Mammary Parenchyma Research Articles

Gene expression adjustment of inflammatory mechanisms in dairy cow mammary gland parenchyma during host defense against staphylococci

Abstract The aim of the study was to identify differences in the expression of splice variants of the PRMT2, LTF and C4A genes in the mammary glands of healthy dairy cows and those infected with staphylococci. An expression study was conducted on 38 Polish Holstein-Friesian dairy cows who were removed from the herd owing to subclinical and chronic mastitic or reproductive issues. Two days before slaughter, milk samples were taken for microbiological analysis and examined for the presence of bacteria. The mammary gland parenchyma samples with a predominance of secretory tissue were taken; these were divided into three groups according to the health status of the mammary gland: H (without pathogenic bacteria in milk), CoNS (with coagulase-negative staphylococci in milk), and CoPS (with coagulase-positive staphylococci in milk). Two of the investigated genes, LTF and C4A, demonstrated variants unequivocally expressed in infected tissue. Two LTF gene variants were found to be associated with cow health status, and with the type of bacteria causing mastitis (CoPS or CoNS). In addition, the expression of C4A isoforms differed with regard to mastitis etiology groups. The comprehensive evaluation of PRMT2 transcript suggested that the gene may also be involved in course of mastitis: two of four PRMT2 transcripts showed increased expression in the mammary gland of the CoPS group compared to controls. The obtained results are important for the knowledge on the etiology of bovine mastitis. The effects of the identified mastitis-relevant splice variants need to be further explored on the protein level to verify the suitability of splice variants and recognize their contribution towards the disease phenotypes and course.

Growth performance, antibody response, and mammary gland development in New Zealand dairy replacement bovine heifers fed low or high amounts of unpasteurized whole milk.

This study evaluated the influence of feeding low and high preweaning allowances of unpasteurized whole milk (MA) on intake, selected blood metabolites, antibody response, mammary gland growth, and growth of New Zealand (NZ) dairy heifers to 7 mo of age. At 10 ± 2 d of age (study day 0), group-housed (six·pen-1) heifer calves (Holstein-Friesian × Jersey) were allocated to low (4 L whole milk·calf-1·d-1; n = 7 pens) or high (8 L whole milk·calf-1·d-1; n = 7 pens) MA for the next 63 d. Calves were gradually weaned between days 63 ± 2 and 73 ± 2. Calves in each pen had ad-libitum access to clean water, pelleted calf starter, and chopped grass hay from day 1 to 91 ± 2 d. At 92 ± 2 d, all calves were transferred to pasture, grazed in a mob, and their growth and selected blood metabolites were measured until day 209. All animals were weighed weekly during the indoor period (to day 91) and then at days 105, 112, 128, 162, 184, and 209. Skeletal growth measurements and blood samples to analyze selected metabolites were collected at the start of the experiment, weaning, and then postweaning on day 91, and day 201. Specific antibodies against Leptospira and Clostridia were quantified in weeks 7, 13, and 27. Mammary glands were scanned using ultrasonography at the start of the experiment, weaning, and day 201. Feeding high vs. low amounts of MA increased the preweaning growth in heifer calves (P = 0.02) without negatively affecting postweaning average daily gain (ADG) (P = 0.74). Compared with heifers fed with low MA, high MA fed heifers had a greater increase in antibodies against Leptospira and Clostridia by 13 wk of age (P = 0.0007 and P = 0.06, respectively). By 27 wk of age, the antibody response was the same in heifers offered low or high MA. There was no effect of MA on the total size of the mammary gland, measured by ultrasonography, at weaning and 7 mo of age. However, the greater MA was associated with more mammary parenchyma (P = 0.01) and less mammary fat pad (P = 0.03) in back glands at 7 mo of age compared with heifers fed lower MA. In conclusion, feeding a high vs. a low amount of unpasteurized whole milk increased the preweaning growth of New Zealand replacement heifers without negatively affecting their ADG during postweaning under grazing conditions. Feeding more (8 vs. 4 L·d-1) unpasteurized whole milk positively affected antibody responses early in life and mammary gland composition by 7 mo of age in dairy heifers reared for pasture-based dairy systems.

