O-glycan structures in apo(a) subunit of human lipoprotein(a) suppresses the pro-angiogenic activity of galectin-1 on human umbilical vein endothelial cells.

Apolipoprotein(a) [apo(a)] is a highly polymorphic O-glycoprotein circulating in human plasma as lipoprotein(a) [Lp(a)]. The O-glycan structures of apo(a) subunit of Lp(a) serve as strong ligands of galectin-1, an O-glycan binding pro-angiogenic lectin abundantly expressed in placental vascular tissues. But the pathophysiological significance of apo(a)-galectin-1 binding is not yet been revealed. Carbohydrate-dependent binding of galectin-1 to another O-glycoprotein, neuropilin-1 (NRP-1) on endothelial cells activates vascular endothelial growth factor receptor 2 (VEGFR2) and mitogen-activated protein kinase (MAPK) signaling. Using apo(a), isolated from human plasma, we demonstrated the potential of the O-glycan structures of apo(a) in Lp(a) to inhibit angiogenic properties such as proliferation, migration, and tube-formation in human umbilical vein endothelial cells (HUVECs) as well as neovascularization in chick chorioallantoic membrane. Further, in vitro protein-protein interaction studies have confirmed apo(a) as a superior ligand to NRP-1 for galectin-1 binding. We also demonstrated that the protein levels of galectin-1, NRP-1, VEGFR2, and downstream proteins in MAPK signaling were reduced in HUVECs in the presence of apo(a) with intact O-glycan structures compared to that of de-O-glycosylated apo(a). In conclusion, our study shows that apo(a)-linked O-glycans prevent the binding of galectin-1 to NRP-1 leading to the inhibition of galectin-1/neuropilin-1/VEGFR2/MAPK-mediated angiogenic signaling pathway in endothelial cells. As higher plasma Lp(a) level in women is an independent risk factor for pre-eclamsia, a pregnancy-associated vascular complication, we propose that apo(a) O-glycans-mediated inhibition of the pro-angiogenic activity of galectin-1 may be one of the underlying molecular mechanism of pathogenesis of Lp(a) in pre-eclampsia.

Dietary Interventions for the Treatment of Inflammatory Bowel Diseases: An Updated Systematic Review and Meta-analysis.

BACKGROUND & AIMS: This study aimed (1) to systematically review controlled trials of solid food diets for the treatment of inflammatory bowel disease (IBD); and (2) to grade the overall quality of evidence. METHODS: Systematic review of prospective controlled trials of solid food diets for the induction or maintenance of remission in IBD. Two authors independently performed study selection, data extraction, and assessment of certainty of evidence. Meta-analyses were performed on studies with quantitative data on response, remission, and relapse. RESULTS: There were 27 studies for meta-analysis. For induction of remission in Crohn's disease (CD), low refined carbohydrate diet and symptoms-guided diet outperformed controls, but studies had serious imprecision and very low certainty of evidence. The Mediterranean diet was similar to the Specific Carbohydrate Diet (low certainty of evidence), and partial enteral nutrition (PEN) was similar to exclusive enteral nutrition (very low certainty of evidence). PEN reduced risk of relapse (very low certainty of evidence), whereas reduction of red meat or refined carbohydrates did not (low certainty of evidence). For ulcerative colitis, diets were similar to controls (very low and low certainty of evidence). CONCLUSIONS: Among the most robust dietary trials in IBD currently available, certainty of evidence remains very low or low. Nonetheless, emerging data suggest potential benefit with PEN for induction and maintenance of remission in CD. Reduction of red meat and refined carbohydrates might not reduce risk of CD relapse. As more dietary studies become available, the certainty of evidence could improve, thus allowing for more meaningful recommendations for patients.

Ironing It All Out: A Comprehensive Review of Iron Deficiency Anemia in Inflammatory Bowel Disease Patients.

