Distinctive gut microbiomes of ankylosing spondylitis and inflammatory bowel disease patients suggest differing roles in pathogenesis and correlate with disease activity.

BACKGROUND: Multiple studies have confirmed dysbiosis in ankylosing spondylitis (AS) and inflammatory bowel disease (IBD); however, due to methodological differences across studies, it has not been possible to determine if these diseases have similar or different gut microbiomes. RESULTS: In this study, faecal and intestinal biopsies were obtained from 33 Australian AS patients (including 5 with concomitant IBD, 'AS-IBD'), 59 IBD patients and 105 healthy controls. Stool samples were also obtained from 16 Italian AS patients and 136 Swedish AS patients. Focusing on the Australian cohort, AS, AS-IBD and IBD patients differed from one another and from healthy controls in both alpha and beta diversity. AS patients with and without clinical IBD could be distinguished from one another with moderate accuracy using stool microbiome (AUC=0.754). Stool microbiome also accurately distinguished IBD patients from healthy controls (AUC=0.757). Microbiome composition was correlated with disease activity measured by BASDAI and faecal calprotectin (FCP) levels. Enrichment of potentially pathogenic Streptococcus was noted in AS, AS-IBD and IBD patients. Furthermore, enrichment of another potentially pathogenic genus, Haemophilus, was observed in AS, AS-IBD, IBD, AS patients with increased BASDAI, and IBD patients with faecal calprotectin >100 µg/mg. Apart from these genera, no other taxa were shared between AS and IBD patients. CONCLUSIONS: In conclusion, the distinct gut microbiome of AS and AS-IBD patients compared to IBD patients and healthy controls is consistent with immunological and genetic evidence suggesting that the gut plays a different role in driving AS compared with IBD. However, enrichment of two potentially pathogenic genera in both diseases suggests that the presence of a shared/common microbial trigger of disease cannot be discounted.

Transcriptome analysis of ankylosing spondylitis patients before and after TNF-α inhibitor therapy reveals the pathways affected.

Tumor necrosis factor-α (TNF-α) inhibitors are highly effective in suppressing inflammation in ankylosing spondylitis (AS) patients, and operate by suppression of TFN-α and downstream immunological pathways. To determine the mechanisms of action of TNF-α inhibitors in AS patients, we used transcriptomic and bioinformatic approaches on peripheral blood mononuclear cells from AS patients pre and post treatment. We found 656 differentially expressed genes, including the genome-wide significant AS-associated genes, IL6R, NOTCH1, IL10, CXCR2 and TNFRSF1A. A distinctive gene expression profile was found between male and female patients, mainly because of sex chromosome-linked genes and interleukin 17 receptor C, potentially accounting for the differences in clinical manifestation and treatment response between the genders. In addition to immune and inflammation regulatory pathways, like intestinal immune network for IgA production, cytokine-cytokine receptor interaction, Ras signaling pathway, allograft rejection and hematopoietic cell lineage, KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analyses revealed that infection-associated pathways (influenza A and toxoplasmosis) and metabolism-associated pathways were involved in response to TNF-α inhibitor treatment, providing insight into the mechanism of TNF-α inhibitors.

Epigenetic and gene expression analysis of ankylosing spondylitis-associated loci implicate immune cells and the gut in the disease pathogenesis.

Ankylosing spondylitis (AS) is a common immune-mediated arthropathy primarily affecting the spine and pelvis. Most AS patients have subclinical intestinal inflammation, suggesting the gut microbiome and the immune response play a role in pathogenesis. Susceptibility to AS is primarily genetic, and at least 114 susceptibility variants have been identified to date. We applied bioinformatic methods utilizing epigenetic and gene and protein expression data to identify the cell types through which AS-associated variants operate. Variants were enriched in transcriptionally regulated regions in monocytes, CD4+ and CD8+ T cells, natural killer cells, regulatory T cells and B cells and mucosa from the small intestine, sigmoid colon and rectum. Weak signals were detected in bone cells, consistent with bone disease being a secondary manifestation. RNA sequencing of blood cells from AS patients and controls identified differentially expressed genes. Interrogation of expression databases showed that the upregulated genes were enriched in monocytes and downregulated genes were enriched in CD8+ T cells and natural killer cells. Gene Ontology term enrichment analysis identified microbes and the gut in the aetiology of AS. These findings identify the key immune cell types that drive the disease, and further highlight the involvement of the gut microbiome in the pathogenesis of AS.

Disease-associated polymorphisms in ERAP1 do not alter endoplasmic reticulum stress in patients with ankylosing spondylitis.

