Reducing gut microbiome-driven adipose tissue inflammation alleviates metabolic syndrome.

BACKGROUND: The gut microbiota contributes to macrophage-mediated inflammation in adipose tissue with consumption of an obesogenic diet, thus driving the development of metabolic syndrome. There is a need to identify and develop interventions that abrogate this condition. The hops-derived prenylated flavonoid xanthohumol (XN) and its semi-synthetic derivative tetrahydroxanthohumol (TXN) attenuate high-fat diet-induced obesity, hepatosteatosis, and metabolic syndrome in C57Bl/6J mice. This coincides with a decrease in pro-inflammatory gene expression in the gut and adipose tissue, together with alterations in the gut microbiota and bile acid composition. RESULTS: In this study, we integrated and interrogated multi-omics data from different organs with fecal 16S rRNA sequences and systemic metabolic phenotypic data using a Transkingdom Network Analysis. By incorporating cell type information from single-cell RNA-seq data, we discovered TXN attenuates macrophage inflammatory processes in adipose tissue. TXN treatment also reduced levels of inflammation-inducing microbes, such as Oscillibacter valericigenes, that lead to adverse metabolic phenotypes. Furthermore, in vitro validation in macrophage cell lines and in vivo mouse supplementation showed addition of O. valericigenes supernatant induced the expression of metabolic macrophage signature genes that are downregulated by TXN in vivo. CONCLUSIONS: Our findings establish an important mechanism by which TXN mitigates adverse phenotypic outcomes of diet-induced obesity and metabolic syndrome. TXN primarily reduces the abundance of pro-inflammatory gut microbes that can otherwise promote macrophage-associated inflammation in white adipose tissue. Video Abstract.

Location-Specific ASPECTS Paradigm in Acute Ischemic Stroke: A Systematic Review and Meta-Analysis.

BACKGROUND: Weighting neuroimaging findings based on eloquence can improve the predictive value of ASPECTS, possibly aiding in informed treatment decisions for acute ischemic stroke. PURPOSE: Our aim was to study the contribution of region-specific ASPECTS infarction to acute ischemic stroke outcomes. DATA SOURCES: We searched MEDLINE and EMBASE for reports on ASPECTS in patients with acute ischemic stroke from 2000 to March 2019. STUDY SELECTION: Two investigators independently reviewed articles and extracted data. Three-month poor functional outcome defined as mRS >2 was the primary end point. DATA ANALYSIS: A random-effects meta-analysis was performed to compare the association between infarct and mRS >2 among ASPECTS regions. Subanalyses included the following: laterality of stroke (left/right), imaging technique (NCCT or advanced imaging with DWI, CTP, or CTA), and interventional technique (IV-tPA/conservative management or mechanical thrombectomy). DATA SYNTHESIS: M6 infarct was most associated with poor functional outcome (OR = 3.26; 95% CI, 2.21-4.80; P < .001). Pair-wise comparisons of ASPECTS regions regarding the association between infarct and mRS >2 were not significant, with the exception of M6 versus lentiform (P = .009). However, pair-wise comparisons among ASPECTS regions were not significant among subgroup analyses. LIMITATIONS: Limitations were the heterogeneity of time points, neuroimaging modalities, and interventional techniques; limited studies for inclusion; publication bias among some comparisons; and the retrospective nature of included studies. CONCLUSIONS: Our study indicated an unequal impact of some ASPECTS subregions in predicting outcomes of patients with acute ischemic stroke. Stroke laterality, imaging technique, and interventional technique subgroup analyses showed no differences among ASPECTS regions in predicting outcome. Investigation in larger cohorts is required to assess the association of ASPECTS with acute ischemic stroke outcome.

Chromosome 2 sequence of the human malaria parasite Plasmodium falciparum.

Chromosome 2 of Plasmodium falciparum was sequenced; this sequence contains 947,103 base pairs and encodes 210 predicted genes. In comparison with the Saccharomyces cerevisiae genome, chromosome 2 has a lower gene density, introns are more frequent, and proteins are markedly enriched in nonglobular domains. A family of surface proteins, rifins, that may play a role in antigenic variation was identified. The complete sequencing of chromosome 2 has shown that sequencing of the A+T-rich P. falciparum genome is technically feasible.

Cerebrotendinous xanthomatosis: a defect in mitochondrial 26-hydroxylation required for normal biosynthesis of cholic acid.

