Computer-Aided Detection of Quantitative Signatures for Breast Fibroepithelial Tumors Using Label-Free Multi-Photon Imaging.

Fibroadenomas (FAs) and phyllodes tumors (PTs) are major benign breast tumors, pathologically classified as fibroepithelial tumors. Although the clinical management of PTs differs from FAs, distinction by core needle biopsy diagnoses is still challenging. Here, a combined technique of label-free imaging with multi-photon microscopy and artificial intelligence was applied to detect quantitative signatures that differentiate fibroepithelial lesions. Multi-photon excited autofluorescence and second harmonic generation (SHG) signals were detected in tissue sections. A pixel-wise semantic segmentation method using a deep learning framework was used to separate epithelial and stromal regions automatically. The epithelial to stromal area ratio and the collagen SHG signal strength were investigated for their ability to distinguish fibroepithelial lesions. An image segmentation analysis with a pixel-wise semantic segmentation framework using a deep convolutional neural network showed the accurate separation of epithelial and stromal regions. A further investigation, to determine if scoring the epithelial to stromal area ratio and the SHG signal strength within the stromal area could be a marker for differentiating fibroepithelial tumors, showed accurate classification. Therefore, molecular and morphological changes, detected through the assistance of computational and label-free multi-photon imaging techniques, enable us to propose quantitative signatures for epithelial and stromal alterations in breast tissues.

Non-traumatic bilateral rectus sheath hematoma during septic disseminated intravascular coagulation.

A non-traumatic abdominal wall hematoma is rare, and occurs occasionally due to coughing, physical activity, or antithrombotic/anticoagulant therapy. The condition is usually unilateral; however, rare bilateral cases have been reported. Here, we report a rare case of a non-traumatic bilateral rectus sheath hematoma. The patient was a 60-year-old woman who was urgently admitted to our hospital due to the occurrence of pneumonia during postoperative chemotherapy for breast cancer. Because she exhibited disseminated intravascular coagulation, a therapy with antibacterial agents, thrombomodulin alpha, and catecholamines was initiated. During hospitalization, hemorrhagic shock due to hematomas in both rectus abdominis muscles was observed without any discernible cause. Subsequent emergency angioembolization was successful, and abdominal computed tomography performed 3 months after the onset of the rectus sheath hematoma confirmed a reduction in the hematoma size.

[A Case of Breast Cancer during Pregnancy Treated with Neoadjuvant Chemotherapy during Pregnancy].

Although breast cancer during pregnancy is relatively rare, the number of such cases has risen in recent years owing to an increase in mean childbirth age and the increasing prevalence of breast cancer. Here we report the case of a 37-year-old breast cancer patient who received neoadjuvant chemotherapy during pregnancy. The woman previously consulted an outside physician after noting a mass in her right breast at 25 weeks' gestation. Breast ultrasonography revealed a right breast tumor and axillary lymphadenopathy. A histopathological examination indicated right breast cancer and axillary lymph node metastasis. She was referred to our department for pregnancy management. Chest X-rays and abdominal ultrasonography were utilized in the search for metastases. She received 2 courses of doxorubicin and cyclophosphamide(AC)therapy during pregnancy and gave birth via cesarean section at 35 weeks' gestation. After delivery, the AC was resumed. The patient completed a total of 4 courses of AC followed by 4 courses of docetaxel (dosed every 3 weeks). She underwent total right mastectomy and axillary dissection; because the tumor was BRCA2 mutation-positive, a risk-reducing salpingo- oophorectomy was also performed. Adjuvant therapy included radiotherapy and tamoxifen but no luteinizing hormone- releasing hormone agonists. At the time of this writing more than 1 year post-surgery, she has not experienced recurrence; although the infant has a congenital clubfoot, she suffers from no other cognitive or developmental delays.

A Case of a Rapidly Growing Granulocyte Colony-Stimulating Factor-Producing Squamous Cell Carcinoma of the Breast.