Iron deficiency anemia affects approximately 45% of patients with inflammatory bowel disease (IBD), negatively impacts the quality of life in this patient population, and significantly burdens our healthcare system. The pathogenesis of iron deficiency in IBD patients is multifactorial, including intestinal bleeding, malabsorption, and inadequate oral intake. Regular screening and diagnosis in these patients are imperative, and often patients have mixed iron deficiency anemia and anemia of chronic disease, especially in those with active inflammation. Iron may be replenished either orally or intravenously. While oral iron is safe, affordable, and easy to administer, patients often suffer from intolerable gastrointestinal side effects, and particularly in IBD patients, oral iron may increase inflammation and contribute to flares. Therefore, although it is substantially underused, intravenous (IV) iron is considered first-line treatment for patients with active disease, severe anemia, oral iron intolerance, and erythropoietin requirements. Several IV iron formulations are available, and iron sucrose and ferric carboxymaltose are the most frequently used and well studied in patients with IBD. However, iron isomaltoside could potentially become a popular choice among providers given its safety, efficacy, and convenience. Overall, screening, diagnosis, and treatment of iron deficiency anemia are important in patients with IBD. Individual patient characteristics, risks, and benefits, and advantages and disadvantages, should be considered when determining the best route and formulation for iron repletion.

Superior Efficacy of Infliximab Versus Adalimumab for First-Line Treatment of Crohn's Perianal Fistulae.

BACKGROUND: Crohn's disease perianal fistulae (CD-PAF) occur in 25% of patients and are notoriously challenging to manage. Tumor necrosis factor inhibitors are first line agents. AIMS: The aim of this study was to compare infliximab (IFX) versus adalimumab (ADA) efficacy in CD-PAF healing over time. METHODS: A retrospective study at two large-tertiary medical centers was performed. Inclusion criteria were actively draining CD-PAF and initial treatment with IFX or ADA following CD-PAF diagnosis. The primary endpoints were perianal fistula response and remission at 6 and 12 months. Secondary endpoints included biologic persistence over time and dose escalation at 6 and 12 months. RESULTS: Among 151 patients included in the study, 92 received IFX and 59 received ADA as first line agents after CD-PAF diagnosis. At 6 months, the 64.9% of the IFX group and 34.8% of the ADA group demonstrated CD-PAF clinical improvement (p < 0.01). Univariate and multivariate analyses demonstrated significant differences among the IFX and ADA groups for clinical response at 6-months and 12-months (p = 0.002 and p = 0.042, respectively). There were no factors that predicted response, with the exception of concomitant immunomodulator affecting the 6-month clinical response (p = 0.021). Biologic persistence, characterized by Kaplan Meier methods, was significantly longer in the IFX group compared to the ADA group (Log-rank p = 0.01). CONCLUSION: IFX induction and maintenance is associated with higher rates of response and remission in CD-PAF healing as well as higher treatment persistence compared to ADA. Additionally, our study supports the use of concomitant immunomodulator therapy for CD-PAF healing and remission.

Adipocytes utilize sucrose as an energy source-Effect of different carbohydrates on adipocyte differentiation.

Adipose tissue is a complex and heterogenic tissue exhibiting high variability and appears to have multiple functions, especially in metabolic regulation. Change in carbohydrate source is reported to have a profound effect in the regular functioning of adipocytes. Here, we analyzed the role of two monosaccharides namely, glucose (5.2 and 25 nM), galactose (25 mM), and two disaccharides namely, lactose and sucrose (both at 25 mM) in the adipocyte differentiation process and its utilization by adipocytes as an energy source. The change in cell morphology, adipocyte-specific gene expression, and protein levels were analyzed at three different time points: 2, 6, and 48 hr. Oil Red O staining at Day 8 of differentiation showed that no other carbohydrates were able to increase lipid content as better as 25 mM glucose. Gene expression pattern was altered by the change in glucose concentration and sucrose was able to mimic the effect of glucose even though, the lipid synthesis was solely promoted by high glucose levels. Galactose and lactose did not show any effect in promoting adipocyte differentiation. The expression of PPAR γ was high in the presence of sucrose and galactose, possibly of adipogenic cocktail in enhancing the expression rather than the effect of carbohydrate. Acarbose, a potent glucosidase inhibitor was able to inhibit the lipid content in adipocytes grown with sucrose as a carbohydrate source and shows the possibility of its direct utilization. Lactate production by cells upon differentiation also proved the possible uptake of glucose after sucrose cleavage.

