weeks. At the end of the experiment, serum and urine parameters were examined. Antioxidant enzymes and lipid peroxidation levels in the kidney were also determined along with its histological examination. The expression levels of tumor necrosis factor-alpha and Transforming growth factor beta genes were also evaluated. Glucose, cholesterol, triglyceride, and HbA1c concentrations were significantly reduced in the asafoetida 50. On the other hand, in the treatment group, serum creatinine, urea, and albumin levels decreased and increased in urine. Antioxidant enzymes in the kidney improved significantly, and the expression of tumour necrosis factor-alpha and transforming growth factor-beta genes decreased. Histopathological examination also showed that necrosis, epithelial damage, and leukocyte infiltration increased in the diabetic and the treatment group. The result of biochemical analysis, enzymatic, and histological examinations showed that asafoetida may delay the progression of diabetic nephropathy due to the presence of anti-inflammatory and antioxidant activities.

Genetic Polymorphisms of COL1A1 Promoter Region (rs1800012) and TGFB1Signal Peptide (rs1800471): Role in Cervical Insufficiency Susceptibility?

A structural or functional cervix problem prevents a woman from carrying a full-term pregnancy, which leads to the disease known as cervical insufficiency. Cervical insufficiency is partially inherited, and in certain situations, variations in genes related to connective tissue metabolism may be involved. The main objective of this investigation was to describe the collagen type I alpha 1 chain (COL1A1) gene rs1800012 polymorphism and the transforming growth factor beta 1 (TGFB1) gene rs1800471 polymorphism in a cohort of patients suffering from cervical insufficiency. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) assays have been used to analyze the DNAs of 93 patients with cervical insufficiency and 103 healthy controls. The chi-square test was used for statistical analysis. There were significant differences in the genotype frequencies of the COL1A1 gene rs1800012 (G > T) and TGFB1 gene rs1800471 (G > C) polymorphisms between the patient and the control groups (p = 0.049 and p = 0.049, respectively). Also, the C allele of the TGFB1 rs1800471 polymorphism was significantly higher in the patient group than the control group (p = 0.016). Following clinical assessment, the COL1A1 rs1800012 polymorphism was found to be connected to the history of cerclage (p = 0.010). Additionally, the frequency of the TT/GG composite genotype of COL1A1 rs1800012/TGFB1 rs1800471 polymorphisms was significantly lower in the patient group than the control group (p = 0.049). The TT genotype of COL1A1 rs1800012 polymorphism was found to be protective against cervical insufficiency, while the C allele of TGFB1 rs1800471 polymorphism was found to predispose to the disease. It appears that the TT/GG composite genotype of COL1A1 rs1800012/TGFB1 rs1800471 polymorphisms protects against cervical insufficiency.

Nephroprotective effect of remote ischemic conditioning on type 2 diabetic rats.

Diabetic nephropathy is one of the main causes of kidney failure in the end stage of diabetes worldwide. The present study was conducted with the aim of using the remote ischemic conditioning (RIC) method to prevent diabetic nephropathy. Diabetes was induced by high-fat diet (60%) and streptozotocin injection (35 mg/kg) in rats. RIC was performed by tightening a tourniquet around the upper thigh and releasing it for three cycles of 5 min of ischemia and 5 min of reperfusion daily for an 8-week duration. At the end of the experiment, serum and urine parameters were examined. Anti-oxidant enzymes and lipid peroxidation levels in the kidney were also determined along with histological examination. The expression levels of tumor necrosis factor-alpha and transforming growth factor beta genes were also evaluated. Glucose, cholesterol, triglyceride, and HbA1c concentrations were not significantly reduced in the RIC group. On the other hand, serum creatinine, urea, and albumin levels decreased and increased in urine. Anti-oxidant enzymes did improve in the kidney significantly and the expression of tumor necrosis factor-alpha and transforming growth factor beta genes decreased significantly. Histopathological examination also showed that necrosis, epithelial damage, and leukocyte infiltration increased in the diabetic group and improved in the treatment group. The results of biochemical analysis, and enzymatic and histological examinations showed that although RIC could not reduce blood glucose and lipids, nevertheless it may delay the progression of diabetic nephropathy due to the presence of anti-inflammatory and anti-oxidant activities.

Anti-Fibronectin Aptamer Modifies Blood Clot Pattern and Stimulates Osteogenesis: An Ex Vivo Study.

