Breast cancer medical oncology summary 2021.

Today, breast cancer has actually become merely an umbrella term that encompasses several cancers with different types of genesis, different genomic and phenotypic characteristics, different needs for systemic treatment and different prognosis. Early diagnosis and good multidisciplinary cooperation, choice of a proper treatment sequence, good supportive treatment and psychological support of the patient are crucial for a therapeutic success. The surgeon plays an important role in the treatment plan of patients with breast cancer; nevertheless, breast cancer is a systemic disease and thus as a rule, surgery alone is usually not sufficient to manage even early stages of the tumor. Surgeons as members of a multidisciplinary team need to know basic information on systemic treatment of breast cancer and have an understanding of how oncologists think; only then it will possible to achieve multidisciplinary consensus as painlessly as possible in each individual case.

Association of triple positivity with prognostic parameters and overall survival in a population-based study of 6,122 HER2-positive breast cancer patients: analysis of real-world clinical practice based on a research database.

Triple-positive breast cancer (TPBC), i.e. HER2-positive (HER2+) and hormone receptors-positive breast cancer, is a specific subgroup of breast cancers. TPBC biology is characterized by strong mutual interactions between signaling pathways stimulated by estrogens and HER2 amplification. The present study aims to carry out a population-based analysis of treatment outcomes in a cohort of hormone receptor (HR) positive and negative breast cancer patients who were treated with anti-HER2 therapy in the Czech Republic. The BREAST research database was used as the data source for this retrospective analysis. The database covers approximately 95% of breast cancer patients treated with targeted therapies in the Czech Republic. The analysis included 6,122 HER2-positive patients. The patients were divided into two groups, based on estrogen receptor (ER) or progesterone receptor (PR) positivity: hormone receptor negative (HR-) patients had both ER- and PR-negative tumors (n=2,518), unlike positive (HR+) patients (n=3,604). HR+ patients were more often diagnosed premenopausal at the time of diagnosis, presented more often at stage I or II and their tumors were less commonly poorly differentiated. The overall survival (OS) was significantly higher in subgroups of HR+ patients according to treatment setting. When evaluated by stages, significantly higher OS was observed in HR+ patients diagnosed at stages II, III, and IV and regardless of tumor grade.

Molecular and IHC analysis of head and neck carcinomas associated with HPV infection.

Head and neck squamous cell carcinomas (HNSCC) are a highly heterogenous disease which can be induced by two main carcinogens - tobacco and/or alcohol, or by HR HPV infection. This work examined 60 paraffin-embedded biopsies of head and neck carcinomas after histological verification. HPV infection, including its specific types in various HNSCC areas, was studied using multiplex qPCR. Expression levels of p16INK4A and p53 were detected by subsequent IHC analysis as being potential diagnostic markers. Based on the assumption that patients with HNSCC could benefit from anti-EGFR therapy (cetuximab), but the predictors are not yet defined, analyses of point mutations of ras genes (Kras, Nras) were carried out using multiplex qPCR and sequence analysis of the Braf gene. All statistical data were processed by Chí-x2 test.HPV infection was detected in 23.34 % of cases with HNSCC, of which 100 % were HPV 16, which is the most frequently infection found in the oropharyngeal region. Using IHC analysis, a positive expression of P16INK4A was detected in 100 % of HPV-positive HNSCC while this expression was discovered to be highly correlated with HPV infection. Furthermore, a correlation between p53 and HPV-negative HNSCC was proved. The mutation incidence was the highest in the Kras gene (codon 12 and codon 146), Nras (codon 12) and Braf. A correlation between tumor location in the oropharyngeal region and Kras mutations was proved. The HPV infection correlated with Kras mutations in case of codon 146 but on the grounds of low amount of output data, these figures could be irrelevant. In one case, c.1808 G>A, protein 603 Arg>Gln mutation was found in the Braf gene but its correlation with head and neck carcinomas has not been described yet (Tab. 2, Fig. 2, Ref. 24). Keywords: head and neck carcinomas, biopsy, HPV types, PCR, p16INK4A, p53, molecular predictors, Kras, Nras, Braf.

The use of Human Inflammatory Response and Autoimmunity RT2 lncRNA PCR Array for plasma examination in breast cancer patients prior to therapy.