Oxidation of side chain of 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol was studied in a patient with cerebrotendinous xanthomatosis (CTX) and in control subjects, using various subcellular fractions of liver homogenate and a method based on isotope dilution-mass spectrometry. In the control, 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol was converted into 5 beta-cholestane-3 alpha,7 alpha,12 alpha,26-tetrol and 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid by the mitochondrial fraction, and into 5 beta-cholestane-3 alpha,7 alpha,12 alpha,-25-tetrol by the microsomal fraction. In the CTX patient, liver mitochondria were completely devoid of 26-hydroxylase activity. The same mitochondrial fraction catalyzed 25-hydroxylation of vitamin D3. The microsomal fraction of liver of the subject with CTX contained more than 50-fold the normal amount of 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol. The basic metabolid defect in CTX appears to be a lack of the mitochondrial 26-hydroxylase. The excretion in the bile of 5 beta-cholestane-3 alpha,7 alpha,12 alpha,25-tetrol and 5 beta-cholestane-3 alpha,7 alpha,12 alpha,24 alpha,25-pentol observed in CTX patients may be secondary to the accumulation of the major substrate for the 26-hydroxylase, i. e., 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol, and exposure of this substrate to the normally less active microsomal 25-and 24-hydroxylases. It is concluded that the major pathway in the biosynthesis of cholic acid in human liver involves a mitochondrial C27-steroid 26-hydroxylation.

Regulation by homeoproteins: a comparison of deformed-responsive elements.

Homeotic genes of Drosophila melanogaster encode transcription factors that specify segment identity by activating the appropriate set of target genes required to produce segment-specific characteristics. Advances in understanding target gene selection have been hampered by the lack of genes known to be directly regulated by the HOM-C proteins. Here we present evidence that the gene 1.28 is likely to be a direct target of Deformed in the maxillary segment. We identified a 664-bp Deformed Response Element (1.28 DRE) that directs maxillary-specific expression of a reporter gene in transgenic embryos. The 1.28 DRE contains in vitro binding sites for Deformed and DEAF-1. The Deformed binding sites do not have the consensus sequence for cooperative binding with the cofactor Extradenticle, and we do not detect cooperative binding to these sites, though we cannot rule out an independent role for Extradenticle. Removing the four Deformed binding sites renders the 1.28 DRE inactive in vivo, demonstrating that these sites are necessary for activation of this enhancer element, and supporting the proposition that 1.28 is activated by Deformed. We show that the DEAF-1 binding region is not required for enhancer function. Comparisons of the 1.28 DRE with other known Deformed-responsive enhancers indicate that there are multiple ways to construct Deformed Response Elements.

Hypermagnesemia as a cause of refractory hypotension, respiratory depression, and coma.

Hypermagnesemia developed in a patient as a result of excess antacid ingestion, bowel obstruction, and renal failure. Before the diagnosis was considered, refractory hypotension, respiratory depression, and coma developed, all of which were eventually reversed through the lowering of the serum magnesium concentration by hemodialysis.

Pulmonary cytomegalovirus infection: detection by Gallium 67 imaging in the transplant patient.

Cytomegalovirus (CMV) infeciton is a frequent complication during the first few months following renal transplantation. The diagnosis is sometimes difficult but may be made by viral culture, a fourfold rise in the CMV antibody titer, or by demonstration of the CMV inclusions in the affected tissue. An increased pulmonary uptake of galliulm citrate Ga 67 has been demonstrated following renal transplantation in two patients, each of whom had a fourfold rise in CMV complement fixing antibody titer, one of whom additionally had CMV inclusion bodies in a lung biopsy specimen prior to clinical or radiological demonstration of the pulmonary involvement. Gallium imaging, therefore, appears to be a valuable noninvasive test for early diagnosis of CMV pulmonary infections.

Interruptions of high-dose radiation therapy decrease long-term survival of favorable patients with unresectable non-small cell carcinoma of the lung: analysis of 1244 cases from 3 Radiation Therapy Oncology Group (RTOG) trials.