Scaffold (SCA) functionalization with aptamers (APT) provides adsorption of specific bioactive molecules on biomaterial surfaces. The aim of this study was to observe if SCA enriched with anti-fibronectin APT can favor coagulum (PhC) and osteoblasts (OSB) differentiation. 20 μg of APT was functionalized on SCA by simple adsorption. For PhC formation, SCAs were inserted into rat calvaria defects for 17 h. Following proper transportation (buffer solution PB), OSBs (UMR-106 lineage) were seeded over PhC + SCAs with and without APT. Cells and PhC morphology, PhC cell population, protein labeling and gene expression were observed in different time points. The APT induced higher alkaline phosphatase and bone sialoprotein immunolabeling in OSB. Mesenchymal stem cells, leukocytes and lymphocytes cells were detected more in the APT group than when scaffolds were not functionalized. Additionally, an enriched and dense fibrin network and different cell types were observed, with more OSB and white blood cells in PhC formed on SCA with APT. The gene expression showed higher transforming growth factor beta 1 (TGF-b1) detection in SCA with APT. The SCA functionalization with fibronectin aptamers may alter key morphological and functional features of blood clot formation, and provides a selective expression of proteins related to osteo differentiation. Additionally, aptamers increase TGF-b1 gene expression, which is highly associated with improvements in regenerative therapies.

Impact of dietary zinc on the growth performance, histopathological analysis, antioxidant capability, and inflammatory response of largemouth bass Micropterus salmoides

Zinc plays a crucial role in the antioxidant capacity, and inflammatory response of aquatic species, but its impact on largemouth bass Micropterus salmoides is rarely reported. Therefore, this paper aimed to investigate the effects of different levels of zinc on the growth performance, histopathology, antioxidant capacity, and inflammatory cytokines of largemouth bass Micropterus salmoides. Fish with an initial weight of 7.84 ± 0.06 g were cultured for 10 weeks. Five experimental diets were prepared with supplemented proteinate Zn (Bioplex Zn, Alltech) (0, 30, 60, 90, and 120 mg/kg), which were named the Zn-42, Zn-73, Zn-103, Zn-133, and Zn-164 groups. No evident difference was found between the dietary zinc level and the survival rate, the crude lipid content of the whole fish, or the visceral somatic index. Weight gain, condition factor, whole-body crude protein content, interleukin-10, and transforming growth factor beta gene expression were gradually enhanced with up to 102.68 mg/kg zinc and decreased at higher levels. The hepatosomatic index, feed conversion ratio, malondialdehyde level in the liver, aspartate aminotransferase, and alanine transaminase activity in the serum, gradually decreased up to 102.68 mg/kg zinc, and gradually increased beyond this. Activation of the nuclear factor erythroid-derived 2-like 2/Kelch-like ECH-associated protein 1 signaling pathway gradually up-regulated the mRNA levels and activities of glutathione peroxidase, total antioxidant capacity, catalase, and superoxide dismutase in the liver, this antioxidant ability was lower when the zinc was greater than 102.68 mg/kg. The gene expressions of nuclear factor-k-gene binding and pro-inflammation cytokines (interleukin-1β, interleukin-15, tumor necrosis factor alpha, and interleukin-8) were up-regulated up to 102.68 mg/kg zinc and then gradually repressed. In conclusion, using broken line analysis to estimate weight gain and Zn proteinate as the zinc source, the recommended dietary zinc for largemouth bass is 66.57 mg/kg zinc.

MicroRNAs Markedly Expressed in Apical Periodontitis Cooperatively Regulate Cytokines and Growth Factors Promoting an Anti-inflammatory Response

