Controlled ovarian stimulation outcomes of fertility preservation procedures in newly diagnosed breast cancer patients: a retrospective study from a single-tertiary-IVF centre.

The aim of this study was to evaluate the effectiveness of controlled ovarian stimulation (COS) using the letrozole-supplemented stimulation protocol in breast cancer (BC) patients prior to their cancer treatment. Sixty-one BC patients (Stages 0-3) who were referred to a university IVF unit for fertility preservation (FP) and underwent embryo and/or oocyte cryopreservation between 2008 - 2020 were included in this retrospective study. Time intervals between breast surgery and initial fertility consultation (IFC)/completion of FP procedures were evaluated. COS outcomes were assessed and compared between the early follicular phase (EFP) and the random-start (RS) protocols. The patients' mean age was 33.3 ± 4.9 years. The mean time interval between breast surgery and IFC was 20.6 ± 11 (day, mean ± SD) and from IFC to completion of FP procedure was 14.7 ± 5.3. Overall, 9.1 ± 5.9 mature oocytes were obtained, with a peak serum oestradiol level of 388 ± 358 pg/mL. The number of oocytes obtained (11.5 ± 9.3 vs. 10.9 ± 6.9, p = .9) and maturation rates (84.3 ± 17.5% vs. 89.2 ± 11.7, p = .5) were not statistically different between the EPF and RS protocols. The study results support that oocyte or embryo freezing can be performed effectively in a limited time period with letrozole-supplemented COS protocols before the initiation of oncological treatments in breast cancer patients.Impact statementWhat is already known on this subject? Currently, embryo and oocyte freezing are considered the most established fertility preservation (FP) methods for newly diagnosed cancer patients.What do the results of this study add? This study reports the COS outcomes of newly diagnosed breast cancer patients for FP over a period of twelve years from a single IVF unit. The results support that a considerable number of oocytes can be harvested with letrozole-supplemented COS protocol, which appears to be an effective protocol for BC patients.What are the implications of these findings for clinical practice and/or further research? There is a need for additional studies evaluating long-term follow-up of patients with their pregnancy outcomes.

The Effect of Primary Surgery in Patients with De Novo Stage IV Breast Cancer with Bone Metastasis Only (Protocol BOMET MF 14-01): A Multi-Center, Prospective Registry Study.

BACKGROUND: More evidence shows that primary surgery for de novo metastatic breast cancer (BC) prolongs overall survival (OS) in selected cases. The aim of this study was to evaluate the role of locoregional treatment (LRT) in BC patients with de novo stage IV bone only metastasis (BOM). METHODS: The prospective, multicenter registry study BOMET MF14-01 was initiated in May 2014. Patients with de novo stage IV BOM BC were divided into two groups: those receiving systemic treatment (ST group) and those receiving LRT (LRT group). Patients who received LRT were further divided into two groups: ST after LRT (LRT + ST group) and ST before LRT (ST + LRT group). RESULTS: We included 505 patients in this study; 240 (47.5%) patients in the ST group and 265 (52.5%) in the LRT group. One hundred and thirteen patients (26.3%) died in the 34-month median follow-up, 85 (35.4%) in the ST group and 28 (10.5%) in LRT group. Local progression was observed in 39 (16.2%) of the patients in the ST group and 18 (6.7%) in the LRT group (p = 0.001). Hazard of death was 60% lower in the LRT group compared with the ST group (HR 0.40, 95% CI 0.30-0.54, p < 0.0001). CONCLUSION: In this prospectively maintained registry study, we found that LRT prolonged survival and decreased locoregional recurrence in the median 3-year follow-up. Timing of primary breast surgery either at diagnosis or after ST provided a survival benefit similar to ST alone in de novo stage IV BOM BC patients.

Comparison of HER2 status determination methods in HER2 (2+) patients: Manual fluorescent in situ hybridization (FISH) vs. dual silver enhanced in situ hybridization (SISH).