PURPOSE: To determine if prolonged treatment time adversely affects survival for patients with inoperable non-small cell carcinoma of the lung. METHODS AND MATERIALS: Patients enrolled on three randomized studies (RTOG 8311, 8321, 8403) between 1983-1989 formed the database. Previous analyses found that the addition of thymosin (8321) or prophylactic cranial irradiation (8403) failed to prolong survival: both studies used thoracic irradiation with standard fractionation to 55-60 Gy in 30 fractions. In 8311, patients were treated by hyperfractionated radiation therapy to randomly assigned total doses of 60.0 Gy, 64.8 Gy, 69.6 Gy, 74.4 Gy or 79.2 Gy, 1.2 Gy twice daily, 5 days per week. Patients analyzed received +/- 4% of the assigned total dose and lived > 90 days (to ensure that all patients would have completed treatment). Completion < 5 days beyond protocol specifications was classified as "per protocol." Elapsed treatment time exceeding specifications by 5-9 days was a minor deviation, 10-13 days was a major deviation-acceptable, and > or = 14 days was a major deviation-unacceptable. Absolute survival was the endpoint to evaluate the effect of delays. The log rank statistic was used to test for survival differences in the univariate setting, the Cox regression model was used in the multivariate setting. RESULTS: Of 293 patients treated with standard fractionation, eight (2.7%) had deviations from the specified treatment time (six minor, two major-acceptable). With hyperfractionation, 90 (15%) patients had deviations (40 minor, 21 major-acceptable, 29 major-unacceptable). As the assigned dose increased, the deviation rate increased (9.7% for 60.0 Gy vs. 20.8% for 79.2 Gy). Survivals for hyperfractionation patients with any deviations in treatment time were significantly shorter than those treated "per protocol" (p = 0.16): estimated 2- and 5-years rates were 24% and 10% versus 13% and 3%, respectively. Multivariate analyses showed the delay effect to be entirely in patients treated with 69.6 Gy or higher; there was also dependence upon the patients' prognosis. In patients with favorable prognosis (KPS 90-100, weight loss < or = 5%, no N3), the difference in survival was pronounced (33% and 15% vs. 14% and 0% at 2- and 5-years, respectively). Such differences were not found in patients with unfavorable prognostic factors. CONCLUSIONS: Interruptions delaying completion of planned radiation therapy were more frequent with higher total doses (> or = 69.6 Gy). Favorable patients (high KPS, little weight loss, < N3 nodal metastasis) had markedly adverse effects on long-term survival associated with delays to completion of the planned total dose.

Outflow facility in acute experimental ciliochoroidal detachment.

Total outflow facility was measured by constant pressure perfusion in six cynomolgus monkey eyes with ciliochoroidal detachment and in fellow eyes following sham operation. Outflow facility in eyes with ciliochoroidal detachment was 0.22 +/- 0.04 microliter/min-1/mmHg-1 (P less than 0.05). Since the spontaneous intraocular pressure following acute ciliochoroidal detachment was lower than the normal episcleral venous pressure, it is concluded that the increase in outflow facility in eyes with ciliochoroidal detachment actually may be a measure of the rate of fluid flow across the sclera out of the suprachoroidal space.

Fluid permeability of monkey ciliary epithelium in vivo.

Four months after total iridectomy, the aqueous humor of each of six cynomolgus monkeys was perfused at the spontaneous intraocular pressure under urethane anesthesia. The perfusate solution consisted of mock aqueous humor containing trace amounts of 14C-dextran (70,000 mol. wt.). The aqueous was stirred to bathe the ciliary epithelium. Aqueous flow was calculated from the tracer dilution. The aqueous humor was then perfused with bovine serum albumin (35 mg/ml) added to the perfusate. A new aqueous flow value was determined. Aqueous flow during mock aqueous perfusion was 1.28 +/- 0.12 microliters/min and 1.64 +/- 0.12 microliters/min during mock aqueous plus albumin perfusion. From the increase in aqueous flow due to the osmotic effect of the albumin solution (13.2 mm Hg), the osmotic fluid permeability of the ciliary epithelium was calculated to be 0.027 +/- 0.007 microliters/min/mm Hg. This permeability value places the ciliary epithelium in the category of membranes with "leaky" tight junctions. However, the low fluid permeability of the ciliary epithelium suggests a minor role for ultrafiltration in aqueous humor formation in the monkey.

Aqueous humor dynamics in experimental iridocyclitis.

Ocular inflammation was induced by intravitreal bovine serum albumin (BSA) injection in one eye of each of six cynomolgus monkeys. The fellow eyes were injected with sterile saline alone. The intraocular pressure decreased by 12.2 +/- 1.3 mmHg (mean +/- SE) 2 days after BSA injection and 4.0 +/- 1.1 mmHg after saline injection. Aqueous flow and uveoscleral outflow were determined with fluorescein isothiocyanate (FITC) dextran 70. Aqueous flow in inflamed eyes averaged 0.32 +/- 0.04 ul/min, less than half the rate of control eyes (0.77 +/- 0.08 ul/min, P = 0.01). The facility of uveoscleral outflow in inflamed eyes was four times that of control eyes (0.2 +/- 0.03 vs 0.05 +/- 0.01 ul/min/mmHg, respectively, P = 0.009). Fluorescence microscopic examination revealed intense fluorescence of the edematous ciliary body muscle and of the suprachoroidal space extending to the posterior pole. These findings indicate that BSA-induced ocular inflammation causes a simultaneous reduction in aqueous humor flow and an increase in uveoscleral outflow, resulting in ocular hypotony.

