Preparation of Polyclonal Antibodies Against Testis-specific Protease 50 and Characterization of Antibody Specificity

Introduction TheTSP50genehasbeenidentifiedasoneofthe testis specificoncogenes,whichisexpressedathigh levelsinapproximately92%ofhumanbreastcancer samples,makingitanattractivemolecularmarkerand apotentialtargetfordiagnosisandtherapy[1].Itisho mologoustomany…     

Toward breast cancer resistance protein (BCRP) inhibitors: design, synthesis of a series of new simplified fumitremorgin C analogues

In this study, we report the design and synthesis of a series of new simplified fumitremorgin C analogues. The preliminary biological study indicated some of these simplified fumitremorgin C might be developed into breast cancer resistance inhibitors.     

BRCA1 mutation screening using restriction endonuclease fingerprinting-single-strand conformation polymorphism in an automated capillary electrophoresis system

Efficient mutation scanning techniques are needed for the rapid detection of novel disease-associated mutations and rare-sequence variants of putative importance. The large size of the breast cancer 1 gene (BRCA1) and the many mutations found throughout its entire coding sequence make screening for mutations in this gene particularly challenging. We have developed a method for screening exon 11 of the BRCA1 gene based on restriction enzyme digestion of fluorescence-labeled polymerase chain reaction (PCR) products followed by single-strand conformation polymorphism (SSCP) using an automated capillary electrophoresis system, denoted capillary restriction endonuclease fingerprinting (REF)-SSCP electrophoresis. Using this strategy on a control set of samples, we were able to detect 17 of 18 known sequence alterations. The method was then applied to screen 73 Norwegian females with family histories of breast and/or ovarian cancer. A total of 172 sequence alterations were detected, including substitutions, insertions, and deletions. One novel substitution of unknown function was identified. Sequencing of all samples negative in the capillary REF-SSCP system gave no additional mutations confirming the high sensitivity of the described methodology. Capillary REF-SSCP electrophoresis appeared as a technically convenient technique, requiring amplification of fewer PCR fragments than traditional SSCP. The novel strategy allows high-throughput mutation scanning without radioactive labeling and polyacrylamide gel electrophoresis (PAGE).     

GeneMCL in microarray analysis

Accurately and reliably identifying the actual number of clusters present with a dataset of gene expression profiles, when no additional information on cluster structure is available, is a problem addressed by few algorithms. GeneMCL transforms microarray analysis data into a graph consisting of nodes connected by edges, where the nodes represent genes, and the edges represent the similarity in expression of those genes, as given by a proximity measurement. This measurement is taken to be the Pearson correlation coefficient combined with a local non-linear rescaling step. The resulting graph is input to the Markov Cluster (MCL) algorithm, which is an elegant, deterministic, non-specific and scalable method, which models stochastic flow through the graph. The algorithm is inherently affected by any cluster structure present, and rapidly decomposes a graph into cohesive clusters. The potential of the GeneMCL algorithm is demonstrated with a 5,730 gene subset (IGS) of the Van't Veer breast cancer database, for which the clusterings are shown to reflect underlying biological mechanisms.     

乳腺癌与血清微量元素关系的分析

经临床、病理、放射或CT检查确诊的22名乳腺癌患者。在放疗前后分析血清中Fe、Zn、Cu、Mn、Se、Cr、Co、Ni的含量。结果显示：1．乳腺癌患者与健康人比较Cu、Ni含量增加，而Fe、Zn含量减少；2．健康人锌与其他元素的比值与乳腺癌患者比较，Fe／Zn上升，而Cu／Zn下降；3．乳腺癌患者放疗后与放疗前比较，Zn、Fe、Se、Co含量降低，而Cu、Ni含量增加，因此放疗时要针对降低元素给予合理的补充。     

Comparative study of the instrumental couplings of high performance liquid chromatography with microwave-assisted digestion hydride generation atomic fluorescence spectrometry and inductively coupled plasma mass spectrometry for chiral speciation of selenomethionine in breast and formula milk

