ABSTRACT
Primary production is the basis for energy and biomolecule flow in food webs. Nutritional importance of terrestrial and plastic carbon via mixotrophic algae to upper trophic level is poorly studied. We explored this question by analysing the contribution of osmo- and phagomixotrophic species in boreal lakes and used 13 C-labelled materials and compound-specific isotopes to determine biochemical fate of carbon backbone of leaves, lignin-hemicellulose and polystyrene at four-trophic level experiment. Microbes prepared similar amounts of amino acids from leaves and lignin, but four times more membrane lipids from lignin than leaves, and much less from polystyrene. Mixotrophic algae (Cryptomonas sp.) upgraded simple fatty acids to essential omega-3 and omega-6 polyunsaturated fatty acids. Labelled amino and fatty acids became integral parts of cell membranes of zooplankton (Daphnia magna) and fish (Danio rerio). These results show that terrestrial and plastic carbon can provide backbones for essential biomolecules of mixotrophic algae and consumers at higher trophic levels.
Subject(s)
Carbon , Plastics , Animals , Carbon/metabolism , Lignin , Polystyrenes , Food Chain , Fatty AcidsABSTRACT
Chemotaxonomic biomarkers are needed to monitor and evaluate the nutritional quality of phytoplankton communities. The biomolecules produced by different phytoplankton species do not always follow genetic phylogeny. Therefore, we analyzed fatty acids, sterols, and carotenoids from 57 freshwater phytoplankton strains to evaluate the usability of these biomolecules as chemotaxonomic biomarkers. We found 29 fatty acids, 34 sterols, and 26 carotenoids in our samples. The strains were grouped into cryptomonads, cyanobacteria, diatoms, dinoflagellates, golden algae, green algae, and raphidophytes, and the phytoplankton group explained 61%, 54%, and 89% of the variability of fatty acids, sterols, and carotenoids, respectively. Fatty acid and carotenoid profiles distinguished most phytoplankton groups, but not flawlessly. For example, fatty acids could not distinguish golden algae and cryptomonads, whereas carotenoids did not separate diatoms and golden algae. The sterol composition was heterogeneous but seemed to be useful for distinguishing different genera within a phytoplankton group. The chemotaxonomy biomarkers yielded optimal genetic phylogeny when the fatty acids, sterols, and carotenoids were used together in multivariate statistical analysis. Our results suggest that the accuracy of phytoplankton composition modeling could be enhanced by combining these three biomolecule groups.
Subject(s)
Diatoms , Phytoplankton , Phytoplankton/genetics , Social Identification , Diatoms/genetics , Fatty Acids/analysis , Sterols , Carotenoids , BiomarkersABSTRACT
Carcinosarcoma (CS) of the breast is a rare entity (less than 0.2% of breast malignancies), characterized by the presence of a biphasic pattern of malignant epithelial and mesenchymal elements, and with a high risk of loco-regional recurrence. The diagnosis of CS of the breast is difficult and needs detailed histological investigations to differentiate it from other malignant breast tumors. Expertise and evidence-based information on optimal treatment is very limited due to the low incidence and inconsistent classification. The principles of treatment modalities seem to be similar to others breast malignancies. CS has a different biologic behavior from others breast cancers, being very aggressive in keeping with its high-grade mesenchymal stroma. Still many questions remain about its origin and optimal treatment modalities for better outcome. We report the case of CS of the breast without local or regional recurrence after six years of follow-up in an 82-year-old woman.
Subject(s)
Breast Neoplasms/pathology , Carcinosarcoma/pathology , Aged, 80 and over , Breast Neoplasms/surgery , Carcinosarcoma/surgery , Disease-Free Survival , Female , Humans , Mastectomy, Modified Radical , Treatment OutcomeABSTRACT
Chlamydophila abortus and Coxiella burnetii are one of the major pathogens implicated in abortion in cattle. Their characteristic of obligate intracellular bacteria, and of zoonotic agents, makes their culture difficult in diagnostic laboratories, and the traditional tools of diagnosis (detection of sera antibodies by ELISA, Stamp's coloration) encounter specificity, sensitivity and interpretability limits. Individual PCR have recently been developed. Nevertheless, their income/cost is a limiting factor for breeders. As the symptoms are not specific, the request for analysis often concerns the two valences. Consequently, the development and the validation of an internal multiplex PCR appears to be a suitable solution.
