Miracle-dz/UniFAR-FTUs huggingface dataset:
Cell Culture and Coculture for Oncological Research in Appropriate Microenvironments

With the increase in knowledge on the importance of the tumor microenvironment, cell culture models of cancers can be adapted to better recapitulate physiologically relevant situations. Three main microenvironmental factors influence tumor phenotype: the biochemical components that stimulate cells, the fibrous molecules that influence the stiffness of the extracellular matrix, and noncancerous cells like epithelial cells, fibroblasts, endothelial cells, and immune cells. Here we present methods for the culture of carcinomas in the presence of a matrix of specific stiffness, and for the coculture of tumors and fibroblasts as well as epithelial cells in the presence of matrix. Information is provided to help with choice and assessment of the matrix support and in working with serum‐free medium. Using the example of a tissue chip recapitulating the environmental geometry of carcinomas, we also highlight the development of engineered platforms that provide exquisite control of cell culture parameters necessary in research and development. © 2019 by John Wiley & Sons, Inc.
Perivascular Niche of Postnatal Mesenchymal Stem Cells in Human Bone Marrow and Dental Pulp

Mesenchymal stem cell populations have previously been identified in adult bone marrow and dental pulp that are capable of regenerating the bone marrow and dental pulp microenvironments, respectively. Here we show that these stem cell populations reside in the microvasculature of their tissue of origin. Human bone marrow stromal stem cells (BMSSCs) and dental pulp stem cells (DPSCs) were isolated by immunoselection using the antibody, STRO‐1, which recognizes an antigen on perivascular cells in bone marrow and dental pulp tissue. Freshly isolated STRO‐1 positive BMSSCs and DPSCs were tested for expression of vascular antigens known to be expressed by endothelial cells (von Willebrand factor, CD146), smooth muscle cells, and pericytes (α‐smooth muscle actin, CD146), and a pericyte‐associated antigen (3G5), by immunohistochemistry, fluorescence‐activated cell sorting (FACS), and/or immunomagnetic bead selection. Both BMSSCs and DPSCs lacked expression of von Willebrand factor but were found to be positive for α‐smooth muscle actin and CD146. Furthermore, the majority of DPSCs expressed the pericyte marker, 3G5, while only a minor population of BMSSCs were found to be positive for 3G5. The finding that BMSSCs and DPSCs both display phenotypes consistent with different perivascular cell populations, regardless of their diverse ontogeny and developmental potentials, may have further implications in understanding the factors that regulate the formation of mineralized matrices and other associated connective tissues.
Abnormal Cortical and Trabecular Bone in Youth With Type 1 Diabetes and Celiac Disease

OBJECTIVE This study compared bone health in youth with type 1 diabetes and celiac disease (CD) versus type 1 diabetes alone. RESEARCH DESIGN AND METHODS This was a case-control study of 42 youth with coexisting type 1 diabetes and CD and 40 with type 1 diabetes matched for age, sex, diabetes duration, and HbA1c. Bone mineral density (BMD), bone mineral content (BMC), and BMC-to-lean tissue mass (LTM) ratio were measured using DXA and reported as z-scores for height. Total, trabecular, and cortical bone and muscle parameters were measured using peripheral quantitative computed tomography (pQCT) and reported as z-scores for age. RESULTS Mean age at assessment was 14.3 ± 3.1 years; diabetes duration, 8.0 ± 3.5 years; HbA1c, 8.2 ± 1.5% (66 ± 5 mmol/mol); and 25-hydroxy vitamin D, 71 ± 21 nmol/L. Comparing youth with coexisting CD versus type 1 diabetes alone, DXA showed lower BMC-to-LTM ratio (0.37 ± 1.12 vs. 0.73 ± 2.23, P = 0.007) but no difference in total BMD. Youth with coexisting CD also had lower BMC-to-LTM ratio versus the general population (P = 0.04). Radial pQCT showed lower total BMC (−0.92 ± 1.40 vs. −0.26 ± 1.23, P = 0.03) despite similar bone and muscle cross-sectional area. In multivariable linear regression, lower BMC was associated with higher insulin dose (P = 0.03) but not HbA1c. CONCLUSIONS Youth with both type 1 diabetes and CD have lower BMC relative to LTM and lower BMC, indicating abnormal trabecular and cortical bone development despite similar bone and muscle size. These findings suggest that the two conditions confer a lower bone turnover state. We recommend further examination of bone health in this population; future research should examine early interventions to improve bone health.
Knowledge and Attitude Among Lebanese Pregnant Women Toward Cord Blood Stem Cell Storage and Donation