The mechanism by which human leukocyte antigen B27 (HLA-B27) contributes to ankylosing spondylitis (AS) remains unclear. Genetic studies demonstrate that association with and interaction between polymorphisms of endoplasmic reticulum aminopeptidase 1 (ERAP1) and HLA-B27 influence the risk of AS. It has been hypothesised that ERAP1-mediated HLA-B27 misfolding increases endoplasmic reticulum (ER) stress, driving an interleukin (IL) 23-dependent, pro-inflammatory immune response. We tested the hypothesis that AS-risk ERAP1 variants increase ER-stress and concomitant pro-inflammatory cytokine production in HLA-B27(+) but not HLA-B27(-) AS patients or controls. Forty-nine AS cases and 22 healthy controls were grouped according to HLA-B27 status and AS-associated ERAP1 rs30187 genotypes: HLA-B27(+)ERAP1(risk), HLA-B27(+)ERAP1(protective), HLA-B27(-)ERAP1(risk) and HLA-B27(-)ERAP1(protective). Expression levels of ER-stress markers GRP78 (8 kDa glucose-regulated protein), CHOP (C/EBP-homologous protein) and inflammatory cytokines were determined in peripheral blood mononuclear cell and ileal biopsies. We found no differences in ER-stress gene expression between HLA-B27(+) and HLA-B27(-) cases or healthy controls, or between cases or controls stratified by carriage of ERAP1 risk or protective alleles in the presence or absence of HLA-B27. No differences were observed between expression of IL17A or TNF (tumour necrosis factor) in HLA-B27(+)ERAP1(risk), HLA-B27(+)ERAP1(protective) and HLA-B27(-)ERAP1(protective) cases. These data demonstrate that aberrant ERAP1 activity and HLA-B27 carriage does not alter ER-stress levels in AS, suggesting that ERAP1 and HLA-B27 may influence disease susceptibility through other mechanisms.

Comparison of elemental carbon in lake sediments measured by three different methods and 150-year pollution history in Eastern China.

Concentrations of elemental carbon (EC) were measured in a 150 yr sediment record collected from Lake Chaohu in Anhui Province, eastern China, using three different thermal analytical methods: IMPROVE_A thermal optical reflectance (TOR), STN_thermal optical transmittance (TOT), and chemothermal oxidation (CTO). Distribution patterns for EC concentrations are different among the three methods, most likely due to the operational definition of EC and different temperature treatments prescribed for each method. However, similar profiles were found for high-temperature EC fractions among different methods. Historical soot(TOR) (high-temperature EC fractions measured by the IMPROVE_A TOR method) from Lake Chaohu exhibited stable low concentrations prior to the late 1970s and a sharp increase thereafter, corresponding well with the rapid industrialization of China in the last three decades. This may suggest that high-temperature thermal protocols are suitable for differentiating between soot and other carbon fractions. A similar soot(TOR) record was also obtained from Lake Taihu (~200 km away), suggesting a regional source of soot. The ratio of char(TOR) (low-temperature EC fraction measured by the IMPROVE_A TOR method, after correction for pyrolysis) to soot(TOR) in Lake Chaohu shows an overall decreasing trend, consistent with gradual changes in fuel use from wood burning to increasing fossil fuel combustions. Average higher char(TOR)/soot(TOR) was observed in Lake Taihu than in Lake Chaohu in the past 150 years, consistent with the longer and more extensive industrialization around the Taihu region.

Distribution and ecotoxicological significance of trace element contamination in a ~150 yr record of sediments in Lake Chaohu, Eastern China.

This paper presents the concentrations, vertical profiles and possible sources of selected major and trace elements in a sediment core covering ∼150 years of sedimentation in Lake Chaohu, eastern China. Element concentrations were measured by portable X-ray Fluorescence Spectroscopy (XRF) and were used to evaluate possible environmental consequences of the recent industrialization in China. Statistical analyses identify four groups: (1) organic carbon (OC), total nitrogen (TN), Pb, Zn, and As associated with the use of chemical fertilizers and pesticides; (2) Mn, Cr, Ni and Cu from industrial and mining activities; (3) Fe, Rb, K, Co, Ti and Ca influenced by post-depositional processes and land exploitation; and (4) Zr and Sr from the soil. The vertical profiles of elements placed in the first two groups show distinct increases in concentrations above depths of 20 cm (~1978), coincident with the timing of industrialization in China, and the anthropogenic-derived fluxes are higher than the lithogenic-derived fluxes over the last three decades. With the exception of Zr and Sr, association of the measured metals with organic carbon and nitrogen suggests that organic matter may act as a carrier phase. The geoaccumulation index (I(geo)) reveals increased contamination from elements in the first two groups in recent years. Element concentrations, compared with the Effects Range-Low (ERL) and Effects Range-Median (ERM) levels set by NOAA, suggest that adverse biological effects from Ni contamination are very likely.

A potent nonporphyrin class of photodynamic therapeutic agent: cellular localisation, cytotoxic potential and influence of hypoxia.