HER2 amplification has been demonstrated in 15-25% of invasive breast carcinomas and can be assessed using immunohistochemical and in situ hybridization methods. Here, we compared the accuracy of dual SISH to manual FISH in HER2 (2+) breast carcinoma and evaluated the feasibility of dual SISH method in routine practice. Sixty HER2 (2+) consecutive tumor samples diagnosed between January 2009 and February 2013 were selected. Demographic, histological and immunohistochemical features and FISH results were recruited from patient records and compared to dual SISH results. Nine (15%) of the 60 tumor samples were excluded from statistical analysis due to lack of interpretable SISH signals. HER2 staining percentages by immunohistochemistry differed between 20 and 80%. HER2 amplification was shown in 7 (13.7%) and 8 (15.7%) patients by FISH and SISH, respectively. Very good agreement was observed between FISH and SISH methods (kappa value: 0.92). Significant correlation was found between HER2 staining percentage and FISH positivity, in contrast to SISH positivity (p=0.012 vs. p=0.069). Our results are consistent with previously reported literature, indicating SISH can be used to determine HER2 status. However, preanalytical and analytical problems may cause inadequate or uncountable signals, making interpretation impossible for the pathologist and highlighting the importance of standardization and quality control programs.

Orbital Metastasis from Breast Cancer: Three Cases and Brief Review of the Literature.

Metastatic disease is a relatively rare cause of orbital tumors. While many different types of primary malignancies have been documented, lung and breast cancers are the most prevalent ones among them. Herein, three cases of orbital metastasis from breast cancer are reported. The first patient had no history of primary malignancy, and the initial presentation was orbital metastasis from advanced breast cancer. The second patient had a history of neo-adjuvant chemotherapy, mastectomy, and adjuvant radiotherapy. The recurrence of the disease was diagnosed via symptomatic metastasis to orbit involving the lateral rectus muscle. The third patient had a history of mastectomy, adjuvant radiotherapy, and hormone therapy. Considering that even patients without a diagnosis of primary malignancy may present with orbital metastasis, ophthalmologists' awareness of this issue is critical.

Blood and tissue levels of persistent organic pollutants and genetic susceptibility in patients with breast cancer.

We investigated possible associations between the internal concentrations of POPs and correlations between blood and tumor tissue concentrations in patients who underwent surgery for breast cancer and breast reduction as controls. Genetic variations in CYP1A1, GSTP1, GSTM1, and GSTT1 and hOGG1 were evaluated to determine whether they represent risk factors for breast cancer. Certain POPs have been found to be associated with breast cancer development. GST-P1 polymorphism represented a significant risk for breast cancer with unadjusted OR. However, the GSTT1 null polymorphism represented a significant risk for breast cancer when OR adjusted for age and smoking status. CYP1A1 polymorphism was a significant risk factor for breast cancer, regardless of whether the OR was adjusted. These results suggest that exposure to certain POPs, GSTT1 and CYP1A1 polymorphisms, age, and smoking status are risk factors for breast cancer. In addition, the blood concentrations of some POPs represent surrogates for breast tissue concentrations.

Prediction of malignancy upgrade rate in high-risk breast lesions using an artificial intelligence model: a retrospective study

PURPOSE: High-risk breast lesions (HRLs) are associated with future risk of breast cancer. Considering the pathological subtypes, malignancy upgrade rate differs according to each subtype and depends on various factors such as clinical and radiological features and biopsy method. Using artificial intelligence and machine learning models in breast imaging, evaluations can be made in terms of risk estimation in different research areas. This study aimed to develop a machine learning model to distinguish HRL cases requiring surgical excision from lesions with a low risk of accompanying malignancy. METHODS: A total of 94 patients who were diagnosed with HRL by image-guided biopsy between January 2008 and March 2020 were included in the study. A structured database was created with clinical and radiological characteristics and histopathological results. A machine learning prediction model was created to make binary classifications of lesions as malignant or benign. Random forest, decision tree, K-nearest neighbors, logistic regression, support vector machine (SVM), and multilayer perceptron machine learning algorithms were used. Among these algorithms, SVM was the most successful. The estimations of malignancy for each case detected by artificial intelligence were combined and statistical analyses were performed. RESULTS: Considering all cases, the malignancy upgrade rate was 24.5%. A significant association was observed between malignancy upgrade rate and lesion size (P = 0.004), presence of mammography findings (P = 0.022), and breast imaging-reporting and data system category (P = 0.001). A statistically significant association was also found between the artificial intelligence prediction model and malignancy upgrade rate (P < 0.001). With the SVM model, an 84% accuracy and 0.786 area-underthe- curve score were obtained in classifying the data as benign or malignant. CONCLUSION: Our artificial intelligence model (SVM) can predict HRLs that can be followed up with a lower risk of accompanying malignancy. Unnecessary surgeries can be reduced, or second line vacuum excisions can be performed in HRLs, which are mostly benign, by evaluating on a case-by-case basis, in line with radiology-pathology compatibility and by using an artificial intelligence model.

