Breast Samples Research Articles

Abstract PD4-08: PD4-08 A Novel Single Cell Model of Tamoxifen Response in Primary Human Breast Tumors

Abstract A Novel Single Cell Model of Tamoxifen Response in Primary Human Breast Tumors Austin Whitman, Hyunsoo Kim, Kamila Wisniewska, Rasha Kakati, Susana Garcia Recio, Hector Franco, Charles Perou, Philip Spanheimer Background: Resistance to endocrine therapy is a primary cause of treatment failure and death in patients with estrogen receptor (ER)-positive breast cancer. Intratumor heterogeneity is associated with resistance to therapy across tumors, and specifically in ER+/HER2- breast cancer, heterogeneity in ER and PR expression is associated with a worse response to endocrine therapy. We hypothesize that subpopulations within and across ER+/HER2- human breast tumors have distinct responses to tamoxifen and that discerning heterogeneity in response will improve understanding of inherent and emerging resistance to endocrine therapy. Methods: We developed an operating room-to-laboratory pipeline immediately after surgical resection for studies using alive tissue. Tissue samples were obtained and single cell suspensions created using physical and enzymatic dissociation. Cells were treated with tamoxifen (10 M) or control media for 12 hours in suspension and single cell RNA libraries generated using the 10X Genomics droplet-based kit and sequenced using the Illumina NextSeq2000. Results: We obtained normal breast tissue from 2 women undergoing reduction mammoplasty and tumor tissue from 10 women with ER+/HER2- invasive breast carcinoma. In tamoxifen treated and control matched pairs, a total of 22,195 cells from normal breast and 94,558 cells from tumor samples were sequenced. Computational analysis using consensus clustering was performed and cell types assigned using canonical correlation. Both tumor and normal samples identified clustering by cell type and not by patient revealing significant variability in cell type abundance between samples. In the normal breast samples, we performed differentially expressed genes (DEG) analysis comparing tamoxifen treatment to control for each cell type (Immune cells, fibroblasts, basal epithelial cells, luminal progenitor cells, and mature luminal cells) and enrichment analysis of up- and down-regulated genes performed. Strong depletion of estrogen induced genes was observed in tamoxifen-treated normal luminal progenitor and mature luminal cells, but not in basal epithelial cells or fibroblasts, demonstrating distinct, subpopulation-specific response to tamoxifen. In the 10 tumor matched pairs, 4 had a high epithelial proportion and tumor cells identified using inferred copy number variation. Tumor cells in 3 of these 4 samples showed significant down regulation of estrogen response genes with tamoxifen treatment. Using scBCSubtype to assign PAM50 subtype to individual tumor cells, the 3 responsive tumors were comprised primarily of LumA cells while the unresponsive tumor was predominantly LumB. Finally, we developed a novel score to quantify responsiveness at the single cell level based on downregulation of estrogen response genes with tamoxifen treatment relative to matched cluster-specific untreated expression. This analysis demonstrated heterogeneity in response to tamoxifen in tumor cells and identified distinct subpopulations of responsive and unresponsive tumor cells to tamoxifen treatment. Conclusion: We developed a novel ex vivo model to determine heterogeneity in therapeutic response to tamoxifen in normal human breast tissue and primary human breast tumors. We demonstrate differences in tamoxifen response by cell type and identify distinctly responsive and resistant subpopulations within human tumors. This provides a foundation to define features of responsive and resistant populations on the individual cell and specimen basis, and should allow us to develop precise, single cell-based predictors of response to endocrine therapy, and to identify genes and pathways driving resistance to therapy. Citation Format: Hyunsoo Kim, Austin Whitman, Kamila Wisniewska, Susana Garcia-Recio, Rasha Kakati, Hector L. Franco, Charles M. Perou, Philip Spanheimer. PD4-08 A Novel Single Cell Model of Tamoxifen Response in Primary Human Breast Tumors [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr PD4-08.

Menstrual Phase and Menopausal Status Classification of Benign Breast Tissue Using Hormone-Regulated Gene Expression and Histomorphology: A Validation Study.