A 34-year-old woman with a rapidly growing right breast mass visited our hospital. The mass was diagnosed as a right breast cancer (cT3N1M0 stage ⦀A). Her serum leucocyte count and C-reactive protein levels were high, and she had persistent fever. However, serum procalcitonin and ß-D-glucan levels were normal, and no apparent infection focus was detected, although her serum granulocyte colony-stimulating factor (G-CSF) level was markedly elevated to 42.7 pg/mL. Therefore, a G-CSF-producing breast cancer was suspected. A pathological analysis of the surgical specimen revealed a squamous cell carcinoma of the breast (pT2N0 [i+] M0 stage ∥A). Right mastectomy (with the resection of the pectoralis major muscle), axillary lymph node dissection, and split layer grafting were performed. The leucocyte count and serum G-CSF level decreased on postoperative day (POD) 1 and normalized on POD 6. As adjuvant chemotherapy, 4 cycles of a combination chemotherapy with adriamycin and cyclophosphamide and 12 cycles of weekly paclitaxel were administered. After chemotherapy, the patient also underwent postmastectomy radiotherapy. Currently, 30 months after surgery, the patient is alive and well with neither progression nor distant metastasis. G-CSF-producing breast cancers tend to rapidly grow such as in the current case; thus, surgery should be performed immediately, followed by appropriate adjuvant treatment.

CNKSR1 serves as a scaffold to activate an EGFR phosphatase via exclusive interaction with RhoB-GTP.

Epidermal growth factor receptor (EGFR) and human EGFR 2 (HER2) phosphorylation drives HER2-positive breast cancer cell proliferation. Enforced activation of phosphatases for those receptors could be a therapeutic option for HER2-positive breast cancers. Here, we report that degradation of an endosomal small GTPase, RhoB, by the ubiquitin ligase complex cullin-3 (CUL3)/KCTD10 is essential for both EGFR and HER2 phosphorylation in HER2-positive breast cancer cells. Using human protein arrays produced in a wheat cell-free protein synthesis system, RhoB-GTP, and protein tyrosine phosphatase receptor type H (PTPRH) were identified as interacting proteins of connector enhancer of kinase suppressor of Ras1 (CNKSR1). Mechanistically, constitutive degradation of RhoB, which is mediated by the CUL3/KCTD10 E3 complex, enabled CNKSR1 to interact with PTPRH at the plasma membrane resulting in inactivation of EGFR phosphatase activity. Depletion of CUL3 or KCTD10 led to the accumulation of RhoB-GTP at the plasma membrane followed by its interaction with CNKSR1, which released activated PTPRH from CNKSR1. This study suggests a mechanism of PTPRH activation through the exclusive binding of RhoB-GTP to CNKSR1.

Metaplastic carcinoma of the breast and BRCA1 germline mutation: a case report and review.

BACKGROUND: Metaplastic carcinoma of the breast consists of both invasive ductal carcinoma and metaplastic carcinoma. This rare subtype of cancer has a poor prognosis. The development of metaplastic breast cancer and relationship with BRCA1 are not well known. Here, we report a rare case of germline BRCA1 mutation-positive breast cancer with chondroid metaplasia. CASE PRESENTATION: A 39-year-old Japanese woman with a family history of breast cancer in her mother and ovarian cancer in her maternal grandmother consulted at our hospital with a left breast mass. Needle biopsy for the mass was performed, leading to a diagnosis of invasive breast cancer with chondroid metaplasia. We performed left mastectomy + sentinel lymph node biopsy + tissue expander insertion and replaced with a silicone implant later. Pathological examination revealed that the patient had triple-negative breast cancer. Four courses of doxorubicin+ cyclophosphamide therapy were performed as adjuvant therapy after surgery. We performed genetic counseling and genetic testing, and the results suggested the germline BRCA1 mutation 307 T> A (L63*). She has currently lived without a relapse for 2 years post-surgery. CONCLUSIONS: There have been only 6 cases of metaplastic breast carcinoma with germline BRCA1 mutations including our case. Patients with BRCA1 mutations may develop basal-like subtypes or M type of triple-negative breast cancer besides metaplastic breast cancers.

Dual-energy computed tomography for evaluation of breast cancer: value of virtual monoenergetic images reconstructed with a noise-reduced monoenergetic reconstruction algorithm.