Breast Hemangioma Evaluation with Magnetic Resonance Imaging: A Rare Case Report.

Vascular tumors are rare in the breasts, and the most common forms include hemangiomas and angiosarcomas. Hemangiomas are rare benign vascular tumors. Most of them are asymptomatic and nonpalpable clinically, and the vast majority of such lesions are detected incidentally by mammography. Breast hemangiomas are difficult to diagnose using conventional imaging modalities since their imaging findings are variable. The following is a case presentation of an asymptomatic forty-five-year old female patient who was diagnosed with a rare hemangioma. Physical examination, ultrasonography (US) and mammographic examination were normal. Dynamic contrast enhanced magnetic resonance imaging (MRI) showed a non-mass pathological enhancement. After a short-term follow up, a comparative MRI was obtained and biopsy was planned, due to the heterogeneous non-mass enhancement on MRI. Needle core biopsy with US guidance was performed, resulting in benign findings. However, because of the discordance between imaging and histopathology, an MRI-guided wire localization followed by open surgical biopsy was performed. Histopathologic evaluation reported capillary hemangioma.The imaging findings, including US, mammography and MRI, of hemangioma are reviewed and described in this case report.

Immunogenicity of a xenogeneic multi-epitope HER2+ breast cancer DNA vaccine targeting the dendritic cell restricted antigen-uptake receptor DEC205.

Breast cancer was ranked first in global cancer incidence in 2020, and HER2 overexpression in breast cancer accounts for 20-30% of breast cancer patients. Current therapeutic strategies increase the survival rate, but resistance to them occurs frequently, and there is an urgent need to develop novel treatments such as DNA vaccines which can induce a specific and long-lasting immune response against HER2 antigens. To enhance the immunogenicity of DNA vaccines, dendritic cells (DCs) can be targeted using multi-epitope proteins that provide accurate immune focusing. For this purpose, we generated a DNA vaccine encoding a fusion protein composed of 1) in silico discovered antigenic epitopes of human and rat HER2 proteins (MeHer2) and 2) a single-chain antibody fragment (ScFv) specific for the DC-restricted antigen-uptake receptor DEC205 (ScFvDEC). The xenogeneic multi-epitope DNA vaccine (pMeHer2) encodes three only T-cell epitopes, two only B-cell epitopes, and two T and B cell epitopes, and pScFvDEC-MeHer2 vaccine additionally encodes ScFvDEC introduced at the N terminus of the MeHer2. Then, mouse groups were immunized with pScFvDEC-MeHer2, pMeHer2, pScFvDEC, pEmpty, and PBS to determine the elicited immune response. pScFvDEC-MeHer2 vaccinated mice showed a strong IgG response (P < 0.0001) and pScFvDEC-MeHer2 induced a significant IgG2a increase (P < 0.01). The percentages of both IFN-γ secreting CD4 and CD8 T cells were higher in mice immunized with pScFvDEC-MeHer2 compared with the pMeHer2. pScFvDEC-MeHer2 and pMeHer2 secreted significantly higher levels of extracellular IFN-γ compared with to control groups (P < 0.0001). In addition, the IFN-γ level of the pScFvDEC-MeHer2 vaccine group was approximately two times higher than the pMeHer2 group (P < 0.0001). Overall, this study identified the pScFvDECMeHer2 construct as a potential DNA vaccine candidate, supporting further studies to be conducted on HER2+ animal models.

Abdominal cocoon syndrome as a rare cause of mechanical bowel obstruction: report of two cases.