The validation of breast cancer risk biomarkers in benign breast samples (BBS) is a long-sought goal, hampered by the fluctuation of gene and protein expression with menstrual phase (MP) and menopausal status (MS). Previously, we identified hormone-related gene expression and histomorphology parameters to classify BBS by MS/MP. We now evaluate both together, to validate our prior results. BBS were obtained from consenting women (86 premenopausal, 55 postmenopausal) undergoing reduction mammoplasty (RM) or contralateral unaffected breast (CUB) mastectomy. MP/MS was defined using classical criteria for menstrual dates and hormone levels on the day of surgery. BBS gene expression was measured with reverse transcription quantitative polymerase chain reaction (RT-qPCR) for three luteal phase (LP) genes (TNFSF11, DIO2, MYBPC1) and four menopausal genes (PGR, GREB1, TIFF1, CCND1). Premenopausal samples were classified into LP or non-LP, using published histomorphology parameters. Logistic regression and receiver-operator curve analysis was performed to assess area under the curve (AUC) for prediction of MP/MS. In all 131 women, menopausal genes plus age>50 years predicted true MS [AUC 0.93, 95% confidence interval (CI) 0.89, 0.97]. Among premenopausal women, high TNFSF11 expression distinguished non-LP from LP samples (AUC 0.80, 95% CI 0.70, 0.91); the addition of histomorphology improved the prediction nonsignificantly (AUC 0.87, 95% CI 0.78, 0.96). In premenopausal subsets, addition of histomorphology improved LP prediction in RM (AUC 0.95, 95% CI 0.87, 1.0), but not in CUB (0.84, 95% CI 0.72, 0.96). Expression of five-gene set accurately predicts menopausal status and menstrual phase in BBS, facilitating the development of breast cancer risk biomarkers using large, archived sample repositories.

Non-Destructive Detection of Chicken Freshness Based on Electronic Nose Technology and Transfer Learning

As a non-destructive detection method, an electronic nose can be used to assess the freshness of meats by collecting and analyzing their odor information. Deep learning can automatically extract features and uncover potential patterns in data, minimizing the influence of subjective factors such as selecting features artificially. A transfer-learning-based model was proposed for the electronic nose to detect the freshness of chicken breasts in this study. First, a 3D-printed electronic nose system is used to collect the odor data from chicken breast samples stored at 4 °C for 1–7 d. Then, three conversion to images methods are used to feed the recorded time series data into the convolutional neural network. Finally, the pre-trained AlexNet, GoogLeNet, and ResNet models are retrained in the last three layers while being compared to classic machine learning methods such as K Nearest Neighbors (KNN), Random Forest (RF), and Support Vector Machines (SVM). The final accuracy of ResNet is 99.70%, which is higher than the 94.33% correct rate of the popular machine learning model SVM. Therefore, the electronic nose combined with conversion to images shows great potential for using deep transfer learning methods for chicken freshness classification.

Dietary inositol-stabilized arginine silicate numerically reduced woody breast severity in male Ross 708 broilers without altering growth

The woody breast (WB) myopathy is an unintended outcome of fast broiler chicken growth and high breast muscle yields. Myodegeneration and fibrosis in the living tissue are results of hypoxia and oxidative stress driven by lack of blood supply to muscle fibers. The study aim was to titrate a vasodilator ingredient, inositol-stabilized arginine silicate (ASI), as a feed additive to improve blood flow and ultimately, breast meat quality. A total of 1,260 male Ross 708 broilers were assigned to: 1) a control basal diet, or the control diet plus increasing ASI: 2) 0.025% ASI, 3) 0.05% ASI, 4) 0.10% ASI, or 5) 0.15% ASI. At d 14, 28, 42, and 49, growth performance was measured on all broilers and serum from 12 broilers/diet was analyzed for creatine kinase and myoglobin presence. On d 42 and 49, 12 broilers/diet were measured for breast width, then left breast fillets were excised, weighed, palpated for WB severity, and visually scored for degree of white striping (WS). At 1 d postmortem, 12 raw fillets/treatment underwent compression force analysis, and at 2d postmortem, the same fillets were analyzed for water-holding capacity. mRNA was isolated from 6 right breasts/diet at both d 42 and 49 for qPCR quantification of myogenic gene expression. Birds fed the lowest dose of 0.025% ASI had a 5-point/3.25% feed conversion ratio reduction compared to birds fed 0.10% ASI over wk 4 to 6 and reduced serum myoglobin at 6-wk of age compared to the control. Breasts from birds fed 0.025% ASI received 42% greater normal WB scores at d 42 compared to control fillets. At d 49, breasts from broilers fed 0.10 and 0.15% ASI received 33% normal WB scores. At d 49, 0.025% AS-fed broiler breasts showed no severe WS. Increased myogenin expression was observed in 0.05 and 0.10% ASI breast samples on d 42 and myoblast determination protein-1 expression was upregulated in breasts from birds fed 0.10% ASI on d 49 compared to the control. Therefore, a dietary inclusion of 0.025, 0.10, or 0.15% ASI was beneficial in reducing WB and WS severity and promoting muscle growth factor gene expression at age of harvest without diminishing bird growth or breast muscle yields.