Long non-coding RNAs (lncRNAs) are defined as RNA molecules longer than 200 nucleotides with poor protein-coding capacity and key functions in regulation of gene expression. Dysregulations of lncRNAs (e.g. HOTAIR and MALAT I) were detected in plasma of breast cancer (BC) patients. Plasma samples are examined as liquid biopsies for purposes of non-invasive diagnostics therefore the research of plasma lncRNAs as potential plasma biomarkers became highly topical. 84 lncRNAs were profiled in 18 plasma samples - 9 BC patients and 9 age-matched healthy - using Human Inflammatory Response & Autoimmunity RT2 lncRNA PCR Array. Total RNA from plasma samples was isolated using miRNeasy Serum/Plasma Kit. Although a pre-amplification recommended for quantification from small starting RNA amounts was used, only 3 lncRNAs (A2ML1-AS1, GAS5 and SNHG5) were detected in all plasma samples. A total of 72 lncRNAs (e.g. HOTAIR or MALAT I) were detected only in some samples and 9 lncRNAs were not detected in any samples. No significant differences were observed in levels of plasma lncRNAs between the BC patients and healthy controls despite the fact that our panel contained also the lncRNAs whose levels were previously reported as significantly different in plasma or cancer tissues (e.g. GAS5, HOTAIR, MALAT I) in BC patients. Detection of lncRNAs in plasma is due to their low concentrations quite difficult as compared with tissues. Our findings suggest that analysis of plasma lncRNAs using this technology is not suitable for use as non-invasive diagnostic tool in BC patients.

The potential roles of vesicle-enclosed miRNAs in communication between macrophages and cancer cells in tumor microenvironment.

Functional microRNA (miRNA) molecules are transported in extracellular vesicles among tumor cells and cells of the immune system. Macrophages as integral components of tumor microenvironment are known as potential contributors to tumor growth and progression. We searched for studies which could provide a direct link between the particular miRNAs transported between cancer cells and macrophages and experimental evidence of subsequent alterations in biological functions of target cells. The validated targets of such microRNAs were found using miRWalk database. These targets were further subjected to analysis by DAVID (Database for Annotation, Visualization and Integrated Discovery) to find the most prominent cellular events that could be potentially regulated in macrophages by miRNAs originated from cancer cells and vice versa. We found that the 5 miRNAs (let-7b, miR-21, miR-29a, miR-222-3p, miR-451) derived from cancer cells may together regulate 2304 target genes in macrophages. The genes involved in regulation of apoptosis, regulation of gene expression and protein transport were significantly overrepresented in this set. Four of the five sets of target genes for these individual miRNAs overlap in MYC oncogene. MYC dependent transcriptional program is responsible for cell cycle entry and regulates the inflammatory response in macrophages.Both miRNAs for which the functional transports from macrophages to cancer cells were experimental proven (miR-223, miR-142-3p) target total 684 genes including some well-known tumor suppressors like TP53 or APC. Suppression of tumor suppressor genes by miRNAs derived from macrophages may eventually contribute to cancer cell proliferation.Due to the complexity of tumor microenvironment, the altered expression profiles of its components affected by miRNA uptake from extracellular vesicles could contribute to the outcome of carcinogenesis therefore the vesicular transport of miRNAs should be studied more extensively in this context.

[Pregnancy-associated Breast Cancer]. / S tehotenstvím spojený karcinom prsu.

Pregnancy-associated breast cancer is defined as carcinoma diagnosed during pregnancy or breastfeeding, up to a year after delivery. Pregnant patients with breast cancer can be treated using procedures for non-pregnant patients, with some modifications designed to avoid damage to the fetus. Breastfeeding breast cancer patients need to stop immediately. Abortion before start of treatment or during therapy does not increase survival of these patients. Chemotherapy of breast cancer may start roughly from the second month of pregnancy. Effective treatment with anthracyclines, taxanes, and cisplatin is relatively safe for both mother and fetus. During pregnancy, patients can undergo surgery but are unable to undergo radiotherapy. Also, hormonal therapy and trastuzumab treatment is not safe for pregnant women with breast cancer. Prognosis of pregnant breast cancer patients is similar to non-pregnant patients. Worse prognosis was noted for breast cancer patients diagnosed during breastfeeding. After the cancer treatment is finished, breastfeeding is completely safe, but technical reasons it can usually only be done using the contra-lateral breast. Children of mothers who were treated for breast cancer during pregnancy do not show any worsening of physical and psychiatric parameters of development. Pregnancy following treatment for breast cancer does not affect the patients prognosis in a negative way, according to clinical studies, not even in patients who suffered from a hormonal-dependent carcinoma.Key words: breast cancer - pregnancy - breastfeeding - chemotherapy - radiotherapy - biological treatment - hormonal therapyThe author declares she has no potential conflicts of interest concerning drugs, products, or services used in the study.The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 10. 7. 2016Accepted: 20. 7. 2016.