Background: Umbilical cord blood (UCB) used to be considered waste material and was discarded at birth. However, cord blood is now considered a rich source of adult stem cells that can be used to treat many conditions and diseases. This study was performed to determine pregnant women’s knowledge and attitudes toward cord blood stem cell banking and donation in Lebanon. Methods: A descriptive study was conducted in antenatal clinics in Beirut and data were collected using a questionnaire distributed to pregnant women after provision of informed consent. A total of 244 women responded. Results: Less than half of the women (46%) reported knowledge about cord blood banking. However, participants with university and secondary education had significantly higher odds of considering UCB storage compared to those with primary education (odds ratio (OR) 8.62, 95% confidence interval (CI) 2.74–27.15 and OR 21.23,95% CI 6.55–68.86, respectively). Older pregnant women were less likely to think about UCB stem cells storage (OR 0.92, 95% CI 0.85–0.98). Conclusion: Respondents who had an existing knowledge about UCB stem cells banking in general were more likely to consider storing UCB in blood banks if affordable (45.9%). Therefore, it is necessary to inform pregnant women about stem cell banking so that they can make the appropriate decisions for themselves.
Prevalence of suicide attempts in individuals with schizophrenia: a meta-analysis of observational studies

Abstract Aims Suicide attempt is an important indicator of suicide and potential future mortality. However, the prevalence of suicide attempts has been inconsistent across studies. This meta-analysis aimed to examine the prevalence of suicide attempts in individuals with schizophrenia and associated correlates. Methods Relevant publications in Embase, PsycINFO, PubMed, Web of science and Cochrane were systematically searched. Data on the prevalence of suicide attempts in individuals with schizophrenia were pooled using a random-effects model. Results Thirty-five studies with 16 747 individuals with schizophrenia were included. The pooled lifetime prevalence of suicide attempts was 26.8% (95% CI 22.1–31.9%; I2 = 97.0%), while the 1-year prevalence, 1-month prevalence and the prevalence of suicide attempts from illness onset were 3.0% (95% CI 2.3–3.7%; I2 = 95.6%), 2.7% (95% CI 2.1–3.4%; I2 = 78.5%) and 45.9% (95% CI 42.1–49.9%; I2 = 0), respectively. Earlier age of onset (Q = 4.38, p = 0.04), high-income countries (Q = 53.29, p < 0.001), North America and Europe and Central Asia (Q = 32.83, p < 0.001) were significantly associated with a higher prevalence of suicide attempts. Conclusions Suicide attempts are common in individuals with schizophrenia, especially those with an early age of onset and living in high-income countries and regions. Regular screening and effective preventive measures should be implemented as part of the clinical care.
GluN2A-NMDA receptor–mediated sustained Ca2+ influx leads to homocysteine-induced neuronal cell death

Homocysteine, a metabolite of the methionine cycle, is a known agonist of N-methyl-d-aspartate receptor (NMDAR), a glutamate receptor subtype and is involved in NMDAR-mediated neurotoxicity. Our previous findings have shown that homocysteine-induced, NMDAR-mediated neurotoxicity is facilitated by a sustained increase in phosphorylation and activation of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK MAPK). In the current study, we investigated the role GluN1/GluN2A-containing functional NMDAR (GluN2A-NMDAR) and GluN1/GluN2B-containing functional NMDAR (GluN2B-NMDAR) in homocysteine-induced neurotoxicity. Our findings revealed that exposing primary cortical neuronal cultures to homocysteine leads to a sustained low-level increase in intracellular Ca2+. We also showed that pharmacological inhibition of GluN2A-NMDAR or genetic deletion of the GluN2A subunit attenuates homocysteine-induced increase in intracellular Ca2+. Our results further established the role of GluN2A-NMDAR in homocysteine-mediated sustained ERK MAPK phosphorylation and neuronal cell death. Of note, the preferential role of GluN2A-NMDAR in homocysteine-induced neurotoxicity was distinctly different from glutamate-NMDAR–induced excitotoxic cell death that involves overactivation of GluN2B-NMDAR and is independent of ERK MAPK activation. These findings indicate a critical role of GluN2A-NMDAR–mediated signaling in homocysteine-induced neurotoxicity.
Effects of Corticosterone and Microgravity on Inflammatory Cell Production of Superoxide

