Search

Background Encapsulated pancreatic fluid collection (PFC) is a requisite for endoscopic drainage procedures. The 4-week threshold for defining walled-off necrosis does not capture the dynamic process of encapsulation. We aim to investigate the changes in the wall characteristics of PFC in acute necrotizing pancreatitis (ANP) by comparing baseline contrast-enhanced computed tomography (CECT) with follow-up CT scans.

Background The role of dual-modality drainage of walled-off necrosis (WON) in patients with acute pancreatitis (AP) is established. However, there are no data on the association of clinical outcomes with the timing of percutaneous catheter drainage (PCD). We investigated the impact of the timing of PCD following endoscopic drainage of WON on clinical outcomes in AP.

Summary: Brain metastasis cancer-associated fibroblasts (bmCAFs) are emerging as crucial players in the development of breast cancer brain metastasis (BCBM), but our understanding of the underlying molecular mechanisms is limited. In this study, we aim to elucidate the pathological contributions of fucosylation (the post-translational modification of proteins by the dietary sugar L-fucose) to tumor-stromal interactions that drive the development of BCBM. Here, we report that patient-derived bmCAFs secrete high levels of polio virus receptor (PVR), which enhance the invasive capacity of BC cells. Mechanistically, we find that HIF1α transcriptionally upregulates fucosyltransferase 11, which fucosylates PVR, triggering its secretion from bmCAFs. Global phosphoproteomic analysis of BC cells followed by functional verification identifies cell-cell junction and actin cytoskeletal signaling as modulated by bmCAF-secreted, -fucosylated PVR. Our findings delineate a hypoxia- and fucosylation-regulated mechanism by which bmCAFs contribute to the invasiveness of BCBM in the brain.

Abstract Introduction Aggressive cancers commonly ferment glucose to lactic acid at high rates, even in the presence of oxygen. This is known as aerobic glycolysis, or the “Warburg Effect.” It is widely assumed that this is a consequence of the upregulation of glycolytic enzymes. Oncogenic drivers can increase the expression of most proteins in the glycolytic pathway, including the terminal step of exporting H+ equivalents from the cytoplasm. Proton exporters maintain an alkaline cytoplasmic pH, which can enhance all glycolytic enzyme activities, even in the absence of oncogene-related expression changes. Based on this observation, we hypothesized that increased uptake and fermentative metabolism of glucose could be driven by the expulsion of H+ equivalents from the cell. Results To test this hypothesis, we stably transfected lowly glycolytic MCF-7, U2-OS, and glycolytic HEK293 cells to express proton-exporting systems: either PMA1 (plasma membrane ATPase 1, a yeast H+-ATPase) or CA-IX (carbonic anhydrase 9). The expression of either exporter in vitro enhanced aerobic glycolysis as measured by glucose consumption, lactate production, and extracellular acidification rate. This resulted in an increased intracellular pH, and metabolomic analyses indicated that this was associated with an increased flux of all glycolytic enzymes upstream of pyruvate kinase. These cells also demonstrated increased migratory and invasive phenotypes in vitro, and these were recapitulated in vivo by more aggressive behavior, whereby the acid-producing cells formed higher-grade tumors with higher rates of metastases. Neutralizing tumor acidity with oral buffers reduced the metastatic burden. Conclusions Therefore, cancer cells which increase export of H+ equivalents subsequently increase intracellular alkalization, even without oncogenic driver mutations, and this is sufficient to alter cancer metabolism towards an upregulation of aerobic glycolysis, a Warburg phenotype. Overall, we have shown that the traditional understanding of cancer cells favoring glycolysis and the subsequent extracellular acidification is not always linear. Cells which can, independent of metabolism, acidify through proton exporter activity can sufficiently drive their metabolism towards glycolysis providing an important fitness advantage for survival.