A Hybrid Machine Learning Approach to Screen Optimal Predictors for the Classification of Primary Breast Tumors from Gene Expression Microarray Data.

The high dimensionality and sparsity of the microarray gene expression data make it challenging to analyze and screen the optimal subset of genes as predictors of breast cancer (BC). The authors in the present study propose a novel hybrid Feature Selection (FS) sequential framework involving minimum Redundancy-Maximum Relevance (mRMR), a two-tailed unpaired t-test, and meta-heuristics to screen the most optimal set of gene biomarkers as predictors for BC. The proposed framework identified a set of three most optimal gene biomarkers, namely, MAPK 1, APOBEC3B, and ENAH. In addition, the state-of-the-art supervised Machine Learning (ML) algorithms, namely Support Vector Machine (SVM), K-Nearest Neighbors (KNN), Neural Net (NN), Naïve Bayes (NB), Decision Tree (DT), eXtreme Gradient Boosting (XGBoost), and Logistic Regression (LR) were used to test the predictive capability of the selected gene biomarkers and select the most effective breast cancer diagnostic model with higher values of performance matrices. Our study found that the XGBoost-based model was the superior performer with an accuracy of 0.976 ± 0.027, an F1-Score of 0.974 ± 0.030, and an AUC value of 0.961 ± 0.035 when tested on an independent test dataset. The screened gene biomarkers-based classification system efficiently detects primary breast tumors from normal breast samples.

Isoproterenol-induced Upregulation of HPSE Accelerates Triple-negative Breast Cancer Cell Proliferation and Migration through Enhancing the Transcriptional Activity of HIF-1α.

Triple-negative breast cancer (TNBC) is considered to be the most malignant subtype of breast cancer (BC). Heparanase (HPSE) has been reported to contribute to tumor development, but its potential function in TNBC is not clear. The intention of this study was to investigate whether HPSE affects TNBC progression and to explore the possible mechanisms. Bioinformatics analyses were applied to analyze the expression of HPSE in TNBC samples and normal breast samples. The mRNA and protein levels of HPSE in TNBC cells were detected by RT-qPCR and western blot. Function assays, including CCK-8 assay, colony formation assay, transwell assay and wound healing assay, were conducted to validate the effects of HPSE silencing on TNBC cell proliferation and migration. Mechanism experiments were performed to explore the upstream molecular mechanism of HPSE in TNBC cells. Silencing of HPSE suppressed the proliferation and migration of TNBC cells. Moreover, hypoxia-inducible factor-1 alpha (HIF-1α) interacted with the HPSE promoter and promoted the transcription of HPSE. Isoproterenol (ISO), a pharmacological substitute for chronic stress-induced sympathetic activation, was proven to induce HIF-1α upregulation, so as to transcriptionally activate HPSE in TNBC cells. Furthermore, it manifested that ISO facilitated TNBC cell proliferation and migration in an HPSE-dependent way. HPSE activated by ISO-induced HIF-1α promoted TNBC cell proliferation and migration, which might offer a novel insight for TNBC treatment.

ABCB1 Amplicon Contains Cyclic AMP Response Element-Driven TRIP6 Gene in Taxane-Resistant MCF-7 Breast Cancer Sublines.

A limited number of studies are devoted to regulating TRIP6 expression in cancer. Hence, we aimed to unveil the regulation of TRIP6 expression in MCF-7 breast cancer cells (with high TRIP6 expression) and taxane-resistant MCF-7 sublines (manifesting even higher TRIP6 expression). We found that TRIP6 transcription is regulated primarily by the cyclic AMP response element (CRE) in hypomethylated proximal promoters in both taxane-sensitive and taxane-resistant MCF-7 cells. Furthermore, in taxane-resistant MCF-7 sublines, TRIP6 co-amplification with the neighboring ABCB1 gene, as witnessed by fluorescence in situ hybridization (FISH), led to TRIP6 overexpression. Ultimately, we found high TRIP6 mRNA levels in progesterone receptor-positive breast cancer and samples resected from premenopausal women.