An abdominal cocoon is an extremely rare condition, and has been reported mainly in young adolescent women as a cause of small bowel obstruction. In these patients, the small bowel is encased in a fibrous sac called an abdominal cocoon. We hereby present two cases who were diagnosed only by laparotomy and their correlation with the literature. They both received early intervention, thus preventing the need for bowel resection. The pathology of both membranes showed inflammation.

Clinical Contribution of Next-Generation Sequencing Multigene Panel Testing for BRCA Negative High-Risk Patients With Breast Cancer.

BACKGROUND: Breast cancer is the most common malignancy in women and thought to be hereditary in 10% of patients. Recent next-generation sequencing studies have increased the detection of pathogenic or likely pathogenic (P/LP) variants in genes other than BRCA1/2 in patients with breast cancer. This study evaluated pathogenic variants, likely pathogenic variants, and variants of unknown significance in 18 hereditary cancer susceptibility genes in patients with BRCA1/2-negative breast cancer. PATIENTS AND METHODS: This retrospective study included 188 high-risk BRCA1/2-negative patients with breast cancer tested with a multigene cancer panel using next-generation sequencing. RESULTS: Among 188 proband cases, 18 variants in 21 patients (11.1%) were classified as P/LP in PALB2 (n = 6), CHEK2 (n = 5), MUTYH (n = 4), ATM (n = 3), TP53 (n = 2), BRIP1 (n = 1), and MSH2 (n = 1). Three novel P/LP variants were identified. An additional 28 variants were classified as variants of unknown significance and detected in 30 different patients (15.9%). CONCLUSION: This is one of the largest study from Turkey to investigate the mutation spectrum in non-BRCA hereditary breast cancer susceptibility genes. A multigene panel test increased the likelihood of identifying a molecular diagnosis in patients with BRCA 1/2-negative breast cancer at risk for a hereditary breast cancer syndrome. More studies are needed to enable the clinical interpretation of these P/LP variants in hereditary patients with breast cancer.

CRISPR-dCas9 powered impedimetric biosensor for label-free detection of circulating tumor DNAs.

Label-free biosensors which can be integrated into lab-on-a-chip platforms have the advantage of using small volumes for rapid and inexpensive measurements contrary to label-based technologies which are often more costly and time-consuming. In this study, graphene oxide screen printed electrodes (GPHOXE) were modified by deactivated Cas9 (dCas9) proteins and synthetic guide RNA (sgRNA) as the biorecognition receptor for label-free detection of circulating tumor DNAs (ctDNA). This was achieved by detection of a tumor related mutation (PIK3CA exon 9 mutation) via sequence-specific recognition followed by electrochemical impedance spectroscopy (EIS) analysis. The biosensor showed high specificity as there was no impedance signal for other ctDNA sequences, even the single nucleotide mismatch. dCas9-sgRNA modified biosensor demonstrated linear detection limits between 2 and 20 nM for 120 bp ctDNA's in 40 s. The calibration curve showed good linearity, LOD was calculated as 0.65 nM and LOQ was calculated as 1.92 nM. Selectivity and repeatability studies were carried out in real blood samples and the recovery was higher than 96%. In conclusion, dCas9-sgRNA was effectively immobilized and optimized on GPHOXE as the selective biorecognition receptor of this ultrafast impedimetric biosensor. The CRISPR-dCas9 powered impedimetric system showed good selectivity, high repeatability and good recovery properties. This is the first literature to report the use of CRISPR/Cas technology as a label-free tool that can be used in an impedimetric system for detection of ctDNA's.

Toxoplasma gondii destroys Her2/Neu-expressing mammary cancer cells in vitro using a continuous feed medium approach.

