Monitoring developmental force distributions in reconstituted embryonic epithelia.

The way cells are organized within a tissue dictates how they sense and respond to extracellular signals, as cues are received and interpreted based on expression and organization of receptors, downstream signaling proteins, and transcription factors. Part of this microenvironmental context is the result of forces acting on the cell, including forces from other cells or from the cellular substrate or basement membrane. However, measuring forces exerted on and by cells is difficult, particularly in an in vivo context, and interpreting how forces affect downstream cellular processes poses an even greater challenge. Here, we present a simple method for monitoring and analyzing forces generated from cell collectives. We demonstrate the ability to generate traction force data from human embryonic stem cells grown in large organized epithelial sheets to determine the magnitude and organization of cell-ECM and cell-cell forces within a self-renewing colony. We show that this method can be used to measure forces in a dynamic hESC system and demonstrate the ability to map intracolony protein localization to force organization

Reduction of frontal plane knee load caused by lateral trunk lean depends on step width

The internal knee abduction moment (KAM) in osteoarthritis is reduced by increased lateral trunk lean (TL). Mechanistically, this occurs as the Centre of Mass (COM) moves further over the stance leg. Since the size of the base of support constrains the COM, an associated increase in step width (SW) would be expected to maintain stability. This study tested the effects of TL on SW and KAM in healthy participants (n = 21) who performed normal and 6° TL walks. The latter was controlled via audio-visual biofeedback. We found two distinct gait strategies in TL walk: widening the step width substantially (>50%) to permit an increase in the COM displacement (WSW, n = 13), or maintaining a baseline SW and minimally displacing the COM by moving the hip/pelvic complex in the opposite direction (NSW, n = 8). WSW doubled SW (11.3 ± 2.4 v. 24.7 ± 5.5 cm, p  0.05). These two distinct gait strategies resulted in unique patterns of KAM reduction across the stance phase. NSW reduced KAM impulse significantly in the initial half (0.08 ± 0.02 v. 0.06 ± 0.02, p = .04) but not in the later stance phase (0.07 ± 0.02 v. 0.07 ± 0.04, p > 0.05). WSW reduced KAM significantly in both initial (0.11 ± 0.03 v. 0.08 ± 0.04, p < 0.001) and later stance phase (0.09 ± 0.02 v. 0.06 ± 0.03, p < 0.001). KAM peak results followed the pattern of impulse. This study has revealed two distinct mechanisms for increasing lateral trunk lean which can be used to explain discrepancies in past research and in the future could be used to individualise gait re-training strategies

Integrin-mediated traction force enhances paxillin molecular associations and adhesion dynamics that increase the invasiveness of tumor cells into a three-dimensional extracellular matrix.

Metastasis requires tumor cells to navigate through a stiff stroma and squeeze through confined microenvironments. Whether tumors exploit unique biophysical properties to metastasize remains unclear. Data show that invading mammary tumor cells, when cultured in a stiffened three-dimensional extracellular matrix that recapitulates the primary tumor stroma, adopt a basal-like phenotype. Metastatic tumor cells and basal-like tumor cells exert higher integrin-mediated traction forces at the bulk and molecular levels, consistent with a motor-clutch model in which motors and clutches are both increased. Basal-like nonmalignant mammary epithelial cells also display an altered integrin adhesion molecular organization at the nanoscale and recruit a suite of paxillin-associated proteins implicated in invasion and metastasis. Phosphorylation of paxillin by Src family kinases, which regulates adhesion turnover, is similarly enhanced in the metastatic and basal-like tumor cells, fostered by a stiff matrix, and critical for tumor cell invasion in our assays. Bioinformatics reveals an unappreciated relationship between Src kinases, paxillin, and survival of breast cancer patients. Thus adoption of the basal-like adhesion phenotype may favor the recruitment of molecules that facilitate tumor metastasis to integrin-based adhesions. Analysis of the physical properties of tumor cells and integrin adhesion composition in biopsies may be predictive of patient outcome

THE OAK ORCHARD SOIL WATER ASSESSMENT TOOL A decision support system for watershed management Part 1: Calibration and Validation

