Breast tissue regeneration is driven by cell-matrix interactions coordinating multi-lineage stem cell differentiation through DDR1.

Mammary morphogenesis is an orchestrated process involving differentiation, proliferation and organization of cells to form a bi-layered epithelial network of ducts and lobules embedded in stromal tissue. We have engineered a 3D biomimetic human breast that makes it possible to study how stem cell fate decisions translate to tissue-level structure and function. Using this advancement, we describe the mechanism by which breast epithelial cells build a complex three-dimensional, multi-lineage tissue by signaling through a collagen receptor. Discoidin domain receptor tyrosine kinase 1 induces stem cells to differentiate into basal cells, which in turn stimulate luminal progenitor cells via Notch signaling to differentiate and form lobules. These findings demonstrate how human breast tissue regeneration is triggered by transmission of signals from the extracellular matrix through an epithelial bilayer to coordinate structural changes that lead to formation of a complex ductal-lobular network.

A unifying fractional wave equation for compressional and shear waves.

This study has been motivated by the observed difference in the range of the power-law attenuation exponent for compressional and shear waves. Usually compressional attenuation increases with frequency to a power between 1 and 2, while shear wave attenuation often is described with powers less than 1. Another motivation is the apparent lack of partial differential equations with desirable properties such as causality that describe such wave propagation. Starting with a constitutive equation which is a generalized Hooke's law with a loss term containing a fractional derivative, one can derive a causal fractional wave equation previously given by Caputo [Geophys J. R. Astron. Soc. 13, 529-539 (1967)] and Wismer [J. Acoust. Soc. Am. 120, 3493-3502 (2006)]. In the low omegatau (low-frequency) case, this equation has an attenuation with a power-law in the range from 1 to 2. This is consistent with, e.g., attenuation in tissue. In the often neglected high omegatau (high-frequency) case, it describes attenuation with a power-law between 0 and 1, consistent with what is observed in, e.g., dynamic elastography. Thus a unifying wave equation derived properly from constitutive equations can describe both cases.

Suckling in newborn rats: eliminated by nipple lavage, reinstated by pup saliva.

Chemical lavage of the nipples of anesthetized maternal rats virtually eliminated suckling by their 4- to 5-day-old pups. Normal suckling was immediately reinstated, however, by painting a vacuum distillate of the wash or of pup saliva onto the nipples. Thus, a substance necessary to direct and release suckling, possibly rat pup saliva, appears to coat the nipple surface.

Influence of cell adhesive molecules attached onto PEG-lipid-modified fluid surfaces on cell adhesion.

The involvement of intercellular interactions in various biological events indicates the importance of studying cell-cell interactions using fluid model surfaces. Here, we propose a fluid surface composed of a self-assembled monolayer (SAM) and poly(ethylene glycol)-conjugated phospholipid (PEG-lipid) derivatives, which can be an alternative to supported lipid membranes. The modification of SAM surfaces with PEG-lipids carrying functional peptides resulted in the formation of the fluid surfaces with different mobility, which was quantitatively determined by quartz crystal microbalance with dissipation (QCM-D) and fluorescence recovery after photobleaching (FRAP). Different types of fluid surfaces with calculated diffusion coefficients between 0.9 ± 0.25 and 0.16 ± 0.03 µm2/sec for PEG-lipids derivatives were fabricated, onto which arginylglycylaspartate (RGD) peptides were immobilized for cell adhesion, and compared to solid surfaces with the same surface density of RGD peptides. The fluid surfaces revealed that cell adhesions of epithelial cells (MCF-10 A) and human umbilical vein endothelial cells (HUVEC) could not be established on the surfaces with higher fluidity, while cells could adhere onto surfaces with lower fluidity, where the lateral diffusion of PEG-lipids was approximately 20 times lower, and solid surfaces. Interestingly, cells that adhered onto the surface with lower fluidity proliferated at a normal rate while maintaining their round morphology, which was a different shape from that observed on solid surfaces. Thus, the scaffold fluidity greatly influenced cell adhesion behaviors, demonstrating that it is an important parameter for designing novel biomimetic scaffolds for biomedical applications.

Effects of breast stimulation for spontaneous onset of labor on salivary oxytocin levels in low-risk pregnant women: A feasibility study.

