INTRODUCTION:Burn injuries present a significant challenge for both military personnel and civilians, accounting for 5%-20% of military casualties and contributing to substantial medical costs and poor clinical outcomes. Medical technologies used in multidomain operations against near-peer adversaries are often heavy, difficult to use, immunogenic, and are not able to support prolonged casualty care in the field. To address these limitations, SiOxMed has developed a novel silica-based fiber (SBF) universal combat matrix with hemostatic and wound repair properties. These studies assess the effects of SBF on immune modulation, extracellular matrix (ECM) remodeling, and cellular proliferation in a porcine full thickness burn model. MATERIALS AND METHODS:Specific pathogen-free Yorkshire pigs were subjected to full-thickness burns using a modified branding iron. Twenty-four hours post-injury, wound beds were debrided before SBF matrix or standard gauze dressing treatment (n = 4 wounds per group). Scheduled bandage changes were followed by euthanasia and necropsy on day 25. Tissue biopsies were collected for histological and immunohistochemical analysis at days 4, 11, 18, and 25 to compare epithelialization, dermal collagen remodeling, and immune modulation among treatment groups. RESULTS:Histological evaluation using H&E (hematoxylin and eosin) and Masson's trichrome staining revealed enhanced rete ridge formation and improved collagen remodeling in SBF-treated wounds compared to control. Immunofluorescent staining for T-cell markers (CD4, CD8) and macrophage markers (CD68) demonstrated a reduced inflammatory response in the SBF group. Ki-67 staining indicated comparable cellular proliferation between SBF-treated and control wounds. CONCLUSIONS:Silica-based fiber treatment improved epidermal rete ridge formation, enhanced ECM collagen remodeling, and reduced inflammatory cell infiltration while maintaining normal cellular proliferation. These findings indicate that SBF not only minimizes inflammation compared to standard gauze treatment but also provides an optimal scaffold environment for tissue regeneration. Given its unique mechanism of action, ultralight weight, stability under ambient conditions, and user-friendly design, SBF shows strong potential for military and civilian applications, particularly in austere prehospital care and mass casualty scenarios.

Chemo-immunotherapy is central to the treatment of small cell lung cancer (SCLC). Despite modest progress made with the addition of immunotherapy, current cytotoxic regimens display minimal survival benefit and new treatments are needed. Thymidylate synthase (TS) is a well-validated anti-cancer drug target, but conventional TS inhibitors display limited clinical efficacy in refractory or recurrent SCLC. We performed RNA-Seq analysis to identify gene expression changes in SCLC biopsy samples to provide mechanistic insight into the potential utility of targeting pyrimidine biosynthesis to treat SCLC. We identified systematic dysregulation of pyrimidine biosynthesis, including elevated TYMS expression that likely contributes to the lack of efficacy for current TS inhibitors in SCLC. We also identified E2F1-3 upregulation in SCLC as a potential driver of TYMS expression that may contribute to tumor aggressiveness. To test if TS inhibition could be a viable strategy for SCLC treatment, we developed patient-derived organoids (PDOs) from human SCLC biopsy samples and used these to evaluate both conventional fluoropyrimidine drugs (e.g., 5-fluorouracil), platinum-based drugs, and CF10, a novel fluoropyrimidine polymer with enhanced TS inhibition activity. PDOs were relatively resistant to 5-FU and while moderately sensitive to the front-line agent cisplatin, were relatively more sensitive to CF10. Our studies demonstrate dysregulated pyrimidine biosynthesis contributes to drug resistance in SCLC and indicate that a novel approach to target these pathways may improve outcomes.

Glioblastoma multiforme (GBM) is the most aggressive glioma of the primary central nervous system. Due to the lack of effective treatment options, the prognosis for patients remains bleak. Fibroblast activation protein alpha (FAP), a 170 kDa type II transmembrane serine protease was observed to be expressed on glioma cells and within the glioma tumor microenvironment. To understand the utility of targeting FAP in this tumor type, the immuno-PET radiopharmaceutical [89Zr]Zr-Df-Bz-F19 mAb was prepared and Lindmo analysis was used for its in vitro evaluation using the U87MG cell line, which expresses FAP endogenously. Lindmo analysis revealed an association constant (Ka) of 10−8 M−1 and an immunoreactivity of 52%. Biodistribution studies in U87MG tumor-bearing mice revealed increasing radiotracer retention in tumors over time, leading to average tumor-to-muscle ratios of 3.1, 7.3, 7.2, and 8.3 at 2, 24, 48 and 72 h, respectively. Small animal PET corroborated the biodistribution studies; tumor-to-muscle ratios at 2, 24, 48, and 72 h were 2.0, 5.0, 6.1 and 7.8, respectively. Autoradiography demonstrated accumulated activity throughout the interior of FAP+ tumors, while sequential tumor sections stained positively for FAP expression. Conversely, FAP− tissues retained minimal radioactivity and were negative for FAP expression by immunohistochemistry. These results demonstrate FAP as a promising biomarker that may be exploited to diagnose and potentially treat GBM and other neuroepithelial cancers.

