Carbohydrate Elimination or Adaptation Diet for Symptoms of Intestinal Discomfort in IBD: Rationales for “Gibsons' Conundrum”

Therapeutic use of carbohydrates in inflammatory bowel diseases (IBDs) is discussed from two theoretical, apparent diametrically opposite perspectives: regular ingestion of prebiotics or withdrawal of virtually all carbohydrate components. Pathogenesis of IBD is discussed connecting microbial flora, host immunity, and genetic interactions. The best studied genetic example, NOD2 in Crohn's disease, is highlighted as a model which encompasses these interactions and has been shown to depend on butyrate for normal function. The role of these opposing concepts in management of irritable bowel syndrome (IBS) is contrasted with what is known in IBD. The conclusion reached is that, while both approaches may alleviate symptoms in both IBS and IBD, there is insufficient data yet to determine whether both approaches lead to equivalent bacterial effects in mollifying the immune system. This is particularly relevant in IBD. As such, caution is urged to use long-term carbohydrate withdrawal in IBD in remission to control IBS-like symptoms

SINGLE-CELL ELECTROPORATION USING ELECTROLYTE-FILLED CAPILLARIES -EXPERIMENTAL AND MODELING INVESTIGATIONS

Electrolyte-filled capillaries (EFCs) with fine tips provide a highly concentrated electric field for local single-cell electroporation (SCEP) with high spatial resolution. A complete circuit for SCEP experiments was built that consisted of a test circuit and an electroporation circuit, with the ability to monitor electrically the electroporation pulses. SCEP itself was monitored in real time by observing the loss of a fluorescent adduct of glutathione (Thioglo-1-GSH) from the intracellular space. SCEP can be applied for transfection of individual adherent cells. We hypothesize that transfection of single cells can be accomplished with the plasmid contained in a single capillary. During SCEP, electroosmotic flow can pump electrolyte out of the capillary enhancing plasmid transfer into cells. This was confirmed from both simulation and transfection experiments. Cells were successfully transfected with EGFP-C2 plasmid when the loss of Thioglo-1-GSH upon SCEP (ΔF) is larger than 10% and its mass transfer rate (M) through the membrane exceeds 0.03 s-1. A series of SCEP experiments has been carried out on PC-3 cells (with 2-µm tip opening) and A549 cells (with 4~5-µm tip opening) to investigate how the parameters such as cell-to-tip distance (dc), cell size (dm) and shape, temperature, current, and the cell cycle affect SCEP outcomes (M and resealing rate α) via statistical analysis. A good linear regression is achieved only at a low temperature of 15℃. The main factors affecting small molecule transport across cell membrane are dc, dm and electric current. A range of M (0.03 s-1 ~ 0.4 s-1 for PC-3 cells, or 0.03 s-1 ~ 0.5 s-1 for A549 cells) gives the best linear regressions. M is also affected by the cell cycle of A549 cells, and correlated with cell roundness only for PC-3 cells. Cells reseal faster at higher temperature; while lower temperature provides better survivability with identical ΔF. Lastly, numerical models were elaborated as a platform for better understanding of the SCEP process and prediction of the trends of SCEP under various experimental conditions. A mass transport model involving potential distribution, diffusion, convection and electrokinetic flow was extended to study mass transport at a buffer-filled pipette tip/porous medium interface

