Ewing Sarcoma-Chest Wall Reconstruction Following Resection of Rare Primary Chest Wall Tumor.

Ewing sarcoma (ES) represents the second most common primary osseous malignancy in children and young adults, most often occurring in the diaphysis of the long bones. While rare, ES can present as an osseous tumor of the ribs and/or chest wall. These tumors are known as Askin's tumors and most commonly present with symptoms resembling pneumonia. We report the case of a 26-year-old man who was found to have a right lung mass extending into his anterolateral chest wall after presenting to the hospital for evaluation of unremitting chest pain. Biopsy was performed and the patient diagnosed with ES. After completion of neoadjuvant chemotherapy, the patient underwent resection of the right chest wall mass. The chest wall was reconstructed in a novel fashion with titanium plates and a reinforced tissue matrix patch. Due to a paucity of cases, no treatment or reconstruction algorithm currently exists for management of these malignancies.

Surely you can't B. cereus: Bacillus cereus infection resulting in spontaneous pseudoaneurysm of a nonaccessed arteriovenous graft.

Vascular graft infections are a well-described complication of loop arteriovenous grafts (AVGs) placed for hemodialysis access and are reported to occur in 0.5% to 6.0% of AVGs. The most common microorganisms implicated in these infections are the Staphylococcus species. We present a case of a chronically nonaccessed graft rupture caused by an indolent B. cereus colonization, which is usually a foodborne contaminant. The finding of this organism as the causal agent in an AVG infection warrants further research into the potential emergence of the Bacillus species as a contributing factor in the morbidity and mortality resulting from AVG infection.

Wnt5a Regulates the Assembly of Human Adipose Derived Stromal Vascular Fraction-Derived Microvasculatures.

Human adipose-derived stromal vascular fraction (hSVF) cells are an easily accessible, heterogeneous cell system that can spontaneously self-assemble into functional microvasculatures in vivo. However, the mechanisms underlying vascular self-assembly and maturation are poorly understood, therefore we utilized an in vitro model to identify potential in vivo regulatory mechanisms. We utilized passage one (P1) hSVF because of the rapid UEA1+ endothelium (EC) loss at even P2 culture. We exposed hSVF cells to a battery of angiogenesis inhibitors and found that the pan-Wnt inhibitor IWP2 produced the most significant hSVF-EC networking decrease (~25%). To determine which Wnt isoform(s) and receptor(s) may be involved, hSVF was screened by PCR for isoforms associated with angiogenesis, with only WNT5A and its receptor, FZD4, being expressed for all time points observed. Immunocytochemistry confirmed Wnt5a protein expression by hSVF. To see if Wnt5a alone could restore IWP2-induced EC network inhibition, recombinant human Wnt5a (0-150 ng/ml) was added to IWP2-treated cultures. The addition of rhWnt5a significantly increased EC network area and significantly decreased the ratio of total EC network length to EC network area compared to untreated controls. To determine if Wnt5a mediates in vivo microvascular self-assembly, 3D hSVF constructs containing an IgG isotype control, anti-Wnt5a neutralizing antibody or rhWnt5a were implanted subcutaneously for 2w in immune compromised mice. Compared to IgG controls, anti-Wnt5a treatment significantly reduced vessel length density by ~41%, while rhWnt5a significantly increased vessel length density by ~62%. However, anti-Wnt5a or rhWnt5a did not significantly affect the density of segments and nodes, both of which measure vascular complexity. Taken together, this data demonstrates that endogenous Wnt5a produced by hSVF plays a regulatory role in microvascular self-assembly in vivo. These findings also suggest that manipulating Wnt signaling could enhance control of hSVF vascularization in tissue engineering applications.

Restoration of Physiologically Responsive Low-Density Lipoprotein Receptor-Mediated Endocytosis in Genetically Deficient Induced Pluripotent Stem Cells.