IntroductionMicroRNAs have been shown to play a role in the pathogenesis of apical periodontitis. Upregulation of miR-10a-5p and downregulation of miR-891a-5p were previously reported in apical periodontitis samples. This study aims to perform a functional characterization of miR-10a-5p, investigating its capacity to regulate the expression of inflammatory cytokines and growth factors, as well as a possible co-regulation mechanism with miR-891a-5p in the development of apical periodontitis. MethodsmiR-10a-5p mimics/controls and miR-891a-5p inhibitors/controls were introduced to human K-562 cells in the presence or absence of lipopolysaccharide. Total RNA was extracted from cell lysates, and target genes were examined via quantitative reverse transcription-polymerase chain reaction. Cell lysates were also subjected to proteomics analysis. Furthermore, mimics of miR-10a-5p and inhibitors of miR-891a-5p were co-transfected into K-562 cells. RNA sequencing and quantitative reverse transcription-polymerase chain reaction were carried out to examine their target genes. ResultsOverexpression of miR-10a-5p led to downregulation of tumor necrosis factor-alpha and interleukin-1 beta mRNA and upregulation of transforming growth factor-beta 1 (TGFB1) mRNA expression, whereas interleukin 3 and TGF-β1 proteins were upregulated. Simultaneous overexpression of miR-10a-5p and inhibition of miR-891a-5p further increased TGFB1 mRNA transcript levels. RNA sequencing revealed that genes co-regulated by miR-10a-5p and miR-891a-5p may be involved in apical periodontitis-related pathways such as tumor necrosis factor, transient receptor potential, and vascular endothelial growth factor signaling pathways. ConclusionsmiR-10a-5p may modulate the expression of multiple inflammatory cytokines and growth factors such as tumor necrosis factor-alpha, IL-1β, interleukin 3, and TGF-β1. In addition, miR-10a-5p and miR-891a-5p cooperatively regulate TGFB1 gene expression, and the gene network of this co-regulation is integrated with many pathways in apical periodontitis.

The rs1800469 T/T and rs1800470 C/C genotypes of the TGFB1 gene confer protection against diabetic retinopathy in a Southern Brazilian population.

The transforming growth factor beta 1 (TGFB1) is a pro-inflammatory cytokine that plays a key role in the mechanisms of angiogenesis and breakdown of the blood-retina barrier, which are implicated in the pathogenesis of diabetic retinopathy (DR). Polymorphisms in the TGFB1 gene have been associated with DR; however, results are still contradictory. Therefore, the aim of this study was to investigate the potential association between two TGFB1 polymorphisms and DR. This study included 992 patients with diabetes mellitus (DM): 546 patients with DR (cases) and 446 patients without DR and with ≥10 years of DM (controls). The TGFB1 rs1800469 and rs1800470 polymorphisms were genotyped by real-time PCR. Frequency of rs1800469 T/T genotype was higher in controls compared to DR cases (18.3% vs. 12.7%, P= 0.022). This genotype remained associated with protection for DR, adjusting for covariables (OR= 0.604; 95% CI 0.395 - 0.923; P= 0.020, recessive model). The rs1800470 C/C genotype was observed in 25.4% of the controls and 18.0% of the cases (P= 0.015); thus, being associated with protection against DR under the recessive model (OR= 0.589; 95% CI 0.405 - 0.857; P= 0.006), adjusting for covariables. In conclusion, the TGFB1 rs1800469 and rs1800470 polymorphisms are associated with protection against DR in DM patients from Southern Brazil.

Influence of photo biomodulation using 980 nm diode laser and exsosomes derived from dental pulp stem cells on pulp regeneration of dogs’ teeth

Recently , photobiomodulation has been shown to positively modulate the properties of dental pulp stem cells (DPSCs) in regenerative endodontics which is considered a promising solution to standard therapies for preserving pulp vitality and avoiding more extensive treatments such as endodontic therapy and tooth extraction. Therefore, the present study evaluated the effects of photobiomodulation and exsosomes of DPSCs for pulp regeneration in dogs . Five healthy mature mongrel dogs were divided into four groups ;group I, negative control; group II; Positive control(exposed pulp covered by A sterile piece of polytetrafluoroethylene (PTFE, Teflon tape) , group III: PBM group with 980 nm diode laser with output power 100 mw for one minute , group IV; exosomes, group V: exosomes + PBM.. Each dog represented one group. Blood samples were collected from dogs after 14dayspostoperatively. Biochemicals as total and ionized calcium, phosphorus, and alkaline phosphatase levels and the gene expression levels of matrix metallopeptidase 9, osteocalcin, transforming growth factor beta genes were measured with real-time quantitative polymerase chain reaction. Statistical analysis showed that there were significance differences between PBM, exsosomes and combination group of PBM and exsosomes compared to control group in different assessed biochemicals and genes expression’s levels. PBM combined with exsosomes enhanced the mineral content like total and ionized calcium , phosphorus level and alkaline phosphatase (ALP) as well as increased gene expression of Matrix metallopeptidase 9 (MMP 9), Transforming growth factor beta (TGF- β), and Osteocalcin OCN during pulp regeneration in dogs.