SINGLE-CELL ELECTROPORATION USING ELECTROLYTE-FILLED CAPILLARIES WITH MICRO-SCALE TIPS

Single-cell electroporation (SCEP) is a recently developing powerful technique for cell analysis and cell manipulation. In the first chapter of this thesis, a review about the theories and techniques is fulfilled, including a detailed description of the factors affecting SCEP, and a discussion about how to optimize SCEP for high efficiency and survivability. Based on the previous experimental results and numerical simulation, a hypothesis is proposed which leads us to find that small tips could be a solution to electroporate small cells with simultaneous maximization of electroporation efficiency and survivability when using electrolyte-filled capillaries (EFC) with pulled tips.In the second chapter, an integrated circuit for SCEP and controlling is demonstrated. EFCs with 2 &mu tips are constructed and used for SCEP of A549 cells with an extremely high spatial resolution. Distance between tip and cell is revealed to be vital in SCEP because of its direct control of the local electric field distribution and strength; to control distance precisely, a current measurement method inspired by tip-cell giga-seals is applied. High temporal resolution videos hint an abrupt intracellular fluorescence loss at the time scale of pulse duration followed by recovery in the small portion of cell membrane facing the tips. Viability of cells is highly related to the fluorescence loss, fluorescence exposure and dye types. Comsol simulation using the real shape capillaries helps to guide the electroporation throughout our experiments.However, this protocol evokes overcoming technical difficulties in terms of getting high survivability and decreasing variance, which are our two main aims. The advantage of small tips and the hypothesis are still to be examined. This is referred in the third chapter, as well as other following-up future work

Incidence and predictors of left ventricular function change following ST-segment elevation myocardial infarction

AimThe purpose of the study was to assess the incidence and predictors of left ventricular function change in patients with ST-segment elevation myocardial infarction (STEMI) undergoing primary PCI.Methods312 patients with STEMI who received primary percutaneous coronary intervention (PCI) between January 2015 and December 2016 were consecutively enrolled in this study. Multiple logistic regression analysis was used to evaluate independent predictors of left ventricular ejection fraction (LVEF) improvement after long-term follow-up.ResultsWe finally analyzed the LVEF change in 186 patients from baseline to follow-up. The mean age was 61.3 ± 12.5 years, with 78.5% being male. The median duration of follow-up after STEMI was 1,021 (389–1,947) days. 54.3% had a decrease in LVEF and 45.7% experienced an improvement in LV function after primary PCI through long-term follow-up. Logistic regression analysis showed lower peak troponin I, non-anterior STEMI, lower baseline LVEF, and no previous myocardial infarction history were independently associated with LVEF improvement.Conclusion54.3% of patients with STEMI undergoing primary PCI had a decrease in LVEF during long-term follow-up. LVEF recovery can be predicted by baseline characteristics

Motivational orientation and risk taking in elite winter climbers: A qualitative study

Objectives: High risk sports participants have typically been viewed as a homogenous group despite variability in performance characteristics and the level of risk undertaken. Prolonged engagement high risk sports such as winter climbing are relatively underserved within current literature. Elite winter climbers attempt climbs that are outside the scope of the current ‘known’ i.e. unclimbed routes. The majority of the current understanding of motivation in high risk sports is based on quantitative research and the methodologies and instruments used. The purpose of this study was to explore the experiences of elite winter climbers and gain a richer understanding of their motivational orientation and risk taking behaviour. Design: Qualitative – inductive. Method: Four elite male winter climbers (aged 42-49 years old) took part in semi-structured interviews and explore their motivational orientation and risk taking behaviour. A thematic analysis was used. Results: Two super-ordinate themes of enactive mastery and engendered disinhibition emerged from the data. Enactive mastery was interpreted as a composite of two higher order themes; task mastery and self-mastery. Engendered disinhibition was interpreted as a composite of two higher order themes; social cognitive appraisal and self-perception. Conclusion: Enactive mastery and engendered disinhibition emerged as key behavioural and psychological determinants that influenced individuals to attempt more difficult and riskier forms of winter climbing. Goal achievement was their primary motive which was set within a confidence frame encapsulated within these super-ordinate themes

Alterations in erythrocyte fatty acid composition in preclinical Alzheimer\u27s disease