In this investigation we studied the effects of corticosterone and microgravity on Propionibacterium acnes‐induced inflammatory cells ability to produce superoxide (O2 ‐). We found in vitro and in vivo exposure of murine peritoneal inflammatory cells to corticosterone did not inhibit the O2 ‐ response. We also found that in microgravity P. acnes‐induced inflammatory cells were capable of producing four times as much O2 ‐ as at 1g. Therefore, neither corticosterone nor microgravity experienced during parabolic flight prevents an O2 ‐ response by inflammatory cells.
Total Ankle Arthroplasty Survival and Risk Factors for Failure

Background: Total ankle arthroplasty (TAA) is an increasingly selected treatment for end-stage ankle arthritis; however, failure and revision of the tibial and talar components remains an issue. Although multiple risk factors have been shown to contribute to early component revision, no study has looked at combining such risk factors into a predictive model that could potentially decrease revision rates and improve implant survival. This study aimed to develop a predictive model for TAA failure based on patient characteristics, patient-reported outcomes (PROs), and immediate postoperative radiographs. Methods: A retrospective review of a single-site ankle arthritis database was conducted. All patients with current-generation ankle replacements including the Hintegra and Infinity prostheses implanted between 2004 and 2015 and with complete postoperative radiographs taken between 6 and 12 weeks postoperatively were included. Eight coronal and sagittal radiographic parameters were assessed and performed twice by 2 independent orthopedic surgeons on included TAAs. These radiographic parameters were then analyzed in association with patient demographics and PRO. Advanced statistical methods including survival analysis were used to construct a predictive model for TAA survival. A total of 107 patients were included and analyzed with a median clinical follow-up of 49 months (minimum 24 months). Results: A predictive model was created, with 4 parameters identified as being statistically associated with TAA metal-component revision: diabetes mellitus, poor baseline Ankle Osteoarthritis Scale (AOS) score, excessively dorsiflexed talar component, and an anteriorly/posteriorly translated talus relative to the tibial axis. The presence of 3 parameters predicted TAA survival of 0.60 whereas presence of all 4 parameters predicted survival of only 0.13 in the period studied. Conclusion: Our predictive model is based on a combination of patient factors, PROs, and radiographic TAA alignment. We believe it can be used by surgeons to predict failure in their TAA patients, thereby optimizing postoperative outcomes by improving patient selection and modifiable outcome-specific parameters. Level of Evidence: Level III, retrospective cohort study using prospectively collected data.
Angiopoietin 1 expression levels in the myocardium direct coronary vessel development

Mutational studies in genetically engineered mice have shown that the angiopoietin/Tie2(Tek) signaling pathway is indispensable for vascular development. To further investigate the role of Angiopoietin 1 in heart development, we developed transgenic mice that express Angiopoietin 1 under control of doxycycline in cardiac myocytes. Ninety percent of all transgenic mice die between embryonic day 12.5 and 15.5. Beginning at embryonic day 12.5, transgenic mice exhibit dilated atria, a significant thinning of the myocardial wall, and eventual outflow tract collapse. In addition, hearts of the most severely affected transgenic embryos have no coronary arteries as a result of the defective development and maintenance of the epicardium. The subsequent lack of blood and nutrient delivery to the developing heart may account for decreases in N‐cadherin expression and subsequent loss of cell–cell contact leading to cell death, and ultimately the collapse and hemorrhage of the heart. These results suggest a pivotal role for Angiopoietin 1 in cardiovascular development, specifically the development of the epicardium and coronary vasculature. Developmental Dynamics 229:500–509, 2004. © 2004 Wiley‐Liss, Inc.
Genome-wide association study identiﬁed novel candidate loci affecting wood formation in Norway spruce

SUMMARY Norway spruce is a boreal forest tree species of signiﬁcant ecological and economic importance. Hence there is a strong imperative to dissect the genetics underlying important wood quality traits in the species. We performed a functional genome-wide association study (GWAS) of 17 wood traits in Norway spruce using 178 101 single nucleotide polymorphisms (SNPs) generated from exome genotyping of 517 mother trees. The wood traits were deﬁned using functional modelling of wood properties across annual growth rings. We applied a Least Absolute Shrinkage and Selection Operator (LASSO-based) association mapping method using a functional multilocus mapping approach that utilizes latent traits, with a stability selection probability method as the hypothesis testing approach to determine a signiﬁcant quantitative trait locus. The analysis provided 52 signiﬁcant SNPs from 39 candidate genes, including genes previously implicated in wood formation and tree growth in spruce and other species. Our study represents a multilocus GWAS for complex wood traits in Norway spruce. The results advance our understanding of the genetics inﬂuencing wood traits and identiﬁes candidate genes for future functional studies.
Pathogenesis of Respiratory Syncytial Virus Infection in BALB/c Mice Differs Between Intratracheal and Intranasal Inoculation