The Effect of Antibiotics Used Throughout Chicken Cycle on the Bacterial Count of Their Meat

There is no doubt that poultry industry is a very useful to our society Therefore poultry meat has its own value even economic value or nutrional value. The feature of poultry meat helps the consumer to obtain the more nutritional value such as Protein, Low fat, Carbs and vitamins. The contamination of poultry meat effects on the quality of meat and its content. Therefore it effects the population and can cuase infectious diseases .The present experiment was conducted on the broilers that obtained from private farm located at Egypt-Alexandria road. The broiler chicks were fed on well-balanced diet. Starter diet was given till the 20th day of age after that chicks were fed on finisher diet which was given from the 21st day till the end of the experiment. Water and food were offered adlibitium. The chicks were housed in clean well ventilated separate experimental rooms, previously fumigated with formalin and potassium permanganate. The floor was bedded by fresh clean chopped wheat straw forming litter of 3.5 cm depth. Each room was provided by suitable numbers of feeders and water utensils. Each group of bird was provided by suitable feeder and water. This study aimed to investigate the effect of antibiotics administrated during broilers chicken cycle on the bacteriological count of their meat through study of: 1- The level of mesophilc, aerobicenterobacteriacae and coliform counts among the breast and thigh samples at 15 day and 35 days of production cycle. 2- The level of mould and yeast counts among the breast and thigh samples at 15 day and 35 days of production cycle. 3- Identification of some isolated mould and yeast.4- Determination of the antibiotic residues among the breast and thigh samples at 15 day and 35 days of production cycle. The Staistical analytical results of Mesophilc aerobic bacterial counts isolated from breast and thigh of broilers at different periods of experiment was 34.15 X 103 ± 14.28 X 103 in breast at day 15 and was 38.64 X 103 ±19.88 X 103 in thigh at day 15. But in day 35 was 82.15 X 103 ± 24.51 X 103 in breast and was 89.91 X 103 ±28.99 X 103 in thigh at day 35 .The results of Enterobacteriaceae bacterial counts isolated from breast and thigh of broilers at different periods of experiment was 4.20 X 102±1.70 X 102 in breast at day 15 and was 1.51 X 103±4.70 X 102 in thigh at day 15 . But in day 35 was 9.04 X 103±3.93 X 103 n breast and was 3.21 X 103±1.32 X 103 in thigh at day 35.The results of Coliforms bacterial counts isolated from breast and thigh of broilers at different periods of experiment was 1.64 X 103 ± 1.13 X 103 in breast at day 15 and was 3.30 X 102± 6 X 10 in thigh at day 15. But in day 35 was 4.5 X 102± 1.7 X 102 in breast and was 7 X 10 ± 2 X 10 in thigh at day 35. Tht results of Moulds counts isolated from breast and thigh of broilers at different periods of experiment was 2 X 10 ± 1.0 X 10 in breast at day 15 and was 2.0 X 10 ± 1.0 X 10 in thigh at day 15. But at day 35 was 3.0 X 10 ± 1.0 X 10 in breast and was 3 X 10 ± 1.0 X 10 in thigh at day 35.The results of Yeast counts isolated from breast and thigh in different time of experiment was 0.04 X 103± 0.02 X 103 in breast at day 15 and was 0.11 X 103 ± 0.05 X 103 in thigh at day15. But at day 35 was 0.03 X 103 ± 0.01 X 103 in breast and was 0.03 X 103± 0.01 X 103 in thigh at day 35.

Perfil de ácidos graxos de frangos de corte de crescimento lento suplementados com minerais biocomplexados

ABSTRACT: This study was conducted to evaluate the effects of dietary supplementation with biocomplexed selenium and chromium on the fatty acid profile of the breast and thigh of males and females broilers of the Label Rouge lineage. The experimental design was completely randomized arranged in a 3 x 2 factorial scheme corresponding to three diets (control; 0.40 ppm of chromium; 0.50 ppm of selenium) and two sexes. Each treatment consisted of three plots of five broilers. The fatty acid profile was evaluated in the breast and thigh samples. The addition of biocomplexed selenium in the broilers diets increased the content of long chain fatty acid of the omega 3 and 6 series (docosahexaenoic - DHA and arachidonic acids, respectively), the total of omega 6 and the stearic acid content on the breast (pectoralis major muscle). In addition, selenium supplementation caused a reduction in oleic acid content and in total of monounsaturated in the same muscle. Males had higher contents of linoleic and linolenic acids, and total of polyunsaturated; and lower content of oleic acid, total of monounsaturated and saturated in the thigh. The broilers supplemented with biocomplexed selenium and males presented better lipid quality in the muscle by breast and thigh, respectively.