PURPOSE: To evaluate the image quality and lesion visibility of virtual monoenergetic images (VMIs) reconstructed using a new monoenergetic reconstruction algorithm (nMERA) for evaluation of breast cancer. MATERIALS AND METHODS: Forty-two patients with 46 breast cancers who underwent 4-phasic breast contrast-enhanced computed tomography (CT) using dual-energy CT (DECT) were enrolled. We selected the peak enhancement phase of the lesion in each patient. The selected phase images were generated by 120-kVp-equivalent linear blended (M120) and monoenergetic reconstructions from 40 to 80 keV using the standard reconstruction algorithm (sMERA: 40, 50, 60, 70, 80) and nMERA (40 +, 50 +, 60 +, 70 +, 80 +). The contrast-to-noise ratio (CNR) was calculated and objectively analyzed. Two independent readers subjectively scored tumor visibility and image quality each on a 5-point scale. RESULTS: The CNR at 40 + and tumor visibility scores at 40 + and 50 + were significantly higher than those on M120. The CNR at 50 + was not significantly different from that on M120. However, the overall image quality score at 40 + was significantly lower than that at 50 + and on M120 (40 + vs M120, P < 0.0001 and 40 + vs 50 +, P = 0.0001). CONCLUSIONS: VMI reconstructed with nMERA at 50 keV is preferable for evaluation of patients with breast cancer.

L233P mutation in the bovine leukemia virus Tax protein depresses endothelial cell recruitment and tumorigenesis in athymic nude mice.

Bovine leukemia virus (BLV) can be divided into two categories based on the amino acid at position 233 in the Tax protein, which probably plays a crucial role in leukemogenesis. We show here that a rat fibroblast cell line stably expressing L233-Tax formed significantly larger tumors than P233-Tax-expressing cells in a murine xenograft study. Although the microvessel density was comparable in both tumors, visible blood vessel invasion was observed only on tumors from L233-Tax-expressing cells. Endothelial cell tube formation assays using human umbilical vein endothelial cells showed no significant difference in angiogenic activity between conditioned medium from L233- and P233-Tax-expressing cells, whereas in vitro chemotaxis assays revealed that only L233-Tax-expressing cells produced a chemoattractant for endothelial cells. Since pathological neovascularization can occur from the recruitment of endothelial progenitor cells, these results suggest that L233-Tax-expressing cells recruit murine endothelial progenitor cells and promote neovascularization to support tumor growth. BLV-infected lymphoma cells may also recruit bovine endothelial progenitor cells to promote neovascularization. The findings of this study are consistent with our previous observation that BLV carrying P233-Tax has a significantly longer incubation period for developing tumors than the virus carrying L233-Tax and provide insight into the function of Tax in leukemogenesis by BLV.

Patient-related Risk Factors for Nausea and Vomiting with Standard Antiemetics in Patients with Breast Cancer Receiving Anthracycline-based Chemotherapy: A Retrospective Observational Study.

PURPOSE: The aim of this study was to identify a high-risk or low-risk population for chemotherapy-induced nausea and vomiting among patients with breast cancer treated with a current standard 3-drug antiemetic regimen and receiving anthracycline. METHODS: We analyzed data from chemotherapy-naive Japanese patients with breast cancer, who had received the first cycle of anthracycline-based regimen and were treated with a 3-drug combination of aprepitant, palonosetron, and dexamethasone. This study was carried out at Ehime University Hospital (Toon, Japan) using electronic medical records from May 2011 to June 2017. The primary end point was complete response (CR), which was defined as no emesis and no use of rescue medication. FINDINGS: A total of 103 patients were included in this study. The percentages of patients who had a CR in the overall, acute, and delayed phases were 35.0%, 40.8%, and 50.5%, respectively. Multivariate logistic regression analysis revealed that age <55 years and body mass index <27.5 kg/m2 were significantly associated with an increased risk for CR failure in the overall and acute phases. In contrast, a history of alcohol habit was significantly associated with a decreased risk for CR failure in overall phase. IMPLICATIONS: The present findings suggest that, among patients with breast cancer receiving anthracycline and treated with aprepitant, palonosetron, and dexamethasone, patients younger than 55 years and having a body mass index <27.5 kg/m2 are high-risk populations for chemotherapy-induced nausea and vomiting, whereas those with a history of habitual alcohol consumption is a low-risk one.