Brain and blood fatty acids (FA) are altered in Alzheimer’s disease and cognitively impaired individuals, however, FA alterations in the preclinical phase, prior to cognitive impairment have not been investigated previously. The current study therefore evaluated erythrocyte FA in cognitively normal elderly participants aged 65 – 90 years via trans-methylation followed by gas chromatography. The neocortical beta-amyloid load (NAL) measured via positron emission tomography (PET) using ligand 18F-Florbetaben, was employed to categorise participants as low NAL (standard uptake value ratio; SUVR \u3c 1.35, N = 65) and high NAL or preclinical AD (SUVR ≥ 1.35, N = 35) wherein, linear models were employed to compare FA compositions between the two groups. Increased arachidonic acid (AA, p \u3c 0.05) and decreased docosapentaenoic acid (DPA, p \u3c 0.05) were observed in high NAL. To differentiate low from high NAL, the area under the curve (AUC) generated from a ‘base model’ comprising age, gender, APOEε4 and education (AUC = 0.794) was outperformed by base model + AA:DPA (AUC = 0.836). Our findings suggest that specific alterations in erythrocyte FA composition occur very early in the disease pathogenic trajectory, prior to cognitive impairment. As erythrocyte FA levels are reflective of tissue FA, these alterations may provide insight into the pathogenic mechanism(s) of the disease and may highlight potential early diagnostic markers and therapeutic targets

Alterations in serum kynurenine pathway metabolites in individuals with high neocortical amyloid-β load: A pilot study

The kynurenine pathway (KP) is dysregulated in neuroinflammatory diseases including Alzheimer\u27s disease (AD), however has not been investigated in preclinical AD characterized by high neocortical amyloid-β load (NAL), prior to cognitive impairment. Serum KP metabolites were measured in the cognitively normal KARVIAH cohort. Participants, aged 65-90 y, were categorised into NAL+ (n = 35) and NAL- (n = 65) using a standard uptake value ratio cut-off = 1.35. Employing linear models adjusting for age and APOEϵ4, higher kynurenine and anthranilic acid (AA) in NAL+ versus NAL- participants were observed in females (kynurenine, p = 0.004; AA, p = 0.001) but not males (NALxGender, p = 0.001, 0.038, respectively). To evaluate the predictive potential of kynurenine or/and AA for NAL+ in females, logistic regressions with NAL+/- as outcome were carried out. After age and APOEϵ4 adjustment, kynurenine and AA were individually and jointly significant predictors (p = 0.007, 0.005, 0.0004, respectively). Areas under the receiver operating characteristic curves were 0.794 using age and APOEϵ4 as predictors, and 0.844, 0.866 and 0.871 when kynurenine, AA and both were added. Findings from the current study exhibit increased KP activation in NAL+ females and highlight the predictive potential of KP metabolites, AA and kynurenine, for NAL+. Additionally, the current study also provides insight into he influence of gender in AD pathogenesi

Polyphenol-rich potato extracts exert sex-dimorphic protective effects on the ozone-induced pulmonary inflammatory response in C57BL/6 mice