Uveoscleral outflow: diffusion or flow?

Shallow peripheral ciliochoroidal detachments with 10(-4) M fluorescein isothiocyanate dextran 70 were created in cynomolgus monkey eyes. Anterior chamber fluorophotometric readings were taken for 6 hr. From the anterior chamber fluorescence values, the rate of tracer movement from the supraciliary space into the anterior chamber was calculated. The rate of movement was 0.003 microliter/min, expressed in equivalent volumes of tracer solution. This value is more than 200 times lower than the rate of tracer movement from the anterior chamber to the supraciliary space. It is concluded that tracer movement from the anterior chamber to the supraciliary space (uveoscleral route) results from fluid flow rather than diffusion.

Permeability of the isolated dog retinal pigment epithelium to carboxyfluorescein.

Outward (retina to choroid) and inward (choroid to retina) permeabilities of carboxyfluorescein and fluorescein in the isolated dog retinal pigment epithelium (RPE)-choroid were determined. Outward permeability was 9 and 47 times larger than inward permeability for carboxyfluorescein and fluorescein, respectively. The outward permeability of carboxyfluorescein was seven times lower than that of fluorescein, whereas there was no statistical difference between the inward permeabilities. Carboxyfluorescein is thus distinguished from fluorescein by its low affinity to the outwardly directed organic anion transport system. 10(-4) M probenecid caused greater than 98% inhibition of the outward transport of 6 X 10(-5) M carboxyfluorescein and 6 X 10(-6) M fluorescein.

Volume flow across the isolated retinal pigment epithelium of cynomolgus monkey eyes.

The retinal pigment epithelium (RPE)-choroid was isolated from cynomolgus monkey eyes with experimental retinal detachments and the volume flow was determined in vitro using Ussing-type chambers. With zero pressure difference across the membrane, retina-to-choroid volume flow was 5.0 microliter/hr/cm2 in eyes with subacute retinal detachments (1-2 weeks). In eyes with chronic retinal detachment (8-20 months), the flow was 7.3 microliter/hr/cm2. Volume flow was not affected by the elimination of ambient bicarbonate. Transepithelial potential difference and resistance were 8.9 mV, retinal side positive, and 339 ohm-cm2, respectively, in chronic retinal detachments. It is concluded that there is a posteriorly directed flow of fluid across the RPE in cynomolgus monkey eyes with chronic retinal detachments.

Effect of plasma osmolality and intraocular pressure on fluid movement across the blood-retinal barrier.

The inward permeability of the blood-retinal barrier to carboxyfluorescein was determined in monkey eyes with and without rhegmatogenous retinal detachment (RD). In the absence of changes in the diffusional permeability of the retinal pigment epithelium (RPE), inward permeability changes reflect changes in fluid flow across the RPE. Intravenous injection of mannitol resulted in a 15 mosmol/kg increase in plasma osmolality which decreased inward permeability 37% in eyes with RD and 21% in eyes with vitrectomy alone. When the intraocular pressure was raised 20 mm Hg above normal, inward permeability decreased 29% in eyes with RD and 32% in normal eyes. It is concluded that fluid flow across the blood-retinal barrier is influenced by both plasma osmolality and intraocular pressure.

Episcleral venous pressure: a comparison of invasive and noninvasive measurements.

Noninvasive (pressure chamber) measurements of episcleral venous pressure were followed by invasive (direct cannulation) measurements at the same venous site in seven eyes of four anesthetized rhesus monkeys. There were three definite effects on the vein caused by the pressure chamber that could be used as endpoints: (1) slight indentation; (2) intermittent collapse; and (3) sustained collapse of the vein lumen. The mean pressure in the chamber corresponding to these endpoints was 9.9 +/- 0.9, 23.5 +/- 2.9, and 31.4 +/- 4.0 mmHg (+/- SE), respectively. After the chamber was removed, the pressure in the veins determined by cannulation measurements at the same sites was 11.3 +/- 0.5 mmHg (+/- SE). Therefore, the first endpoint with the pressure chamber (slight indentation) correlates most closely and slightly underestimates the cannulated pressure. Endpoints defined by partial or complete venous collapse overestimates the venous pressure. Simultaneous measurements with the chamber and a cannula show a rise of local venous pressure caused by the chamber.