INTRODUCTION: Toxoplasma gondii is an opportunistic protozoan and can be grown using several human cell lines. Breast cancer is the second leading cause of cancer death in women. Her2/Neu-expressing mammary cancer cell lines called TUBO can be grown in vitro. In recent years, protozoan parasites have become popular means of use in cancer therapy research. In this study, we analyzed whether T. gondii tachyzoites can destroy TUBO cells using a novel continuous feed medium approach. METHODOLOGY: Two sets of flasks (each containing four groups) containing TUBO cells were inoculated with T. gondii Ankara strain tachyzoites. First set containing 5×106 TUBO cells were inoculated with TUBO-tachyzoite ratios of 1:2, 1:1, 2:1, and 4:1 and second set containing 1×106 TUBO cells were inoculated with TUBO-tachyzoite ratios of 10:1, 100:1, 1000:1, and 2000:1. Thereafter, culture supernatants were harvested at various days until TUBO cells were destroyed and tachyzoites were counted. RESULTS: In the first and second sets of flasks, TUBO cells were destroyed between days 8 to 12 and 12 to 25, respectively. In addition, the amount of tachyzoites increased 7- 43 and 595 to 112500 times in the first and second set of flasks, respectively. CONCLUSIONS: These results show that T. gondii tachyzoites successfully destroy Her2/Neu-expressing mammary cancer cells using a continuous feed medium approach. Although this idea may be too premature for the moment, the approach defined herein may support future researchers investigating the relationship between cancer and parasites which can make important progress toward saving cancer patient lives.

Surgical Trends in Breast Cancer in Turkey: An Increase in Breast-Conserving Surgery.

PURPOSE: Breast cancer is the most frequent cancer in women, and there is a great variability in surgical practice for treating that cancer in different countries. The aims of this study were to analyze the effect of guidelines from the Turkish Federation of Breast Diseases Societies on academic institutions that have breast centers and to evaluate surgical practice in Turkey in 2018. PATIENTS AND METHODS: Between January and March 2019, a survey was sent to breast surgeons who were working in breast centers in academic institutions. The sampling frame included 24 academic institutions with breast centers in 18 cities in Turkey to evaluate interdisciplinary differences among breast centers and seven regions in Turkey regarding patients' choices, surgical approaches, and academic institutions. RESULTS: All surgeons responded to the survey, and all 4,381 patients were included. Most of the surgeons (73.9%) were working in a breast center. Multidisciplinary tumor boards were performed in 87% of the breast centers. The average time between clinical evaluation and initiation of treatment was 29 days; the longest time was in Southeast Anatolia (66 days). Only 6% of patients had ductal carcinoma in situ. Sentinel lymph node biopsy was available in every region across the country and was performed in 64.5% of the patients. In 2018, the overall breast-conserving surgery rate was 57.3% in Turkey, and it varied from 72.2% in the Black Sea region to 33.5% in Central Anatolia (P < .001). Oncoplastic breast surgery options were available at all breast centers. However, 25% of the breast centers from the Black Sea region and half the breast centers from Eastern Anatolia and the Mediterranean region did not perform this type of surgery. CONCLUSION: Increasing rates of nonpalpable breast cancer and decreasing rates of locoregional recurrences favored breast-conserving surgery, especially in developed countries. Guidelines from the Turkish Federation of Breast Diseases Societies resulted in more comprehensive breast centers and improved breast health in Turkey.

Severe decrease in SpO2 and methemoglobinemia following subareolar isosulfan blue administration and clinical relevance.

The level of axillary lymph node involvement in breast cancer is a critical decision factor for adjuvant therapy and the most important indicator of prognosis and survival. Sentinel lymph node biopsy is a minimally invasive technique with low morbidity in axillary staging of breast cancer. Radiocolloid substances (Technetium-99m) and/or blue dyes such as methylene blue or isosulfan blue are used during sentinel lymph node biopsy. Isosulfan blue stain is frequently used in sentinel lymph node biopsy and rarely causes complications. The present case report presents a severe decrease in SpO2 due to methemoglobinemia following isosulfan blue administration as well as skin and urine signs and inconsistency with clinical picture in a 67-year-old, 77 kg, ASA II female case who underwent sentinel lymph node biopsy under general anesthesia.

The value of 18F-FDG PET/CT imaging in breast cancer staging.