Dietary polyphenols have been shown to improve diet-induced metabolic symptoms and pollution-related airway inflammation, yet very few studies have examined both contexts together. The current study investigated the efficacy of a polyphenolic-rich potato extract (PE) in ozone-exposed male (M) and female (F) C57BL/6 mice fed a high-fat Western-style diet (WD) based adiposity and glucose intolerance, and lung health. Mice were fed ad libitum with WD supplemented with either a 20 PE (chlorogenic acid, 40 mg/kg diet; ferulic acid, 1.2 mg/kg diet) or 100 PE (chlorogenic acid, 200 mg/kg diet; ferulic acid, 6 mg/kg diet). After 4 wk on the diets, animals were exposed to 0.8 ppm ozone or air by inhalation for 4 h and euthanized 24 h post exposure. Independent of ozone exposure, supplementation with 100 PE attenuated weight gain (37% M and 48% F) and significantly reduced (p < 0.01) adiposity and total fat mass relative to the untreated controls in both sexes. Both 20 and 100 PE treatments restored fasting blood glucose levels (p < 0.001) and improved insulin sensitivity (p < 0.05). Based on cellular evaluation from bronchoalveolar lavage fluid (BALF), ozone-induced pulmonary inflammation/injury was suppressed by 100 PE treatment as indicated by a reduction in alveolar macrophage cell counts (464 ± 12.1 F and 457 ± 16.7 M) below than the respective air-exposed group under the same treatment condition (490 ± 5.3 M and 477 ± 7.4 F). Additionally, a decrease in neutrophil cell counts (9 ± 0.7 M and 7 ± 0.9 F) relative to the ozone-exposed untreated group (13 ± 1.7 M and 11 ± 1.9 F) was observed. The 100 PE dose also reduced total protein concentrations in BALF (p < 0.001) in the ozone-exposed groups (106.09 ± 1.10 vs. 95.61 ± 0.83 µg/mL M and 82.45 ± 1.47 vs. 74.56 ± 0.68 µg/mL F). Similarly, epithelial counts in BALF were decreased due to 100 PE in the ozone-exposed groups (129 ± 20.5 vs. 75 ± 14.3 M and 137 ± 28.2 vs. 67 ± 18.2 F). All these markers signify acute lung injury. The lung inflammatory markers (IL-4, IL-6, IL-13, KC, MCP-1, MIP-1β, RANTES, and TNF-α) measured from BALF exhibited some changes as well. Although the BALF cytokines were in general increased with ozone exposure, these levels were lowered consistently in male mice with 20 PE treatment without reaching statistical significance. Pro-inflammatory and antioxidant gene expression profiles of the air- and ozone-exposed groups were compared to evaluate the extent of transcriptional changes based on stratification of the PE dietary treatments. A sex dimorphic response, particularly evident in the changes in mRNA abundance of ET1, GPX1, GSTM1, HMOX1, and SOD2 where males varied in their response to PE treatment and ozone exposure, while females displayed consistent response to PE treatment regardless of ozone exposure. For MT2, there appeared to be two opposing effects between the sexes, with 100 PE group in males showing significant suppression (p = 0.05) relative to air-exposed group yet the opposite effect occurred among the females in both treatments (p = 0.05 and p < 0.05, respectively). With IL6 both sexes displayed significant elevation (p < 0.05) compared to air control in both PE-treated groups. These findings ascertain that dietary PE supplementation improves adiposity and glucose intolerance associated with consumption of WD, and protects against lung injury. However, there is sex dimorphism in the PE-mediated protection against ozone-induced lung inflammation. The work presented in this thesis is the first to document the inhibitory effects of PE on pulmonary inflammation in relation to their ability to mitigate ozone-induced toxicity in a sex-dependent manner.Les polyphénols alimentaires ont démontré des capacités à améliorer les symptômes métaboliques causées par l'alimentation et l'inflammation des voies respiratoires liées à la pollution, mais très peu d'études ont examiné les deux contextes ensemble. L'étude actuelle a étudié l'efficacité d'un extrait de pomme de terre riche en polyphénol (PE) dans des souris C57BL/6 mâle (M) et femelle (F) exposées à de l'ozone et soumises à un régime de type occidental qui est riche en matières grasses (WD) l'adiposité et une intolérance au glucose, ainsi que la santé pulmonaire. Les souris ont été nourris ad libitum avec le WD supplémenté avec soit 20 PE (acide chlorogénique, 40 mg/kg d'aliment, l'acide férulique, 1,2 mg/kg