Subject(s)
Cattle Diseases/diagnosis , Cattle Diseases/microbiology , Chlamydophila Infections/veterinary , Chlamydophila/isolation & purification , Coxiella burnetii/isolation & purification , Polymerase Chain Reaction/methods , Q Fever/veterinary , Abortion, Veterinary/microbiology , Animals , Cattle , Chlamydophila/genetics , Chlamydophila Infections/diagnosis , Chlamydophila Infections/microbiology , Coxiella burnetii/genetics , Polymerase Chain Reaction/veterinary , Q Fever/diagnosis , Q Fever/microbiology , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
BACKGROUND: HER-2 amplification is an important prognostic biomarker and treatment determinant in breast carcinoma. AIMS: To correlate immunocytochemical (ICC) expression of HER-2 and gene amplification determined by chromogenic in situ hybridisation (CISH) using liquid based cytology (LBC) with immunohistochemistry (IHC) and CISH using histological samples of the same breast carcinomas. METHODS: Frozen sections and cytobrushings of 103 breast carcinomas were analysed. Four techniques were performed on each tumour: two on LBC samples (ICC, and CISH, both graded as positive, indeterminate, or negative) and two on histological samples (IHC and CISH). Two cell lines (MCF-7, negative; BT 474, positive) were used as controls for cytological analysis. A complementary fluorescence in situ hybridisation technique was carried out in histological samples with low amplification (4-10 dots/nucleus). RESULTS: Interobserver agreement for the four techniques calculated by the kappa coefficient indicated a substantial agreement. Nine cases failed in cytology because of poor cellularity. Among 94 cases, 19 were amplified; 73, 12, and 9 tumours were scored 0 or 1+, 2+, and 3+, respectively by IHC and 75, 13, and 6, respectively, by ICC. CISH found no amplification in 72 tumours. Correlations between the IHC and CISH results in the histological and cytological samples were always significant. CONCLUSIONS: Her-2 status could be determined in LBC samples and correlated well with reference histological methods using in situ hybridisation. ICC was less reliable because of the presence of the cytoplasmic membrane. However, these results should be confirmed by a large multicentre study.
Subject(s)
Biomarkers, Tumor/metabolism , Breast Neoplasms/metabolism , Receptor, ErbB-2/metabolism , Adult , Aged , Aged, 80 and over , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Carcinoma, Ductal/genetics , Carcinoma, Ductal/metabolism , Carcinoma, Ductal/pathology , Female , Genes, erbB-2 , Humans , Immunoenzyme Techniques , In Situ Hybridization , In Situ Hybridization, Fluorescence , Middle Aged , Neoplasm Proteins/metabolism , Prognosis , Prospective Studies , Tumor Cells, CulturedABSTRACT
AIMS: HER2 protein is over-expressed in 15-30% of breast carcinomas. Immunohistochemistry (IHC) is a common and inexpensive method able to specifically detect HER2 protein. However, lack of standardization of IHC has been considered responsible for discrepancies in HER2 status assessment performed by IHC and fluorescence in-situ hybridization (FISH). This prompted us to perform a multicentric IHC calibration test to achieve a maximum accuracy of HER2-IHC compared with HER2-FISH taken as the reference method. METHODS AND RESULTS: Twelve French laboratories participated in this study, including 119 cases of invasive breast carcinomas for which both fixed and frozen tissues were available. HER2 expression was determined in fixed tissues by individual in-house IHC techniques, using either CB11 (Novocastra, Newcastle, UK) or A0485 (Dako, Glostrup, Denmark) anti-HER2 antibodies. Two cut-off values were used: 10% and 60% of immunostained cells. In 116 of the 119 cases, HER2 gene status could also be determined by FISH on frozen sections, performed in a single laboratory. Results were centralized and compared. When suboptimal concordance between IHC and FISH was observed, IHC was calibrated and a second run was performed. The specificity, sensitivity and accuracy of IHC compared with FISH were noted before and after calibration. Forty-four out of 116 (38%) tumours showed HER2 gene amplification. Accuracy of IHC was complete in the first run for 6/12 laboratories. Calibration, necessary for the six others, relied mainly on the combination of a heat-induced epitope retrieval step with an increase of dilution of the primary antibody. In the second run, HER2 over-expression was found in 46 (40%) and 44 (38%) of the 116 cases, using 10% or 60% of stained cells as cut-offs, respectively. The corresponding accuracy rates were 93% and 95%. CONCLUSIONS: This study showed that a high accuracy of IHC could be obtained for the determination of HER2 status in all laboratories using their in-house IHC technique, provided that a calibration process was performed. Antigen retrieval procedure, high dilutions of anti-HER2 antibody and the use of specific controls were crucial for HER2-IHC calibration. A 95% accuracy rate of IHC, using FISH as gold standard, was obtained by considering immunolabelling HER2-IHC results as a continuous variable, and taking 60% invasive stained cells as the cut-off for HER2 over-expression.