Cell annotation using scRNA-seq data: A protein-protein interaction network approach

Pathway analysis is an important step in the interpretation of single cell transcriptomic data, as it provides powerful information to detect which cellular processes are active in each individual cell. We have recently developed a protein-protein interaction network-based framework to quantify pluripotency associated pathways from scRNA-seq data. On this occasion, we extend this approach to quantify the activity of a pathway associated with any biological process, or even any list of genes. A systems-level characterization of pathway activities across multiple cell types provides a broadly applicable tool for the analysis of pathways in both healthy and disease conditions. Dysregulated cellular functions are a hallmark of a wide spectrum of human disorders, including cancer and autoimmune diseases. Here, we illustrate our method by analyzing various biological processes in healthy and cancer breast samples. Using this approach we found that tumor breast cells, even when they form a single group in the UMAP space, keep diverse biological programs active in a differentiated manner within the cluster.•We implement a protein-protein interaction network-based approach to quantify the activity of different biological processes.•The methodology can be used for cell annotation in scRNA-seq studies and is freely available as R package.

Are both distinct epithelial and stromal cells molecular analysis from phyllodes tumors versus fibroadenoma components affected in breast fibroepithelial progression?

To determine molecular events involved in the tumorigenesis of phyllodes tumors (PT) and the role of each stromal (SC) and epithelial (EC) cell. Frozen breast samples enriched with epithelial and stromal cells from three fibroadenomas and 14 PT were retrieved and laser microdissected. Sanger and polymerase chain reaction-based sequencing of exon 2 MED12 and TERT promoter hotspot mutations were performed; 44K microarray platform was used to analyze gene expression. All three fibroadenomas (FAs) presented mutations in MED12, but not in TERT, whose mutation was observed in five of the 14 PTs. EC and SC of each affected tumor displayed identical alterations. Of the total differentially expressed genes (DEG) (EC = 1,543 and SC = 850), 984 were EC-eDEGs and 291 were SC-eDEGs. We found a high similarity of diseases and functions enriched by both cell types, but dissimilarity in the number of enriched canonical pathways. Three signaling canonical pathways overlapping with EC and SC were predicted to be activated in one cell type and inactivated in the other, while no overlap in eDEGs was assigned to them. We also identified 13 EC-eDEGs and five SC-eDEGs enriched networks, in which the SC-eDEGs were able to segregate FA from PT samples. Identical TERT mutations from both SC and ES origins might affect the PTs tumorigenesis. Gene expression differences suggest coordinated molecular processes between these components with determinant differences acquired by SC, able to fully distinguish PTs from FAs lesions.

Microwave Breast Lesion Classification – Results from Clinical Investigation of the SAFE Microwave Breast Cancer System

Microwave breast cancer imaging (MWI) is an emerging non-invasive technology used to clinically assess the internal breast tissue inhomogeneity. MWI utilizes the variance in dielectric properties of healthy and cancerous tissue to identify anomalies inside the breast and make further clinical predictions. In this study, we evaluate our SAFE MWI system in a clinical setting. Capability of SAFE to provide breast pathology is assessed. Patients with BI-RADS category 4 or 5 who were scheduled for biopsy were included in the study. Machine learning approach, more specifically the Adaptive Boosting (AdaBoost) model, was implemented to determine if the level of difference between backscattered signals of breasts with the benign and malignant pathological outcome is significant enough for quantitative breast health classification via SAFE. A dataset of 113 (70 benign and 43 malignant) breast samples was used in the study. The proposed classification model achieved the sensitivity, specificity, and accuracy of 79%, 77%, and 78%, respectively. The non-ionizing and non-invasive nature gives SAFE an opportunity to impact breast cancer screening and early detection positively. Device classified both benign and malignant lesions at a similar rate. Further clinical studies are planned to validate the findings of this study.

Evaluation of the Prevalence of Staphylococcus aureus in Chicken Fillets and Its Bio-Control Using Different Seaweed Extracts.