A case of idiopathic granulomatous mastitis that canmimic a breast cancer

Introduction: Idiopathic granulomatous mastitis (IGM) is an uncommon and persistent inflammatory breast disease with no identified cause. It affects women of reproductive age who have previously breastfed. Diagnosis of IGM is difficult because clinical and radiological features might mimic breast cancer. We report an unusual case of IGM in a breastfeeding mother mimicking breast cancer. Observation: A 32-year-old female patient, with no particular pathological history, has been presenting for 3 months mass of the right breast with erythema. A few weeks later, she presented a suppuration and ulceration of the right breast. The breast examination revealed a painless and poorly marginated mass, mobile in relation to the muscle plane and adherent to the skin at the junction of the external quadrants measuring 40*40mm. Breast ultrasound showed a heterogeneous hypoechoic irregular mass in the outer quadrants with surrounding edema, and dilatation of the milk ducts, classified as BI-RADS 4. Histologically, there was an epithelioid and gigantocellular granulomatous reaction of the mammary parenchyma, with a polymorphic inflammatory infiltrating of lymphocytes, plasma cells, and neutrophils. The patient received corticosteroids, bromocriptine, amoxicillin and clavulanic acid associated with topical care of the lesion. The evolution was favorable with the regression of the mass after 18 months. Discussion: IGM is an uncommon and benign condition that is frequently misdiagnosed. It mostly occurs in young women during the genital period. IGM does not have any pathognomonic mammographic characteristics. Breast ultrasound can show a nodular hypoechoic image with irregular contours with a long axis parallel to the skin. IGM is characterized by lobulocentric non-caseating granulomas made up of epithelioid histiocytes mixed with Langhan's multinucleate giant cells on histological examination. The treatment of IGM is not well established. The management of the disease is based on its clinical form. IGM is a gradual and recurrent disease that has a major impact on the quality of life and requires close surveillance. Conclusion: IGM is an uncommon and persistent inflammatory disorder of the breast that can be difficult to diagnose. Breast cancer is the predominant differential diagnosis, with the possibility of an association. Increased awareness of this disorder will lead to more accurate and timely diagnosis and treatment.

A deep learning algorithm predicts milk yield and production stage of dairy cows utilizing ultrasound echotexture analysis of the mammary gland

For decades now, ultrasonography is being utilized in dairy animals’ udder health care. Recent studies have employed echotexture analysis for the examination of the mammary parenchyma and the investigation of pathological conditions. The present study aimed to investigate whether a deep neural network could predict the cows’ daily milk yield and production stage, utilizing features from echotexture analysis of mammary gland sonograms. A dataset of 3072 B-mode sonograms was created, after the ultrasonographic examination of Holstein dairy cows at successive stages of their production cycle from 2019 to 2021. Echotexture analysis was applied to these sonograms providing a dataset of features. This feature set was utilized as input for the construction of 5 models, based on the same deep neural network architecture. The first model aimed to binary classify the daily milk yield of dairy cows between “High Lactation” and “No Lactation”. The other four models aimed to binary classify the production stage of the cows between four pairs of stages: “Dry Period” – “Peak Lactation”, “Late Lactation” – “Peak Lactation”, “Late Lactation” – “Fresh Cow” and “Peri-Partum” – “Peak Lactation”. The “High Lactation – No Lactation” model achieved high performance, with the best execution reaching 91.3% Accuracy and 90.91% F1-score. Among the four production stage classification models, the “Dry Period” – “Peak Lactation” achieved the highest Accuracy and F1- score. The “Late Lactation” – “Peak Lactation”, “Late Lactation” – “Fresh Cow” and “Peri-Partum” – “Peak Lactation” models also presented promising results. These results indicate that a deep neural network can utilize features from mammary sonograms’ echotexture analysis to predict a cow’s daily milk yield and production stage. A larger dataset could lead to better performance of the models. Further progress could make the qualitative assessment of the mammary parenchyma possible and aid on-site diagnoses of pathological conditions, in the direction of smart dairy farming.

