In-depth characterization of a new patient-derived xenograft model for metaplastic breast carcinoma to identify viable biologic targets and patterns of matrix evolution within rare tumor types.

Metaplastic breast carcinoma (MBC) is a rare breast cancer subtype with rapid growth, high rates of metastasis, recurrence and drug resistance, and diverse molecular and histological heterogeneity. Patient-derived xenografts (PDXs) provide a translational tool and physiologically relevant system to evaluate tumor biology of rare subtypes. Here, we provide an in-depth comprehensive characterization of a new PDX model for MBC, TU-BcX-4IC. TU-BcX-4IC is a clinically aggressive tumor exhibiting rapid growth in vivo, spontaneous metastases, and elevated levels of cell-free DNA and circulating tumor cell DNA. Relative chemosensitivity of primary cells derived from TU-BcX-4IC was performed using the National Cancer Institute (NCI) oncology drug set, crystal violet staining, and cytotoxic live/dead immunofluorescence stains in adherent and organoid culture conditions. We employed novel spheroid/organoid incubation methods (Pu·MA system) to demonstrate that TU-BcX-4IC is resistant to paclitaxel. An innovative physiologically relevant system using human adipose tissue was used to evaluate presence of cancer stem cell-like populations ex vivo. Tissue decellularization, cryogenic-scanning electron microscopy imaging and rheometry revealed consistent matrix architecture and stiffness were consistent despite serial transplantation. Matrix-associated gene pathways were essentially unchanged with serial passages, as determined by qPCR and RNA sequencing, suggesting utility of decellularized PDXs for in vitro screens. We determined type V collagen to be present throughout all serial passage of TU-BcX-4IC tumor, suggesting it is required for tumor maintenance and is a potential viable target for MBC. In this study we introduce an innovative and translational model system to study cell-matrix interactions in rare cancer types using higher passage PDX tissue.

Feeding low dietary levels of organic trace minerals improves broiler performance and reduces excretion of minerals in litter.

1. This experiment was conducted to evaluate the effect of organic and inorganic trace minerals in diets fed to 1872, one-day-old male Ross 708 broiler chicks. Two sources of supplemental trace minerals (inorganic or organic) and four trace minerals levels (12.5%, 25.0%, 37.5% or 50.0%) plus a positive control containing inorganic minerals at 100% of recommended levels according to Brazilian industry standards, were used in the diets. 2. When analysed with mineral source as the main factor, broilers fed diets containing organic minerals had better feed conversion (FCR) throughout the trial, until 48 d of age (1.754 organic vs. 1.783 inorganic; P < 0.05), improved liveability at 48 d (95.8% organic vs. 93.6% inorganic; P < 0.05) and higher weight gain at 48 d (3.941 kg organic vs. 3.881 kg inorganic; P < 0.05) compared to the inorganic trace mineral diets. At lower dietary inclusion levels (12.5% and 25%) the organic mineral-fed birds had a four-point improvement in FCR (P < 0.05). 3. Feeding organic minerals improved tibial ash at 48 d from 49.5% to 51.25% (P < 0.05). The P and Ca in the tibia at 48 d were significantly (P < 0.05) higher for the birds fed organic minerals (Ca: 17.4% vs. 18.7%; P: 8.8% vs. 9.4%, respectively). The lower levels of trace minerals in feed reduced the mineral concentration of Zn, Cu and Mn in the faeces and, hence, in the litter (P < 0.05). 4. Feeding organic minerals promoted a small, but statistically significant, drop in pH of breast meat compared to inorganic minerals at 48 d (5.83 vs. 5.86, respectively; P < 0.05). The organic mineral diets reduced drip loss in breast meat compared to inorganic trace mineral supplementation at 32 d of age, but not in older birds (2.46% vs. 3.77%, respectively; P < 0.05). 5. Oxidation status, measured by superoxide dismutase activity in breast muscle at 48 d of age, significantly increased with mineral inclusion levels (P < 0.001), irrespective of source. 6. It was concluded that the use of organic minerals, even at low levels in the diet, is beneficial for broiler performance and reduces the excretion of minerals in litter.

Short- and long-term effects of furosemide on lung function in infants with bronchopulmonary dysplasia.

Although furosemide improves lung mechanics in some infants with broncho-pulmonary dysplasia (BPD), this may not be important unless gas exchange also improves. To determine the relationship between improvement in mechanics and improvement in gas exchange, the short- and long-term effects of furosemide therapy were studied in 16 spontaneously breathing infants with severe BPD who were both oxygen dependent and hypercarbic (mean PCO2 54 +/- 11 torr). Each infant was examined at least three times: before furosemide therapy, 1 hour after the first dose of furosemide, and after a 6- to 10-day course. Ten of the 16 infants were also examined three times during a 7-day control period. Transcutaneous PO2 and PCO2, esophageal pressure, air flow, and tidal volume were measured. Pulmonary resistance, lung compliance, and the alveolar to skin PO2 difference were calculated. After a single dose of furosemide, only compliance significantly improved. After prolonged therapy, compliance, resistance, and oxygenation significantly improved in the group as a whole, but better oxygenation was achieved in only six of 16 infants. tcPCO2 was unaffected by long-term furosemide therapy, but in all infants with decreased tcPCO2 1 hour after a single dose, there was sustained decrease in PCO2 after prolonged therapy. Changes in gas exchange were not explained by changes in lung mechanics. These data indicate that long-term diuretic therapy can improve the mechanical properties of the lungs of spontaneously breathing infants with BPD, but that gas exchange is usually unaffected.