Bioprinting has become an essential tool of biomedical engineering and regenerative medicine. However, current biologically relevant bioinks, i.e. printing materials made from materials found within the extracellular matrix, are unable to support themselves and require a sacrificial support material to create large structures. To address this issue, we have developed a novel thixotropic bioink made from methacrylated collagen I, thiolated hyaluronic acid, and gelatin nanoparticles which is self-supporting and capable of creating large hollow structures through bioprinting. Once printed, this material can be photo-crosslinked to finalize the structure and can be immerse in media for culture with minimal swelling. An intersecting tubular structure 5 mm in diameter with a 1 mm wall thickness was able to be printed longitudinally without support material using a CellInk BIOX bioprinter. This bioink was also able to support HepG2 cells in bioprinted organoids, with adenosine triphosphate assays and immunohistochemistry showing proliferation. Responses to acetaminophen and troglitazone were as expected, proving that the new bioink does not significantly alter HepG2 drug response. With great mechanical properties as well as good biocompatibility, this new biologically relevant bioink is a great improvement over current bioinks.

You have accessJournal of UrologyStem Cell Research: Stem Cell Research II1 Apr 2018MP81-20 NON-INVASIVE, DUAL-LABELING TECHNOLOGY FOR CELL TRACKING IN STEM CELL THERAPY FOR ERECTILE DYSFUNCTION Ethan Matz, Lei Dou, Fangpeng Shu, Xin Gu, Ryan Terlecki, Jennifer Paxton, Ting Long, Lysette Mutkus, Frank Marini, James Yoo, Anthony Atala, John Jackson, and Yuanyuan Zhang Ethan MatzEthan Matz More articles by this author , Lei DouLei Dou More articles by this author , Fangpeng ShuFangpeng Shu More articles by this author , Xin GuXin Gu More articles by this author , Ryan TerleckiRyan Terlecki More articles by this author , Jennifer PaxtonJennifer Paxton More articles by this author , Ting LongTing Long More articles by this author , Lysette MutkusLysette Mutkus More articles by this author , Frank MariniFrank Marini More articles by this author , James YooJames Yoo More articles by this author , Anthony AtalaAnthony Atala More articles by this author , John JacksonJohn Jackson More articles by this author , and Yuanyuan ZhangYuanyuan Zhang More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.2728AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Intracavernosal administration of stem cells has demonstrated efficacy in animal models of erectile dysfunction (ED). The fate of grafted cells, however, is poorly characterized, and it is unknown if labeling technology impacts cellular function. Thus, alongside human placental stem cells (hPSC) we aim to use a novel dual-labeling technique featuring a red fluorescent protein variant mKATE and a new codon-optimized luciferase from Renilla Reniformis (mKATE-renLUC) using a lentivirus vector. We aim to determine the effect of labeling on in vitro function in preparation for future bioluminescence-tracking following intracavernosal administration. METHODS To optimize the transfection protocol, different hPSC concentrations, vectors (mKATE-renLUC, GFP-FLUC), and lentivirus titers were tested. Cell viability, proliferation, migration, and paracrine effects of hPSC expressing mKATE-renLUC were examined, and compared to non-labeled cells. Paracrine effects of labeled cells were examined using an angiogenesis array. Cell survival and distribution of hPSC expressing mKATE-renLUC were monitored by IVIS imaging system in vitro. RESULTS Cell viability, morphology, migration, and cell proliferation of mKATE-renLUC labeled hPSC were similar to those of non-labeled cells in vitro. Cell survival and migration of renLUC-labeled hPSC were efficiently monitored using IVIS imaging system. The angiogenesis array demonstrated similar expression of factors from both cell types. CONCLUSIONS This study demonstrates that dual-labeling technology using mKATE-renLUC provides a safe and effective cell tracking approach with brighter fluorophores and codon optimized luciferase. This development is promising for longer and more effective in vivo stem cell tracking after intracavernosal administration in models of ED. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e1104 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Ethan Matz More articles by this author Lei Dou More articles by this author Fangpeng Shu More articles by this author Xin Gu More articles by this author Ryan Terlecki More articles by this author Jennifer Paxton More articles by this author Ting Long More articles by this author Lysette Mutkus More articles by this author Frank Marini More articles by this author James Yoo More articles by this author Anthony Atala More articles by this author John Jackson More articles by this author Yuanyuan Zhang More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