Increased sensitivity of Treg cells from patients with PBC to low dose IL-12 drives their differentiation into IFN-γ secreting cells.

IL-12 is a pro-inflammatory cytokine that induces the production of interferon-γ (IFNγ) and favours the differentiation of T helper 1 (Th1) cells. In the presence of IL-12 human Treg cells acquire a Th1-like phenotype with reduced suppressive activity in vitro. Primary biliary cholangitis (PBC) is an autoimmune cholestatic liver disease characterised by high Th1 and Th17 infiltrating cells, reduced frequencies of Treg cells, and a genetic association with IL-12 signalling. Herein, we sought to evaluate the IL-12 signalling pathway in PBC pathology, by studying human samples from patients with PBC, alongside those with primary Sjögren's syndrome (pSS)(autoimmune disease with IL-12 signalling gene association), primary sclerosing cholangitis (PSC) (cholestatic liver disease without IL-12 gene association) and healthy individuals. Our data revealed that TLR stimulation of PBC (n = 17) and pSS monocytes (n = 6) resulted in significant induction of IL12A mRNA (p < 0.05, p < 0.01, respectively) compared to PSC monocytes (n = 13) and at similar levels to HC monocytes (n = 8). PSC monocytes expressed significantly less IL-12p70 (108 pg/ml, mean) and IL-23 (358 pg/ml) compared to HC (458 pg/ml and 951 pg/ml, respectively) (p < 0.01, p < 0.05). Treg cells from patients with PBC (n = 16) and pSS (n = 3) but not PSC (n = 10) and HC (n = 8) responded to low dose (10 ng/ml) IL-12 stimulation by significant upregulation of IFNγ (mean 277 and 254 pg/ml, respectively) compared to PSC and HC Treg cells (mean 22 and 77 pg/ml, respectively)(p < 0.05). This effect was mediated by the rapid and strong phosphorylation of STAT4 on Treg cells from patients with PBC and pSS (p < 0.05) but not PSC and HC. In the liver of patients with PBC (n = 7) a significantly higher proportion of IL-12Rß2+Tregs (16% on average) was detected (p < 0.05) compared to other liver disease controls (5%)(n = 18) which also showed ex vivo high IFNG and TBET expression. CONCLUSION: Our data show an increased sensitivity of PBC and pSS Treg cells to low dose IL-12 stimulation, providing ongoing support for the importance of the IL12-IL-12Rß2-STAT4 pathway on Treg cells in disease pathogenesis and potentially treatment.

Time-dependent alterations in mRNA, protein and microRNA during in vitro adipogenesis.

Adipogenesis is a complex biological process involving synchronised interplay of different nuclear receptors. Aberration in the process leads to obesity and associated disorders. Addressing the complexity of molecular mechanisms, we worked on characterising the changes in NR1C3/PPARγ-, NR1H3/LXRα- and NCoAs/SRCs-associated microRNA, genes and proteins during different time points of adipogenesis. Glucose uptake of differentiating cells was checked at selected time points with FACS. Observations on gene expression pattern pointed a correlation in adipogenic-related genes and increased expression of PPARγ, but not LXRα. Western blot experiments also supported the gene expression pattern. MicroRNAs that vary during adipogenesis was selected using bioinformatics tools and database. Real-time PCR-based experiments showed a change in the expression of mmu-mir-23a-3p, 206-3p, 17-3p, 126a-3p and 1a-3p. Mmu-mir-23a-3p showed a gradual decrease in expression corresponding to the progression of adipogenesis. MicroRNA 23a-3p and 1a-3p showed positive association to the mRNA levels of NCoA1 and 3. Overall, the study elaborates time-dependent variations in nucleic acid and protein expression during adipogenesis in accordance to fatty acid and glucose metabolism.

Academic Stress May Contribute to the Onset of Pediatric Inflammatory Bowel Diseases.