Contribution of ABCB1 and CYP2D6 genotypes to the outcome of tamoxifen adjuvant treatment in premenopausal women with breast cancer.

Recent pre-clinical evidence suggests that the active metabolite of tamoxifen, endoxifen, is a substrate for efflux pump P-glycoprotein. The aim of our study was to evaluate, if the polymoprhisms within ABCB1 gene alter tamoxifen adjuvant treatment efficacy in premenopausal women. Totally 71 premenopausal women with estrogen receptor positive breast cancer indicated for tamoxifen adjuvant treatment were followed retrospectively for median period of 56 months. The gentic polymorphisms of CYP2D6 and ABCB1 were analyzed and potential covariates as tumor grading, staging, age at the diagnosis, comedication, quantitative positivity of ER or PR were also evaluated. Cox proportional-hazards regression model indicated that patients carrying at least one variant allele in ABCB1 rs1045642 had significantly longer time to event survival compared to wild type subjects. Non-significant trend was noted for better treatment outcome of patients carrying at least one variant allele in the SNP rs2032582, while for the CYP2D6 polymorphism poor metabolizer phenotype resulted in worse outcome in comparison to extensive metabolizers subjects with HR of 4.04 (95 % CI 0.31-52.19). Similarly, patients using CYP2D6 inhibitors had non-significantly shorter time-to-event as compared to never users resulting in hazard ratio of 2.06 (95 % CI 0.40-10.63). ABCB1 polymorphisms may affect outcome of tamoxifen adjuvant treatment in premenopausal breast cancer patiens. This factor should be taken into account in addition to the CYP2D6 polymorphism or phenotypic inhibition of CYP2D6 activity.

RAGE and its ligands in cancer - culprits, biomarkers, or therapeutic targets?

Receptor for advanced glycation end products (RAGE) plays a central role in the regulation of tissue homeostasis, regeneration and resolution of inflammation, but under pathological conditions RAGE-mediated pathways may induce diminished apoptosis, but enhanced autophagy and cell necrosis. These mechanisms may contribute to malignant transformation, cancer progression and metastases. Soluble RAGE may bind natural RAGE ligands and counteract some of the RAGE-mediated effects. Activation of RAGE was demonstrated in different types of cancer (including colon, pancreatic and breast cancer). Expression of RAGE and serum levels of soluble RAGE may serve as cancer biomarkers and strategies aimed at interfering with RAGE signaling might be promising anticancer drugs.

Predictive factors for the presence of tumor cells in bone marrow and peripheral blood in breast cancer patients.