A hydrologic model (SWAT) was developed and calibrated for the Oak Orchard watershed to evaluate sources and sinks of sediment and nutrients. The model included the most important anthropogenic features that impacted water flow and nonpoint source pollution in the watershed. These features included reservoirs at the Iroquois National Wildlife Refuge, Waterport and Medina; point sources such as the Erie Canal, US Gypsum, Allen Canning, wastewater treatment plants at Medina, Oakfield and Elba, and tiledrains at the mucklands, an intensely farmed area that was drained to combat malaria in the 19th century. The model included point sources for every subbasin so that the effects of future point sources can be evaluated. The model was calibrated for waterflow and sediment using observed loading data collected by Makarewicz and Lewis (2000, 2009). To achieve the proper water balance observed at the watershed, seasonal inputs of water had to be added from the Erie Canal and the Onondaga escarpment. This water came from outside of the watershed. The resulting calibration had a Nash-Sutcliffe (NS) prediction efficiency of 0.81 for the calibration period (1997-1999). The total cumulative sediment loading was within 2%, of observed and the monthly sediment loads fell within the uncertainty of the observed data (NS=0.31). Cumulative total phosphorous loads were within 2% of observed and the NS prediction efficiency was 0.91. The model validated very poorly in the 2008 time period primarily because of inaccurate precipitation data and incorrect groundwater fluxes from the escarpment. Further research needs to evaluate the timing and amount of groundwater flow from the escarpment because it has a significant impact on monthly flows in this watershed. It is likely that other watersheds that are nestled against the Onondaga escarpment are impacted by spring flows from this geologic feature

Diminished salivary epidermal growth factor secretion : a link between Sjogren's syndrome and autoimmune gastritis?

Objectives: Healthy human labial salivary glands produce epidermal growth factor (EGF). In Sjogren's syndrome (SS), EGF staining is diminished. SS is also associated with chronic autoimmune corpus gastritis. We therefore hypothesized that EGF secretion would be diminished in SS and that this could affect gastric target cells.Methods: Salivary EGF secretion in SS was compared to that in healthy controls using an enzyme-linked immunosorbent assay (ELISA). EGF receptor (EGFR) immunoreactive cells in the gastric corpus of healthy human subjects were analysed using immunostaining.Results: Salivary secretion of EGF was diminished in SS patients (232.4, range 52.6-618.4, vs. 756.6, range 105.3-1631.6 pg/min, p=0.002). Proton-pump positive parietal cells were mostly EGFR immunoreactive whereas very few pepsinogen I (PGI)-positive cells were EGFR positive.Conclusions: As EGF is relatively acid resistant, salivary gland-derived EGF might participate in an exo/endocrine mode of parietal cell maintenance in the gastric corpus. Deficiency of salivary gland-derived EGF in SS patients may cause impairment of gastric parietal cells resulting in exposure of immunogenic cryptic antigens and loss of immunological self-tolerance.Peer reviewe

Diminished salivary epidermal growth factor secretion : a link between Sjogren's syndrome and autoimmune gastritis?

Objectives: Healthy human labial salivary glands produce epidermal growth factor (EGF). In Sjogren's syndrome (SS), EGF staining is diminished. SS is also associated with chronic autoimmune corpus gastritis. We therefore hypothesized that EGF secretion would be diminished in SS and that this could affect gastric target cells.Methods: Salivary EGF secretion in SS was compared to that in healthy controls using an enzyme-linked immunosorbent assay (ELISA). EGF receptor (EGFR) immunoreactive cells in the gastric corpus of healthy human subjects were analysed using immunostaining.Results: Salivary secretion of EGF was diminished in SS patients (232.4, range 52.6-618.4, vs. 756.6, range 105.3-1631.6 pg/min, p=0.002). Proton-pump positive parietal cells were mostly EGFR immunoreactive whereas very few pepsinogen I (PGI)-positive cells were EGFR positive.Conclusions: As EGF is relatively acid resistant, salivary gland-derived EGF might participate in an exo/endocrine mode of parietal cell maintenance in the gastric corpus. Deficiency of salivary gland-derived EGF in SS patients may cause impairment of gastric parietal cells resulting in exposure of immunogenic cryptic antigens and loss of immunological self-tolerance.Peer reviewe

Exclusive neuronal expression of SUCLA2 in the human brain

SUCLA2 encodes the ATP-forming subunit (A-SUCL-) of succinyl-CoA ligase, an enzyme of the citric acid cycle. Mutations in SUCLA2 lead to a mitochondrial disorder manifesting as encephalomyopathy with dystonia, deafness and lesions in the basal ganglia. Despite the distinct brain pathology associated with SUCLA2 mutations, the precise localization of SUCLA2 protein has never been investigated. Here we show that immunoreactivity of A-SUCL- in surgical human cortical tissue samples was present exclusively in neurons, identified by their morphology and visualized by double labeling with a fluorescent Nissl dye. A-SUCL- immunoreactivity co-localized >99% with that of the d subunit of the mitochondrial F0-F1 ATP synthase. Specificity of the anti-A-SUCL- antiserum was verified by the absence of labeling in fibroblasts from a patient with a complete deletion of SUCLA2. A-SUCL- immunoreactivity was absent in glial cells, identified by antibodies directed against the glial markers GFAP and S100. Furthermore, in situ hybridization histochemistry demonstrated that SUCLA2 mRNA was present in Nissl-labeled neurons but not glial cells labeled with S100. Immunoreactivity of the GTP-forming subunit (G-SUCL-) encoded by SUCLG2, or in situ hybridization histochemistry for SUCLG2 mRNA could not be demonstrated in either neurons or astrocytes. Western blotting of post mortem brain samples revealed minor G-SUCL- immunoreactivity that was however, not upregulated in samples obtained from diabetic versus non-diabetic patients, as has been described for murine brain. Our work establishes that SUCLA2 is expressed exclusively in neurons in the human cerebral cortex