Subject(s)
Adenocarcinoma/metabolism , Breast Neoplasms/metabolism , Genes, erbB-2 , Immunohistochemistry/methods , Receptor, ErbB-2/metabolism , Adenocarcinoma/genetics , Adenocarcinoma/secondary , Adult , Aged , Aged, 80 and over , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Female , Humans , Immunohistochemistry/standards , In Situ Hybridization, Fluorescence , Middle Aged , Neoplasm Staging , Receptor, ErbB-2/genetics , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
The HER2 proto-oncogene encodes a transmembrane protein, which is considered to function as a growth factor receptor. Overexpression of this protein found by immunohistochemistry in about 20% of infiltrating breast carcinomas, has a predictive value of response to treatment by trastuzumab, an anti-HER2 humanized monoclonal antibody. Search for HER2 gene amplification is necessary to adapt the immunohistochemical technique quality and also in the cases of delicate analysis or weak overexpression. It is usually carried out by Fluorescence In Situ Hybridization (FISH). A more recent hybridization technique, named CISH because of its chromogenic revelation is an alternative method, which gives highly correlated results with FISH. We present details of this technique, which may be more familiar for the pathologists than FISH, because reading analysis is similar to that of immunohistochemical staining.
Subject(s)
Chromogenic Compounds/analysis , Genes, erbB-2 , In Situ Hybridization/methods , Nucleic Acid Amplification Techniques , Breast Neoplasms/chemistry , Breast Neoplasms/genetics , Carcinoma, Ductal, Breast/chemistry , Carcinoma, Ductal, Breast/genetics , Chromosomes, Human, Pair 17/genetics , DNA Probes , Digoxigenin/analysis , Female , Humans , In Situ Hybridization, Fluorescence , Proto-Oncogene Mas , Specimen HandlingABSTRACT
Does radical mastectomy for cancer remain needed? Breast conserving treatment may be achieved by surgery, primary chemotherapy followed by radiotherapy and surgery. This article attempts at defining (according to clinical and pathological parameters of the tumor and patient's characteristics), when conservative treatment is not allowed and radical mastectomy must be performed. Mastectomy must be performed first when there are multiple tumors or a tumor too large with respect to the breast volume or diffuse microcalcifications on mammograms.ometimes the stage of pregnancy, a personal history of collagen vascular disease or prior radiotherapy or the willing of the patient lead to perform radical mastectomy.econdary mastectomy is necessary in case of failure of conservative treatment or recurrence after breast conserving treatment.
ABSTRACT
Does radical mastectomy for cancer remain needed? Breast conserving treatment may be achieved by surgery, primary chemotherapy followed by radiotherapy and surgery. This article attempts at defining (according to clinical and pathological parameters of the tumor and patient's characteristics), when conservative treatment is not allowed and radical mastectomy must be performed. Mastectomy must be performed first when there are multiple tumors or a tumor too large with respect to the breast volume or diffuse microcalcifications on mammograms. Sometimes the stage of pregnancy, a personal history of collagen vascular disease or prior radiotherapy or the willing of the patient lead to perform radical mastectomy. Secondary mastectomy is necessary in case of failure of conservative treatment or recurrence after breast conserving treatment.
Subject(s)
Breast Neoplasms/surgery , Mastectomy, Simple , Breast Neoplasms/pathology , Carcinoma, Ductal, Breast/pathology , Carcinoma, Ductal, Breast/surgery , Female , Humans , Neoplasm Recurrence, Local/surgery , Neoplasms, Multiple Primary/pathology , Neoplasms, Multiple Primary/surgeryABSTRACT
In a previous study, we reported that repeated combined chemotherapy and radiotherapy, and a single combined treatment had different consequences. In this study the effect of the chronology of the repeated combined treatments was tested, i.e. it was determined whether the treatment is more efficient when the first treatment is Fluorouracil or irradiation, or when they are given simultaneously. It was first demonstrated that, under our conditions, neither radiotherapy nor chemotherapy were chronodependent. The combined treatments were more efficient that the single treatment although their chronology had no significant consequences. Nevertheless, the simultaneous treatment appeared slightly better than the administration of Fluorouracil 6 h before or 6 h after irradiation. These results confirm our in vitro experiments.