Acquiring sufficient amounts of high-quality cells remains an impediment to cell-based therapies. Induced pluripotent stem cells (iPSC) may be an unparalleled source, but autologous iPSC likely retain deficiencies requiring correction. We present a strategy for restoring physiological function in genetically deficient iPSC utilizing the low-density lipoprotein receptor (LDLR) deficiency Familial Hypercholesterolemia (FH) as our model. FH fibroblasts were reprogrammed into iPSC using synthetic modified mRNA. FH-iPSC exhibited pluripotency and differentiated toward a hepatic lineage. To restore LDLR endocytosis, FH-iPSC were transfected with a 31 kb plasmid (pEHZ-LDLR-LDLR) containing a wild-type LDLR (FH-iPSC-LDLR) controlled by 10 kb of upstream genomic DNA as well as Epstein-Barr sequences (EBNA1 and oriP) for episomal retention and replication. After six months of selective culture, pEHZ-LDLR-LDLR was recovered from FH-iPSC-LDLR and transfected into Ldlr-deficient CHO-a7 cells, which then exhibited feedback-controlled LDLR-mediated endocytosis. To quantify endocytosis, FH-iPSC ± LDLR were differentiated into mesenchymal cells (MC), pretreated with excess free sterols, Lovastatin, or ethanol (control), and exposed to DiI-LDL. FH-MC-LDLR demonstrated a physiological response, with virtually no DiI-LDL internalization with excess sterols and an ~2-fold increase in DiI-LDL internalization by Lovastatin compared to FH-MC. These findings demonstrate the feasibility of functionalizing genetically deficient iPSC using episomal plasmids to deliver physiologically responsive transgenes.

Hazard Analysis and Control of Roast Beef Jus Preparation in Foodservice Establishments.

Roast beef and its jus prepared in foodservice establishments are often implicated as vehicles of foodborne illness. Preparation practices that could contribute to survival or growth of foodborne disease bacteria were examined. Temperatures were reached during cooking that would kill vegetative forms of these organisms. Prolonged holding of cooked jus on ranges with the heat turned off or on table tops created conditions in which spores could germinate and vegetative cells multiply. Conditions prevailed during cooling that could promote bacterial growth. Reheating jus to the boiling point would kill any vegetative forms that had multiplied during storage.

Hazard Analysis of Frozen Dinners Prepared at a Catering Establishment 1.

Frozen dinners prepared by a caterer were reported to have been spoiled. Microbiological testing of samples was performed, and a survey of time-temperature exposures (during preparation, storage, delivery and reheating) was conducted of procedures duplicating those at the time the spoiled food was prepared. Growth of spoilage bacteria was not inhibited by freezing a customer's week's supply of packaged meals in cardboard boxes. This growth continued during transit in a precooled insulated truck. Meals reheated in a plywood box oven (furnished by the caterer for this purpose) failed to reach lethal temperatures for vegetative mesophilic bacteria within a practicable time (90 min). Holding the meals in wire baskets or in shallow metal trays during freezing and reheating the frozen meals to 74 C (165 F) in conventional domestic ovens prevented spoilage of the meals.

Hazard Analysis and Control of Roast Beef Preparation in Foodservice Establishments 1.

Roast beef preparation practices were examined in eight foodservice establishments for the likelihood of contamination and the possibilities of survival or growth during each step of the operations. Clostridium perfringens was isolated from raw beef, equipment, and cooked beef. Staphylococcus aureus was isolated from raw beef, equipment, workers' hands, and cooked beef. Salmonellae were isolated from neither meat nor equipment. Numerous opportunities were observed for contamination of cooked beef during operations in most of the establishments. No opportunities for multiplication of foodborne disease bacteria were observed during thawing of frozen beef. From recorded time-temperature data, it was calculated that vegetative foodborne pathogens could survive in 76% of the geometric centers and on 5% of the surfaces of beef during cooking. Survival of these organisms could occur in 36% of the geometric centers and on 11% of the surfaces of the cooked beef during post-oven temperature rise periods. These organisms could have survived in 25% of the geometric centers and on 33% of the surfaces of the cooked roasts during hot holding; they could have multiplied on 25% and 27%, respectively. During cooling, the potential for multiplication of vegetative cells of foodborne pathogens existed in 83% of the geometric centers and 79% of the surfaces of the roasts. During reheating, these organisms would have survived in 71% of the geometric centers and on 13% of the surfaces of the roasts. Recommendations are given for hot holding, cooling, and reheating so as to minimize microbiological problems.