Acetazolamide effect on the inward permeability of the blood-retinal barrier to carboxyfluorescein.

The inward permeability of the blood-retinal barrier to carboxyfluorescein was determined in 12 cynomolgus monkeys. Probenecid (175 mg/kg), an inhibitor of active outward transport of carboxyfluorescein, did not affect the inward permeability, indicating that the inward permeability is independent of the active outward transport system. However, acetazolamide (20 mg/kg), which causes increased outward fluid movement across the retinal pigment epithelium (RPE), significantly reduced the inward permeability. Thus, inward diffusion of carboxyfluorescein interacts with outward fluid flow across the RPE. Since carboxyfluorescein has low lipid solubility and remains extracellular, it is concluded that the pathway of fluid movement and carboxyfluorescein diffusion across the RPE is paracellular.

Permeability of the blood-retinal barrier to carboxyfluorescein in eyes with rhegmatogenous retinal detachment.

Outward and inward permeability of carboxyfluorescein across the blood-retinal barrier were measured fluorophotometrically in seven cynomolgus monkey eyes with experimental rhegmatogenous retinal detachment. Probenecid was used to inhibit outward transport of carboxyfluorescein. The outward permeability was 1.98 +/- 0.31 microliter/min in eyes with retinal detachment and 0.84 +/- 0.15 microliter/min in control eyes with vitrectomy alone (P less than 0.01). The inward permeability, determined separately following intravenous injection, was significantly lower than the outward permeability: 0.14 +/- 0.02 microliter/min for eyes with retinal detachment and 0.04 +/- 0.01 microliter/min for control eyes. Since the outward permeability minus the inward permeability in the presence of probenecid represents that fraction of tracer moving due to fluid flow, it may be concluded that outward flow of fluid across the blood-retinal barrier is a substantial contributor to carboxyfluorescein loss from the vitreous cavity following intravitreal injection.

Effect of intraocular pressure on uveoscleral outflow following cyclodialysis in the monkey eye.

Cyclodialysis was performed in both eyes of five cynomolgus monkeys. Two days later the intraocular pressure (IOP) had fallen from 17.7 +/- 0.8 to 7.1 +/- 1.4 mm Hg (P less than 0.001). At that time, both eyes were perfused for 30 min with fluorescein-isothiocyanate (FITC) dextran (MW 70,000), one at 35 mm Hg and the other at 4 mm Hg. Four pairs of control eyes (without cyclodialysis) were perfused in the same manner. At 4 mm Hg, uveoscleral outflow was 0.02 +/- 0.02 microliter/min in control eyes and 0.05 +/- 0.04 microliter/min in eyes following cyclodialysis. However, at 35 mm Hg, uveoscleral outflow in eyes with cyclodialysis increased to 2.13 +/- 0.47 microliters/min compared to 0.32 +/- 0.10 microliter/min in control eyes. Thus the "facility" of uveoscleral outflow in control eyes is 0.01 microliter/min/mm Hg and in eyes following cyclodialysis is 0.07 microliter/min/mm Hg. It is concluded that cyclodialysis results in a pressure-dependent increase in uveoscleral outflow.

Uveoscleral outflow following cyclodialysis in the monkey eye using a fluorescent tracer.

Cyclodialysis was performed in one eye of each of eight cynomolgus monkeys. Two days later, the intraocular pressure was 1.6 +/- 0.7 mmHg in eyes with cyclodialysis and 12.0 +/- 0.7 mmHg in fellow control eyes. 10(-4) M fluorescein-isothiocyanate dextran (70,000 molecular weight) was perfused into the anterior chamber of each eye for 30 min. The eyes were enucleated and dissected into sclera, choroid, retina, iris, and ocular fluid. Samples were homogenized and centrifuged, and the fluorescence of the supernatant was measured. Expressed as equivalent volumes of aqueous, the rate of anterior chamber movement of tracer via uveoscleral pathways was 1.40 +/- 0.17 microliter/min in cyclodialysis eyes and 0.34 +/- 0.10 microliter/min in control eyes. Cyclodialysis results in a fourfold increase in uveoscleral outflow, contributing to the observed hypotony.