[A Treatment Strategy against Double Presentation of Breast Cancer and Malignant Lymphoma].

Case 1 involved a 75-year-old woman with breast cancer and diffuse large B-cell lymphoma(DLBCL).Although we initially administered the R-CHOP regimen, the breast tumor increased in size and surgery had to be performed.After surgery, the R-CHOP regimen was re-initiated and DLBCL achieved clinical complete response.Case 2 involved a 74-year-old woman with breast cancer and gastric MALT lymphoma.After administration of rituximab and H. pylori eradication, a therapeutic effect was achieved in the lymphoma.A docetaxel and FEC regimen was continuously administered and surgery was performed. Case 3 involved a 62-year-old woman with breast cancer and follicular lymphoma.She presented with a history of DLBCL treatment.We performed mastectomy and sentinel lymph node biopsy, which revealed metastasis of breast cancer, and axillary lymph node dissection was subsequently performed.Considering the pathological stage, adjuvant chemotherapy was needed.We selected the TCH regimen based on her past treatment.In conclusion, it is necessary to treat patients with double presentation of breast cancer and malignant lymphoma through cooperation with different departments.

[A Case of Microangiopathic Hemolytic Anemia with Bone Marrow Carcinomatosis from Breast Cancer].

We report a case of microangiopathic hemolytic anemia(MHA)caused by metastatic breast cancer treated with weekly paclitaxel. A 58-year-old woman was diagnosed with metastatic breast cancer 2 years earlier. She was treated with various chemotherapy regimens and hormonal therapy, before being switched to fulvestrant 3 months earlier. She presented with severe anemia, and was diagnosed with MHA with bone marrow carcinomatosis following bone marrow biopsy. She was treated with weekly paclitaxel and recovered successfully. A subsequent biopsy showed that the bone marrow carcinomatosis had decreased. MHA due to breast cancer is rare and is associated with poor prognosis; however, rapid initiation of chemotherapy may be effective.

[A Case of Stage IV Breast Cancer with Long-Term Survival by Chemotherapy Developing Pulmonary Tumor Thrombotic Microangiopathy during the Treatment Course].

Pulmonary tumor thrombotic microangiopathy(PTTM)caused by pulmonary artery microscopic tumor emboli and fibrocellular and/or fibromuscular proliferation leads to progressive pulmonary hypertension and respiratory failure.The prognosis is extremely poor and most patients die shortly after onset.We report a patient with Stage IV breast cancer and long-term survival who developed PTTM during chemotherapy treatment.A 63-year-old woman with multiple metastases in her cerebellum, bone, lung, and lymph node after left breast conserving surgery started to experience dyspnea and malaise 7 years after the surgery.Two months later, she was urgently admitted to hospital because of respiratory failure and was diagnosed with pulmonary hypertension.However, pulmonary thrombosis and tumor thrombus were not observed.We clinically diagnosed her with PTTM and administered chemotherapy in addition to treatment for pulmonary hypertension.Her medical condition improved gradually and she survived for the subsequent 2 years.When observing progressive hypoxia and pulmonary hypertension without obvious pulmonary embolism findings on imaging, PTTM should be considered.Early diagnosis and immediate induction of chemotherapy for primary disease can improve the survival of patients with PTTM.

A pectic polysaccharide isolated from the roots of Bupleurum falcatum L. stimulates the tyrosine phosphorylation of lipid rafts of murine B cells.