UNLABELLED: Simultaneous detection of disseminated tumor cells (DTCs) and circulating tumor cells (CTCs) was shown to be associated with an especially poor prognosis and increased incidence of disease-related deaths in non-metastatic breast cancer patients. We analyzed the occurance of DTCs and CTCs in patients with primary breast cancer and evaluated the correlation of their presence with other prognostic markers and investigated the changes in DTCs/CTCs number at different time points during treatment.Blood of 50 patients with primary breast cancer were used for immunomagnetic separation and detection of circulating tumor cells using the commercial available system the AdnaTest Breast Cancer™ (AdnaGen GmbH, Langenhagen, Germany). Bone marrow aspirates from 50 patients were analyzed for DTCs by immunocytochemistry using the pan-cytokeratin antibody conjugated with FITC (Monoclonal Anti-Cytokeratin antibody F3418, Sigma Aldrich).DTCs were identified in 30% (15/50) and CTCs in 22% (11/50) of patients. We found that DTC positivity could point to a significantly high risk of larger primary tumor size (p-value 0.011) and significantly higher risk of lymph node involvement (p-value 0.002). For CTC positivity, no such relationship was proven. DTCs have shown significantly higher prevalence in ER/PR-negative females and in HER2-positive cases. CTCs were equally prevalent in patients with the presence and absence of standard prognostic and predictive markers such as ER, PR and HER2. We found no correlation between CTCs and DTCs findings (r = -0.097, p = 0.504). We used DTCs/CTCs analysis for therapy monitoring in a small group of 29 patients, who underwent neoadjuvant chemotherapy (NACT). We find out no significant correlation between DTCs/CTCs detection and the primary tumor response to NACT. A pathologic complete response (pCR) was achieved by 31% (9/29) of the patients in our study, however, no association was observed between pCR and the detection of DTCs after NACT.These results support the use of DTCs/CTCs analysis in early breast cancer to generate clinically useful prognostic information. The study of these cells apart from the impact on refining prognosis, has the exciting potential of individualising treatment for women with breast cancer. KEYWORDS: breast cancer, disseminated tumor cells, circulating tumor cells, bone marrow aspiration, prognostic/predictive markers, therapy monitoring.

[OPERa Study]. / Studie OPERa.

BACKGROUND: On the whole, most European and international guidelines recommend prophylactic use of granulocyte-colony stimulating factors (G-CSFs) when the risk of chemotherapy-induced febrile neutropenia (FN) in cancer patients exceeds 20%. In patients treated with intermediate-risk chemotherapy regimens the recent EORTC guidelines recommend to consider supplementary patient-related adverse risk factors such as elderly age ( 65 years) prior to administrating each cycle of chemotherapy. The primary objective of our study is to describe the most important FN risk factors that underlie the use of pegfilgrastim PP in daily practice in the Czech Republic; secon-dary endpoints include FN incidence, chemotherapy dose intensity, anti-infective agents admini-stration, hospitalization length and safety of chemotherapy regimens. PATIENTS AND METHODS: This prospective, multicenter, non-interventional study enrolled patients receiving a chemotherapy with high FN risk ( 20% according to EORTC guidelines) based on investigators` assessment. RESULTS: Data were collected on a total of 333 patients treated for breast cancer (69%), lymphoma (20%), ovarian (5%), lung (4%) and testicular cancer (1%). The most frequent indications for G-CSF prophylaxis were myelotoxic chemotherapy regimen (96%), elderly age (36%), advanced stage disease (35%), female gender (30%), cancer type (15%) and previous FN episode (12%). The overall FN incidence was 3% in patients receiving primary pegfilgrastim prophylaxis (n = 210) and 12% in patients with no pegfilgrastim PP (n = 123). CONCLUSION: The myelotoxicity of a chemotherapeutic regimen was the most significant FN risk factor identified by the inquired physicians. The second most compelling FN risk factor was elderly age and advanced stage disease. FN incidence in patients who received pegfilgrastim PP was relatively low in comparison to the commonly expected FN incidence in a population of patients receiving a chemotherapy regimen with high risk of FN.

Endotelial activation and flow-mediated vasodilation in young patients with breast cancer.

Endothelial activation and dysfunction may play a significant role in the progression of breast cancer. In our study we examined markers of endothelial activation (soluble ICAM-1, P-selectin, E-selectin) in 98 young patients with breast cancer (< 40 years). In 50 of them (and 20 age-matched controls) we also measured flow mediated vasodilation. Patients with breast cancer had significantly higher serum levels of soluble E-selectin, P-selectin and ICAM-1, P-selectin was higher in patients with larger tumors, node involvement and seemed to be apredictor of poor outcome. We were unable to find significant difference in the parameters of flow mediated vasodilation between patients with breast cancer and healthy subjects, although both peak blood flow (PBF) and flow mediated vasodilation (FMD) seemed to be skewed compared to healthy subjects toward mean and lower levels. Cluster analysis enabled us to distinguish several larger groups of patients with different degree of endothelial activation and function and different outcome. Group of patients with high E-selectin, high ICAM-1 (higher endothelial activation) and low VEGF (putative endothelial damage) had more frequently negative estrogen receptors and had worse outcome compared to the group of patients with lower E-selectin, lower ICAM-1 and mostly positive estrogen receptors. Further studies of larger groups of patients should help to identify the panel of endothelial markers which could help in predicting the outcome of young patients with breast cancer.