266 Bentley Lecture: Feeding and Management of Gilts to Maximize Lactation Performance

Abstract This presentation will provide an overview of nutritional and management strategies that can be used in gilts to maximize their future milking potential. Increasing sow milk yield is most critical because of the current use of hyperprolific sow lines. One important factor affecting milk yield that is often overlooked is mammary development. In swine, there are three periods of rapid mammary development, namely, from 3months of age until puberty, from 90days of gestation until farrowing, and during lactation. It is only during these periods that one can attempt to stimulate mammary development. From 90days of age until puberty, a 20% feed restriction drastically reduces mammary tissue mass whereas decreasing dietary crude protein from 18.7% to 14.4% has no effect on mammary development, and feeding the phytoestrogen genistein increases mammary cell number. During late gestation, feeding very high energy levels (10.5 Mcal ME/d) may have detrimental effects on mammary development and subsequent milk production. On the other hand, increasing SID Lys intake from 18.6g/d to 26.0g/d (via the inclusion of additional soybean meal) led to a 44% greater mass of mammary parenchyma. Increasing concentrations of the growth factor IGF-1 via porcine somatotropin injections from days 90 to 110 of gestation increased mammary parenchymal weight by 22%. Feed intake throughout gestation is important to consider because it affects body condition, and gilts that are too thin (<16mm backfat thicknes) in late gestation have reduced mammary development. Management of primiparous lactating sows is also important. During the first 2days of lactation, special care should be taken to ensure that all teats are being suckled because teats that were not previously suckled will produce less milk in second parity.

Relationship Between Somatic Cell Counts and Mammary Gland Parenchyma Ultrasonography in Buffaloes.

The aim of the present study was to determine whether the echotextural features of the mammary gland parenchyma in buffaloes during lactation at different somatic cell levels could be used to diagnose mastitis. This study was divided into two parts. In the first experiment, experimental buffaloes (n = 65) with somatic cell counts (SCC) tests (n = 94) in different seasons, including spring (n = 22), summer (n = 24), autumn (n = 37), and winter (n = 11), were used to obtain ultrasonic variables for each quarter of mammary gland that could best explain the corresponding somatic cell level. In the second part of the study, the first part's experimental results were verified by subjecting at least one-quarter udder of eight buffaloes to ultrasonography seven times during mid-July to mid-August for obtaining ultrasonic values at different somatic cell levels. The echo textural characteristics [mean numerical pixel values (NPVs) and pixel heterogeneity (pixel standard deviation, PSD)] were evaluated using 16 ultrasonographic images of each buffalo with Image ProPlus software. The effects of SCC, days in milk (DIM), scanning order (SO), season, as well as the scanning plane and udder quarter (SP + UQ) on both the PSD and NPVs of the mammary gland were significant (p < 0.05). The correlation coefficient between pre-milking sagittal PSD and somatic cell score (SCS) was the highest (r = 0.4224, p < 0.0001) with fitted linear model: y = 0.19445x (dependent variable: SCS, independent variables: pre-milking sagittal PSD; R2 = 0.84, p < 0.0001). In addition, SCC and ultrasonic of udder quarter were followed for 1 month, confirming that pre-milking sagittal PSD of mammary gland value could explain the SCC variation in milk. The current study demonstrated that the ultrasonographic examination of the udder could be one of the complementary tools for diagnosing subclinical mastitis in buffaloes.

Morphology of the mammary gland in romanian buffalo.

The gross morphology of the mammary gland in buffalo is the foundation that is necessary for the process to improve the species for agricultural use. This study determined the mammary gland's anatomical, histological and imaging structure in the Romanian buffalo. The research material was represented by ten udders, five udders from adult buffaloes and five udders from young females slaughtered in the abattoir. The udders were collected immediately after slaughtering the animals, then transported to USAMV Cluj-Napoca, Romania, examined macroscopically, histologically and through imagistic methods. The morphological examination tracked the topography, external conformation, vascularization and innervation of the mammary gland in the buffalo. The histological examination highlighted differences between the ductoacinary structures of the mammary gland parenchyma in adult buffaloes compared with young females. The imaging examination showed the detachment of the middle mammary artery and the arterial anastomoses established between the mammary arteries on the left side of the udder and those on the right side and between the arteries of the quarters on the same side. This research can be theoretical and practical support to other studies in establishing the most accurate diagnosis and therapeutic protocol in various clinical pathologies of the udder in buffalo.