This study aims to assess the occurrence of Staphylococcus aureus in chicken fillets and to control its growth using various lyophilized seaweed extracts (i.e., Halimeda opuntia (HO), Actinotrichia fragilis, and Turbinaria turbinata) by an agar disk diffusion assay in vitro. Results showed that prevalence of S. aureus in breast and thigh samples reached of 92% and 84%, respectively. Lyophilized HO extract was the only seaweed that showed the antibacterial activity against S aureus with a significant difference at p < 0.05. The minimum inhibitory concentration (MIC) of HO extract was 1.5%, with an inhibition zone of 8.16 ± 0.73 mm. Regarding 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, IC50 was recorded at 55.36 μg/mL, whereas cytotoxic IC50 of the lyophilized HO extract on peripheral blood mononuclear cells (PBMCs) was 33.7 µg/mL; a higher IC50 of HO extracts permits their use as a safe food additive in meat products. Moreover, total phenolic compounds and total flavonoids compounds recorded 20.36 ± 0.092 and 16.59 ± 0.029 mg/mL, respectively. HPLC analyses of phenolic compounds profiles exhibited many bioactive substances and the higher ratio was daidzein with 10.84 ± 0.005 µg/mL and followed by gallic acid with a value of 4.06 ± 0.006 µg/mL. In a challenge study, chicken fillet (CHF) experimentally inoculated with S. aureus (ST) and treated with the lyophilized HO algal extract at 4% and 6% (CHF/ST/HO) showed a complete reduction of S. aureus count on the 6th and 4th days in chicken fillet stored at 4 °C, respectively. Moreover, CHF/ST/HO at 4% and 6% of HO extract enhanced the sensory attributes of grilled un-inoculated chicken fillet. Thus, lyophilized HO extracts are promising antibacterial and antioxidant candidates in the chicken meat industry.

Prevalence of Imaging-Detected Silent Female Breast Cancer in Autopsy Specimens: A Study Using Image-Guided Biopsies.

Background This study was conducted to evaluate the prevalence of imaging-detected silent breast cancer in females, with the hypothesis that the incidence of imaging-detected silent breast cancer in females is greater than the true disease incidence. The main purpose of this study is the attempt to prove whether breast imaging can identify silent breast cancers that apparently are common inserial histology analysis. Methodology A series of 217 consecutive medicolegal autopsies on fresh Portuguese cadavers were performed from July 2016 to December 2019 at the National Institute of Legal Medicine and Forensic Science, Lisbon, Portugal. The criteria for exclusion were age younger than 40 years, the autopsy performed in less than 48 hours after death, any major injury to one or both breasts, and known or clinically evident breast cancer. Once the eligibility criteria were met, and the sample collection authorization was obtained, a bilateral subcutaneous modi-fied radical mastectomy was performed in each fresh cadaver at the National Institute of Legal Medicine and Forensic Science. Mammography, ecography, and excisionalbiopsies of suspect areas were conducted on the collected samples. Results The indication for excisional biopsy by imaging was assigned in eight cases, and no breast cancer was discovered in the excised specimens. Conclusions In light of the findings, it cannot be concluded that the imaging-detected silent breast cancer prevalence is higher than the actual incidence of the disease, so the author's initial hypothesis was rejected. Mammography does not overdiagnose breast cancer.Benign breast alterations are common, accounting for 43.6% of the corpses collected, while low-suspicion alterations were discovered in 1.84% of breast samples.The objective examination, which included inspection and palpation, missed 37.5% of the biopsied breast changes. This finding indicated that an objective examination leads to a significant number of false-negative results which cannot be used as a screening method.

Differences in bedding material could alter the growth performance of White Pekin ducks raised for 42 days.