Bupleuran 2IIc, a pectic polysaccharide isolated from the roots of Bupleurum falcatum L., was previously characterized as a T cell-independent B cell mitogen. The endo-(1-->4)-alpha-D-polygalacturonase-resistant moiety of bupleuran 2IIc (bupleuran 2IIc/PG-1) was the active site for expression of the activity, and expression of the cyclin D2 gene by bupleuran 2IIc/PG-1 may be mediated via activation of Src family tyrosine kinase, phosphatidylinositol 3-kinase (PI 3-K) and phospholipase C (PLC)-gamma followed by activation of protein kinase C (PKC) and calcium mobilization (Matsumoto et al., Int. Immunopharmacol., 5, 1373-1386 (2005)). Plasma membrane microdomains (lipid rafts) are enriched in signaling molecules and suggested to be involved in numerous cell functions, including membrane traffic and signaling. When B cells were stimulated with bupleuran 2IIc/PG-1, clustering of membrane lipid rafts was observed. To consider whether lipid rafts are implicated in bupleuran 2IIc/PG-1-mediated B cell proliferation, we analyzed the phosphorylation of tyrosine residues of proteins in lipid rafts. When murine B cells were stimulated with bupleuran 2IIc/PG-1, tyrosine phosphorylation of proteins in lipid rafts fraction was observed within 5 min. Tyrosine phosphorylation in lipid rafts fraction by bupleuran 2IIc/PG-1 was inhibited by the Src-family tyrosine kinase inhibitor, PP2. Together with previously published data, the results presented in this study suggest that activation of signaling molecules in lipid rafts by stimulation of bupleuran 2IIc/PG-1 contributes to B cell proliferation as the membrane-proximal signaling event.

Suppression of Fas-mediated apoptosis of keratinocyte cells by chikusetsusaponins isolated from the roots of Panax japonicus.

Fas-mediated apoptotic cell death of a human keratinocyte cell line, HaCaT cells, and of a murine keratinocyte cell line, Pam212 cells, was suppressed by pre-treatment with a methanol extract from the roots of Panax japonicus C.A. Meyer. Activity-guided fractionation led to the isolation of chikusetsusaponins IV, IVa, V and polysciasaponin P5 as the active ingredients. Of these compounds, chikusetsusaponin IV, was most active when applied at a concentration of 12.5 microg/mL. The intracellular hallmark events of apoptosis such as DNA fragmentation and chromatin condensation were significantly reduced by the treatment with chikusetsusaponin IV. The apoptotic cell death of Jurkat cells was also suppressed by treatment with the active saponins. These results suggest that the use of chikusetsusaponins IV, IVa, V, polysciasaponin P5, or a crude extract of P. japonicus containing these saponins is expected to relieve cutaneous symptoms caused by excessive apoptotic cell death in the skin through the Fas/FasL pathway.

A possible signal transduction pathway for cyclin D2 expression by a pectic polysaccharide from the roots of bupleurum falcatum L. in murine B cell.

Bupleuran 2IIc, a pectic polysaccharide isolated from the roots of bupleurum falcatum L., was previously characterized as a T-cell-independent B cell mitogen. This study focuses on elucidating the mechanism by which bupleuran 2IIc induces cyclin D2 production for inducing mitogenesis in murine B cells. Bupleuran 2IIc was digested with endo-alpha-(1-->4)-D-polygalacturonase and the resulting bupleuran 2IIc/PG-1 ("ramified" region) strongly stimulated cyclin D2 expression. When murine B cells were stimulated with bupleuran 2IIc/PG-1, phosphorylation of tyrosine residues of a number of proteins was observed. Cyclin D2 expression by bupleuran 2IIc/PG-1 was inhibited by the tyrosine kinase inhibitors, genistein and herbimycin A, and the Src family tyrosine kinase inhibitor, PP2, suggesting a possible role for tyrosine kinases. The stimulation by bupleuran 2IIc/PG-1 of cyclin D2 expression was significantly decreased by inhibitors, PI 3-kinase (LY294002 and Wortmannin), PLCgamma (U73122), PKC (H-7), receptor-operated calcium entry inhibitor (SK&F 96365), and calcineurin (FK506). Both PD98059 and U0126, highly selective inhibitors of MEK1 and MEK1/2, respectively, did not strongly suppress the expression of cyclin D2 after stimulation by bupleuran 2IIc/PG-1. The results suggest that (1) bupleuran 2IIc/PG-1 is the active site for induction of cyclin D2 by bupleuran 2IIc, (2) the expression of the cyclin D2 gene by bupleuran 2IIc/PG-1 may be mediated via the activation of PI 3-kinase and PLCgamma followed by activation of PKC and calcium mobilization, and (3) the ERK1/2 cascade is not a central signaling pathway for bupleuran 2IIc/PG-1-induced cyclin D2 expression.