It is currently unclear whether seasonality affects the onset of inflammatory bowel diseases (IBDs: Crohn disease and ulcerative colitis) in children. Here, we examined the records of pediatric patients with IBD diagnosed between 2009 and 2015 in a discovery cohort of 169 cases and a validation cohort of 122 subjects, where the month of symptoms onset could be determined. No seasonal patterns could be identified in respect to conception, birth, and disease onset. An annual rhythm of symptomatic onset, however, correlating with academic semesters was identified. IBD symptoms in the discovery cohort presented significantly more (P = 0.0218) during 5 months including the initiation (August, September, January) and the termination of academic semesters (December, May) compared to any other 5 months of the year. This observation was validated in the independent cohort (P < 0.0001). Our findings imply that academic stress may contribute to disease onset in pediatric IBD, which may improve timely diagnosis.

Effect on oxidative stress, glucose uptake level and lipid droplet content by Apigenin 7, 4'-dimethyl ether isolated from Piper longum L.

Piper longum L. (Family: Piperaceae), is a widely used herb in several Ayurvedic formulations prescribed for various diseases. Potential of the plant material as an antidiabetic and cardio protective agent has not been evaluated so far. In the study, we designed experiments to evaluate antioxidant, glucose uptake potential and lipid content regulating potential of extracts and compound from P. longum fruits. Solvent extracts from Piper longum fruits using hexane, ethyl acetate, methanol, 70 % methanol-water were taken and apigenin 7, 4'-dimethyl ether (ADE) was isolated from ethyl acetate extract. Antioxidant activity, glucose uptake potential and adipocyte differentiation assay was performed with extract and pure compound. Antioxidant activity in terms of TRP (196.03 µg/mg GAE), DPPH assay (IC50-173.09 µg/mL), hydroxyl radical scavenging assay (IC50-20.42 µg/mL), inhibiting LDL oxidation (IC50-51.99 µg/mL) and to enhance SOD activity (25.3 %) was higher in ethyl acetate extract (EAP). Phenolic and flavonoid content was measured and showed a positive correlation with antioxidant activity. Presence of apigenin 7, 4'-dimethyl ether (ADE) and piperine (Pip) in EAP was determined by HPTLC analysis and was isolated. ADE inhibited α-glucosidase and α-amylase enzymes and enhanced 2-NBDG uptake in L6 cells. Hypolipidemic effect of ADE on mouse pre-adipocyte (3T3L1) cell lines also showed a dose dependent reduction on lipid droplet content and effective concentration range was determined as 1-2.5 µg/mL. The results suggested that Piper longum fruits can provide a natural source of antioxidants with antidiabetic and anti obesity potential.

Machine Learning-based Characterization of Longitudinal Health Care Utilization Among Patients With Inflammatory Bowel Diseases.

BACKGROUND: Inflammatory bowel disease (IBD) is associated with increased health care utilization. Forecasting of high resource utilizers could improve resource allocation. In this study, we aimed to develop machine learning models (1) to cluster patients according to clinical utilization patterns and (2) to predict longitudinal utilization patterns based on readily available baseline clinical characteristics. METHODS: We conducted a retrospective study of adults with IBD at 2 academic centers between 2015 and 2021. Outcomes included different clinical encounters, new prescriptions of corticosteroids, and initiation of biologic therapy. Machine learning models were developed to characterize health care utilization. Poisson regression compared frequencies of clinical encounters. RESULTS: A total of 1174 IBD patients were followed for more than 5673 12-month observational windows. The clustering method separated patients according to low, medium, and high resource utilizers. In Poisson regression models, compared with low resource utilizers, moderate and high resource utilizers had significantly higher rates of each encounter type. Comparing moderate and high resource utilizers, the latter had greater utilization of each encounter type, except for telephone encounters and biologic therapy initiation. Machine learning models predicted longitudinal health care utilization with 81% to 85% accuracy (area under the receiver operating characteristic curve 0.84-0.90); these were superior to ordinal regression and random choice methods. CONCLUSION: Machine learning models were able to cluster individuals according to relative health care resource utilization and to accurately predict longitudinal resource utilization using baseline clinical factors. Integration of such models into the electronic medical records could provide a powerful semiautomated tool to guide patient risk assessment, targeted care coordination, and more efficient resource allocation.

Zingiber officinale extract exhibits antidiabetic potential via modulating glucose uptake, protein glycation and inhibiting adipocyte differentiation: an in vitro study.