d'aliment) ou 100 PE (acide chlorogénique, 200 mg /kg d'aliment, l'acide férulique, 6 mg/kg d'aliments). Après 4 semaines sur le régime alimentaire, les animaux ont été exposés à l'ozone de 0,8 ppm ou à l'air pendant 4 h et euthanasiés 24 h après l'exposition. Indépendamment de l'exposition à l'ozone, la supplémentation avec 100 PE a atténué le gain de poids (37% M et 48% F) et a réduit de manière significative (p < 0,01) l'adiposité et la masse graisseuse totale par rapport aux témoins non traités dans les deux sexes. Les deux traitements 20 et 100 PE ont restaurés les niveaux de glucose sanguin à jeun (p < 0,001) et amélioré la sensibilité à l'insuline (p < 0,05). Sur la base de l'évaluation cellulaire du liquide de lavage bronchoalvéolaire (BALF), l'inflammation pulmonaire/lésion induite par l'ozone a été supprimée par le traitement 100 PE, comme indiqué par la réduction du nombre de cellules de macrophages alvéolaires (464 ± 12,1 F et 457 ± 16,7 M) au-dessous du groupe respectif exposé à l'air dans les mêmes conditions de traitement (490 ± 5,3 M et 477 ± 7,4 F). En outre, une diminution du nombre de neutrophiles (9 ± 0,7 M et 7 ± 0,9 F) par rapport au groupe exposée à l'ozone non traité (13 ± 1,7 M et 11 ± 1,9 F) a été observée. Les doses de 100 PE ont aussi réduites les concentrations de protéines totales dans BALF (p < 0,001) dans les groupes exposés à l'ozone. De même, le nombre de cellules épithéliales dans BALF ont diminué à cause du traitement 100 PE dans les groupes exposés à l'ozone. Tous ces marqueurs signifient une lésion pulmonaire aiguë. Les marqueurs d'inflammation du poumon (IL-4, IL-6, IL-13, KC, MCP-1, MIP-1β, RANTES, et TNF-α) mesurés à partir de BALF montrent des changements. Bien que les cytokines de BALF aient en général augmenté avec l'ozone, avec les traitements 20 PE ces niveaux ont baissés régulièrement chez les souris mâles sans différence statistique. Les profils d'expression des gènes pro-inflammatoires et anti-oxydants des groupes exposés à l'air et à la couche d'ozone ont été comparés pour évaluer l'ampleur des changements de transcription basés sur la stratification des traitements alimentaires PE. Un dimorphisme sexuel, particulièrement évident dans les changements de la quantité d'ARNm de ET1, GPX1, GSTM1, HMOX1, et SOD2 où les réponses au traitement PE et à l'ozone étaient variés chez les mâles, tandis que les femelles affichés une réponse cohérente au traitement PE indépendamment de l'exposition à l'ozone. Pour MT2, il semble y avoir deux effets opposés entre les sexes, avec le groupe de 100 PE chez les males présentant une suppression significative (p = 0,05) par rapport au groupe exposé à l'air tandis que l'effet inverse s'est produit chez les femelles dans les deux traitements (p = 0,05 et p < 0,05, respectivement). Avec IL6 les deux sexes affichaient une élévation significative (p < 0,05) par rapport à l'air dans les deux groupes traités avec PE. Ces résultats montrent que la supplémentation PE améliore l'adiposité et l'intolérance au glucose associées à la consommation de WD, et protège contre les lésions pulmonaires. Cependant, Il y a un dimorphisme sexuel dans la protection médié PE contre l'inflammation pulmonaire induite par l'ozone

Single-variant association test results for CHD GWAS data analysis, the National Birth Defects Prevention Study

SLR_phase1_results.csv are the single locus association test results in phase I of NBDPS GWAS data analysis. SLR_phase2_results.csv are the single locus association test results in phase II of NBDPS GWAS data analysis. SLR_fisher_combined_p_values.csv are Fisher's combination results combining the ANOVA p-values in phase I and II that evaluate the joint effects of maternal and fetal genetic variants and their interaction. meta_fetal_effects.csv is the meta-analysis results of the fetal variant effects in phase I and II. meta_maternal_effects.csv is the meta-analysis results of the maternal variant effects in phase I and II. meta_interaction_effects.csv is the meta-analysis results of the interaction effects in phase I and II. </p

The effect of chloramphenicol treatment on ferrochelatase activity in dogs

Chloramphenicol fed to dogs at a dose of 100 mg/kg/day resulted in a decrease in bone marrow ferrochelatase activity to 5–35% of control levels. Chloramphenicol in concentrations of 25 and 50 μg/ml had no effect on ferrochelatase activity in vitro