Pathological evaluation of mastitis in goats and changes in milk composition

A study was aimed to evaluate the pathological alterations in caprine mastitis, its associated risk factors along with compositional changes in milk. Based on history, clinical signs, alteration in milk composition and somatic cell count (SCC), 246 lactating goats (of various breeds and ages) from 14 field flocks situated at the outskirts of Bhubaneswar (Khordha) district of Odisha were screened for mastitis. The overall prevalence of mastitis was 69.9% with 63.0% for sub-clinical mastitis (SCM) and 6.9% for clinical mastitis (CM). Risk factor analysis pertaining to proportionate distribution among mastitis affected goats (n=172) showed a higher prevalence of mastitis in nondescript goats (−48%), 3 to 4 yr age group (56%), rainy season (57%), goats in larger flocks (85%), cement concrete floor (88%), 3rd lactation (51%), goats with heavy body weight (88%) and goats with history of multiple kidding (55%). There were neutrophilia and lymphopenia along with significant (p<0.05) decrease in sodium, chloride, total protein, globulin and glucose concentrations in mastitis does. Histopathology revealed both interlobular and intralobular fibrosis, replacement of mammary parenchyma with connective tissue proliferation and atrophy of glandular tissue. Alterations in milk composition showed a decrease in fat, solid-not-fat (SNF), lactose, acidity and specific gravity values in mastitis affected goat milk. The results of present study will be helpful for the clinician in assessing the disease severity and application of suitable disease management.

Classification of Breast Cancer in Mammograms with Deep Learning Adding a Fifth Class

Breast cancer is one of the diseases of most profound concern, with the most prevalence worldwide, where early detections and diagnoses play the leading role against this disease achieved through imaging techniques such as mammography. Radiologists tend to have a high false positive rate for mammography diagnoses and an accuracy of around 82%. Currently, deep learning (DL) techniques have shown promising results in the early detection of breast cancer by generating computer-aided diagnosis (CAD) systems implementing convolutional neural networks (CNNs). This work focuses on applying, evaluating, and comparing the architectures: AlexNet, GoogLeNet, Resnet50, and Vgg19 to classify breast lesions after using transfer learning with fine-tuning and training the CNN with regions extracted from the MIAS and INbreast databases. We analyzed 14 classifiers, involving 4 classes as several researches have done it before, corresponding to benign and malignant microcalcifications and masses, and as our main contribution, we also added a 5th class for the normal tissue of the mammary parenchyma increasing the correct detection; in order to evaluate the architectures with a statistical analysis based on the received operational characteristics (ROC), the area under the curve (AUC), F1 Score, accuracy, precision, sensitivity, and specificity. We generate the best results with the CNN GoogLeNet trained with five classes on a balanced database with an AUC of 99.29%, F1 Score of 91.92%, the accuracy of 91.92%, precision of 92.15%, sensitivity of 91.70%, and specificity of 97.66%, concluding that GoogLeNet is optimal as a classifier in a CAD system to deal with breast cancer.

Expression of cytokines in dairy cattle mammary gland parenchyma during chronic staphylococcal infection

The study aim was to determine the expression of genes potentially related to chronic mastitis at the mRNA and protein levels, viz. chemokine C–C motif receptor 1 (CCR1), C–C motif chemokine ligand 2 (CCL2), C–C motif chemokine ligand 5 (CXCL5), tumor necrosis factor α (TNFα), interleukin 1β (IL-1β), interleukin 6 (IL-6), interleukin 8 (IL-8), interleukin 18 (IL-18), in bovine mammary gland parenchyma. The study examines the differences in expression of selected genes between cows with chronic mastitis caused by coagulase-positive (CoPS) or coagulase-negative staphylococci (CoNS) and those with healthy udders (H). Samples were collected from the udder quarters from 40 Polish Holstein–Friesian cows; 54 of these samples were chosen for analysis based on microbiological analysis of milk taken two days before slaughter. They were categorized into three groups: CoPS (N = 27), CoNS (N = 14) and H (N = 13). The RNA expression was analyzed by RT-qPCR and protein concentration by ELISA. No differences in the mRNA levels of seven genes (TNFα, IL-18, CCR1, IL-1β, CCL2, IL-8, IL-6) and four proteins (TNFα, IL-18, CCR1, IL-1β) were identified between the CoPS and H groups. Higher transcript levels of CXCL5 (p ≤ 0.05) gene were noted in CoPS than in H. Compared to H, higher concentrations of IL-8 and CXCL5 (p ≤ 0.05) were observed in CoPS (0.05 < p < 0.1) and CCL2 (0.05 < p < 0.1) in CoNS, while lower levels of Il-6 were found in CoPS. This may suggest that during chronic mastitis the organism stops producing pro-inflammatory cytokines, probably to protect the host tissues against their damage during prolonged infection.