Dynamics of sexual development in teleosts with a note on Mugilcephalus

The process of sex determination (SD) and sex differentiation (Sd) with associated sex-specific behaviour in teleosts is brought about by genetic factors and environmental factors under the influence of chemical messengers. SD is initiated by inherited genes, which in turn influence Sd by the production of hormones. To understand the plasticity of SD in fishes, the functional role of genes must be clearly elucidated. During early development, the dimorphic expression of male and female are mediated by the differentially sexualized brain. In mammals, SD region (SDR) on the Y chromosome (SRY) can be considered as male-specific copy of SOX3 and in the absence of SRY in teleosts suggests SD might be regulated by alternate genes. DM (doublesex/mab)-related genes, AMH (anti-mullerian hormone), TGF-b (transforming growth factor beta), GSDF (gonadal soma derived growth factor) and other genes (around 18) located in chromosomes of teleosts are found to be responsible for SD. The SD gene, sdY (sexually dimorphic on the Y chromosome) has been detected in more than 15 salmonid species, and in medaka and tilapia SD system is governed by heterogametic mechanism. In mullet, Mugil cephalus Dor et al. identified 27 SDR genes and suggested to be potential candidate genes for SD. Recently whole-genome sequencing data were produced from mullet and assembled into a draft genome sequence in which >30 loci are potentially associated with SD. Further analysis is required to know the involvement of each sex-biased gene for its functional influence on sex. Investigators attempted to know the sex-linkage group in tilapia and found that 2 linkage group i.e. LG1 and LG22 show SD loci, whereas in mullet it was documented that LG8 is the SD loci. Rigorous study with all male population(YY) or knockout of male marker gene, might give conclusive understanding on SD in future. The review highlights the absence of genomic understanding of SD and SDR genes for futuristic direction of research.

Transcription regulators differentiate mesenchymal stem cells into chondroprogenitors, and their in vivo implantation regenerated the intervertebral disc degeneration.

BACKGROUNDIntervertebral disc degeneration (IVDD) is the leading cause of lower back pain. Disc degeneration is characterized by reduced cellularity and decreased production of extracellular matrix (ECM). Mesenchymal stem cells (MSCs) have been envisioned as a promising treatment for degenerative illnesses. Cell-based therapy using ECM-producing chondrogenic derivatives of MSCs has the potential to restore the functionality of the intervertebral disc (IVD). AIMTo investigate the potential of chondrogenic transcription factors to promote differentiation of human umbilical cord MSCs into chondrocytes, and to assess their therapeutic potential in IVD regeneration.METHODSMSCs were isolated and characterized morphologically and immunologically by the expression of specific markers. MSCs were then transfected with Sox-9 and Six-1 transcription factors to direct differentiation and were assessed for chondrogenic lineage based on the expression of specific markers. These differentiated MSCs were implanted in the rat model of IVDD. The regenerative potential of transplanted cells was investigated using histochemical and molecular analyses of IVDs. RESULTSIsolated cells showed fibroblast-like morphology and expressed CD105, CD90, CD73, CD29, and Vimentin but not CD45 antigens. Overexpression of Sox-9 and Six-1 greatly enhanced the gene expression of transforming growth factor beta-1 gene, BMP, Sox-9, Six-1, and Aggrecan, and protein expression of Sox-9 and Six-1. The implanted cells integrated, survived, and homed in the degenerated intervertebral disc. Histological grading showed that the transfected MSCs regenerated the IVD and restored normal architecture.CONCLUSIONGenetically modified MSCs accelerate cartilage regeneration, providing a unique opportunity and impetus for stem cell-based therapeutic approach for degenerative disc diseases.

Risk and protection strategies of Amolops wuyiensis intestine against gastrointestinal nematode (Cosmocercoides wuyiensis n. sp.) infection.