Proteinuria and hypertension in patients treated with inhibitors of the VEGF signalling pathway--incidence, mechanisms and management.

Anti-VEGF therapy dramatically improved the outcome of patients with renal cancer and other advanced malignancies, but may be complicated by proteinuria and hypertension. VEGF is indispensable for the normal development of glomerulus and preservation of glomerular filtration barrier. Interference with its action may result in damage to glomerular endothelial cells and (in severe cases) in renal thrombotic microangiopathy. Blood pressure and proteinuria (using dipstick) should be assessed in all patients before starting anti-VEGF therapy and regularly monitored during the treatment. Patients with severe proteinuria and/or impaired renal function should be referred to the nephrologist for further work-up. Hypertension caused by anti-VEGF therapy can be effectively treated; progression of proteinuria and/or renal dysfunction may require tapering, or even withdrawal of anti-VEGF treatment.

Disseminated and circulating tumour cells and their role in breast cancer.

Metastatic spread of the primary tumour is responsible for the vast majority of cancer-related deaths. Detection of disseminated tumour cells in the bone marrow and circulating tumour cells in the peripheral blood is correlated with early metastatic relapse in breast cancer. Positive detection of disseminated tumour cells was associated with poor overall survival of patients. Current research has been focused on integrating minimal residual disease as a prognostic and predictive tool in the management of breast cancer. Detection of disseminated tumour cells/circulating tumour cells is not yet standardized in clinical practice because of using different enrichment and detection methods. Therefore, standardization of the used methods is necessary in the future. Previous achieved findings must be verified in larger prospective multicentre studies. Further characterization of disseminated tumour cells/circulating tumour cells will be essential for developing and monitoring the efficacy of new therapeutic concepts. The aim of this review was to provide a short survey of the metastatic cascade and cancer stem cell theory, and data on the molecular and functional characterization of disseminated tumour cells/circulating tumour cells. Finally, we discuss the potential clinical impact of disseminated tumour cells/circulating tumour cells and results of several recent studies.

[Ductal approaches in mammary diagnostics]. / Intraduktální prístupy v mamární diagnostice.

OBJECTIVE: Information about new possibilities of early diagnostics in mammary lesions. TYPE OF STUDY: Review. SETTING: Gynecology-Obstetrics Clinic, 1st Medical Faculty and General Teaching Hospital, Prague. SUBJECT AND METHODS: Most malignant tumors of the breast originate from ductal epithelium. A direct examination of the ductal system, could significantly improve diagnostics of breast cancer as well as its preinvasive stages (DCIS) and to influence mortality. The concept of ductal approaches includes several techniques and ductal lavage and duscoscopy. CONCLUSIONS: Ductal approaches represent an attractive area for minimal load upon the patients. Specificity and sensitivity of these methods have some limits, which will be subject to change in relation to understanding of carcinogenesis and in a close relation to the knowledge of biomarkers, genomics and proteomics. Ductoscopy appears to be the ideal method for the future due to possibilities of direct visualization of epithelium in combination with biopsy and ductal lavage. It other advantages include minimal invasiveness, minimal risk and the origin of possible complications for the patient.

Carbonyl and oxidative stress in patients with breast cancer--is there a relation to the stage of the disease?