Human respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract disease requiring hospitalization in infants. There are no market-approved vaccines or antiviral agents available, but a growing number of vaccines and therapeutics are in (pre)clinical stages of development. Reliable animal models are crucial to evaluate new vaccine concepts, but in vivo RSV research is hampered by the lack of well-characterized animal models that faithfully mimic the pathogenesis of RSV infection in humans. Mice are frequently used in RSV infection and vaccination studies. However, differences in the use of mouse strains, RSV subtypes, and methodology often lead to divergent study outcomes. To our knowledge, a comparison between different RSV inoculation methods in mice has not been described in the literature, even though multiple methods are being used across different studies. In this study, we evaluated various pathological and immunological parameters in BALB/c mice after intratracheal or intranasal inoculation with RSV-A2. Our study reveals that intranasal inoculation induces robust pathology and inflammation, whereas this is not the case for intratracheal inoculation. As immunopathology is an important characteristic of RSV disease in infants, these data suggest that in mice intranasal inoculation is a more appropriate method to study RSV infection than intratracheal inoculation. These findings will contribute to the rational experimental design of future in vivo RSV experiments.
Quantifying Retinal Pigment Epithelium Dysmorphia and Loss of Histologic Autofluorescence in Age-Related Macular Degeneration

Purpose Lipofuscin and melanolipofuscin organelles in retinal pigment epithelium (RPE) cells are signal sources for clinical fundus autofluorescence (AF). To elucidate the subcellular basis of AF imaging, we identified, characterized, and quantified the frequency of RPE morphology and AF phenotypes in donor eyes with age-related macular degeneration (AMD). Methods In 25 RPE–Bruch's membrane flat mounts from 25 eyes, we analyzed 0.4-μm z-stack epifluorescence images of RPE stained with phalloidin for actin cytoskeleton. Using a custom ImageJ plugin, we classified cells selected in a systematic unbiased fashion in six phenotypes representing increasing degrees of pathology. For each cell, area, AF intensity, and number of Voronoi neighbors were compared with phenotype 1 (uniform AF, polygonal morphology) via generalized estimating equations. We also analyzed each cell's neighborhood. Results In 29,323 cells, compared with phenotype 1, all other phenotypes, in order of increasing pathology, had significantly larger area, reduced AF, and more variable number of neighbors. Neighborhood area and AF showed similar, but subtler, trends. Cells with highly autofluorescent granule aggregates are no more autofluorescent than others and are in fact lower overall in AF. Pre-aggregates were found in phenotype 1. Phenotype 2, which exhibited degranulation despite normal cytoskeleton, was the most numerous nonhealthy phenotype (16.23%). Conclusions Despite aggregation of granules that created hyperAF aggregates within cells, overall AF on a per cell basis decreased with increasing severity of dysmorphia (abnormal shape). Data motivate further development of subcellular resolution in clinical fundus AF imaging and inform an ongoing reexamination of the role of lipofuscin in AMD.
Risk factors for group B streptococcus early-onset disease: an Italian, area-based, case-control study

Abstract Purpose: Intrapartum antibiotic prophylaxis (IAP) prevents group B streptococcus (GBS) early-onset disease (EOD). No European study evaluates the relative impact of risk factors (RFs) for EOD after a screening-based strategy and widespread IAP use We aimed to evaluate the risks of EOD in an Italian region where a screening-based strategy for preventing EOD was implemented. Materials and methods: Cases of EOD born at or above 35 weeks’ gestation were reviewed and matched with controls. Results: There were 109 cases of EOD among 532,154 live births. Most cases had negative GBS prenatal screening (56/91, 61.5%) and were unexposed to IAP (86/109, 78.9%). At multivariate analysis, GBS bacteriuria (OR = 6.99), positive prenatal screening (OR = 13.7) and maternal intrapartum fever (OR = 188.3) were associated with an increased risk of EOD, whereas intrapartum beta-lactam antibiotics were associated with a decreased risk of EOD (≥4 h: OR = 0.008; <4 h: OR = 0.04). Neonates born to nonfebrile, GBS positive pregnant women, receiving beta-lactam antibiotics had very low probability of EOD, particularly if IAP was adequate. Conclusions: GBS positive prenatal screening, GBS bacteriuria and intrapartum fever are associated with EOD. Intrapartum beta-lactam antibiotics reduce the probability of EOD in neonates born to nonfebrile mothers.
Altered Bile Transporter Expression and Cholesterol Metabolism in Children with Cholesterol and Pigment Gallstones.