The tumor microenvironment is a heterogeneous population of cells consisting of the tumor bulk plus supporting cells. It is becoming increasingly evident that these supporting cells are recruited by cancer cells from nearby endogenous host stroma and promote events such as tumor angiogenesis, proliferation, invasion, and metastasis, as well as mediate mechanisms of therapeutic resistance. In addition, recruited stromal cells range in type and include vascular endothelial cells, pericytes, adipocytes, fibroblasts, and bone-marrow mesenchymal stromal cells. During normal wound healing and inflammatory processes, local stromal cells change their phenotype to become that of reactive stroma. Under certain conditions, however, tumor cells can co-opt these reactive stromal cells and further transition them into tumor-associated stromal cells (TASCs). These TASCs express higher levels of proteins, including alpha-smooth muscle actin, fibroblast activating protein, and matrix metalloproteinases, compared with their normal, non-reactive counterparts. TASCs are also known to secrete many pro-tumorigenic factors, including IL-6, IL-8, stromal-derived factor-1 alpha, vascular endothelial growth factor, tenascin-C, and matrix metalloproteinases, among others, which recruit additional tumor and pro-tumorigenic cells to the developing microenvironment. Here, we review the current literature pertaining to the origins of recruited host stroma, contributions toward tumor progression, tumor-associated stromal cells, and mechanisms of crosstalk between endogenous host stroma and tumor cells.

Abstract: High demand of neoplastic tissues for glutamine (Gln) is met by its active transport across cell membranes. Chronic treatment with acrylamide in rodents is associated with an increased incidence of neoplasms, including astrocytomas. In this study, 24‐h acrylamide treatment significantly increased the initial rate of l‐[G‐3H]glutamine uptake in astrocyte cultures derived from the acrylamide‐sensitive Fischer 344 rat, and this effect could be fully inhibited by histidine, a model substrate for the amino acid transport system N. RT‐PCR analysis revealed that acrylamide treatment caused a significant increase in the astrocytic expression of the mRNA coding for the major system N protein, SNAT3, which is specifically overexpressed in malignant gliomas in situ. The acrylamide‐induced upregulation of astrocytic Gln transport via system N is likely to affect Gln homeostasis in these cells and may be causally related to the increased astrocytoma incidence observed in Fischer 344 rats.

OBJECTIVETo analyze the CARD15 gene in families with heritable multi-organ granulomatoses, including the original Blau syndrome kindred as well as other families with related granulomatous conditions.METHODSLinkage mapping was performed in 10 families. Observed recombination events were used to exclude regions centromeric or telomeric to 16q12.1, and the Blau gene critical region was refined to <3 cM, corresponding to a physical distance of 3.5 megabasepairs. Based on its known biochemical function, CARD15 was analyzed as a positional candidate for the Blau syndrome susceptibility gene, by direct DNA sequencing.RESULTSThese studies resulted in the identification, in 5 of the families, of 2 sequence variants at position 334 of the gene product (R334W and R334Q). Affected family members from the original Blau syndrome kindred were heterozygous for the R334W missense mutation; mutations at the same position were also observed in several unrelated Blau syndrome families, some of whose phenotypes included large-vessel arteritis and cranial neuropathy. The missense mutations segregated with the disease phenotype in the families, and were not seen in 208 control alleles.CONCLUSIONThese findings demonstrate that CARD15 is an important susceptibility gene for Blau syndrome and for other familial granulomatoses that display phenotypic traits beyond those of classic Blau syndrome.

Maintenance of the ionic and osmotic composition of the extracellular fluid (ECF) is essential for the optimal functioning of the central nervous system (CNS). Changes in ion and neurotransmitter levels in the cerebrospinal fluid (CSF) can have profound effects on the processing and transmission of neuronal signals. Cell swelling during correction of isotonic imbalances can produce a series of events leading to inappropriate release of excitatory amino acids (EAA). Given the osmoregulatory demands of the CNS, it is not surprising that it possesses well-developed osmoregulatory mechanisms capable of maintaining both extracellular and intracellular ionic composition and volume within narrow limits, despite large fluctuations in the ionic composition and osmolarity of the plasma. We have undertaken a series of studies to test the hypothesis that ethanol (EtOH) acts as an osmotic stressor and stimulates osmoregulatory processes in astrocytes. In the course of these studies, we have investigated the effects of acute and chronic exposure to EtOH on cell volume, as well as uptake and release of amino acids in neonatal rat primary astrocyte cultures.