Long-term in vitro 3D hydrogel co-culture model of inflammatory bowel disease

The in vitro study of the pathogenesis of inflammatory bowel disease (IBD) requires a cell model which closely reflects the characteristics of the in vivo intestinal epithelium. This study aimed to investigate the application of L-pNIPAM hydrogel as a scaffold to develop a long-term 3D co-culture model of Caco-2 and HT29-MTX cells under conditions analogous to inflammation, to determine its potential use in studying IBD. Monocultures and co-cultures were layered on L-pNIPAM hydrogel scaffolds and maintained under dynamic culture conditions for up to 12 weeks. Treatments with IL-1β, TNFα, and hypoxia for 1 week were used to create an inflammatory environment. Following prolonged culture, the metabolic activity of Caco-2 monoculture and 90% Caco-2/10% HT29-MTX co-cultures on L-pNIPAM hydrogels were increased, and finger-like structures, similar in appearance to villi were observed. Following treatment with IL-1β, TNFα and hypoxia, ALP and ZO-1 were decreased, MUC2 increased, and MUC5AC remained unchanged. ADAMTS1 was increased in response to hypoxia. Caspase 3 expression was increased in response to TNFα and hypoxic conditions. In conclusion, L-pNIPAM hydrogel supported long-term co-culture within a 3D model. Furthermore, stimulation with factors seen during inflammation recapitulated features seen during IBD

Maternal urinary metabolic signatures of fetal growth and associated clinical and environmental factors in the INMA study

Background Maternal metabolism during pregnancy is a major determinant of the intra-uterine environment and fetal outcomes. Herein, we characterize the maternal urinary metabolome throughout pregnancy to identify maternal metabolic signatures of fetal growth in two subcohorts and explain potential sources of variation in metabolic profiles based on lifestyle and clinical data. Methods We used 1H nuclear magnetic resonance (NMR) spectroscopy to characterize maternal urine samples collected in the INMA birth cohort at the first (n = 412 and n = 394, respectively, in Gipuzkoa and Sabadell cohorts) and third trimesters of gestation (n = 417 and 469). Metabolic phenotypes that reflected longitudinal intra- and inter-individual variation were used to predict measures of fetal growth and birth weight. Results A metabolic shift between the first and third trimesters of gestation was characterized by 1H NMR signals arising predominantly from steroid by-products. We identified 10 significant and reproducible metabolic associations in the third trimester with estimated fetal, birth, and placental weight in two independent subcohorts. These included branched-chain amino acids; isoleucine, valine, leucine, alanine and 3 hydroxyisobutyrate (metabolite of valine), which were associated with a significant fetal weight increase at week 34 of up to 2.4 % in Gipuzkoa (P < 0.005) and 1 % in Sabadell (P < 0.05). Other metabolites included pregnancy-related hormone by-products of estrogens and progesterone, and the methyl donor choline. We could explain a total of 48–53 % of the total variance in birth weight of which urine metabolites had an independent predictive power of 12 % adjusting for all other lifestyle/clinical factors. First trimester metabolic phenotypes could not predict reproducibly weight at later stages of development. Physical activity, as well as other modifiable lifestyle/clinical factors, such as coffee consumption, vitamin D intake, and smoking, were identified as potential sources of metabolic variation during pregnancy. Conclusions Significant reproducible maternal urinary metabolic signatures of fetal growth and birth weight are identified for the first time and linked to modifiable lifestyle factors. This novel approach to prenatal screening, combining multiple risk factors, present a great opportunity to personalize pregnancy management and reduce newborn disease risk in later life

Continuous-wave phase-sensitive parametric image amplification

We study experimentally parametric amplification in the continuous regime using a transverse-degenerate type-II Optical Parametric Oscillator operated below threshold. We demonstrate that this device is able to amplify either in the phase insensitive or phase sensitive way first a single mode beam, then a multimode image. Furthermore the total intensities of the amplified image projected on the signal and idler polarizations are shown to be correlated at the quantum level.Comment: 14 pages, 7 figures, submitted to Journal of Modern Optics, Special Issue on Quantum Imagin

