Comparison of five different RNA sources to examine the lactating bovine mammary gland transcriptome using RNA-Sequencing.

The objective of this study was to examine five different sources of RNA, namely mammary gland tissue (MGT), milk somatic cells (SC), laser microdissected mammary epithelial cells (LCMEC), milk fat globules (MFG) and antibody-captured milk mammary epithelial cells (mMEC) to analyze the bovine mammary gland transcriptome using RNA-Sequencing. Our results provide a comparison between different sampling methods (invasive and non-invasive) to define the transcriptome of mammary gland tissue and milk cells. This information will be of value to investigators in choosing the most appropriate sampling method for different research applications to study specific physiological states during lactation. One of the simplest procedures to study the transcriptome associated with milk appears to be the isolation of total RNA directly from SC or MFG released into milk during lactation. Our results indicate that the SC and MFG transcriptome are representative of MGT and LCMEC and can be used as effective and alternative samples to study mammary gland expression without the need to perform a tissue biopsy

Incomplete milkings in automatic milking systems

Ponencia presentada en 2nd International Conference on Precision Dairy Farming. Rochester, Minnesota, 18 al 20 de junio de 2019.Fil: Masía, Fernando Miguel. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Cátedra de Producción de Leche; Argentina.Fil: Masía, Fernando Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.Fil: Lyons, Nicolas A. NSW Government. Department of Primary Industries; Australia.Fil: Piccardi, Mónica Belén. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina.Fil: Piccardi, Mónica Belén. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.Fil: Balzarini, Mónica Graciela. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Cátedra de Estadística y Biometría; Argentina.Fil: Balzarini, Mónica Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Centro Científico Tecnológico (CCT Córdoba); Argentina.Fil: Hovey, Russell C. University of California, Davis. Department of Animal Science; Estados Unidos de América.Fil: Garcia, Sergio C. The University of Sydney. School of Life and Environmental Sciences; Australia.Fil: Garcia, Sergio C. The University of Sydney. Sydney Institute of Agriculture; Australia.Automatic milking systems (AMS) rely on voluntary and distributed attendance of cows to the dairy facility throughout lactation. This generates variation in milking intervals (MI), defined as the period of time that elapses between two consecutive milking events, measured in hours. Farmers operating AMS need to manage variation in MI within and between cows. In AMS a robotic arm locates and attaches a cup onto each individual teat. Success of this task depends on several cow and equipment factors, including localisation and insertion of the teats, which is related to the amount of milk in the udder. Unsuccessful attachment of the cups to one or more teats, and premature cup removal, are some of the causes of incomplete milkings (Lyons et al., 2014). The aim of this work was to cluster cows according to the risk of having incomplete milkings and to characterize the groups regarding MI, peak yield (Lpeak) and days to peak yield (Dpeak).http://www.precisiondairyfarming.com/wp-content/uploads/2019/07/PrecisionDairy2019_Proceedings.pdfFil: Masía, Fernando Miguel. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Cátedra de Producción de Leche; Argentina.Fil: Masía, Fernando Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.Fil: Lyons, Nicolas A. NSW Government. Department of Primary Industries; Australia.Fil: Piccardi, Mónica Belén. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias; Argentina.Fil: Piccardi, Mónica Belén. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET); Argentina.Fil: Balzarini, Mónica Graciela. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Cátedra de Estadística y Biometría; Argentina.Fil: Balzarini, Mónica Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Centro Científico Tecnológico (CCT Córdoba); Argentina.Fil: Hovey, Russell C. University of California, Davis. Department of Animal Science; Estados Unidos de América.Fil: Garcia, Sergio C. The University of Sydney. School of Life and Environmental Sciences; Australia.Fil: Garcia, Sergio C. The University of Sydney. Sydney Institute of Agriculture; Australia

Inhibiting tryptophan metabolism enhances interferon therapy in kidney cancer.

Renal cell carcinoma (RCC) is increasing in incidence, and a complete cure remains elusive. While immune-checkpoint antibodies are promising, interferon-based immunotherapy has been disappointing. Tryptophan metabolism, which produces immunosuppressive metabolites, is enhanced in RCC. Here we show indolamine-2,3-dioxygenase-1 (IDO1) expression, a kynurenine pathway enzyme, is increased not only in tumor cells but also in the microenvironment of human RCC compared to normal kidney tissues. Neither kynurenine metabolites nor IDO inhibitors affected the survival or proliferation of human RCC or murine renal cell adenocarcinoma (RENCA) cells in vitro. However, interferon-gamma (IFNγ) induced high levels of IDO1 in both RCC and RENCA cells, concomitant with enhanced kynurenine levels in conditioned media. Induction of IDO1 by IFNα was weaker than by IFNγ. Neither the IDO1 inhibitor methyl-thiohydantoin-DL-tryptophan (MTH-trp) nor IFNα alone inhibited RENCA tumor growth, however the combination of MTH-trp and IFNα reduced tumor growth compared to IFNα. Thus, the failure of IFNα therapy for human RCC is likely due to its inability to overcome the immunosuppressive environment created by increased IDO1. Based on our data, and given that IDO inhibitors are already in clinical trials for other malignancies, IFNα therapy with an IDO inhibitor should be revisited for RCC