Inverted-T pattern reduction mammoplasty in bilateral breast ptosis: cosmetic and oncological outcomes

Breast ptosis is directly caused by Cooper's ligament laxity, with the decline of nipple areola complex (NAC) and mammary parenchyma. Breast cancer with ptosis is always a knotty problem that can hardly be repaired by classic breast conservation surgery (BCS) ending up with a pleasing appearance. We analyzed our 12 years' experience of performing inverted-T pattern techniques to treat bilateral breast ptosis, with or without breast cancer. One hundred forty-eight breasts in 74 patients undergoing inverted-T pattern reduction mammoplasty were included in this study. Information about patients' clinical and surgical characteristics, complications, NAC sensitivity, cosmetic and oncological outcomes were collected and retrospectively analyzed. In the cohort of 57 patients with pure breast ptosis, the mean body mass index (BMI) was 25.2 kg/m2, and the mean weight of resected tissue from the left and right breast reductions were 744.9 and 756.7 g. In the cohort of 17 patients diagnosed as breast cancer with ptosis, the mean BMI was 25.1 kg/m2, and the mean weight of resected tissue were 504.1 g for left and 535.6 g for right side. The majority of repairs were performed for tumors located in the upper outer (58.8%), mostly with inferior or superomedial pedicles (90%). All the upper inner tumors were repaired with inferior pedicles. Minor complications such as seroma (8.1%), NAC epidermolysis (8.1%), delayed wound healing (4.1%) were detected postoperatively. Partial NAC necrosis occurred in one patient (1.4%). 82.4% of all the patients rated "very satisfied" or "satisfied" as the final cosmetic outcomes. NAC sensitivity was "very high" and "high" in 82.4% patients. No local occurrence, distant metastasis and mortality occurred in tumor patients. The inverted-T pattern reduction mammoplasty is a reliable technique to treat bilateral breast ptosis with a low complication rate. For cases with breast cancer, this technique can achieve both satisfying cosmetic outcomes and oncological safety.

Mammary Development in Gilts at One Week Postnatal Is Related to Plasma Lysine Concentration at 24 h after Birth, but Not Colostrum Dose.

Simple SummaryA relationship exists between a female’s early nutritional environment and her ability to produce milk when she lactates as an adult. Colostrum is the first milk available to neonates after birth. We hypothesized that differing levels of colostrum stimulate differences in very early mammary development. Despite differences in weight at 24 h and 7 days, mammary morphological development and DNA content was not found to be different between gilts fed a high versus low dose of colostrum. The rate of mammary gland protein and DNA synthesis over the first week was not different between the groups. Circulating levels of amino acids were determined after 24 h of colostrum feeding, and levels of circulating lysine were found to be related to average daily gain and mammary DNA synthetic rate. Moreover, the level of lysine was related to a lower ratio of DNA to protein synthesis, suggesting that higher lysine favored cell division versus differentiation (by leaving the cell cycle). Further studies are needed in this area.Perinatal nutrition affects future milk production. The number of mammary epithelial cells affect milk production capacity. Therefore, it was hypothesized that the level of colostrum intake affects the proliferation rate and the total number of mammary epithelial cells in the gland. The ratio of newly synthesized protein to newly synthesized DNA reflects the relative amount of cellular differentiation to cell division. The study objective was to determine the relationship between the level of colostrum intake and 24 h-level of circulating amino acid, glucose and insulin with mammary parenchyma histological features, cell division and protein synthesis over the first week postnatal. One of two standardized doses of a homogenate colostrum sample, 10% (n = 8) and 20% (n = 8) of birth bodyweight, was fed to gilts over the first 24 h postnatal. Gilts were administered deuterium oxide immediately after birth and daily to label newly synthesized DNA and proteins. Gilts were euthanized on postnatal day seven, and DNA and protein were isolated from mammary parenchyma. DNA and protein fractional synthesis (f) and fractional synthetic rate (FSR) were calculated using mass isotopomer distribution analysis. The ratio of protein f and FSR to DNA f and FSR were calculated and used to indicate the relative amounts of differentiation to cell division. Mammary morphological development was also analyzed by measuring the parenchymal epithelial area and the stromal and epithelial proliferation index on postnatal day seven. Colostrum dose was not related to any of the variables used to evaluate mammary development. However, plasma lysine levels at 24 h postnatal were positively related to average daily gain (ADG; r = 0.54, p = 0.05), DNA f (r = 0.57; p = 0.03) and DNA FSR (r = 0.57; p = 0.03) in mammary parenchyma. Plasma lysine was inversely related to the ratio of protein to DNA f and FSR (r = −0.56; p = 0.04). ADG was related to the parenchymal epithelial area and DNA and protein f and FSR (p < 0.05). These relationships support the idea that the nutritional environment affects early mammary development and that higher lysine levels in the perinatal period favored a greater degree of cell division versus differentiation in mammary of neonatal pigs and thus, warrant further investigations.