Subject(s)
Adenocarcinoma/therapy , Fluorouracil/therapeutic use , Mammary Neoplasms, Experimental/therapy , Adenocarcinoma/drug therapy , Adenocarcinoma/radiotherapy , Animals , Cell Line , Combined Modality Therapy/methods , Mammary Neoplasms, Experimental/drug therapy , Mammary Neoplasms, Experimental/radiotherapy , Mice , Mice, Inbred Strains , Time Factors , Tumor Cells, CulturedABSTRACT
Nine pathologists from different institutions reviewed in a double-blind study 16 breast tumors previously indexed as typical medullary carcinoma, atypical medullary carcinoma, or infiltrative ductal carcinoma. A set of 16 slides was circulated two times among the nine pathologists. The diagnoses of typical and atypical medullary carcinomas were based on a definition given by Ridolfi et al. The interobserver and intraobserver agreement was low, with a kappa value of less than .50. The only histological criterion that had more than 50% agreement was the presence or absence of an in situ component in the tumor, assuming that the disagreement of one pathologist is accepted. This study is a snapshot of the problems encountered in the diagnosis of typical medullary carcinoma in a routine context and it shows high levels of variations in diagnostic consistency.
Subject(s)
Breast Neoplasms/pathology , Carcinoma, Medullary/pathology , Female , Humans , Observer VariationABSTRACT
Lumpectomy together with axillary clearance and radiotherapy is a good alternative to Patey's operation for treating early cancers of the breast. In any case it has not been definitely worked out how much should be removed. In certain patients not much needs to be removed, in other larger areas of tissue need to be excised. We present our technique for carrying out limited lumpectomy which is carried out at the same time as radiation therapy making it possible to perform a less radical clearance. The results in the first 17 patients we have followed up with a mean of 4.5 years are very encouraging. There was only one local recurrence; this was some distance away from the area of the lumpectomy. The limits for the method are determined by the examination that is carried out on the margins that have been removed and on the availability of a team comprising surgeon, histopathologist and radiotherapist.
Subject(s)
Brachytherapy/standards , Breast Neoplasms/surgery , Intraoperative Care/standards , Mastectomy, Segmental/standards , Adult , Age Factors , Aged , Breast Neoplasms/epidemiology , Breast Neoplasms/radiotherapy , Combined Modality Therapy , Esthetics , Follow-Up Studies , France/epidemiology , Humans , Lymph Node Excision/standards , Middle Aged , Neoplasm Recurrence, Local , Radiation DosageABSTRACT
In order to test its potential application to thyroid tumour pathology, the monoclonal antibody Ki-67 was used to demonstrate immunohistochemically proliferating cells in 31 non neoplastic and neoplastic lesions of the thyroid. Twenty benign lesions were negative. Of the 11 malignant tumors, 4 papillary, 2 follicular and 1 medullary carcinomas were also negative; only 3 follicular and 1 medullary carcinomas were positive. Moreover, no correlation was found between Ki-67 immunostaining status and histological typing or pTNM pathological classification in the 11 malignant tumors.
Subject(s)
Antibodies, Monoclonal , Nuclear Proteins/analysis , Thyroid Neoplasms/pathology , Humans , Ki-67 Antigen , Thyroid Neoplasms/chemistry , Thyroid Neoplasms/classificationABSTRACT
To evaluate the sensibility and specificity of MRI in the initial staging of carcinoma of the uterine cervix, a retrospective study of 41 patients (48 MR scans) was undertaken. The sensitivity of T2-weighted images in the detection and determination of tumor location was 90% and specificity was 100%. On T2-weighted images, the signal intensity of the tumor was high in 64%. Determination of tumor size was correct in 80%. The ability of MRI to demonstrate the extension to the uterine corpus was acceptable with a sensitivity of 63% and a specificity of 100%. The extra-uterine extension was difficult to appreciate especially for parametrial, vaginal and bladder involvement with a sensitivity of 38%, 43 and 67% respectively. Concerning the lymph nodes involvement, the sensitivity was 38%. The results of this study point out the ability of MRI to evaluate the size and location of the tumor. The accuracy of MRI to appreciate the involvement of the parameters, vagina and bladder is disappointing.
Subject(s)
Magnetic Resonance Imaging , Uterine Cervical Neoplasms/diagnosis , Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Middle Aged , Neoplasm Invasiveness , Neoplasm Staging , Sensitivity and Specificity , Uterine Cervical Neoplasms/pathologyABSTRACT
Melanin production by two medullary carcinomas of the thyroid is reported and discussed. In both tumours, melanin and calcitonin could be detected in the same cells.