UNLABELLED: Oxidative and carbonyl stress may, on one hand, contribute to the progression of cancer, on the other hand, they may have some antiproliferative effects. We examined serum levels of AGEs (advanced glycation end-products), CML (carboxymethyllysine) and AOPP (advanced oxidation protein products) in 86 patients with breast cancer subdivided based on the clinical stage (TNM classification), histologic grading, expression of hormonal and C-erb B2 receptors and in 14 healthy age-matched women as controls. Breast cancer patients had higher serum concentrations of AGEs (325,581 +/- 66,037 vs. 271,322 +/- 34,826 AU, p < 0.01) even in the early stage of the disease; patients with advanced breast cancer (stage III and IV) had significantly higher both AGEs and AOPP (113.0 +/- 44.9 vs. 78.1 +/- 28.4 micromol/l, p < 0.05) levels, not only compared to controls, but also compared to stages I and II. Serum levels of AOPP were higher in patients having only weakly positive expression of C-erb 2/Her-neu compared to controls and the patients having the highest C-erb2/Her-neu expression. Serum concentrations of AGEs in patients with breast cancer correlated with the age and also with the serum concentration of AOPP. IN CONCLUSION: breast cancer patients had an early increase of AGEs (marker of the carbonyl stress) followed by further increase of AGEs and elevation of AOPP (marker of oxidative stress) in patients with progressive disease. As the clinical significance of these observations is currently uncertain further studies are clearly warranted, especially with respect to their potential therapeutic implications.

Mammaglobin A, a novel marker of minimal residual disease in early stages breast cancer.

Mammaglobin A, in contrast to other factors, is a breast specific member of uteroglobin gene family. Expression is restricted to normal and neoplastic breast epithelium. A highly homologous mammaglobin B is not specific to breast tissue. In this pilot feasibility study we examined expression of both markers for minimal residual disease in the bone marrow of patients with breast cancer. We obtained bone marrow aspirates of 34 patients with stage I (41%), II (56%) and III (3%) breast cancer who underwent either immediate complete resection of the tumor or neoadjuvant therapy with subsequent curative surgery. mRNA was isolated using QIAamp RNA blood mini kit (Qiagen). Subsequently two-step nested RT-PCR for the expression of mammaglobin A and mammaglobin B was performed. Mammaglobin A was detected in samples from 4 (12%) out of 34 patients. None of the specimens was positive for mammaglobin B. With a median follow-up of 21 month we observed only 2 recurrences, one in patient with mammaglobin A positive bone marrow.RT-PCR assay for mammaglobin A may be a useful tool for detection of occult breast cancer cells in the bone marrow. Clinical and prognostic relevance of minimal residual disease should be further investigated.

[Acute phase proteins in female patients with breast carcinoma]. / Proteiny akutní fáze u pacientek s karcinomem mammy.

BACKGROUND: Serum levels of C-reactive protein (CRP) and orosomucoid are increased in more than one half of patients with metastatic breast cancer. The information concerning their levels in non-metastatic breast cancer is inconclusive. The aim of our study was to examine some acute phase proteins (CRP, orosomucoid, prealbumin, alpha 2-macroglobulin and transferrin) in patients with various clinical stages of breast cancer before hormonal and/or chemotherapy and 3 and 12 months after its institution and to assess the influence of their levels on the outcome of patients. METHODS AND RESULTS: Altogether 59 women with breast cancer in clinical stage 0-IV, median age 62 years (47 of them after menopause) were examined. One patient was in stage 0, ten patients in stage I, 24 patients in stage IIA, 11 patients in stage IIB, five patients in stage III and eight patients in stage IV. Acute phase proteins (CRP, orosomucoid, transferrin, prealbumin and alpha 2-macroglobulin) were examined in the sera samples using microturbidimetry. Serum levels of CRP and orosomucoid were higher in patients with breast cancer in all stages compared to controls. Serum levels of CRP (resp. of orosomucoid) higher than mean +2 SD had 30.5% (resp. 39%) of patients with breast cancer. One year after the beginning of therapy serum levels of CRP and orosomucoid significantly decreased, however, in case of orosomucoid they remain higher compared to controls. Serum levels of CRP and orosomucoid correlated before therapy in patients with breast cancer one to another and also with serum levels of soluble TNF (tumour necrosis factor) receptor type I and soluble ICAM-1. IN CONCLUSION: Patients with breast cancer before hormonal therapy and/or chemotherapy had compared to controls increased serum levels of CRP and orosomucoid, however, there was no difference between stages I-III. Observed correlation between serum levels of CRP and soluble TNF receptors suggests the important role of proinflammatory cytokines in stimulating their hepatic synthesis also in patients with breast cancer. Putative prognostic role of persistently increased levels of orosomucoid in patients with non-metastatic breast cancer warrants further investigation.

