Takayasu arteritis (TAK) is an autoimmune large vessel vasculitis that affects the aorta and its major branches, eventually leading to the development of aortic aneurysm and vascular stenosis or ...occlusion. This retrospective and prospective study aimed to investigate whether the gut dysbiosis exists in patients with TAK and to identify specific gut microorganisms related to aortic aneurysm formation/progression in TAK.
We analysed the faecal microbiome of 76 patients with TAK and 56 healthy controls (HCs) using 16S ribosomal RNA sequencing. We examined the relationship between the composition of the gut microbiota and clinical parameters.
The patients with TAK showed an altered gut microbiota with a higher abundance of oral-derived bacteria, such as Streptococcus and Campylobacter, regardless of the disease activity, than HCs. This increase was significantly associated with the administration of a proton pump inhibitor used for preventing gastric ulcers in patients treated with aspirin and glucocorticoids. Among patients taking a proton pump inhibitor, Campylobacter was more frequently detected in those who underwent vascular surgeries and endovascular therapy for aortic dilatation than in those who did not. Among the genus of Campylobacter, Campylobacter gracilis in the gut microbiome was significantly associated with clinical events related to aortic aneurysm formation/worsening in patients with TAK. In a prospective analysis, patients with a gut microbiome positive for Campylobacter were significantly more likely to require interventions for aortic dilatation than those who were negative for Campylobacter. Furthermore, patients with TAK who were positive for C. gracilis by polymerase chain reaction showed a tendency to have severe aortic aneurysms.
A specific increase in oral-derived Campylobacter in the gut may be a novel predictor of aortic aneurysm formation/progression in patients with TAK.
Pulmonary arterial hypertension (PAH) is characterized by stenosis and occlusions of small pulmonary arteries, leading to elevated pulmonary arterial pressure and right heart failure. Although ...accumulating evidence shows the importance of interleukin (IL)-6 in the pathogenesis of PAH, the target cells of IL-6 are poorly understood. Using mice harboring the
allele of
, a subunit of the IL-6 receptor, we found substantial Cre recombination in all hematopoietic cell lineages from the primitive hematopoietic stem cell level in
mice. We also revealed that a CD4
cell-specific gp130 deletion ameliorated the phenotype of hypoxia-induced pulmonary hypertension in mice. Disruption of IL-6 signaling via deletion of
in CD4
T cells inhibited phosphorylation of signal transducer and activator of transcription 3 (STAT3) and suppressed the hypoxia-induced increase in T helper 17 cells. To further examine the role of IL-6/gp130 signaling in more severe PH models, we developed
knockout (KO) rats using the CRISPR/Cas9 system and showed that IL-6 deficiency could improve the pathophysiology in hypoxia-, monocrotaline-, and Sugen5416/hypoxia (SuHx)-induced rat PH models. Phosphorylation of STAT3 in CD4
cells was also observed around the vascular lesions in the lungs of the SuHx rat model, but not in
KO rats. Blockade of IL-6 signaling had an additive effect on conventional PAH therapeutics, such as endothelin receptor antagonist (macitentan) and soluble guanylyl cyclase stimulator (BAY41-2272). These findings suggest that IL-6/gp130 signaling in CD4
cells plays a critical role in the pathogenesis of PAH.
Abstract
Fucoidan is a series of sulfated polysaccharides derived from brown algae that mainly consist of L-fucose. Fucoidan have various biological activities, and our investigation with ...experimental animal models and a healthy human trial demonstrated fucoidan from Cladosiphon okamuranus and Undaria pinnatifida effectively augmented anti-tumor immunity in combination with agaricus mycelium extract. In this context, we have reported that feeding of the fucoidan-agaricus mix (FAM) protected mice from immunosuppressive effect of 5-fluorouracil (5-FU). In current study, we confirmed dose-dependent immunoprotective effects of the dietary FAM (at 20 and 40 mg/day) by observing recovery in spleen weight, NK cell activity and IFN-γ production in 5-FU-treated mice. Furthermore, we investigated in vitro effects of FAM against 5-FU on growth and function of primary cultured immune-related cells. As the results with thioglycollate-induced peritoneal exudates cells (PEC), 5-FU dose-dependently suppressed the growth whereas FAM exerted no effects. However, production of cytokines such as IL-12p40 and TNF-α in PEC were stimulated by FAM treatment even in the presence of 5-FU. In the case of bone marrow derived cells differentiated with GM-CSF and M-CSF, FAM markedly enhanced the growth and IL-12 production of these cells. From these results, it was suggested that FAM contributed to amelioration of decline in anti-tumor immune function by 5-FU treatment through activation of innate immune cells.