This study was completed in effort to characterize the notch sensitivity of additively manufactured (AM) Inconel 718 produced by laser powder bed fusion (L-PBF). Three different root radii on V-notched test specimens and smooth specimens were evaluated under tensile conditions for specimens built in vertical and horizontal orientations. Both the total axial strain and localized notch diametral strain were measured. Finite element analysis (FEA) was completed on each specimen geometry to confirm the actual strain measurements near the notch. Test results showed the tensile strength of the notched specimens were larger than the tensile strength values of the smooth specimens. These tensile results equate to a notch-sensitivity ratio (NSR) greater than one, indicating that the L-PBF Inconel 718 material is a notch-strengthened material. It is suspected that the notch strengthening is a result of increased triaxial stress produced near the notch tip causing added material constraints, resulting in higher strength values for the notched specimens. Fractography analysis was completed on the various fracture surfaces and identified a dominate ductile failure mode within all of the specimens; however, the amount of ductility reduced with smaller notch root radii. While this study provides the initial notch responses of the L-PBF Inconel 718, further research must be completed in regard to the impact of notches on more complex loading behaviors, such as fatigue and stress-rupture conditions.

Purpose Chronic pelvic pain (CPP) in women is often associated with marked emotional distress and disability, with particular impairments in sexual functioning. Research supports the efficacy of interdisciplinary chronic pain rehabilitation programs (ICPRPs) in treating chronic pain, however less is known about their utility in CPP. Methods This retrospective study examined pain-related sexual impairment, emotional symptoms, and pain severity in CPP patients before and after completing a 3–4 week ICPRP. Predictors of post-treatment sexual impairment were also investigated. Participants included 58 female CPP patients and 58 age-matched females with non-pelvic chronic pain (NPCP). Results All participants reported robust improvements across outcome measures. Women with CPP reported greater pre- and post-treatment impairment in sexual function than NPCP patients, despite significant treatment-related improvements. In contrast, CPP patients also reported higher levels of depression at baseline but showed greater treatment related-improvements. In participants with CPP, treatment-related improvements in depression, alexithymia, and pain severity significantly explained decreases in pain-related sexual impairment following treatment, whereas none of these variables explained sexual impairment outcomes in women with NPCP. Conclusion Results demonstrate that ICPRPs can effectively treat CPP, particularly through changes in depression and alexithymia. Future research should examine whether specific interventions can be added in ICPRPS to address CPP-related sexual impairment.

Many cancer cell lines are aneuploid and heterogeneous, with multiple karyotypes co-existing within the same cell line. Karyotype heterogeneity has been shown to manifest phenotypically, thus affecting how cells respond to drugs or to minor differences in culture media. Knowing how to interpret karyotype heterogeneity phenotypically would give insights into cellular phenotypes before they unfold temporally. Here, we re-analyzed single cell RNA (scRNA) and scDNA sequencing data from eight stomach cancer cell lines by placing gene expression programs into a phenotypic context. Using live cell imaging, we quantified differences in the growth rate and contact inhibition between the eight cell lines and used these differences to prioritize the transcriptomic biomarkers of the growth rate and carrying capacity. Using these biomarkers, we found significant differences in the predicted growth rate or carrying capacity between multiple karyotypes detected within the same cell line. We used these predictions to simulate how the clonal composition of a cell line would change depending on density conditions during in-vitro experiments. Once validated, these models can aid in the design of experiments that steer evolution with density-dependent selection.

Abstract Growing tumors are dynamic and nonlinear ecosystems, wherein cancer cells adapt to their local microenvironment, and these adaptations further modify the environment, inducing more changes. From nascent intraductal neoplasms to disseminated metastatic disease, several levels of evolutionary adaptations and selections occur. Here, we focus on one example of such an adaptation mechanism, namely, “niche construction” promoted by adaptation to acidosis, which is a metabolic adaptation to the early harsh environment in intraductal neoplasms. The avascular characteristics of ductal carcinoma in situ (DCIS) make the periluminal volume profoundly acidic, and cancer cells must adapt to this to survive. Based on discovery proteomics, we hypothesized that a component of acid adaptation involves production of collagen by pre‐cancer cells that remodels the extracellular matrix (ECM) and stabilizes cells under acid stress. The proteomic data were surprising as collagen production and deposition are commonly believed to be the responsibility of mesenchymally derived fibroblasts, and not cells of epithelial origin. Subsequent experiments in 3D culture, spinning disk and second harmonic generation microscopy of DCIS lesions in patients’ samples are concordant. Collagen production assay by acid‐adapted cells in vitro demonstrated that the mechanism of induction involves the RAS and SMAD pathways. Secretome analyses show upregulation of ECM remodeling enzymes such as TGM2 and LOXL2 that are collagen crosslinkers. These data strongly indicate that acidosis in incipient cancers induces collagen production by cancer cells and support the hypothesis that this adaptation initiates a tumor‐permissive microenvironment promoting survival and growth of nascent cancers.