The effect of different commercially available bedding materials on the growth performance and carcass characteristics of ducks for 42 days was investigated. 336 one-day-old White-Pekin ducklings (60.48 ± 0.16 g) were randomly allocated into 24-floor pens with one of the three beddings namely i) coco peat, ii) rice husks, or iii) sawdust. 14 ducklings per pen and 8 replicate pens per bedding material were used. Birds were fed a starter diet from days 1-21 and a grower diet from days 22-42. Weekly growth performance evaluation was conducted for the average body weight, weight gains, daily feed intake, and feed conversion efficiency. One bird per pen was sacrificed on day 42 for the evaluation of carcass characteristics including the carcass, breast, and leg muscle percentages. Breast and leg muscle samples were then collected and analyzed for their proximate and pH values. Higher body weights (p < 0.05) were noticed with rice husks on day 42 only. Improved daily gains (p < 0.05) were also noticed for birds raised with rice husks over the entire period (days 1-42). Concerning feed intake, higher values (p < 0.05) were similarly noted with rice husks for the grower phase (days 22-42), and the entire experimental period (days 1-42). Marginally improved feed intake values were also noted with the use of rice husks as the bedding materials on day 42 (p = 0.092). Improved feed efficiency (p < 0.05) was noticed with rice husks on day 35, the grower period, and the entire 42-day period. However, no significant differences were noticed for most of the carcass characteristics that were evaluated. Nevertheless, higher (p < 0.05) pH values for the breast muscle were noticed with the use of coco peat and sawdust as the bedding. Conclusively, the bedding type could have a significant impact on the growth performance of ducks without adverse effects on carcass characteristics. The use of rice husks as bedding might be advantageous and is therefore recommended.

Oncologic necessity for the complete removal of residual microcalcifications after neoadjuvant chemotherapy for breast cancer

The surgical range of breast cancer that shows pathologic complete response (pCR) without change in microcalcifications after neoadjuvant chemotherapy (NAC) is controversial. This study examined whole breast specimens to evaluate the necessity of mastectomy in those cases. The viability of cancer cells around the residual microcalcification was assessed using prospectively collected breast samples to confirm the presence or absence of cancer cells. A total of 144 patients with breast cancer and diffuse microcalcifications were classified into the reduced mass with no change in residual microcalcification (RESMIN, n = 49) and non-RESMIN (n = 95) groups. Five specimens were prospectively evaluated to assess the presence of viable cancer cells around the microcalcification. Tumor responses to NAC were significantly better with high pCR rates in the RESMIN group (p = 0.005 and p = 0.002). The incidence of human epidermal growth factor receptor 2-positive and triple-negative breast cancers was significantly high in the RESMIN group (p = 0.007). Although five (10.2%) patients had locoregional recurrence in the RESMIN group, no local recurrence in the breast was reported. Although pCR was highly estimated, residual cancers, including ductal carcinoma in situ, remained in 80% cases. Therefore, given the weak scientific evidence available currently, complete removal of residual microcalcifications should be considered for oncologic safety.

Australian Donation and Transplantation Biobank: A Research Biobank Integrated Within a Deceased Organ and Tissue Donation Program.

Feasibility of a research biobank integrated within the deceased organ and tissue donation program was assessed. DonateLife Victoria sought consent for ADTB donation after consent was received for organ donation for transplantation from the donor's senior available next of kin. ADTB samples were collected during donation surgery and distributed fresh to researchers or stored for future research. The main outcome measures were ADTB donation rates, ADTB sample collection, ADTB sample use, and to identify ethical considerations. Over 2 y, samples were collected for the ADTB from 69 donors (28% of 249 donors). Samples were obtained from the spleen (n = 59, 86%), colon (n = 57, 83%), ileum (n = 56, 82%), duodenum (n = 55, 80%), blood (n = 55, 80%), bone marrow (n = 55, 80%), skin (n = 54, 78%), mesenteric lymph nodes (n = 56, 81%), liver (n = 21, 30%), lung (n = 29, 42%), and lung-draining lymph node (n = 29, 42%). Heart (n = 20), breast (n = 1), and lower urinary tract (n = 1) samples were obtained in the second year. Five hundred fifty-six samples were used in 19 ethics-approved research projects spanning the fields of immunology, microbiology, oncology, anatomy, physiology, and surgery. The integration of routine deceased donation and transplantation activities with a coordinated system for retrieval and allocation of donor samples for use in a range of research projects is feasible and valuable.

Thermal inactivation kinetics of uropathogenic Escherichia coli in sous-vide processed chicken breast