BACKGROUND: Ginger, the rhizome of Zingiber officinale Roscoe (Zingiberaceae), a perennial herbaceous plant is native to Southern Asia. Study was aimed to evaluate antioxidant and antidiabetic potential of ginger extract and its characterization. Possible mode of action to elicit antidiabetic activity was also evaluated. METHODS AND RESULTS: Ethyl acetate extract of ginger (EAG) was evaluated for its antioxidant activity in terms of DPPH radical scavenging potential with an IC50 value of 4.59 µg/ml. Antidiabetic activity of EAG was evaluated by estimating antiglycation potential (IC50 290.84 µg/ml). HPLC profiling of EAG revealed the presence of phenolic components, gingerol and shoagol as major constituents. After determining sub-toxic concentration of EAG (50 µg/ml), efficacy of extract to enhance glucose uptake in cell lines were checked in L6 mouse myoblast and myotubes. EAG was effective at 5 µg/ml concentration in both cases. Antibody based studies in treated cells revealed the effect of EAG in expressing Glut 4 in cell surface membrane compared to control. CONCLUSION: The antidiabetic effect of ginger was experimentally proved in the study and has concluded that the activity is initiated by antioxidant, antiglycation and potential to express or transport Glut4 receptors from internal vesicles.

Maternal Lactobacillus reuteri supplementation shifts the intestinal microbiome in mice and provides protection from experimental colitis in female offspring.

The purpose of our experiment was to explore how stochastic (inter-individual variation) gut microbiome composition may link to inflammatory bowel disease (IBD) susceptibility and guide the development of a perinatal preventative probiotic. Dextran sodium sulfate (DSS) was introduced to C57BL/BJ mice to induce acute colitis as a model of IBD. Potentially protective bacteria were identified using a discovery-validation cohort approach toward stochastic DSS susceptibility. Lactobacilli (two different cocktails of L. reuteri and L. johnsonii strains) or control media were supplemented by mouth to dams prior to delivery and during lactation (i.e., perinatal probiotic). The pups were evaluated for DSS susceptibility at young adulthood. Fecal Lactobacillus was increased in the DSS-resistant mice in both the discovery and validation cohorts. Maternal supplementation of female offspring with an L. reuteri cocktail (strains 6798-1, 6798-jm, and 6798-cm) induced progressive microbiome separation and protection against colitis by young adulthood. Maternal supplementation of L. reuteri could confer protection against DSS colitis in young adult female mice. This work is the first to exploit stochastic mammalian microbiome variation to guide microbial therapeutic identification. Our findings underscore neonatal microbiome plasticity and set the stage for the potential development of perinatally deliverable protective probiotics against human IBD.

Parental Education May Differentially Impact Pediatric Inflammatory Bowel Disease Phenotype Risk.

BACKGROUND: The incidence of pediatric inflammatory bowel diseases (PIBDs: Crohn's disease [CD], ulcerative colitis [UC]) is on the rise around the world. Yet, the critical risk factors for this rising incidence are not well understood. Demographic characteristics of PIBD may improve our understanding of their developmental origins and aid in prevention. METHODS: Four hundred eighty-eight consecutive PIBD patients diagnosed at Texas Children's Hospital from 13 counties around Houston were studied. An annual incidence map was created by ZIP code of residence at diagnosis by using ArcGIS and the American Community Survey from the US Census Bureau. Correlation between demographic variables and PIBD incidence was examined. A model to explain incidence from different health factors was created in R. RESULTS: Hispanic children were more likely to be diagnosed with UC (P < 0.01) and unclassified IBD (IBD-U) (P < 0.03) compared with other races/ethnicities. A significant positive correlation (r = 0.35, P < 0.0001) between median household income and PIBD incidence was observed (UC: r = 0.23, P < 0.0001; CD: r = 0.22, P = 0.0004). ZIP codes with majority college-educated adults had a higher incidence of PIBD than ZIP codes with majority high school-educated adults (P < 0.0001). Pediatric cases with CD were more common in ZIP codes where the majority of adults were college educated (P < 0.0001). Pediatric cases with UC, however, were more common in ZIP codes where the majority of adults were high school educated (P = 0.0036). CONCLUSIONS: Hispanic children more commonly present with UC and IBD-U in southern USA. Household income and/or adult education-related environmental/dietary differences may be important in the developmental origins of PIBD in large metro areas, such as Houston.