Growth performance and mammary gland development of Holstein calves fed milk with 15% solids and treated with growth promoters.

The objective of this study was to evaluate growth performance, and mammary gland parenchyma (PAR) development in Holstein female calves fed whole milk combined with milk replacer for 15% total solids (15TS) or only milk (11% TS; 11TS). The effects of zilpaterol hydrochloride (ZH) and an estrogenic implant (EI) on growth performance, serum metabolites, and PAR development in post-weaning calves (63-90days of age) fed 15TS or 11TS were also investigated. In the first phase, 78 calves were randomly divided into two groups (n = 39/group), 15TS, or 11TS (5 to 8 L/day; starter ad libitum). After weaning, calves were randomly assigned to four treatments, no EI, EI, ZH, and ZH-EI. In phase 1, 15TS calves had greater (P < 0.01) average body weight at weaning than 11TS calves (82.0 ± 5.8 vs. 74.1 ± 5.7kg). Pre-weaning average daily gain (718 ± 67 vs. 576 ± 64g/day)was higher (P < 0.01)for 15TS calves than 11TS calves. Feed/gain (1.5 ± 0.3 vs. 1.8 ± 0.3kg of DMI/kg of gain) was lower (P < 0.01) for 15TS calves than 11TS calves. PAR area (8.8 ± 3.3 vs. 5.0 ± 1.6 cm2) was greater (P < 0.01) for 15TS calves than 11TS calves. No differences (P > 0.10) in average daily gain (986 ± 79g/day across treatments) were observed in calves receiving ZH, EI, or ZH-EI fed 15TS or 11TS diets previously. However, the PAR area of 15TS calves administered ZH-EI was 2.2 times greater (P < 0.01) than 11TS calves with no ZH and CI. This study demonstrated the benefits of supplying milk with 15% TS for enhancing overall animal growth and PAR development. Further use of ZH and EI during the post-weaning period did improve mammary PAR growth.

Paciente varón con afectación mamaria recidivante por enfermedad de Rosai-Dorfman

Rosai–Dorfman disease is a histiocytic pathology with 423 described cases. It usually affects young women with bilateral cervical lymphadenopathy associated to fever, weight loss, and night sweats. The cutaneous form is the rarest, with 30 cases with breast involvement, 4 of them in men.We present the case of a 50-year-old man with a solid 24mm×18mm nodule in the upper external quadrant of the right breast with skin papules, without other symptoms. Biopsies showed nonspecific inflammatory changes. A limited exeresis was performed with adequate margins. The pathological study showed a lymphoplasmacytic and histiocytic infiltration of the mammary parenchyma, with an intense expression of S-100 and an IgG4/IgG ratio greater than 40% and emperipolesis in the histiocytes, typical findings of Rosai–Dorfman disease. A recurrence was observed at 2 years, and he was treated with methylprednisolone with a complete remission.Rosai–Dorfman disease is a rare histiocytic condition whose treatment depends on the location of the disease and the symptoms. Observation seems appropriate in cutaneous forms. Surgery is recommended in symptomatic cases or diagnostic doubts. Corticosteroid therapy is reserved for symptomatic and recurrent cases.

Long Intergenic Non-Coding RNAs in the Mammary Parenchyma and Fat Pad of Pre-Weaning Heifer Calves: Identification and Functional Analysis.