We have developed a totally new class of nonporphyrin photodynamic therapeutic agents with a specific focus on two lead candidates azadipyrromethene (ADPM)01 and ADPM06. Confocal laser scanning microscopy imaging showed that these compounds are exclusively localised to the cytosolic compartment, with specific accumulation in the endoplasmic reticulum and to a lesser extent in the mitochondria. Light-induced toxicity assays, carried out over a broad range of human tumour cell lines, displayed EC50 values in the micro-molar range for ADPM01 and nano-molar range for ADPM06, with no discernable activity bias for a specific cell type. Strikingly, the more active agent, ADPM06, even retained significant activity under hypoxic conditions. Both photosensitisers showed low to nondeterminable dark toxicity. Flow cytometric analysis revealed that ADPM01 and ADPM06 were highly effective at inducing apoptosis as a mode of cell death. The photophysical and biological characteristics of these PDT agents suggest that they have potential for the development of new anticancer therapeutics.

The distribution and history of nuclear weapons related contamination in sediments from the Ob River, Siberia as determined by isotopic ratios of plutonium and neptunium.

Isotopic ratios of Pu and Np measured in sediment cores from 5 locations in the Ob River drainage basin show clear evidence of input from sources other than global fallout (non-fallout sources). Historical contaminant records obtained by combining isotopic ratio information with chronological information indicate that non-fallout inputs are from several sources that have varied significantly over the past 50 years. Unique isotopic signatures observed in sediments from tributaries that drain areas containing known or suspected sources of non-fallout contamination are used to identify the source of materials in sediments collected at downstream locations. These data can lead to a better understanding of the transport behavior, fate, and relative importance of particle reactive, weapons related contaminants originating from the nuclear facilities Mayak. Tomsk-7, and Semipalitinsk, which lie within the drainage basin. From our work to date, we draw the following conclusions: (1) Persistent non-fallout contamination is observed in the Ob River above its confluence with the Irtysh River, indicating contamination from the Tomsk-7 facility. (2) Non-fallout contamination in the Tobol River above its confluence with the Irtysh River indicates contamination from the Mayak facility. (3) Non-fallout contamination in the Irtysh River above its confluence with the Tobol River indicates contamination from the Semipalitinsk weapons test site. (4) The occurrence of isotopic ratios in Ob Delta sediments that are similar to those observed in source tributaries suggests that contamination from at least two sources has been transported along the length of the river system. (5) Global fallout, a result of high-yield atmospheric weapons tests conducted by the FSU and USA primarily, is the dominant source of Pu and Np to the region; however, there have been brief periods when inputs from non-fallout sources exceeded those from global fallout.

Creatine kinase release not associated with myocardial necrosis after short periods of coronary artery occlusion in conscious baboons.

The effects of 15 minute periods of coronary artery occlusion on plasma creatine kinase (CK) and CK-MB isoenzyme activity, regional myocardial function and subsequent myocardial necrosis were studied in six conscious baboons 2 to 3 weeks after recovery from instrumentation. Mid left anterior descending coronary artery occlusion induced complete loss of systolic wall thickening (ultrasound transit time technique) and decreases in epicardial (-93%) and endocardial (-96%) blood flows (microsphere technique). Reperfusion after 15 minutes resulted in complete recovery of regional function 24 hours later. Serial plasma enzyme activity revealed a significant increase in total CK from 71 +/- 11 to 976 +/- 158 U/liter and in CK-MB from levels that were too low to measure to 21.4 +/- 2.9 U/liter. At autopsy, neither gross pathologic evidence (triphenyltetrazolium chloride staining technique) nor histologic evidence of myocardial necrosis was observed. Thus, in the conscious baboon short episodes of myocardial ischemia are associated with a significant appearance of CK and CK-MB in the blood in the absence of cellular necrosis.

Hepatic gluconeogenic and ketogenic interrelationships in the lactating cow.

Interrelationships between propionate, palmitate, and butyrate metabolism were investigated in vitro with [1-carbon-14] carboxyl substrates. Production of labeled glucose, ketone bodies, and carbon dioxide was used to estimate rates of bovine hepatic gluconeogenesis and ketogenesis. Incubations were with liver slices from eight lactating Holstein cows fed either a control or high concentrate-low fiber diet. Liver samples were acquired by trochar biopsy at 30, 60, 90, and 180 days postpartum. Ketone production from both palmitate and butyrate was highest in liver slices obtained at 30 days. Glucose production from labeled propionate was also highest in early lactation. The higher rates of gluconeogenesis and ketogenesis in early lactation were associated with higher hepatic carnitine palmitoyltransferase (EC 2.3.1.21) activity. Feeding the high concentrate enhanced gluconeogenesis from propionate and decreased ketogenesis from palmitate. Propionate addition (10 mM) to incubation media also decreased the total amount of palmitate oxidized [( carbon-14] dioxide plus [carbon-14] ketones). Diet had no effect on hepatic butyrate metabolism. Results indicated that ketogenesis is regulated via rate of long chain fatty acid transport into the mitochondria. Stage of lactation has a greater influence on long and short chain fatty acid metabolism than does diet composition.