The Effect of Atorvastatin on Breast Cancer Biomarkers in High-Risk Women

Statins have the potential to reduce breast cancer incidence and recurrence as shown in both epidemiologic and laboratory studies. The purpose of this study was to evaluate the effect of a lipophilic statin, atorvastatin, on breast cancer biomarkers of risk [mammographic density (MD) and insulin growth factor 1 (IGF-1)] in high-risk premenopausal women

Editorial: The Mammary Stroma in Normal Development and Function

The mammary gland can no longer be simply viewed as an organ composed of epithelial cells within a passive stromal microenvironment. Many lines of evidence have evolved to reinforce the notion that mammary epithelial cell growth, differentiation, lactation and progression to cancer involves bidirectional interactions between the epithelial population and its surrounding stroma. Within this stroma are numerous systems that are all capable of modulating epithelial function. In this context, the mammary stroma is not simply a depot of adipose tissue in which mammary epithelial cells undertake a unique growth and differentiation process, although adipocytes can impart numerous modulatory signals to epithelial cells, and vice versa. Rather, the stromal environment constitutes and supports a critical vasculature that supplies nutrients and endocrine cues, a lymphatic system that not only removes metabolites but also provides an intimate interface with the immune system, and an extracellular matrix scaffold in which epithelial cells grow, differentiate and regress. Ultimately all of these components play a critical role in directing the epithelial phenotype during normal mammary gland growth and function. An increasing appreciation for these different systems demands a view of mammary epithelial cells in a much different light, and further necessitates the development of model systems that incorporate and integrate increasing complexity

Methods for Collecting Milk from Mice

Mouse models offer unique opportunities to study mammary gland biology and lactation. Phenotypes within the mammary glands, especially those caused by genetic modification, often arise during lactation, and their study requires the collection of adequate volumes of milk. We describe two approaches for collecting milk from lactating mice. Both methods are inexpensive, are easy to use in the laboratory or classroom, are non-invasive, and yield adequate volumes of milk for subsequent analyses

Diverse and Active Roles for Adipocytes During Mammary Gland Growth and Function

The mammary gland is unique in its requirement to develop in close association with a depot of adipose tissue that is commonly referred to as the mammary fat pad. As discussed throughout this issue, the mammary fat pad represents a complex stromal microenvironment that includes a variety of cell types. In this article we focus on adipocytes as local regulators of epithelial cell growth and their function during lactation. Several important considerations arise from such a discussion. There is a clear and close interrelationship between different stromal tissue types within the mammary fat pad and its adipocytes. Furthermore, these relationships are both stage- and species-dependent, although many questions remain unanswered regarding their roles in these different states. Several lines of evidence also suggest that adipocytes within the mammary fat pad may function differently from those in other fat depots. Finally, past and future technologies present a variety of opportunities to model these complexities in order to more precisely delineate the many potential functions of adipocytes within the mammary glands. A thorough understanding of the role for this cell type in the mammary glands could present numerous opportunities to modify both breast cancer risk and lactation performance

Sequencing the transcriptome of milk production: milk trumps mammary tissue

Background: Studies of normal human mammary gland development and function have mostly relied on cell culture, limited surgical specimens, and rodent models. Although RNA extracted from human milk has been used to assay the mammary transcriptome non-invasively, this assay has not been adequately validated in primates. Thus, the objectives of the current study were to assess the suitability of lactating rhesus macaques as a model for lactating humans and to determine whether RNA extracted from milk fractions is representative of RNA extracted from mammary tissue for the purpose of studying the transcriptome of milk-producing cells. Results: We confirmed that macaque milk contains cytoplasmic crescents and that ample high-quality RNA can be obtained for sequencing. Using RNA sequencing, RNA extracted from macaque milk fat and milk cell fractions more accurately represented RNA from mammary epithelial cells (cells that produce milk) than did RNA from whole mammary tissue. Mammary epithelium-specific transcripts were more abundant in macaque milk fat, whereas adipose or stroma-specific transcripts were more abundant in mammary tissue. Functional analyses confirmed the validity of milk as a source of RNA from milk-producing mammary epithelial cells. Conclusions: RNA extracted from the milk fat during lactation accurately portrayed the RNA profile of milk-producing mammary epithelial cells in a non-human primate. However, this sample type clearly requires protocols that minimize RNA degradation. Overall, we validated the use of RNA extracted from human and macaque milk and provided evidence to support the use of lactating macaques as a model for human lactation