Abstract The Socioscientific Issue (SSI) framework was applied to explore how a team of two teachers navigated SSI cases as students in a STEM education graduate program. Using a case study approach, we found a connection between the specific SSI case and levels of scientific argumentation, as well as teachers’ plans for teaching. Our findings suggest successes and challenges for our teachers in incorporating scientific argumentation through SSI cases into his/her intended classroom practices. Specifically, our participating teachers displayed notably higher levels of claims, reasoning, and ability to question the sources of information, but lower levels of evidence and rebuttals. They had difficulty in connecting claims to evidence and reasoning in discussing the SSI cases. Moreover, their intended classroom implementation considered how to connect the lessons to their students’ lives, build scientific knowledge, and provide meaningful context to engage their students in the study of SSI.

Abstract Friedreich ataxia is an autosomal recessive, neurodegenerative disease characterized by the deficiency of the iron‐sulfur cluster assembly protein frataxin. Loss of this protein impairs mitochondrial function. Mitochondria alter their morphology in response to various stresses; however, such alterations to morphology may be homeostatic or maladaptive depending upon the tissue and disease state. Numerous neurodegenerative diseases exhibit excessive mitochondrial fragmentation, and reversing this phenotype improves bioenergetics for diseases in which mitochondrial dysfunction is a secondary feature of the disease. This paper demonstrates that frataxin deficiency causes excessive mitochondrial fragmentation that is dependent upon Drp1 activity in Friedreich ataxia cellular models. Drp1 inhibition by the small peptide TAT‐P110 reverses mitochondrial fragmentation but also decreases ATP levels in frataxin‐knockdown fibroblasts and FRDA patient fibroblasts, suggesting that fragmentation may provide a homeostatic pathway for maintaining cellular ATP levels. The cardiolipin‐stabilizing compound SS‐31 similarly reverses fragmentation through a Drp1‐dependent mechanism, but it does not affect ATP levels. The combination of TAT‐P110 and SS‐31 does not affect FRDA patient fibroblasts differently from SS‐31 alone, suggesting that the two drugs act through the same pathway but differ in their ability to alter mitochondrial homeostasis. In approaching potential therapeutic strategies for FRDA, an important criterion for compounds that improve bioenergetics should be to do so without impairing the homeostatic response of mitochondrial fragmentation.

Early ducts of breast tumors are unequivocally acidic. High rates of glycolysis combined with poor perfusion lead to a congestion of acidic metabolites in the tumor microenvironment, and pre-malignant cells must adapt to this acidosis to thrive. Adaptation to acidosis selects cancer cells that can thrive in harsh conditions and are capable of outgrowing the normal or non-adapted neighbors. This selection is usually accompanied by phenotypic change. Epithelial mesenchymal transition (EMT) is one of the most important switches correlated to malignant tumor cell phenotype and has been shown to be induced by tumor acidosis. New evidence shows that the EMT switch is not a binary system and occurs on a spectrum of transition states. During confirmation of the EMT phenotype, our results demonstrated a partial EMT phenotype in our acid-adapted cell population. Using RNA sequencing and network analysis we found 10 dysregulated network motifs in acid-adapted breast cancer cells playing a role in EMT. Our further integrative analysis of RNA sequencing and SILAC proteomics resulted in recognition of S100B and S100A6 proteins at both the RNA and protein level. Higher expression of S100B and S100A6 was validated in vitro by Immunocytochemistry. We further validated our finding both in vitro and in patients' samples by IHC analysis of Tissue Microarray (TMA). Correlation analysis of S100A6 and LAMP2b as marker of acidosis in each patient from Moffitt TMA approved the acid related role of S100A6 in breast cancer patients. Also, DCIS patients with higher expression of S100A6 showed lower survival compared to lower expression. We propose essential roles of acid adaptation in cancer cells EMT process through S100 proteins such as S100A6 that can be used as therapeutic strategy targeting both acid-adapted and malignant phenotypes.