OBJECTIVES: This preliminary study aimed to 1) determine changes in the salivary oxytocin (OT) level during breast stimulation for promoting the spontaneous onset of labor in low-risk term pregnancies, and 2) clarify the feasibility of the breast stimulation intervention protocol in terms of practicality and acceptability. METHODS: We used a single arm trial design. Sixteen low-risk pregnant women between 38 and 40 weeks of gestation with cephalic presentation participated. They performed breast stimulation for 3 days with an attendant midwife in a single maternity hospital. Each breast was stimulated for 15 minutes for a total of 1 hour per day. Saliva was collected 10 minutes before the intervention and 15, 30, 60, 75, and 90 minutes after the intervention, yielding 18 samples per woman. RESULTS: Among a total of 282 saliva samples from the 16 participants, OT level was measured in 142 samples (missing rate: 49.6%). The median OT level showed the highest values on day 3 of the breast stimulation, with a marked increase 30 min after the intervention. In the mixed models after multiple imputation for missing data, the OT level on the first day of intervention was significantly lower than that on the third day of intervention. Fatigue from breast stimulation decreased on subsequent days, and most of the women (75%) felt no discomfort with the protocol. Uterine hyperstimulation was not observed. CONCLUSION: Following a 3-day breast stimulation protocol for spontaneous onset of labor, the mean OT level showed the highest values on day 3. The breast stimulation intervention protocol showed good feasibility in terms of practicality and acceptability among the pregnant women. Additional large-scale studies are warranted to confirm the protocol's effectiveness.

Decay of photoacoustic signals from biological tissue irradiated by near infrared laser pulses.

We describe the phenomenon of a sudden decrease in the amplitude of photoacoustic signals arising from nanosecond laser pulse irradiation of biological samples, measured in vitro. Several dental enamel and chicken/turkey breast samples are examined. Moderate optical energy densities (i.e., about 300 mJ/cm2) are used, typical of those exploited in photoacoustic investigations. Measurements show a rapid decay of photoacoustic signals within the first few laser pulses absorbed by the sample. This phenomenon indicates that laser irradiation interacts with biological samples, causing long-term physical changes that can be attributed to a reduction of optical absorption within the samples.

Design, fabrication, and characterization of a tissue-equivalent phantom for optical elastography.

We suitably adapt the design of a tissue-equivalent phantom used for photoacoustic imaging to construct phantoms for optical elastography. The elastography phantom we consider should have optical properties such as scattering coefficient, scattering anisotropy factor, and refractive index; mechanical properties such as storage and loss modulus; and acoustic properties such as ultrasound velocity, attenuation coefficient, and acoustic impedance to match healthy and diseased tissues. The phantom is made of poly (vinyl alcohol) (PVA) and its mechanical, optical, and acoustic properties are tailored by physical cross-linking effected through subjecting a suitable mix of PVA stock and water to a number of freeze-thaw cycles and by varying the degree of hydrolysis in the PVA stock. The optical, mechanical, and acoustic properties of the samples prepared are measured by employing different techniques. The measured variations in the values of optical scattering coefficient, scattering anisotropy factor, and refractive index and storage modulus are found to be comparable to those in normal and diseased breast tissues. The acoustic properties such as sound speed, acoustic attenuation coefficient, and density are found to be close to the average values reported in the literature for normal breast tissue.

Novel magnetic heating probe for multimodal cancer treatment.

PURPOSE: Multifunctional materials consisting of polymers and magnetic nanoparticles (MNPs) are highly sought after in the field of biomedical engineering. These materials offer new opportunities for the development of novel cancer treatment modalities that can increase the efficacy of cancer therapy. In this paper, a novel probe for multimodal cancer treatment is proposed and analyzed. The probe is essentially a cannula with two main parts: a distal heat generating tip made of a magnetic nanocomposite and a proximal insulated shaft. METHODS: A description of the concept and functional operations of the probe is presented. In an effort to assess its feasibility, the authors evaluated the ability of probe tip (made of PMMA-Fe3O4 nanocomposite) to generate heat in biological tissue using alternating magnetic field (AMF) parameters (field strength and frequency) that are acceptable for human use. Heat generation by MNPs was determined using the linear response theory. The effects of Fe3O4 volume fraction on heat generation as well as treatment time on the thermal dose were studied. The finite element method model was tested for its validity using an analytical model. RESULTS: Lesions were revealed to have an ellipsoidal shape and their sizes were affected by treatment time. However, their shapes remained unchanged. The comparison with the analytical model showed reasonably a good agreement to within 2%. Furthermore, the authors' numerical predictions also showed reasonable agreement with the experimental results previously reported in the literature. CONCLUSIONS: The authors' predictions demonstrate the feasibility of their novel probe to achieve reasonable lesion sizes, during hyperthermic or ablative heating using AMF parameters (field strength and frequency) that are acceptable for human use.