Abstract
Fucoidan are sulfated polysaccharides derived from brown seaweeds that mainly consist of L-fucose. Peoples in Asian countries eat seaweeds willingly as healthy foods. In previous studies, we ...have demonstrated that fucoidan augment anti-tumor immunity in an experimental mouse model and in healthy human subjects. However, availability of fucoidan to protect immune system from hazardous side effects of cancer chemotherapy has not been verified enough. Therefore, we performed a mouse model experiment in which immunosuppression was induced by subcutaneous 5-fluorouracil (5-FU) injection (1 mg/mouse/day, 5 times on alternate days) and evaluated protective effects of orally administered fucoidan mix (40 mg/mouse/day). The immunosuppression by 5-FU s.c. was confirmed with the loss in spleen weight of 5-FU-injected mice. NK cell activity was lower in 5-FU-injected mice than sham-treated control mice, but the decline was recovered by the fucoidan mix feeding. The intake of fucoidan mix significantly alleviated the reduction in interferon (IFN)-gamma production of splenocytes from 5-FU-injected mice. Furthermore, flow cytometric analyses revealed that specific population such as Th1 and CD11b+ cells were supported by the fucoidan mix feeding even in the 5-FU-injected mice. These results suggested that oral administration of fucoidan mix is effective to maintain tumor immunity during immunosuppressive cancer chemotherapy.
Abstract
Fucoidan is a sulfated polysaccharide derived from brown seaweeds which mainly consist of L-fucose. In Asian countries such as Japan, Korea and China, peoples eat seaweeds willingly as ...nourishing health foods. We previously showed that the tumor formation by transplanted Sarcoma180 cells was suppressed in mice fed 2% fucoidan mix containing diets, and NK cell activity was enhanced in the fucoidan-fed mice. The results suggest that the activated NK cells participated in elimination of the tumor. However, in human, immune enhancing activities of fucoidan have hardly been proved. Therefore, we performed an open-label trial in this study. Five healthy volunteers (47 ± 6 years old, 2 male and 3 female) took 2.5 g fucoidan mix per day for 30 days. There were no abnormal observations in the values of WBC, RBC, platelet count and blood profile. On the other hand, NK cell activity was augmented by 1.42 times in the average at end of the trial than the initial value. The ratio of IFN-gamma-producing (Th1) cells also increased to 114% for the initial value by the day 30 by the fucoidan mix intake, whereas IL-4-producing (Th2) cells decreased to 77%. As a result, the rise of Th1/Th2 ratio by 1.46 times in the average was observed with 4 subjects among 5 examinees. Furthermore, CD8+ cells tended to expand with the all subjects. These results suggest that intake of fucoidan is effective to augment anti-tumor immunity in human.
Immune function is influenced by many exercise conditions, for example, exercise intensity and frequency. In this study, we compared exercise rats that trained for two weeks then exercised ...exhaustively, with sedentary rats that exercised suddenly and exhaustively on immunoglobulin (Ig) productivity. As a result, in exercised groups, IgA production of mesenteric lymph node (MLN) lymphocytes significantly increased. In the sedentary and exercised groups, Ig production of MLN lymphocytes was tended to decrease by exhaustive exercise. But, in the exercised group, Ig production were maintained higher concentrations than that of the sedentary group. Therefore, we expected that antioxidative ability of rat be enhanced by exercise, and increase Ig production. However, in the exercised groups, lipid peroxide concentration of lymphocyte in culture supernatant was higher value than that of the sedentary groups, and glutathione peroxidase activity in blood was significantly decreased by exhaustive exercise. Thus, training exercise increase Ig productivity and suppress the damage in gut immune system. However, that mechanism may be not related to increase of antioxidative function. On the other hand, oxidative stress by exhaustive exercise may relate to the suppression of Ig productivity. Therefore, it is suggested that the reinforcement and suppression of Ig productivity by exercise are controlled by different mechanism, respectively.