Simple SummaryThe development of mammary gland is directly related to the productivity of dairy animals. Some studies showed that feeding the enhanced plane of nutrition at pre-weaning stage are advantageous to the development of mammary gland. However, regulators which are involved in this biological process remain largely unknown. In this work, we have identified some long intergenic non-coding RNAs (lincRNAs) in mammary parenchyma (PAR) and mammary fat pad (MFP) of heifer calves under different levels of nutrition at pre-weaning stage by using the published RNA-seq database. Furthermore, those putative lincRNAs, which were highly correlated with these key protein-coding genes in mammary gland development, were highlighted. Our results not only confirmed the advantages of feeding calves with enhanced feeding plane in pre-weaning stage, but also provided fundamental base for further research on the biological processes of mammary gland development.Enhanced plane of nutrition at pre-weaning stage can promote the development of mammary gland especially heifer calves. Although several genes are involved in this process, long intergenic non-coding RNAs (lincRNAs) are regarded as key regulators in the regulated network and are still largely unknown. We identified and characterized 534 putative lincRNAs based on the published RNA-seq data, including heifer calves in two groups: fed enhanced milk replacer (EH, 1.13 kg/day, including 28% crude protein, 25% fat) group and fed restricted milk replacer (R, 0.45 kg/day, including 20% crude protein, 20% fat) group. Sub-samples from the mammary parenchyma (PAR) and mammary fat pad (MFP) were harvested from heifer calves. According to the information of these lincRNAs’ quantitative trait loci (QTLs), the neighboring and co-expression genes were used to predict their function. By comparing EH vs R, 79 lincRNAs (61 upregulated, 18 downregulated) and 86 lincRNAs (54 upregulated, 32 downregulated) were differentially expressed in MFP and PAR, respectively. In MFP, some differentially expressed lincRNAs (DELs) are involved in lipid metabolism pathways, while, in PAR, among of DELs are involved in cell proliferation pathways. Taken together, this study explored the potential regulatory mechanism of lincRNAs in the mammary gland development of calves under different planes of nutrition.

Primary breast angiosarcoma with disseminated intravascular coagulation is successfully treated with self‑subcutaneous unfractionated heparin calcium injection: A case report

Angiosarcoma is a rare sarcoma with a poor prognosis and is prone to disseminated intravascular coagulation (DIC), where DIC often interferes with chemotherapy. Primary angiosarcoma of the breast (PASB) is a type of angiosarcoma that is located in mammary parenchyma and is not associated with radiation exposure. The current study reported a 47-year-old female with DIC associated with PASB. The DIC of the patient relapsed during mono-chemotherapy with paclitaxel (PTX) after first-line anticoagulant therapy using thrombomodulin-α. The second-line danaparoid sodium therapy, followed by self-subcutaneous injection of unfractionated heparin calcium (UFH), resulted in long-term stabilization of DIC. Under this second-line anticoagulant therapy, the patient continued chemotherapy and chemoradiotherapy for >13 months in the outpatient setting without impairment of quality of life. The present case suggested that self-subcutaneous injections of UFH may be a useful therapeutic option for long-term control of DIC associated with PASB. However, further prospective clinical trails are needed to verify the efficacy of self-subcutaneous injection of UFH in similar settings.

MicroRNA expression profile in bovine mammary gland parenchyma infected by coagulase-positive or coagulase-negative staphylococci

MicroRNAs (miRNAs) are short, non-coding RNAs, 21–23 nucleotides in length which are known to regulate biological processes that greatly impact immune system activity. The aim of the study was to compare the miRNA expression in non-infected (H) mammary gland parenchyma samples with that of glands infected with coagulase-positive staphylococci (CoPS) or coagulase-negative staphylococci (CoNS) using next-generation sequencing. The miRNA profile of the parenchyma was found to change during mastitis, with its profile depending on the type of pathogen. Comparing the CoPS and H groups, 256 known and 260 potentially new miRNAs were identified, including 32 that were differentially expressed (p ≤ 0.05), of which 27 were upregulated and 5 downregulated. Comparing the CoNS and H groups, 242 known and 171 new unique miRNAs were identified: 10 were upregulated (p ≤ 0.05), and 2 downregulated (p ≤ 0.05). In addition, comparing CoPS with H and CoNS with H, 5 Kyoto Encyclopedia of Genes and Genomes pathways were identified; in both comparisons, differentially-expressed miRNAs were associated with the bacterial invasion of epithelial cells and focal adhesion pathways. Four gene ontology terms were identified in each comparison, with 2 being common to both immune system processes and signal transduction. Our results indicate that miRNAs, especially miR-99 and miR-182, play an essential role in the epigenetic regulation of a range of cellular processes, including immunological systems bacterial growth in dendritic cells and disease pathogenesis (miR-99), DNA repair and tumor progression (miR-182).

Does contrast-enhanced mammography have an impact on the detection of cancer in patients with risk of developing breast cancer?