Microbes in beach sands : integrating environment, ecology and public health

Author Posting. © The Author(s), 2014. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Reviews in Environmental Science and Bio/Technology 13 (2014): 329-368, doi:10.1007/s11157-014-9340-8.Beach sand is a habitat that supports many microbes, including viruses, bacteria, fungi and protozoa (micropsammon). The apparently inhospitable conditions of beach sand environments belie the thriving communities found there. Physical factors, such as water availability and protection from insolation; biological factors, such as competition, predation, and biofilm formation; and nutrient availability all contribute to the characteristics of the micropsammon. Sand microbial communities include autochthonous species/phylotypes indigenous to the environment. Allochthonous microbes, including fecal indicator bacteria (FIB) and waterborne pathogens, are deposited via waves, runoff, air, or animals. The fate of these microbes ranges from death, to transient persistence and/or replication, to establishment of thriving populations (naturalization) and integration in the autochthonous community. Transport of the micropsammon within the habitat occurs both horizontally across the beach, and vertically from the sand surface and ground water table, as well as at various scales including interstitial flow within sand pores, sediment transport for particle-associated microbes, and the large-scale processes of wave action and terrestrial runoff. The concept of beach sand as a microbial habitat and reservoir of FIB and pathogens has begun to influence our thinking about human health effects associated with sand exposure and recreational water use. A variety of pathogens have been reported from beach sands, and recent epidemiology studies have found some evidence of health risks associated with sand exposure. Persistent or replicating populations of FIB and enteric pathogens have consequences for watershed/beach management strategies and regulatory standards for safe beaches. This review summarizes our understanding of the community structure, ecology, fate, transport, and public health implications of microbes in beach sand. It concludes with recommendations for future work in this vastly under-studied area.2015-05-0

Early and Long-Term Results of Unprotected Left Main Coronary Artery Stenting The LE MANS (Left Main Coronary Artery Stenting) Registry

ObjectivesThe aim of the study was to evaluate early and late outcomes after percutaneous coronary intervention (PCI) of unprotected left main coronary artery disease (ULMCA) and to compare bare-metal stent (BMS) and drug-eluting stent (DES) subgroups.BackgroundPCI is an increasingly utilized method of revascularization in patients with ULMCA.MethodsThis multicenter prospective registry included 252 patients after ULMCA stenting enrolled between March 1997 and February 2008. Non–ST-segment elevation acute coronary syndrome was diagnosed in 58% of patients; ST-segment elevation myocardial infarction cases were excluded. Drug-eluting stents were implanted in 36.2% of patients.ResultsMajor adverse cardiovascular and cerebral events (MACCE) occurred in 12 (4.8%) patients during the 30-day period, which included 4 (1.5%) deaths. After 12 months there were 17 (12.1%) angiographically confirmed cases of restenosis. During long-term follow-up (1 to 11 years, mean 3.8 years) there were 64 (25.4%) MACCE and 35 (13.9%) deaths. The 5- and 10-year survival rates were 78.1% and 68.9%, respectively. Despite differences in demographical and clinical data in favor of BMS patients, unmatched analysis showed a significantly lower MACCE rate in DES patients (25.9% vs. 14.9%, p = 0.039). This difference was strengthened after propensity score matching. The DES lowered both mortality and MACCE for distal ULMCA lesions when compared with BMS. Ejection fraction <50% was the only independent risk factor influencing long-term survival.ConclusionsStenting of ULMCA is feasible and offers good long-term outcome. Implantation of DES for ULMCA decreased the risk of long-term MACCE, and particularly improved survival in patients with distal ULMCA disease

Measuring Cosmic Rays with the RadMap Telescope on the International Space Station

The RadMap Telescope is a new radiation-monitoring instrument operating in the U.S. Orbital Segment (USOS) of the International Space Station (ISS). The instrument was commissioned in May 2023 and will rotate through four locations inside American, European, and Japanese modules over a period of about six months. In some locations, it will take data alongside operational, validated detectors for a cross-check of measurements. RadMap’s central detector is a finely segmented tracking calorimeter that records detailed depth-dose data relevant to studies of the radiation exposure of the ISS crew. It is also able to record particle-dependent energy spectra of cosmic-ray nuclei with energies up to several hundred MeV per nucleon. A unique feature of the detector is its ability to track nuclei with omnidirectional sensitivity at an angular resolution of two degrees. In this contribution, we present the design and capabilities of the RadMap Telescope and give an overview of the instrument’s commissioning on the ISS