Relation of plasma oxytocin and prolactin concentrations to milk production in mothers of preterm infants: influence of stress.

Responses of oxytocin and PRL to mechanical breast pumping and the influence of physiological indicators of stress were measured at 2, 4, and 6 weeks postpartum to determine potential causes of inadequate milk production in 18 women with prematurely delivered, nonnursing (<1500 g) infants. Median milk production was similar to that reported in breastfeeding mothers, but a third of mothers were producing less than half as much by week 6. Plasma oxytocin was similar to that previously reported for breastfeeding mothers. The oxytocin area under the curve (AUC) for breast-pumping sessions (70 min) was correlated at each occasion (r = 0.37, 0.58, and 0.55, respectively) with milk yield. Unlike reports of PRL levels in breast-feeding women, PRL AUC declined between weeks 2 and 6 weeks postpartum (P = 0.03); significant increases in plasma PRL occurred in response to pumping at 2 and 4 weeks, but not at 6 weeks. Salivary amylase, a measure of alpha-adrenergic activity, was highly negatively correlated on each occasion with PRL AUC (r = -0.58, -0.68, and -0.86, respectively), but not with oxytocin. Salivary cortisol was negatively correlated to a lesser degree. We hypothesize that deficiencies in preterm lactation are mediated in part upon stress-induced suppression of PRL secretion through an adrenergic mechanism.

Developmental aspects of parathyroid hormone-related protein biology.

Parathyroid hormone-related protein (PTHrP) has been discovered as a parathyroid hormone (PTH)-like factor responsible for the humoral hypercalcaemia of malignancies. Further studies revealed that PTHrP is ubiquitously expressed, in mature as well as in developing normal tissues from various species. Although not completely understood, the biological roles of PTHrP concern a variety of domains, including calcium phosphorus metabolism and bone mineralization, smooth muscle relaxation, cell growth and differentiation, and embryonic development. As a poly-hormone, PTHrP is now acknowledged to act via the paracrine, autocrine, and even the intracrine pathways. This review focuses on the main developmental features of the biology of PTHrP. During embryonic development, PTHrP is considered to be involved as a growth factor that promotes cell proliferation and delays cell terminal maturation. PTHrP has been shown to intervene in the development of various tissues and organs such as the skeleton, skin, hair follicles, tooth, pancreas, and the kidney. In addition, through its midregion sequence, which is able to promote an active transplacental calcium transport, PTHrP may intervene indirectly in the mineralization of the foetal skeleton. PTHrP has also been shown to be necessary for the normal development of the mammary gland, while huge amounts of PTHrP are found in the human milk. Finally, observations of physiologic, vasodilating effects of PTHrP in the kidney suggest its involvment in the control of renal hemodynamics, especially in the perinatal period.

Breast implants, Hippocrates, and burden of proof.

The author argues that the burden of proof for the safety and efficacy of some medical devices properly lies with the manufacturers of the devices and with the providers who use them. The controversy over breast implants is used as a case for this shift in the burden of proof of safety and effectiveness.

WAXS fat subtraction model to estimate differential linear scattering coefficients of fatless breast tissue: phantom materials evaluation.

