The murine nonclassical class I major histocompatibility complex-like CD1.1 molecule protects target cells from lymphokine-activated killer cell cytolysis

Classical class I major histocompatibility complex (MHC) molecules, as well as the nonclassical class I histocompatibility leukocyte antigen (HLA)-E molecule, can negatively regulate natural killer (NK) cell cytotoxicity through engagement of NK inhibitory receptors. We show that expression of murine (m)CD1.1, a nonpolymorphic nonclassical MHC class I-like molecule encoded outside the MHC, protects NK-sensitive RMA/S target cells from adherent lymphokine-activated killer cell (A-LAK) cytotoxicity. Passage of effector cells in recombinant interleukin (rIL)-2 enhanced protection by mCD1.1, suggesting an expansion of relevant A-LAK population(s) or modulation of A-LAK receptor expression. Murine CD1. 1 conferred protection from lysis by rIL-2-activated spleen cells of recombination activating gene (Rag)-1(-/-) mice, which lack B and T cells, demonstrating that mCD1.1 can protect RMA/S cells from lysis by NK cells. An antibody specific for mCD1.1 partially restored A-LAK lysis of RMA/S.CD1.1 transfectants, indicating that cell surface mCD1.1 can confer protection from lysis; therefore, mCD1.1 possibly acts through interaction with an NK inhibitory receptor. CD1.1 is by far the most divergent class I molecule capable of regulating NK cell activity. Finally, mCD1.1 expression rendered RMA/S cells resistant to lysis by A-LAK of multiple mouse strains. The conserved structure of mCD1.1 and pattern of mCD1.1 resistance from A-LAK lysis suggest that mCD1.1 may be a ligand for a conserved NK inhibitory receptor.     

Exploring the circularity potential regarding the multiple use of residual material

Clean Technologies and Environmental Policy - Building on recent insights in exploring material circularity, this study attempts to develop a methodological approach toward allocating an available...     

Cell Force-Driven Basement Membrane Disruption Fuels EGF- and Stiffness-Induced Invasive Cell Dissemination from Benign Breast Gland Acini

Local basement membrane (BM) disruption marks the initial step of breast cancer invasion. The activation mechanisms of force-driven BM-weakening remain elusive. We studied the mechanical response of MCF10A-derived human breast cell acini with BMs of tuneable maturation to physical and soluble tumour-like extracellular matrix (ECM) cues. Traction force microscopy (TFM) and elastic resonator interference stress microscopy (ERISM) were used to quantify pro-invasive BM stress and protrusive forces. Substrate stiffening and mechanically impaired BM scaffolds induced the invasive transition of benign acini synergistically. Robust BM scaffolds attenuated this invasive response. Additional oncogenic EGFR activation compromised the BMs’ barrier function, fuelling invasion speed and incidence. Mechanistically, EGFR-PI3-Kinase downstream signalling modulated both MMP- and force-driven BM-weakening processes. We show that breast acini form non-proteolytic and BM-piercing filopodia for continuous matrix mechanosensation, which significantly push and pull on the BM and ECM under pro-invasive conditions. Invasion-triggered acini further shear and compress their BM by contractility-based stresses that were significantly increased (3.7-fold) compared to non-invasive conditions. Overall, the highest amplitudes of protrusive and contractile forces accompanied the highest invasiveness. This work provides a mechanistic concept for tumour ECM-induced mechanically misbalanced breast glands fuelling force-driven BM disruption. Finally, this could facilitate early cell dissemination from pre-invasive lesions to metastasize eventually.     

Firm relocations in the Netherlands: Why do firms move,and where do they go?*

This study analyses determinants of business relocation and identifies regional characteristics which attract relocating firms. Results indicate that the relocation decisions of firms are sector‐dependent, and the migratory behaviour of firms in knowledge‐intensive sectors notably differs from that in less knowledge‐intensive sectors. Predominantly low‐tech and medium‐low‐tech manufacturing and less knowledge‐intensive service firms paying high average salaries have a higher probability to move out of their present location. For less knowledge‐intensive service firms, the average municipal wage negatively affects their propensity to relocate, while those located in municipalities with high sector‐specific wages have an increased likelihood to move. Relocating firms are generally attracted by densely populated municipalities with high wage levels, and primarily service firms are drawn to municipalities which are specialized in the firm's own sector and appeal to individuals. Sector‐specific wages may either attract, or deter firms, suggesting that this variable may capture both the cost and the quality of the locally available workforce.     