Diagnostic delay and colectomy risk in pediatric ulcerative colitis.

BACKGROUND/PURPOSE: Diagnostic delay or time to diagnosis, and its relationship with colectomy risk has been studied in adult Inflammatory Bowel Disease (IBD), but rarely in pediatric IBD (PIBD), especially pediatric ulcerative colitis (P-UC), which often has a more severe course than adult UC. This study compared the relationship between diagnostic delay and colectomy in P-UC. METHODS: The medical records of P-UC patients, ages <18 years, diagnosed at Texas Children's Hospital from 2012 to 2018 were examined. We identified 106 P-UC patients, where the onset of symptoms of IBD (i.e. fever, diarrhea, blood in stool, weight loss, abdominal pain) could be clearly identified. RESULTS: Twenty (20 = 18.9%) patients progressed to colectomy, and 86 did not. There was no significant difference in diagnostic delay between the patients undergoing colectomy with UC (C-UC) and those with no colectomy (NC-UC) (p = 0.2192). The median (C-UC = 7.1 weeks; NC-UC = 11.9 weeks) and mean (C-UC = 16.5 weeks±4.7; NC-UC = 20.1 ±â€¯2.6) diagnostic delay actually tended to be shorter in C-UC compared to NC-UC. Fecal calprotectin levels were significantly higher (p = 0.0228) in C-UC than NC-UC patients at diagnosis. CONCLUSIONS: Shorter time to diagnosis may reflect disease severity at the time of disease onset and also a more aggressive subsequent course of P-UC. The significantly higher level of fecal calprotectin in the C-UC patients at diagnosis provided biologic/biochemical support for our conclusion. LEVELS OF EVIDENCE: Prognosis study, Level III evidence.

Weight loss is a sufficient and economical single outcome measure of murine dextran sulfate sodium colitis.

Inflammatory bowel diseases (IBD: Crohn's disease and ulcerative colitis) are becoming common around the world without a cure. Animal models of colitis have become instrumental in IBD research. The dextran sulfate sodium (DSS) induced murine colitis model is likely the most utilized due to its simplicity and reproducibility with over 4000 publications on PubMed, where weight loss is the most commonly used and reliable positive correlate. We predicted at current state of art, that the DSS colitis model can be optimized by using weight loss as a single cost-saving outcome measure. Twenty recent and consecutive publications using the DSS model in PubMed were selected for review. Guarded cost estimations for additional outcome measures of colitis beyond weight loss were performed. In all manuscripts (100%), weight loss corroborated the conclusions. Average excess cost for examining additional measures of colitis was approximately $6700 per publication. Two studies (10.5%) were estimated to have spent over $20,000 in excess. Additional measures of colitis either supported the final conclusions found with weight loss, or lead to indeterminate results. Potential annual savings from following our guidance were calculated to be over $60,000 for and IBD lab. We conclude that weight loss is a sufficient, objective, and economical outcome measure of DSS-induced colitis in mice.

A lysosome targetable luminescent bioprobe based on a europium ß-diketonate complex for cellular imaging applications.

Herein, we report a novel lysosome targetable luminescent bioprobe derived from a europium coordination compound, namely Eu(pfphOCH3IN)3(DDXPO) 4 [where HpfphOCH3IN = 4,4,5,5,5-pentafluoro-3-hydroxy-1-(1-(4-methoxyphenyl)-1H-indol-3-yl)pent-2-en-1-one and DDXPO = 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene oxide]. Notably, the newly designed europium complex exhibits significant quantum yield (Φoverall = 25 ± 3%) and 5D0 excited state lifetime (τ = 398 ± 3 µs) values under physiological pH (7.2) conditions when excited at 405 nm. Hence the developed europium complex has been evaluated for live cell imaging applications using mouse pre-adipocyte cell lines (3T3L1). Colocalization studies of the designed bio-probe with commercial Lysosome-GFP in 3T3L1 cells demonstrated the specific localization of the probe in the lysosome with a high colocalization coefficient (A = 0.83). Most importantly, the developed bioprobe exhibits good cell permeability, photostability and non-cytotoxicity.