PURPOSE: Develop a method to subtract fat tissue contributions to wide-angle x-ray scatter (WAXS) signals of breast biopsies in order to estimate the differential linear scattering coefficients µ(s) of fatless tissue. Cancerous and fibroglandular tissue can then be compared independent of fat content. In this work phantom materials with known compositions were used to test the efficacy of the WAXS subtraction model. METHODS: Each sample 5 mm in diameter and 5 mm thick was interrogated by a 50 kV 2.7 mm diameter beam for 3 min. A 25 mm(2) by 1 mm thick CdTe detector allowed measurements of a portion of the θ = 6° scattered field. A scatter technique provided means to estimate the incident spectrum N(0)(E) needed in the calculations of µ(s)[x(E, θ)] where x is the momentum transfer argument. Values of [Formula: see text] for composite phantoms consisting of three plastic layers were estimated and compared to the values obtained via the sum [Formula: see text], where ν(i) is the fractional volume of the ith plastic component. Water, polystyrene, and a volume mixture of 0.6 water + 0.4 polystyrene labelled as fibphan were chosen to mimic cancer, fat, and fibroglandular tissue, respectively. A WAXS subtraction model was used to remove the polystyrene signal from tissue composite phantoms so that the µ(s) of water and fibphan could be estimated. Although the composite samples were layered, simulations were performed to test the models under nonlayered conditions. RESULTS: The well known µ(s) signal of water was reproduced effectively between 0.5 < x < 1.6 nm(-1). The [Formula: see text] obtained for the heterogeneous samples agreed with [Formula: see text]. Polystyrene signals were subtracted successfully from composite phantoms. The simulations validated the usefulness of the WAXS models for nonlayered biopsies. CONCLUSIONS: The methodology to measure µ(s) of homogeneous samples was quantitatively accurate. Simple WAXS models predicted the probabilities for specific x-ray scattering to occur from heterogeneous biopsies. The fat subtraction model can allow µ(s) signals of breast cancer and fibroglandular tissue to be compared without the effects of fat provided there is an independent measurement of the fat volume fraction ν(f). Future work will consist of devising a quantitative x-ray digital imaging method to estimate ν(f) in ex vivo breast samples.

Evolution of immune functions of the mammary gland and protection of the infant.

Abstract The evolution of immunological agents in milk is intertwined with the general aspects of the evolution of the mammary gland. In that respect, mammalian precursors emerged from basal amniotes some 300 million years ago. In contrast to the predominant dinosaurs, proto-mammals possessed a glandular skin. A secondary palate in the roof of the mouth that directed airflow from the nostrils to the oropharynx and thus allowed mammals to ingest and breathe simultaneously first appeared in cynodonts 230 million years ago. This set the stage for mammalian newborns to nurse from the future mammary gland. Interplays between environmental and genetic changes shaped mammalian evolution including the mammary gland from dermal glands some 160 millions of years ago. It is likely that secretions from early mammary glands provided nutrients and immunological agents for the infant. Natural selection culminated in milks uniquely suited to nourish and protect infants of each species. In human milk, antimicrobial, anti-inflammatory, and immunoregulatory agents and living leukocytes are qualitatively or quantitatively different from those in other mammalian milks. Those in human milk compensate for developmental delays in the immunological system of the recipient infant. Consequently, the immune system in human milk provided by evolution is much of the basis for encouraging breastfeeding for human infants.

An elastically compressible phantom material with mechanical and x-ray attenuation properties equivalent to breast tissue.

We have developed a novel phantom material: a solution of polyvinyl alcohol (PVAL) in ethanol and water, freeze-thawed to produce a solid yet elastically compressible gel. The x-ray attenuation and mechanical properties of these gels are compared with published measurements of breast tissue. Gels with PVAL concentrations from 5 to 20% w/v were produced. The linear x-ray attenuation coefficients of these gels range from 0.76 to 0.86 cm(-1) at 17.5 keV, increasing with PVAL concentration. These values are very similar to the published values of breast tissue at this energy, 0.8-0.9 cm(-1). Under compression cancerous breast tissue is approximately ten times stiffer than healthy breast tissue. The Young's moduli of the gels increase with PVAL concentration. Varying the PVAL concentration from 7.5 to 20% w/v produces gels with Young's moduli from 20 to 220 kPa at 15% strain. These values are characteristic of normal and cancerous breast tissue, respectively.

Fractional derivative models for ultrasonic characterization of polymer and breast tissue viscoelasticity.

The viscoelastic response of hydropolymers, which include glandular breast tissues, may be accurately characterized for some applications with as few as 3 rheological parameters by applying the Kelvin-Voigt fractional derivative (KVFD) modeling approach. We describe a technique for ultrasonic imaging of KVFD parameters in media undergoing unconfined, quasi-static, uniaxial compression. We analyze the KVFD parameter values in simulated and experimental echo data acquired from phantoms and show that the KVFD parameters may concisely characterize the viscoelastic properties of hydropolymers. We then interpret the KVFD parameter values for normal and cancerous breast tissues and hypothesize that this modeling approach may ultimately be applied to tumor differentiation.

Physiological basis of longitudinal changes in human milk yield and composition.