OBJECTIVES. We elucidated pathophysiology of pediatric gallstone disease by assessing liver expression of bile transporters in relation to bile acids and surrogates of cholesterol absorption and synthesis in serum and gallstones. METHODS. RNA expression of canalicular bile transporters in liver biopsies from 32 pediatric gallstone patients and from six liver donors (controls) was measured by qRT-PCR. Concentrations of cholesterol and precursors, plant sterols and bile acids in gallstones, and in serum of the patients and 82 healthy children were measured. Primary outcomes were the difference in RNA expressions and serum sterol profiles between patients and controls. RESULTS. Cholesterol stones (CS; n = 15) contained cholesterol >42% and pigment stones (PS; n = 17) <9% of weight. CS-patients had markedly lower serum plant sterols (absorption) and higher cholesterol precursors (synthesis) than PS-patients or healthy controls. CS contained several times more cholesterol precursors and less plant sterols relative to cholesterol than PS, which were enriched by primary bile acids (12-5.2 fold, p<0.001). Liver RNA expression of ABCG5/G8 was similarly increased 2.5-1.8 fold (p < 0.002) in CS and PS-patients, while PS-patients had higher ABCB11 expression (p < 0.05). In PS bile acid concentration correlated with gallstone plant sterols (R = 0.83, p < 0.0001), and ABCG5 expression with ACBC11 expression (R2 = 0.27, p = 0.03). CONCLUSIONS In CS, upregulation of ABCG5/G8 expression associates with low absorption and high gallstone content of cholesterol. In PS, activation of bile acid transport by ACBC11 interconnects with hepatic upregulation of ABCG5/G8 enriching PS with bile acids and plant sterols.
Lymphatic endothelial cell immunohistochemical markers for evaluation of the intestinal lymphatic vasculature in dogs with chronic inflammatory enteropathy

Abstract Background Lymphatic endothelial cell (LEC) immunohistochemical markers have identified intestinal lymphatic vasculature abnormalities in humans with inflammatory bowel disease, but have not been used to evaluate intestinal lymphatic vasculature in a group of dogs with chronic inflammatory enteropathy (CIE). Objectives To utilize LEC markers to identify and measure intestinal lymphatic vasculature in endoscopic biopsy samples of CIE dogs. To evaluate whether measured lymphatic vasculature variables correlate with serum albumin concentrations. Animals Twenty‐four dogs with CIE; n = 13, serum albumin concentration <2.5 g/dL (CIE‐protein‐losing enteropathy [PLE]), n = 11, serum albumin concentration ≥2.5 g/dL (CIE‐N). Methods Prospective study. Lymphatic endothelial cell immunolabeling with Prox‐1 and LYVE‐1 performed on endoscopic biopsy samples from 24 dogs with CIE. Duodenal and ileal villous lacteal width (VLW) and proprial mucosal lacteal width (MLW) were determined for each case and analyzed for correlation with serum albumin concentration. Lacteal dilatation scores using routine H&E histopathology were assessed for correlation with immunohistochemistry (IHC)‐calculated VLW and MLW. Results Lower serum albumin concentrations were correlated with increased VLW (rho = −.4644; P = .02) and MLW (rho = −.6514; P < .001) in the ileum. Lymphatic endothelial cell IHC identified presumptive proprial mucosal lymphangiectasia in some dogs that was not recognized with routine H&E staining. Lacteal dilatation scores were correlated with VLW in duodenum (rho = .4634; P = .02) and ileum (rho = .5292; P = .008), but did not correlate with MLW. Conclusions and Clinical Importance Lymphatic endothelial cell immunolabeling identified presumptive proprial mucosal lymphangiectasia in CIE dogs, particularly in the ileum of hypoalbuminemic dogs. Routine evaluation of villous lacteals likely underestimates abnormalities of the lymphatic vasculature in dogs with CIE.