Anuran amphibians are susceptible to infection by intestinal nematodes, but the damage and response mechanisms that occur in their intestines after infection are only partially understood. In this study, the intestinal disruption and response mechanisms in Amolops wuyiensis frogs infected with Cosmocercoides wuyiensis n. sp. were revealed through analysis of the intestinal histopathology, digestive enzyme activity, transcriptome and intestinal microbiota. Tissue section analysis showed histological damage and inflammation in the infected intestine, and the digestive enzyme activity indicated a decrease in digestion and absorption of some nutrients. We found that infection led to differences in the intestinal microbiota composition, including lower diversity and symbiotic relationships. The greater relative abundance of the genera Burkholderia and Rhodococcus may enhance intestinal immunity to resist pathogenic infections. A comparison of the transcriptomes of infected and uninfected intestines revealed 1055 differentially expressed genes. GO enrichment and KEGG pathways analyses suggested that the guts of infected C. wuyiensis n. sp. show enhanced complement activation, cell adhesion molecule function, NOD-like receptor signalling pathway activity and other innate immunity responses. Among the adaptive immune responses, the intestinal immune network for IgA production was significantly enriched, and the expression of IL-17D and transforming growth factor beta-1 genes were upregulated in the infected intestine. These results imply that C. wuyiensis n. sp. infection of A. wuyiensis intestine may trigger innate and adaptive immune responses,which reduce the post-infection burden. Furthermore,the intestine of A. wuyiensis may also respond to C. wuyiensis n. sp. infection by increasing metallocarboxypeptidase activity and accelerating smooth muscle contraction.

Polymorphism -509C/T in TGFB1 Promoter Is Associated With Increased Risk and Severity of Persistent Allergic Rhinitis in a Chinese Population.

Polymorphism -509C/T in the promoter of transforming growth factor beta1 (TGFB1) gene is implicated in the pathogenesis of asthma. This polymorphism might also act to regulate the development of allergic rhinitis (AR). To investigate whether -509C/T is associated with AR susceptibility and severity in a Han Chinese population. The study enrolled 263 patients with persistent AR and 249 healthy controls. AR patients were classified as mild or moderate/severe AR groups according to the Allergic Rhinitis and its Impact on Asthma classification. TGFB1 gene polymorphism -509C/T was genotyped with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Serum total Immunoglobulin E (IgE) and specific IgE levels were determined using an ImmunoCAP. Significant difference was found in the allele frequency of TGFB1 -509C/T between AR patients and healthy controls (P = .027) but not in the genotype frequency (P =.051). However, the genotype frequency of TGFB1 -509C/T showed significant difference between the mild AR group, the moderate/severe AR group, and the control group (P = .012); between the moderate/severe AR group and the control group (P =.036); between the mild AR group and the moderate/severe AR group (P = .038); but not between the mild AR group and the control group (P =.075). TGFB1 promoter polymorphism -509C/T may be associated with the susceptibility and the severity of persistent AR of Han Chinese, but the functional relationship still needs clarification.

Insights Into Management of Camurati-Engelmann Disease: A Case Series of Three Siblings

Camurati-Engelmann disease (CED) is an autosomal dominant skeletal dysplasia characterized by progressive sclerosis of long bones due to a mutation in the transforming growth factor beta-1 gene. Patients experience progressive pain, weakness, and fatigability over time. There are no consensus guidelines for treatment though the use of several types of glucocorticoids, angiotensin II receptor blockers, and other therapies have been described. We present the cases of three siblings with CED managed with different treatment modalities over time (prednisone, losartan, and deflazacort). We provide objective data (pain scores, walk-test results, erythrocyte sedimentation rates) to demonstrate therapeutic efficacy. Prednisone resulted in the greatest improvement in pain; however, its use was limited by significant weight gain. Deflazacort was successful in improving pain and fatigability without the weight gain. Risks and benefits must be considered carefully as the cost of deflazacort is significantly higher than prednisone or losartan. Int J Clin Pediatr. 2020;9(4):130-134 doi: https://doi.org/10.14740/ijcp403

Effects of combination TGF-B1 transfection and platelet rich plasma (PRP) on three-dimension chondrogenic differentiation of rabbit dental pulp-derived mesenchymal stem cells

ABSTRACT Aim: The aim of this study was to evaluate the effects of standard culture medium and chondrogenic differentiation medium with PRP on chondrogenic differentiation of rabbit dental pulp-derived mesenchymal stem cells (rabbit DPSCs) that are transfected with transforming growth factor-beta 1 (TGF-B1) gene, based on the hypothesis of TGF- B1 and PRP can be effective on the chondrogenesis of stem cells. Materials and Methods: Rabbit DPSCs were characterized by using flow cytometry, immunofluorescent staining, quantitative Real Time Polymerase Chain Reaction (qRT-PCR) and differentiation tests. For the characterization, CD29, CD44 and CD45 mesenchymal cell markers were used. Rabbit DPSCs were transfected with TGF-B1 gene using electroporation technique in group 1; with PRP 10% in group 2; with chondrogenic medium in group 3; with both chondrogenic medium and PRP in group 4. DPSCs were cultured in medium with 10% inactive PRP in group 5, chondrogenic medium in group 6, chondrogenic medium with PRP 10% in group 7. SOX9, MMP13 and Aggrecan gene expression levels were evaluated in 3, 6, 12. and 24. days by qRT-PCR. Results: The expression levels of SOX9, MMP13 and Aggrecan were higher in group 2, 3 and group 7 in 3th day however in 24th day group 7 and group 2 were found higher. The expression levels changed by time-dependent. The extracellular matrix of the cells in experimental groups were positively stained with safranin O and toluidine blue. Conclusion: The combination in culture medium of TGF-B1 gene transfection and 10% PRP accelerates the chondrogenic differentiation of DPSCs.