BackgroundContrast-enhanced mammography (CEM) has been discovered to be more sensitive and specific than two-dimensional full-field digital mammography (FFDM) in both screening and diagnostic settings. The aim of the study was to assess the additive role of CEM in the detection and characterization of breast lesions in women with increased risk of developing breast cancer. This prospective study included 283 female patients with increased risk of developing breast cancer (i.e., positive family history of breast cancer, personal history of breast cancer, and heterogeneously dense mammary parenchyma) coming for either screening (n = 127/283 (49.1%)) or diagnostic (n = 156/283 (55.1%)) purpose. All patients had FFDM and CEM done, and the findings were evaluated independently; final Breast Imaging Reporting And Data System (BIRADS) classification was given for each modality. Results were then compared with histopathology or ultrasound findings with routine follow-up for normal and typically benign findings.ResultsIn this study, 283 women with mean age of 48 were enrolled. Among the studied cases regardless to a specific risk factor, 15/283 (5.3%) were diagnosed as normal, 13/283 (4.6%) as inflammatory lesions, 72/283(25.4%) as benign lesions, 6/283 (2.1%) as benign precancerous lesions, and 177/283 (62.5%) as malignant. The overall sensitivity and specificity of the CEM were 92.7 and 71.43 %, respectively, while FFDM were 80.90 and 59.05%, respectively.ConclusionContrast-enhanced mammography is a valuable screening and diagnostic imaging modality in patients with increased risk of developing breast cancer with diagnostic indices higher than mammography resulting in a significantly higher cancer detection rate.

Organization of mammary blood vessels as affected by mammary parenchymal region and estradiol administration in Holstein heifer calves

Mammary blood flow is central to mammary growth, development, and productivity, but the development of the vasculature network is poorly understood. The objective of this study was to determine how the vascular system adapts to mammary growth by inducing different levels of mammary growth and examining 2 regions of mammary parenchymal tissue. Holstein heifer calves (n = 12) received daily injections on the days immediately preceding euthanasia at 82 d of age. Treatments were control (CON), short-term estradiol (STE), and long-term estradiol (LTE). The CON calves received corn oil injections, the STE calves received 9 injections of corn oil followed by 3 injections of estradiol, and the LTE calves received 12 estradiol injections. Mammary tissues were collected from the center and edge parenchymal regions of all right rear mammary glands to quantify the tissue area of various tissue structures, the percentage of proliferating epithelial cells, and the number and form of blood vessels. Results showed that LTE calves had a greater tissue area occupied by epithelium than CON and STE calves, and the epithelial area in CON and STE calves was similar. Edge parenchyma had a greater percentage of proliferating epithelial cells than center parenchyma across all treatment groups. In the edge region, LTE calves had the greatest percentage of proliferating epithelial cells, coinciding with greater epithelial area. The number of blood vessels per unit of tissue area was greater in center than in edge parenchyma; the corresponding vessel surface area per unit of tissue area followed the same pattern. Mammary blood vessel measures were not markedly influenced by estradiol treatment. These results highlight the marked difference in the number and organization of blood vessels in different mammary parenchyma regions but indicate that the effects of estradiol on stimulating mammary epithelial proliferation does not directly translate to increasing numbers of blood vessels.

Transcripts and protein levels of CSN1S1 and CSN3 genes in dairy cattle mammary gland secretory tissue during chronic staphylococcal infection

Our objective was to determine the influence of chronic coagulase-positive staphylococci (CoPS) or coagulase-negative staphylococci (CoNS) infection on the mRNA and protein levels of two main milk proteins responsible for cheese curd quantity and quality, alpha-S1-casein (CSN1S1) and kappa-casein (CSN3). Measurements were made in cow mammary parenchyma with a prevalence of secretory tissue (MGST). Samples of MGST were collected from the separate quarters and divided into CoPS, CoNS and bacteria-free (H) groups according to the microbiological status of the quarter milk. No differences in CSN1S1 and CSN3 mRNA level were found between groups, however, CSN1S1 protein level was significantly higher in the H group than the CoNS group, and CSN3 protein level was significantly higher in H than CoPS group. Hence, while the CSN1S1 and CSN3 genes appear to be constitutively expressed at the mRNA level in dairy cow MGST during mastitis, CoNS infection negatively affected CSN1S1 protein level, and CoPS infection negatively affected CSN3 protein level. The lack of change at the mRNA level suggests that staphylococcal infection may affect the post-transcriptional or post-translational modifications.