The National Comprehensive Cancer Network (NCCN) guidelines recommend assessment with positron emission tomography with 2-deoxy-2-[fluorine-18]fluoro-D-glucose integrated with computed tomography (18F-FDG PET/CT) in staging of breast cancer, starting from the stage IIIA. Previously, PET/CT contributed to the accurate staging from the stage IIB. Our aim is to evaluate the contribution of 18F-FDG PET/CT in staging of breast cancer patients. A total of 234 patients were retrospectively evaluated. PET/CT was performed preoperatively in 114/234 and postoperatively in 120/234 patients. Initial staging was performed based on histopathological results in 125/234 and clinical results in 109/234 patients, according to the American Joint Committee on Cancer (AJCC) classification. All patients had a normal abdominal ultrasound and chest x-ray. Following PET/CT imaging, modification in the staging was performed in patients with the metastatic findings. In 42/234 (17.9%) patients hypermetabolic extra-axillary regional lymph nodes and in 65/234 patients (27.7%) distant metastatic involvement were detected with PET/CT. Modification in the staging was applied in 82/234 (35%) patients. Patient management was changed in 69/234 (29.4%) cases. The percentage of patients with upstaging, according to each stage, was as follows: IIA: 18.6%, IIB: 30%, IIIA: 46.3%, IIIB: 68.8%, and IIIC: 20.8%. In 43/43 patients, 99mTc-methylene diphosphonate (MDP) bone scan did not show additional bone metastasis. In 5/32 patients, metastatic involvement was detected with sentinel lymph node biopsy (SLNB), but preoperative PET/CT scan did not reveal hypermetabolic lymph nodes. Although our study was limited by the referral bias and lack of homogeneity in the referral group, PET/CT still significantly contributed to the accurate staging and management of our breast cancer patients, starting from the stage IIA.

Effects of ropivacaine on pain after laparoscopic cholecystectomy: a prospective, randomized study.

Postoperative pain after laparoscopic cholecystectomy is an ongoing problem. To relieve this pain, practitioners have used many anesthetic and analgesic drugs. This study was undertaken to assess the effects of incisional and intraperitoneal administration of ropivacaine on postoperative pain and stress response in patients undergoing laparoscopic cholecystectomy. In this prospective, single-blinded, randomized study, 45 patients with ASA (American Society of Anesthesiologists) scores I and II who were about to undergo laparoscopic cholecystectomy were divided into 3 groups. After cholecystectomy, a total of 40 mL of 3.75% ropivacaine was administered pre-incisionally and intraperitoneally to patients in group 1 (n=14); pre-incisionally and intraperitoneally to patients in group 2 (n=17); and intraperitoneally and locally at incision sites to patients in group 3 (n=14). Blood levels of epinephrine and norepinephrine were examined preoperatively, 15 min after insufflation, and at the end of the operation. Visual analog pain scale scores and analgesic requirements were used for 24-h postoperative follow-up of pain levels reported by patients. No statistically significant difference was found among the 3 groups with respect to visual analog pain scale scores, total analgesic requirements, and accompanying pain, nausea, and vomiting. The earliest analgesic requirements were seen in group 2 (P<.005), and less shoulder pain was noted in group 3 (P<.005). Norepinephrine and epinephrine levels showed no statistically significant differences between the 3 groups. Administration of ropivacaine preoperatively and postoperatively for laparoscopic cholecystectomy has similar effects on postoperative pain and the stress response of patients.

Omega-3 fatty acids inhibit oxidative stress in a rat model of liver regeneration.

PURPOSE: Lipid peroxidation and consequent reactive oxygen species in the setting of oxidative stress have crucial roles in liver regeneration, which may adversely affect the regeneration itself and lead to liver failure. The aim of the current study is to investigate whether omega-3 fatty acid supplementation inhibits oxidative stress in an experimental model of liver regeneration. METHODS: Forty rats were allocated to four groups. Rats in group A received a sham operation. Rats in group B were subjected to right portal vein ligation (RPVL) and saline infusion. Rats in groups C and D were subjected to RPVL and total parenteral nutrition (TPN) with an all-in-one admixture containing a soybean oil based lipid emulsion. Rats in group D were additionally supplemented with omega-3 fatty acid infusion. Oxidative stresses in the blood and liver were measured by glutathione, superoxide dismutase, catalase, glutathione peroxidase, malondialdehyde, and nitric oxide. RESULTS: Omega-3 supplementation to the TPN solution significantly corrected alterations in the blood and tissue concentrations of oxidants and anti-oxidants during regeneration (P < 0.05). CONCLUSION: Omega-3 fatty acid supplementation to the TPN solution revealed promising results in removal of oxidative stress that emerges during liver regeneration.