Effects of lavender (Lavandula angustifolia) extract inclusion in diet on growth performance, innate immunity, immune-related gene expression, and stress response of common carp, Cyprinus carpio

In the present study, the effects of dietary lavender (Lavandula angustifolia) extract (LE) on growth performance and immune, antioxidant, and stress responses in common carp, Cyprinus carpio were investigated. For this, four diets containing 0, 0.5, 1.0 and 1.5% LE were offered to triplicate groups of fish (25.8 ± 1.66 g) for 70 days, and then the fish were exposed to a 3-h crowding stress. Growth performance, blood leukocyte count, plasma proteins and innate immune parameters, and head kidney cytokines’ gene expression were studied after the 70-day feeding trial (before the crowding stress). Plasma cortisol, glucose, superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) were measured before and after the crowding stress. Results showed that dietary LE had no significant effects on the fish growth performance, blood leukocyte differential count, plasma albumin and total protein, and head kidney interferon gamma-1 gene expression. Blood leukocyte and plasma globulin, alternative complement and total immunoglobulin significantly increased in the fish fed with 1.0–1.5% LE diets compared to the fish fed with the control diet (0% LE). The plasma lysozyme activity significantly increased along with dietary LE supplemental levels. The lowest expression of interlukin-1 beta gene was related to the 1.0% LE treatment. Tumor necrosis factor-alpha gene expression significantly decreased along with increase in dietary LE levels and the lowest expression was related to the fish fed with 1.5% LE diet. The LE administration led to significant increase in interlukine-10 gene expression and the highest expression was related to the fish fed with 1.0% and 1.5% LE diets. The fish fed with 1.5% LE diet had significantly higher transforming growth factor-beta gene expression compared to the control group. Liver SOD and CAT activities showed significant increases along with dietary LE supplementation levels. After the crowding stress, hepatic SOD decreased in the control group, but not the LE-supplemented groups. Stress led to significant decrease in hepatic CAT activities in all treatments, but CAT activity increased along with the LE levels with highest activity in the fish fed with 1.5% LE. The stress led to significant increase in hepatic MDA in the treatments control and 0.5% LE, but not 1.0–1.5% LE groups. The stress led to significant increase in plasma cortisol and glucose levels in all treatments but dietary LE levels significantly decreased these parameters. Dietary LE supplementation at levels of 1.0–1.5% is recommended for common carp to suppress stress, inflammation and oxidative conditions and augment immune responses in the fish.

Mutated CEACAMs Disrupt Transforming Growth Factor Beta Signaling and Alter the Intestinal Microbiome to Promote Colorectal Carcinogenesis