Currently in Western Australia more than 85% of women are breast-feeding after discharge from the hospital. Breast milk is the only fluid consumed by 64% of infants at 6 months of age, and 25% of infants are still breast-fed at 12 months of age. Therefore, many Western Australian mothers have optimized the art of breast-feeding and thereby provide an ideal population for studies of the physiological, pathological, and pharmacological factors affecting lactation. Reports from other countries conclude that the maximum milk yield of well-nourished women is 700-900 ml/24 h. Our studies have found the average milk yield of mothers breast-feeding single infants was in excess of 1100 ml/24 h for the first 6 months of lactation. Furthermore, mothers breast-feeding twins produced in excess of 2100 ml/24 h over this period, which demonstrates that the potential milk yield for mothers is much higher than 700-900 ml/24 h. Substantial alterations in the composition of breast milk occur at the initiation of lactation and after the cessation of suckling. During established lactation it is generally believed that there is only a slight variation in the composition of the milk. However, intensive studies have revealed acute changes in the concentrations of lactose, glucose, sodium, potassium, and chloride 5-6 days before and 6-7 days after ovulation. These findings suggest that unknown hormonal events associated with the reproductive cycle in women also influence breast milk composition.

Mammalian development in a changing environment: exposure to endocrine disruptors reveals the developmental plasticity of steroid-hormone target organs.

Recent findings in the field of environmental endocrine disruption have revealed that developmental exposure to estrogenic chemicals induces morphological, functional, and behavioral anomalies associated with reproduction. The aim of the present study was to determine the effects of in utero exposure to low doses of the estrogenic chemical bisphenol A (BPA) on the development of the female reproductive tissues and mammary glands in CD-1 mice. Humans are exposed to BPA, which leaches from dental materials and plastic food and beverage containers. Here we report that prenatal exposure to BPA induces alterations in tissue organization within the ovaries and mammary glands and disrupts estrous cyclicity in adulthood. Because estrogen receptors are expressed developmentally in these estrogen-target organs, we propose that BPA may directly affect the expression of genes involved in their morphogenesis. In addition, alterations in the sexual differentiation of the brain, and thus the hypothalamic-pituitary-gonadal axis, may further contribute to the observed phenotype. The emerging field of endocrine disruptors promises to provide new insights into the mechanisms underlying the development of hormone-target organs and demonstrates that the environment plays important roles in the making of phenotypes.

A breast pre-cancer test? Preliminary results based on a breast temperature rhythm abnormality during the menstrual cycle.

This study documents the menstrual thermal cycle of 16 breasts considered at normal risk for breast (8 women) and 15 breasts considered at high risk for breast cancer (i.e. ipsilateral N = 7 or contralateral N = 8 to a previously [locally] excised carcinoma). The surface thermometry studies were carried out for 1 1/2 hours each evening for one menstrual cycle using a special automated instrumentation, the 'Chronobra', with the subject at home. The surface temperatures were adjusted to remove the fluctuations in arterial blood temperature during the menstrual cycle, so that they are thought to represent a breast-specific menstrual rhythm. Across subjects, the cycles were synchronised by the day of the progesterone peak obtained by radioimmunoassay of saliva collected daily. Following ovulation, the normal risk breasts exhibited a steady rise of temperature for 14 days to a well defined peak. In contrast the 'high-risk' breasts exhibited a continuous hyperthermia with smaller peaks. Linear discrimination analysis by multiple regression achieved a complete separation of the individual normal and 'high-risk' data sets. The method shows promise for the development of a non-invasive screening test for breast pre-cancer in premenopausal women.

Interactions between tumoral MCF7 cells and fibroblasts on matrigel and purified laminin.

A reconstituted basement membrane (matrigel) and/or fibroblasts promote the growth of human breast tumors in athymic nude mice. We have investigated in vitro the effect of matrigel or purified glycoproteins (laminin and fibronectin) on tumoral MCF7 cells-fibroblasts interactions. In coculture on matrigel, MCF7 cells organized into clusters attached on top of fibroblasts aggregates. During the process resulting in tumor cells-fibroblasts aggregation, fibroblasts actively migrated while MCF7 cells were passively transported. Using purified proteins, specific antibodies and synthetic peptides, we show that cell aggregation induced by immobilized and soluble laminin is antagonized by exogenous fibronectin or fibronectin synthesized by fibroblasts.