Physical disruption of intervertebral disc promotes cell clustering and a degenerative phenotype

© 2019, The Author(s). To test the hypothesis that physical disruption of an intervertebral disc disturbs cell-matrix binding, leading to cell clustering and increased expression of matrix degrading enzymes that contribute towards degenerative disc cell phenotype. Lumbar disc tissue was removed at surgery from 21 patients with disc herniation, 11 with disc degeneration, and 8 with adolescent scoliosis. 5 μm sections were examined with histology, and 30-µm sections by confocal microscopy. Antibodies were used against integrin α5beta1, matrix metalloproteinases (MMP) 1, MMP-3, caspase 3, and denatured collagen types I and II. Spatial associations were sought between cell clustering and various degenerative features. An additional, 11 non-herniated human discs were used to examine causality: half of each specimen was cultured in a manner that allowed free ‘unconstrained’ swelling (similar to a herniated disc in vivo), while the other half was cultured within a perspex ring that allowed ‘constrained’ swelling. Changes were monitored over 36 h using live-cell imaging. 1,9-Di-methyl methylene blue (DMMB) assay for glycosaminoglycan loss was carried out from tissue medium. Partially constrained specimens showed little swelling or cell movement in vitro. In contrast, unconstrained swelling significantly increased matrix distortion, glycosaminoglycan loss, exposure of integrin binding sites, expression of MMPs 1 and 3, and collagen denaturation. In the association studies, herniated disc specimens showed changes that resembled unconstrained swelling in vitro. In addition, they exhibited increased cell clustering, apoptosis, MMP expression, and collagen denaturation compared to ‘control’ discs. Results support our hypothesis. Further confirmation will require longitudinal animal experiments

The BlackHat Library for One-Loop Amplitudes

We present recent next-to-leading order (NLO) results in perturbative QCD obtained using the BlackHat software library. We discuss the use of n-tuples to separate the lengthy matrix-element computations from the analysis process. The use of n-tuples allows many analyses to be carried out on the same phase-space samples, and also allows experimenters to conduct their own analyses using the original NLO computation.Comment: Talk given at ACAT 2013, Beijing, China, May 16--21, 2013; 6 pages, 2 figures; added reference

Universality in W+Multijet Production

We study $W$-boson production accompanied by multiple jets at 7 TeV at the LHC. We study the jet-production ratio, of total cross sections for $W$+$n$- to $W$+($n-1$)-jet production, and the ratio of distributions in the total transverse hadronic jet energy $H_{\rm T}^{\rm jets}$. We use the ratios to extrapolate the total cross section, and the differential distribution in $H_{\rm T}^{\rm jets}$, to $W$+6-jet production. We use the BlackHat software library in conjunction with SHERPA to perform the computations.Comment: Merge of Moriond 2014 and Loops & Legs 2014 proceedings, 6 pages, 2 figure

Next-to-Leading Order Jet Physics with BlackHat

We present several results obtained using the BlackHat next-to-leading order QCD program library, in conjunction with SHERPA. In particular, we present distributions for vector boson plus 1,2,3-jet production at the Tevatron and at the asymptotic running energy of the Large Hadron Collider, including new Z+3-jet distributions. The Z+2-jet predictions for the second-jet P_T distribution are compared to CDF data. We present the jet-emission probability at NLO in W+2-jet events at the LHC, where the tagging jets are taken to be the ones furthest apart in pseudorapidity. We analyze further the large left-handed W polarization, identified in our previous study, for W bosons produced at high P_T at the LHC.Comment: Presented at RADCOR 2009 - 9th International Symposium on Radiative Corrections (Applications of Quantum Field Theory to Phenomenology), October 25 - 30 2009, Ascona, Switzerland}, 12 pages, 9 figures, LaTeX, v2 updated small correction to polarization effect plo