Influence of the Peripheral Nervous System on Murine Osteoporotic Fracture Healing and Fracture-Induced Hyperalgesia

Osteoporotic fractures are often linked to persisting chronic pain and poor healing outcomes. Substance P (SP), α-calcitonin gene-related peptide (α-CGRP) and sympathetic neurotransmitters are involved in bone remodeling after trauma and nociceptive processes, e.g., fracture-induced hyperalgesia. We aimed to link sensory and sympathetic signaling to fracture healing and fracture-induced hyperalgesia under osteoporotic conditions. Externally stabilized femoral fractures were set 28 days after OVX in wild type (WT), α-CGRP- deficient (α-CGRP −/−), SP-deficient (Tac1−/−) and sympathectomized (SYX) mice. Functional MRI (fMRI) was performed two days before and five and 21 days post fracture, followed by µCT and biomechanical tests. Sympathectomy affected structural bone properties in the fracture callus whereas loss of sensory neurotransmitters affected trabecular structures in contralateral, non-fractured bones. Biomechanical properties were mostly similar in all groups. Both nociceptive and resting-state (RS) fMRI revealed significant baseline differences in functional connectivity (FC) between WT and neurotransmitter-deficient mice. The fracture-induced hyperalgesia modulated central nociception and had robust impact on RS FC in all groups. The changes demonstrated in RS FC in fMRI might potentially be used as a bone traumata-induced biomarker regarding fracture healing under pathophysiological musculoskeletal conditions. The findings are of clinical importance and relevance as they advance our understanding of pain during osteoporotic fracture healing and provide a potential imaging biomarker for fracture-related hyperalgesia and its temporal development. Overall, this may help to reduce the development of chronic pain after fracture thereby improving the treatment of osteoporotic fractures.     

Conditionally immortalized murine bone marrow stromal cells mediate parathyroid hormone-dependent osteoclastogenesis in vitro

PTH recruits and activates osteoclasts to cause bone resorption. These actions of PTH are thought to be mediated indirectly via type 1 PTH/PTH-related peptide receptors (PTH1Rs) expressed by adjacent marrow stromal or osteoblastic cells, although some evidence suggests that PTH may act directly on early hematopoietic osteoclast progenitors. We have established clonal, conditionally immortalized, PTH-responsive, bone marrow stromal cell lines from mice that harbor both a transgene encoding a temperature-sensitive mutant of the simian virus 40 large T antigen and deletion of a single allele of the PTH1R gene. Of 60 stromal cell lines isolated, 45 expressed functional PTH1Rs. During coculture with normal murine spleen cells, 5 of 42 such cell lines could support formation of tartrate-resistant acid phosphatase-positive, multinucleated cells (TRAP+ MNCs) in response to 1,25-dihydroxyvitamin D3, but only 2 of these did so in response to PTH. One of these, MS1 cells, expressed numerous cytokines and proteins characteristic of the osteogenic lineage and showed increased production of interleukin-6 in response to PTH. MS1 cells supported dose-dependent induction by rat (r) PTH-(1-34) (0.1-100 nM) of TRAP+ MNCs that expressed calcitonin receptors and formed resorption lacunae on dentine slices. This effect of PTH, which required cell to cell contact between MS1 and spleen cells, was mimicked by coadministration of cAMP analog and phorbol ester but only partially by either agent alone. The carboxyl-terminal fragment rPTH-(53-84) also induced osteoclast-like cell formation, but the maximal effect was only 30% as great as that of rPTH-(1-34). Importantly, rPTH-(1-34) induced TRAP+ MNC formation even when PTH1R-/- osteoclast progenitors (from fetal liver of mice homozygous for ablation of the PTH1R gene) were cocultured with MS1 cells. We conclude that activation of PTH1Rs on cells of the osteoclast lineage is not required for PTH-(1-34)-induced osteoclast formation in the presence of appropriate PTH-responsive marrow stromal cells. MS1 cells provide a useful model for further study of PTH regulation of osteoclastogenesis.     