We studied interactions among proteins of the carcinoembryonic antigen-related cell adhesion molecule (CEACAM) family, which interact with microbes, and transforming growth factor beta (TGFB) signaling pathway, which is often altered in colorectal cancer cells. We investigated mechanisms by which CEACAM proteins inhibit TGFB signaling and alter the intestinal microbiome to promote colorectal carcinogenesis. We collected data on DNA sequences, messenger RNA expression levels, and patient survival times from 456 colorectal adenocarcinoma cases, and a separate set of 594 samples of colorectal adenocarcinomas, in The Cancer Genome Atlas. We performed shotgun metagenomic sequencing analyses of feces from wild-type mice and mice with defects in TGFB signaling (Sptbn1+/- and Smad4+/-/Sptbn1+/-) to identify changes in microbiota composition before development of colon tumors. CEACAM protein and its mutants were overexpressed in SW480 and HCT116 colorectal cancer cell lines, which were analyzed by immunoblotting and proliferation and colony formation assays. In colorectal adenocarcinomas, high expression levels of genes encoding CEACAM proteins, especially CEACAM5, were associated with reduced survival times of patients. There was an inverse correlation between expression of CEACAM genes and expression of TGFB pathway genes (TGFBR1, TGFBR2, and SMAD3). In colorectal adenocarcinomas, we also found an inverse correlation between expression of genes in the TGFB signaling pathway and genes that regulate stem cell features of cells. We found mutations encoding L640I and A643T in the B3 domain of human CEACAM5 in colorectal adenocarcinomas; structural studies indicated that these mutations would alter the interaction between CEACAM5 and TGFBR1. Overexpression of these mutants in SW480 and HCT116 colorectal cancer cell lines increased their anchorage-independent growth and inhibited TGFB signaling to a greater extent than overexpression of wild-type CEACAM5, indicating that they aregain-of-function mutations. Compared with feces from wild-type mice, feces from mice with defects in TGFB signaling had increased abundance of bacterial species that have been associated with the development of colon tumors, including Clostridium septicum, and decreased amounts of beneficial bacteria, such as Bacteroides vulgatus and Parabacteroides distasonis. We found expression of CEACAMs and genes that regulate stem cell features of cells to be increased in colorectal adenocarcinomas and inversely correlated with expression of TGFB pathway genes. We found colorectal adenocarcinomas to express mutant forms of CEACAM5 that inhibit TGFB signaling and increase proliferation and colony formation. We propose that CEACAM proteins disrupt TGFB signaling, which alters the composition of the intestinal microbiome to promote colorectal carcinogenesis.

Angiographic dynamics of coronary flow state after percutaneous coronary intervention in carriers of polymorphic RS1800470 variants of the TGF-β1 gene

Aim. To study the relationship between the angiographic dynamics of the state of the coronary bed in patients undergoing percutaneous coronary intervention (PCI) due to myocardial infarction (MI) and the carriage of polymorphic rs1800470 variants of the transforming growth factor beta-1 gene (TGF-β1).Material and methods. The study included 89 men with MI of the Caucasian race aged 32 to 65 years (52 [45,0-58,0]). Genomic deoxyribonucleic acid (DNA) was extracted from venous blood by phenol-chloroform technique. The rs1800470 polymorphism of the TGF-β1 gene was tested using real-time polymerase chain reaction (PCR) (TaqMan probes, AB 7900HT). Assessment of the severity of coronary lesions was carried out initially according to the standard coronary angiography (CAG) protocol with the calculation of the Gensini score. CAG was also conducted in the dynamics after 5-103 months (42,3±29,5 months) of the study beginning.Results. In male carriers of the rs1800470 A allele of the TGF-β1 gene, the mean values of the Gensini score statistically significantly increased (47,5±34,1 (CAG-1) and 64,5±35,5 (CAG-2), p <0,001) in comparison with carriers of the homozygous GG rs1800470 variant of the TGF-β1 gene (43,5±21,1 (CAG-1) and 46,2±23,2 (CAG2), p=0,066). In patients who had rs1800470 A allele of the TGF-β1 gene, a 20%decrease in the Gensini score was observed after 7 months (p=0,013), and 30% — after 5 months (p=0,003) of the initial CAG. The development of late stent restenoses in carriers of the rs1800470 A allele and the homozygous risk genotype AA of the TGF-β1 gene was noted at an earlier date — 8 (p=0,047) and 12 months (p=0,002), respectively.Conclusion. Currently, the conduct of CAG in dynamics in patients undergoing PCI is recommended as clinically indicated. The exception is the group of patients who underwent endoprosthesis replacement of the left coronary artery or have uncorrected stenosis of a different location from 3 to 12 months after PCI. The presented data show a possible predictor role of rG1800470 polymorphism of the TGF-β1 gene in relation to the progression of coronary atherosclerosis and the development of late stent restenoses.

Observations on the Natural History of Camurati-Engelmann Disease.