Abstract Background Forty years ago the actin cytoskeleton was determined to be disrupted in fibroblasts from persons with DNA repair-defective, hereditary colon cancer, with no clear connection between the cytoskeleton and DNA repair defects at that time. Recently, the large number of sequenced genomes has indicated that mammalian mutagenesis has a large stochastic component. As a result, large coding regions are large mutagen targets. Cytoskeletal protein-related coding regions (CPCRs), including extra-cellular matrix proteins, are among the largest coding regions in the genome and are indeed very commonly mutated in cancer. Methods To determine whether mutagen sensitivity of the actin cytoskeleton could be assessed experimentally, we treated tissue culture cells with 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and quantified overall cytoskeleton integrity with rhodamine-phalloidin stains for F-actin. Results The above approach indicated cytoskeletal degradation with increasing mutagen exposure, consistent with increased mutagenesis of CPCRs in TCGA, smoker samples, where overall mutation rates correlate with CPCR mutation rates (R2 = 0.8694; p

Teacher candidates are called on to create effective lesson plans to implement during their student teaching semester, and certain components of lesson plans are recognized as particularly significant. Viewing lesson plans as the foundation for impactful instruction, it is necessary for teacher candidates to have skills in evaluating their plans before implementing them in the classroom. This article seeks to provide a set of questions teacher candidates may use to self-evaluate their lesson plans based on three major methods to assess lesson plans.

Category: Trauma Introduction/Purpose: Pilon and unstable ankle fractures are often treated initially with an external fixator (ex-fix) due to soft tissue concerns. Typically, the second stage involves open reduction internal fixation of the fracture as the definitive management. Ex-fix application in the emergency department (ED) has been described previously. We aimed to determine if an ex-fix applied in the ED would have any difference in rate of ex-fix revision, deep infection, and hospital length of stay when compared to an ex-fix that is applied in the operating room (OR). Methods: This is a retroscpective, case-cohort study performed at an urban, level-1 trauma center. Using CPT codes in our billing database, we identified all patients from 2011-2015 who had operative fixation of pilon (AO classification 43B or 43C) or unstable ankle fractures (AO classification 44C) and had application of an ex-fix (either ED or OR) prior to definitive fixation. Patients with open injury, those placed in ex-fix at outside institutions and those treated definitively with ex-fix were excluded. Results: Ninety-six patients met the inclusion criteria. Average age was 47 years and 54 (56%) were male. Thirty-three patients had ex-fix placed in the OR and 63 patients had ex-fix placed in the ED. Post-surgical complications (prominent implant, nonunion, deep infection, DVT, loss of reduction) were seen in 6/33 in the ED and 8/63 in the OR group (χ 2 =0.44, p=0.51). Deep infections occurred in 2/33 OR ex-fix and 5/63 ED ex-fix patients (χ 2 =0.137, p=0.71, OR= 1.4 [95% CI = 0.25 to 7.5]). Revision ex-fix for loss of reduction was performed 4/33 in the OR group and 10/63 in the ED ex-fix group (χ 2 =0.296, p=0.59). Mean length of stay was 14 days for the OR and 13 days for the ED group (p=0.35). Conclusion: We found no significant difference in post-surgical complications (surgical or infectious) or ex-fix revision rates for an ex-fix placed in ED compared to one placed in the OR. Total hospital days were similar in both groups. Prospective studies are needed to validate this method of provisional stabilization in the ED; however, these results indicate that the ED ex-fix is equally efficacious to an OR ex-fix.