Generation of intestinal mucosal lymphocytes in SCID mice reconstituted with mature, thymus-derived T cells

Transfer of peripheral lymph node lymphocytes to SCID mice leads to the long term establishment of mucosal T lymphocytes within the epithelium and lamina propria of the small and large intestines. Analysis of engrafted intraepithelial lymphocytes (IEL) showed that they had acquired a surface phenotype that in several respects is typical of IEL. In addition, the functional profile of engrafted IEL derived from lymph node T cells was similar to that of normal IEL; as the donor-derived T cells exhibited a strong cytolytic activity, a poor proliferative response to mitogenic stimuli, and a tendency to home and expand specifically in the intestine upon transfer to secondary SCID recipients. Optimal engraftment of intestinal T cells required bacterial flora, as the number of lymphocytes was greatly reduced in SCID recipients with a reduced flora. These results demonstrate that mature, thymus-derived T cells can migrate to the intestine and become functionally specialized to the intestinal milieu. The acquisition of phenotypic markers characteristic of the intestinal microenvironment by engrafted cells suggests that T cell migration of lymphocytes to the SCID intestine is not aberrant, but it may reflect processes that are ongoing in immunocompetent mice. Furthermore, these data suggest that the homing and/or expansion of typical, thymus-derived T cells in the intestine may be driven by luminal Ags such as those derived from bacterial flora.     

Efficacy of FIT Produce Wash and Chlorine Dioxide on Pathogen Control in Fresh Potatoes

ABSTRACT:  A commercial fresh pack potato operation was used as a model to evaluate FIT fruit and vegetable wash effectiveness in reducing levels of microorganisms on potatoes and in flume water. Fresh potatoes were washed in flume water with or without FIT, or treated with a spray bar utilizing either FIT, 9 ppm chlorine dioxide (ClO₂), or a water control. Both flume treatments were also evaluated for APC and Gram-negatives. There were no significant differences in reduction of these microorganisms on treated or control potatoes. However, levels of Gram-negative bacteria in FIT-amended flume water were reduced by 5.95 log CFU/g, and the APC was reduced by 1.43 log CFU/g. To validate plant trial findings, this test was repeated using solutions of sterile potato flume water from the fresh pack operation, containing a typical level of dissolved and suspended solids. Treatment solutions prepared with flume water or deionized water containing FIT, 9 ppm ClO₂, or a water control were inoculated with E. coli O157:H7, Salmonella Typhimurium, or Pectobacterium carotovorum ssp. carotovorum . FIT and ClO₂ prepared with deionized water reduced levels of microorganisms by >6.1 to 6.6 log CFU/g to below the detection limit. FIT prepared with flume water reduced levels of all organisms by >6.0 to 6.4 log CFU/g to below the detection limit, whereas ClO₂ prepared from flume water reduced bacterial levels of all organisms by only 0.7 to 1.4 log CFU/g. Neither FIT nor ClO₂ was particularly efficacious against E. coli O157:H7, S. Typhimurium, APC, yeasts, or molds on potato surfaces.     

Postintubation arytenoid subluxation

Shunt infection is a major complication of shunt implantation, with Staphylococcus epidermidis found in almost 45% of infected shunts. This pathogen produces an extracellular slime that enables it to adhere to implantable devices and resist antibiotic therapy. Antimicrobial prophylaxis can prevent slime production. In this paper we report the results of a prospective study involving 100 shunt operations. The objective was to evaluate the efficacy of a single preoperative dose of ceftriaxone (2 g, i.v.) in preventing shunt infection. Ceftriaxone was chosen for its pharmacokinetic properties. No shunt infection was observed over a 4-year follow-up period. On the basis of these results we recommend prophylaxis with ceftriaxone as a safe and effective way of preventing shunt infection. In addition, one-shot prophylaxis with ceftriaxone is more cost-effective than multiple-dose antibiotic regimens.