Camurati-Engelmann disease (OMIM 31300) is a rare cranio-tubular bone dysplasia characterized by osteosclerosis of the long bones and skull caused by dominantly-inherited mutations in the transforming growth factor beta 1 (TGFB1) gene. A wide variation in phenotype has been recognized, even within families carrying the same mutation. In addition, aspects of the natural history of the disorder, in particular whether it is always progressive or can remit spontaneously, remain uncertain. In a large kindred carrying a TGFB1 gene mutation (c.653G > A; p.R218H) we have attempted to clarify the extent of phenotypic variability and the natural history of the disease through detailed individual histories of symptoms, and skeletal imaging by both radiography and scintigraphy. Only one subject had the classical childhood onset with bone pain in the legs and gait disturbance. Eight subjects reported the onset of leg pain in their teenage years that, by their early 20s, had either resolved or persisted at a low level. Two of these eight later developed cranial nerve palsies. There was a wide variation in the radiographic appearance in adults, but disease extent and activity in long bones, as assessed by scintigraphy, was inversely correlated with age (p < 0.025). In younger subjects the radiographic and scintigraphic appearances were concordant, but in older subjects the scintigram could be quiescent despite florid radiographic changes. Sequential scintigrams in two subjects showed reduced activity in the later scan. One subject had suffered meningoencephalitis in early childhood that resulted in paresis of one arm. The affected arm showed markedly less disease involvement, implicating mechanical or growth factors in its etiology. Our data suggest that the natural history of Camurati-Engelmann disease can be benign, and that disease activity commonly attenuates in adulthood. Severe cases of childhood onset and/or with cranial nerve involvement, may occur only in a minority of mutation carriers. © 2019 American Society for Bone and Mineral Research.

Receptor tyrosine kinase MerTK suppresses an allogenic type I IFN response to promote transplant tolerance.

Recipient infusion of donor apoptotic cells is an emerging strategy for inducing robust transplantation tolerance. Daily clearance of billions of self-apoptotic cells relies on homeostatic engagement of phagocytic receptors, in particular, receptors of the tyrosine kinase family TAM (Tyro3, Axl, and MerTK), to maintain self-tolerance. However, an outstanding question is if allogeneic apoptotic cells trigger the same receptor system for inducing allogeneic tolerance. Here, we employed allogeneic apoptotic splenocytes and discovered that the efferocytic receptor MerTK on recipient phagocytes is a critical mediator for transplantation tolerance induced by this strategy. Our findings indicate that the tolerogenic properties of allogeneic apoptotic splenocytes require MerTK transmission of intracellular signaling to suppress the production of inflammatory cytokine interferon α (IFN-α). We further demonstrate that MerTK is crucial for subsequent expansion of myeloid-derived suppressor cells and for promoting their immunomodulatory function, including maintaining graft-infiltrating CD4+ CD25+ Foxp3+ regulatory T cells. Consequently, recipient MerTK deficiency resulted in failure of tolerance by donor apoptotic cells, and this failure could be effectively rescued by IFN-α receptor blockade. These findings underscore the importance of the efferocytic receptor MerTK in mediating transplantation tolerance by donor apoptotic cells and implicate MerTK agonism as a promising target for promoting transplantation tolerance.

Nanostructured lipid carriers containing rapamycin for prevention of corneal fibroblasts proliferation and haze propagation after burn injuries: In vitro and in vivo.

Chemical burns are a major cause of corneal haze and blindness. Corticosteroids are commonly used after corneal burns to attenuate the severity of the inflammation-related fibrosis. While research efforts have been aimed toward application of novel therapeutics. In the current study, a novel drug delivery system based nanostructured lipid carriers (NLCs) were designed to treat corneal alkaline burn injury. Rapamycin, a potent inhibitor of mammalian target of rapamycin pathway, was loaded in NLCs (rapa-NLCs), and the NLCs were characterized. Cell viability assay, cellular uptake of NLCs, and in vitro evaluation of the fibrotic/angiogenic genes suppression by rapa-NLCs were carried out on human isolated corneal fibroblasts. Immunohistochemistry (IHC) assayswere also performed after treatment of murine model of corneal alkaline burn with rapa-NLCs. According to the results, rapamycin was efficiently loaded in NLCs. NLCs could enhance coumarin-6 fibroblast uptake by 1.5 times. Rapa-NLCs efficiently downregulated platelet-derived growth factor and transforming growth factor beta genes in vitro. Furthermore, proliferation of fibroblasts, a major cause of corneal haze after injury, reduced. IHC staining of treated cornea with alpha-smooth muscle actin and CD34 + antibodies showed efficient prevention of myofibroblasts differentiation and angiogenesis, respectively. In conclusion, ocular delivery of rapamycin using NLCs after corneal injury may be considered as a promising antifibrotic/angiogenic treatment approach to preserve patient eyesight.