The recent clinical and commercial success of lipid nanoparticles (LNPs) for nucleic acid delivery has incentivized the development of new technologies to manufacture LNPs. As new technologies emerge, researchers must determine which technologies to assess and how to perform comparative evaluations. In this article, we use a quality-by-design approach to systematically investigate how the mixer technology used to form LNPs influences LNPstructure. Specifically, a coaxial turbulent jet mixer and a staggered herringbone microfluidic mixer were systematically compared via matched formulation and process conditions. A full-factorial design-of-experiments study with three factors and three levels was executed for each mixer to compare process robustness in the production of antisense oligonucleotide (ASO) LNPs. ASO-LNPs generated with the coaxial turbulent jet mixer were consistently smaller, had a narrower particle size distribution, and had a higher ASO encapsulation as compared to the microfluidic mixer, but had a greater variation in internal structure with less ordered cores. A subset of the study was replicated for mRNA-LNPs with comparable trends in particle size and encapsulation, but more frequent bleb features for LNPs produced by the coaxial turbulent jet mixer. The study design used here provides a road map for how researchers may compare different mixer technologies (or process changes more broadly) and how such studies can inform process robustness and manufacturing control strategies.

This study explores the experiences of two teachers participating in professional development workshops focused on supporting implementation of SocioScientific Issues (SSI) and aspects of social justice into STEM classrooms. SSI are ill-defined problems, with a basis in science, but necessarily include moral and ethical decisions that cannot be resolved through science alone. These debatable issues can enhance learning of STEM by engaging students in real-world and authentic problems. The "USTRIVE project" was developed to foster STEM learning through integrated professional development workshops and the development of professional learning communities to support teachers in the use of SSI and incorporation of aspects of social justice in their STEM classrooms. Two research questions were investigated: (a) To what extent did teachers implement SSI into their lesson planning during the project and (b) In what ways did teachers' designed lessons change from the beginning of the workshop?

Introduction: Birth weight and placental weight are good indicators of new born growth parameters. Maternal complications are seen in both lean and obese women. The available studies shows correlations of placental parameters with baby anthropometric variables but limited relationship to maternal BMI which will help very much to anticipate the neonatal outcome and complications. Aim: To find out correlation of maternal BMI and neonatal BMI with placental weight in rural South Kerala. Study Design: Prospective cohort study in a rural medical college in South Kerala. Materials and Methods: All singleton neonates 28 weeks of gestational age and above delivered in the institution and their mothers from January 2015 to October 2015. Neonatal BMI, Maternal BMI and placental weight were calculated. Means (x), standard deviation (SD) and frequency (%) was calculated for the statistical analysis. Student’s ‘t’ test was used to compare the mean results of the continuous variables. Pearson correlation was used to find out the correlation between different anthropometric variables. Results: Out of the 920 live single births 463 (50.3%) were male and 457(49.7%) were females. Among the mothers (86.2%) were between the age group of 25-29 years, about 87.7% of the mothers were unemployed. Mean birth weight was 3000gm (SD=500) Mean baby length was 48.9 (SD=2). The mean placental weight was 499gm (SD=78). Mean maternal BMI was 27 (SD 3.9) and the mean neonatal BMI was 12.5 (SD=1.9). It was found that both maternal BMI (r=0.143 p

Abberent protein-protein interactions potentiate many diseases and one example is the toxic, self-assembly of α-Synuclein in the dopaminergic neurons of patients with Parkinson’s disease; therefore, a potential therapeutic strategy is the small molecule modulation of α-Synuclein aggregation. In this work, we develop an Oligopyridylamide based 2-dimensional FragmentAssisted Structure-based Technique to identify antagonists of α-Synuclein aggregation. The technique utilizes a fragment-based screening of an extensive array of non-proteinogenic side chains in Oligopyridylamides, leading to the identification of NS132 as an antagonist of the multiple facets of αSynuclein aggregation. We further identify a more cell permeable analog (NS163) without sacrificing activity. Oligopyridylamides rescue α-Synuclein aggregation mediated Parkinson’s disease phenotypes in dopaminergic neurons in early and post disease Caenorhabditis elegans models. We forsee tremendous potential in our technique to identify lead therapeutics for Parkinson’s disease and other diseases as it is expandable to other oligoamide scaffolds and a larger array of side chains. [ABSTRACT FROM AUTHOR]