[Soluble adhesion molecules in female patients with breast carcinoma]. / Solubilní adhezní molekuly u pacientek s karcinomem mammy.

BACKGROUND: Serum levels of some soluble adhesion molecules (e.g. ICAM-1, or E-selectin) are higher in some patients with breast cancer (especially metastatic) compared to controls. They bring about the increased risk of developing metastases and they can negatively influence the prognosis of patients. The aim of our study was to assess the serum levels of several soluble adhesion molecules (soluble E-selectin, P-selectin, ICAM-1, VCAM-1 and CD44s) in patients with breast cancer in clinical stage 0-IV before the beginning of hormonal therapy and/or chemotherapy. We wanted to find out if these parameters differ between various stages of breast cancer and if their levels are influenced by the therapy. METHODS AND RESULTS: We examined 59 mostly postmenopausal patients with breast cancer in clinical stage 0-IV (mean age was 62 years). One patient was in stage 0, 10 patients were in stage I, 24 patients were in stage IIA, 11 patients were in stage IIB, 5 patients were in stage III and 8 patients were in stage IV. Serum levels of soluble E-selectin, P-selectin, ICAM-1, VCAM-1 and standard form of CD44 were assessed using ELISA in sera of patients with breast cancer before the institution of hormonal therapy and/or chemotherapy and 3 and 12 months after the beginning of therapy. Compared to controls, patients with (metastatic and also non-metastatic) breast cancer before the beginning of therapy had higher serum levels of soluble E-selectin, P-selectin, ICAM-1 and standard form of CD44. Serum levels of E-selectin, ICAM-1 and also VCAM-1 significantly decreased one year after the beginning of therapy. Serum levels of soluble adhesion molecules mutually correlated and they also correlated with serum levels of soluble TNF and IL-2 receptors and some acute phase proteins (CRP and orosomucoid). CONCLUSIONS: Serum levels of some soluble adhesion molecules (E-selectin, P-selectin, ICAM-1 and CD44s) were higher in some patients with breast cancer and decreased again after hormonal therapy and/or chemotherapy. Only larger studies with longer follow-up may imply if any of these parameters (e.g. soluble P-selectin), or their combination may become useful marker of activity and/or outcome.

Salivary proteins and glycoproteins in phlebotomine sandflies of various species, sex and age.

Salivary gland proteins were studied in sandflies (Diptera: Psychodidae: Phlebotominae) by electrophoretic techniques. In Phlebotomus duboscqi Neveu-Lemaire the protein concentration was about 30 times higher in females than in males. SDS PAGE revealed eight major bands of 29-62 kDa in salivary gland extracts (SGE) from females, whereas only one band of 57kDa was detected in males. The number of protein components in SGE gradually increased with the age of females. In P. papatasi (Scopoli) the typical electrophoretic pattern was reached in 3-5 days after imago emergence, depending on the temperature at which females were maintained. All major protein components of the female SGE were present in the content of glands. Female SGE were compared in seven colonies of five Phlebotomus and Lutzomyia species; electrophoretic profiles distinguished between species and even between colonies of different geographical origin. In general, the highest variability of major protein components was observed in the 38-48kDa region. Four colonies of the subgenus Phlebotomus (P. duboscqi and P. papatasi) possessed common mobility polypeptides, the highest similarity was found between two colonies of P. papatasi. Other species tested significantly differed, specific prominent bands of 33, 35 and 38kDa were found in P. halepensis Theodor, P. perniciosus Newstead and Lutzomyia longipalpis (Lutz & Neiva), respectively. Glycoproteins in SGE of Lu. longipalpis and P. duboscqi females were identified and analysed using blotting with five lectin conjugates. Specific reaction of lectins ConA and WGA revealed the complex type of N-glycans in the 48 and 53-54kDa glycoproteins present in both species. Similar glycosylation was detected in species-specific bands of the 57-60 and 65-67 kDa in P. duboscqi and Lu. longipalpis, respectively. The high mannose type of glycosylation was found in the 20 and 39 kDa polypeptides of Lu. longipalpis and the 40-42 kDa polypeptides of P. duboscqi. Innate lectin activity specific for aminosugars was detected in SGE of P. duboscqi females using haemagglutination tests with rabbit erythrocytes.