The effect of exercise on the elevation of HSP70 was studied in rats fed a different diet on the B_6 content (as pyridoxine 1.5 mg/kg diet or 7.5 mg/kg diet). Minimum requirement of B_6 in the diet ...is known to be about 1.5 mg/kg diet in 20 % casein diet. Rats were divided into 4 groups in each series and assigned as Sc (sedentary control), Se (sedentary-exhaustion), Ec (training control), and Ee (trainingexhaustion). HSP70 was determined in the heart, liver and gastrocnemius muscle (red and white portions). HSP70 in the heart was not changed in all groups of both series. In the liver HSP70 was elevated to very high levels in Sc and Ee groups in the rats fed the high level of B_6, and where as a little in Ee group in rats fed low level of B_6. The levels of inductions were in the muscle not so high very similar but as observed in the liver. AST (aspartate aminotransferase) activity was significantly elevated in Ec and Ee for the liver, and in the three groups for the kidney and muscle (red portion) in the rats fed the high level of B_6. On the contrary in all tissues in all groups fed low level of B_6, AST activity was not changed at all. Thus the low induction of HSP70 in the tissues, especially in the liver of rats fed the low B_6 diet, may suggest impairment of amino acid metabolism and, consequently, insufficiency of energy supply.
Recently developed long-read sequencing (LRS) technology has been considered an option for CYP21A2 analysis. However, the clinical use of LRS for CYP21A2 analysis is limited.
This study's objective ...is to develop an efficient and low-cost LRS system for CYP21A2 screening.
A DNA fragment library was prepared in a single polymerase chain reaction (PCR) that covers the entire CYP21A2 gene and all known junctions caused by TNXB gene structural rearrangements, yielding a single 8-kb product of CYP21A2 or CYP21A1P/CYP21A2 chimera. After barcoding, the PCR products were sequenced on a MinION-based platform with Flongle Flow Cell R9.4.1 and R10.4.1.
The reference genotypes of 55 patients with 21-hydroxylase deficiency (21OHD) were established using the conventional method with multiplex ligation-dependent probe amplification (MLPA) and nested PCR. LRS using Flongle Flow Cell R9.4.1 yielded consistent results. Additionally, the recently updated LRS "duplex" analysis with Flongle flow cell R10.4.1 was tested to reveal an advantage of accurately sequencing a variant located on the homopolymer region. By introducing a barcode system, the cost was reduced to be comparable to that of conventional analysis. A novel single-nucleotide variation was discovered at the acceptor site of intron 7, c.940-1G > C. We also identified a subtype of the classical chimeric junction CH2, "CH2a," in the region from the latter part of intron 5 to exon 6.
We successfully established a novel low-cost and highly accurate LRS system for 21OHD genetic analysis. Our study provides insight into the feasibility of LRS for diagnosing 21OHD and other genetic diseases caused by structural rearrangements.
Post-transcriptional modifications at the first (wobble) position of the tRNA anticodon participate in precise decoding of the genetic code. To decode codons that end in a purine (R) (i.e. NNR), ...tRNAs frequently utilize 5-methyluridine derivatives (xm5U) at the wobble position. However, the functional properties of the C5-substituents of xm5U in codon recognition remain elusive. We previously found that mitochondrial tRNAsLeu(UUR) with pathogenic point mutations isolated from MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) patients lacked the 5-taurinomethyluridine (τm5U) modification and caused a decoding defect. Here, we constructed Escherichia coli tRNAsLeu(UUR) with or without xm5U modifications at the wobble position and measured their decoding activities in an in vitro translation as well as by A-site tRNA binding. In addition, the decoding properties of tRNAArg lacking mnm5U modification in a knock-out strain of the modifying enzyme (ΔmnmE) were examined by pulse labeling using reporter constructs with consecutive AGR codons. Our results demonstrate that the xm5U modification plays a critical role in decoding NNG codons by stabilizing U·G pairing at the wobble position. Crystal structures of an anticodon stem-loop containing τm5U interacting with a UUA or UUG codon at the ribosomal A-site revealed that the τm5U·G base pair does not have classical U·G wobble geometry. These structures provide help to explain how the τm5U modification enables efficient decoding of UUG codons.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
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