Chicken is a suspected reservoir of uropathogenic Escherichia coli (UPEC), resulting in foodborne urinary tract infections (UTIs). Sous-vide ready-to-eat (RTE) food products may be associated with microbial hazards due to the low-temperature long-time (LTLT) process. However, little is known regarding the survival of UPEC during sous-vide cooking. The aim of this study was to evaluate the heat resistance of UPEC in chicken breast during sous-vide processing and establish predictive inactivation models. Chicken breast samples were inoculated with a four-strain cocktail of UPEC, including reference strains from UTI patients and chicken isolates. The inoculated samples, with or without 3% NaCl solution for marination, were vacuum sealed in bags, immersed in a temperature-controlled water bath, and cooked at 50 °C, 55 °C, 60 °C, and 63 °C. The change in survival of populations of UPEC was fitted with the linear and Weibull inactivation models to obtain the survival curves at different temperatures; the D- and z-values were also calculated. The goodness-of-fit was evaluated using the root mean square error (RMSE), sum of squared errors (SSE), adjusted R2, and Akaike information criterion (AIC). The results showed that the linear model with tail was better than the Weibull model in terms of fitting performance. With the addition of salt marinade, D-values at 50 °C, 55 °C, 60 °C, and 63 °C determined by the linear model with tail decreased from 299.78 to 166.93 min, 16,60 to 13.87 min, 4.06 to 3.05 min, and 1.05 to 0.87 min, respectively, compared with the controls. The z-values of control and salt-marinated samples were 6.14 °C and 5.89 °C, respectively. The model developed for predicting UPEC survival under sous-vide cooking was validated using an additional survival curve at 58 °C. The validation results showed that the RMSE was 0.122 and 0.133 log CFU/g, and the proportion of relative error was 0.875 and 0.750 in the acceptable prediction zones for the control and salt-marinated samples, respectively. In conclusion, the heat resistance of an emerging foodborne pathogen, UPEC, in sous-vide processed chicken breast was revealed for the first time. Our results showed that salt marinade (3% NaCl) increases the heat sensitivity of UPEC during the sous-vide processing. The developed survival functions based on the linear model with tail can be applied to control the thermal lethality of UPEC.

Abstract IA011: Patient-derived organoids as models for breast cancer prevention and interception

Abstract Organoid culture technology can be an efficient means for propagating breast tissues from patients, and we have previously generated organoid biobanks derived from normal and malignant human breast samples. Non-cancerous organoids, which are grown in a hydrogel that mimics the basement membrane and are bathed in a cocktail of growth factors, vitamins, and substances that substitute for stroma, grow in a polarized fashion to re-form acinar- and duct-like structures, and differentiate into the major mammary epithelial lineages, and we have found that all the major mammary epithelial lineages can be propagated in vitro. We have utilized mass cytometry and single-cell RNA sequencing to identify specific cell subtypes that are associated with breast cancer risk factors in organoids. In parallel, in malignant organoid cultures we have utilized RNA sequencing and compound screening approaches to identify potential strategies to overcome therapeutic resistance. We now present our initial efforts to establish a DCIS organoid biobank. Similar to the normal breast, organoids can be propagated from DCIS cases that are resistant to endocrine therapy with the goal of identifying strategies to overcome the resistant state. Citation Format: Jennifer M. Rosenbluth. Patient-derived organoids as models for breast cancer prevention and interception [abstract]. In: Proceedings of the AACR Special Conference on Rethinking DCIS: An Opportunity for Prevention?; 2022 Sep 8-11; Philadelphia, PA. Philadelphia (PA): AACR; Can Prev Res 2022;15(12 Suppl_1): Abstract nr IA011.

High hydrostatic processing of marinated ground chicken breast: Exploring the effectiveness on physicochemical, textural and sensory properties and microbial inactivation

High hydrostatic pressure (HHP) processing of marinated ground chicken breast meat samples by 200–600 MPa for 5–15 min at ambient temperature revealed that increased magnitude of pressure and treatment time provided significant increase in pH, surface L*, a*, b*, and total color difference (ΔE), inner a*, b* and ΔE color values, marinade uptake, total antioxidant capacity, total phenolic substance content, textural properties of cooking loss, hardness, chewiness, cohesiveness, and gumminess, and sensory properties of appearance and color. A significant decrease was observed in the inner color L* value, and textural properties of springiness, whereas no significant changes were detected on sensory properties of aroma, hardness, and aftertaste. HHP processing at 600MPa/15 min provided maximum 4.73 and 3.89 log inactivations on the initial total mesophilic aerobic bacteria and total mold and yeast counts. The initial Salmonellae and total coliform counts of 3.53 ± 0.12 and 6.59 ± 0.11 log cfu/g were reduced to undetectable level by 400 MPa/15 min and 600 MPa for 5, 10, and 15 min, respectively. The results reveled that HHP is very efficient for diffusion of marinade solution with providing microbial safety without adversely affecting physical, textural and sensory properties of marinated ground chicken breast samples.