A comparison of chiral separation and analysis of selenomethionine in breast and formula milk, using high performance liquid chromatography (HPLC) on a glycopeptide teicoplanin-based chiral stationary phase (Chirobiotic T), coupled to atomic fluorescence spectrometry (AFS) and inductively coupled plasma (ICP) MS detectors has been performed. The coupling HPLC-microwave-assisted digestion hydride generation requires on-line post-column analytes treatment, and a severe sample clean-up for fat and proteins elimination using centrifugation and ultrafiltration. Underivatized -selenomethionine enantiomers were completely resolved in 10 min using unbuffered water mobile phase at 1 ml min⁻¹ flow. Good selectivity and sensitivities (detection limits 3.1 and 3.5 ng ml⁻¹ as Se for - and -selenomethionine, respectively) were obtained, and method robustness and simplicity, together to the low cost of AFS detector, makes it suitable for infant milk routine analysis. HPLC–ICP-MS coupling exhibits very low detection limits (0.9 ng ml⁻¹, as Se) for each -selenomethionine enantiomers, but the method suffers from matrix influence, that produces a poor S/N ratio and low reliability.

The methods were applied to breast and formula milk samples with recoveries of 80% of the total selenium presence, which is attributable to the existence of other unknown species. -Selenomethionine was the only isomer present in breast milk, but a 30% of -selenomethionine was also detected in formula milk.     

HPLC determination of urinary 2,4- and 2,6-toluendiamines as potential degradation products of polyurethane breast implants

Summary A reversed-phase HPLC method has been developed for the urinary determination of mutagenic 2,4- and 2,6-toluendiamines. These amines are degradation products of polyurethane, a material used to cover textured silicone breast implants. FMOC-Cl was used as fluorescent derivatising agent in order to obtain a limit of detection of 15 ng/ml in urine. Pre-treatment of urine samples was by liquid/liquid extraction and urine specimens of patients after surgury were analysed.     

Microfluidic three-dimensional biomimetic tumor model for studying breast cancer cell migration and invasion in the presence of interstitial flow

Cell migration and invasion are critical steps in cancer metastasis, which are the major cause of death in cancer patients. Tumor-associated macrophages(TAMs) and interstitial flow(IF) are two important biochemical and biomechanical cues in tumor microenvironment, play essential roles in tumor progression. However, their combined effects on tumor cell migration and invasion as well as molecular mechanism remains largely unknown. In this work, we developed a microfluidic-based 3 D breast cancer model by co-culturing tumor aggregates, macrophages, monocytes and endothelial cells within 3 D extracellular matrix in the presence of IF to study tumor cell migration and invasion. On the established platform, we can precisely control the parameters related to tumor microenvironment and observe cellular responses and interactions in real-time. When co-culture of U937 with human umbilical vein endothelial cells(HUVECs) or MDA-MB-231 cells and tri-culture of U937 with HUVECs and MDA-MB-231 cells, we found that mesenchymal-like MDA-MB-231 aggregates activated the monocytes to TAM-like phenotype macrophages. MDA-MB-231 cells and IF simultaneously enhanced the macrophages activation by the stimulation of colony-stimulating factor 1(CSF-1). The activated macrophages and IF further promoted vascular sprouting via vascular endothelial growth factor(VEGFα) signal and tumor cell invasion. This is the first attempt to study the interaction between macrophages and breast cancer cells under IF condition. Taken together, our results provide a new insight to reveal the important physiological and pathological processes of macrophages-tumor communication. Moreover, our established platform with a more mimetic 3 D breast cancer model has the potential for drug screening with more accurate results.     

Determination of organochlorine pesticides (OCPs) in breast milk from Rio Grande do Sul,Brazil, using a modified QuEChERS method and gas chromatography-negative chemical ionisation-mass spectrometry

ABSTRACT

The food chain is the main source of exposure to humans by organochlorine pesticides (OCPs) due to the bioaccumulation. Breast milk can accumulate OCPs, so this matrix is often used as an environmental bioindicator. The currently available methods for the determination of several OCPs and metabolites in breast milk involve, in general, multi-step sample preparation and quantification techniques with low selectivity, high cost and much time and labour. Thus, a fast and efficient method based on sample preparation using the quick, easy, cheap, effective, rugged and safe (QuEChERS) method combined with gas chromatography coupled to mass spectrometry with negative chemical ionisation (GC-NCI-MS) was developed, validated and applied for determination of 16 OCPs and metabolites in breast milk samples. The extract was cleaned by dispersive solid-phase extraction (d-SPE) using MgSO₄ and C18, evaporated in a Turbovap® system, redissolved and analysed by GC-NCI-MS. The method was validated showing acceptable recoveries (72–118%) and precision (RSD <19%). Method limits of detection (LODs) and quantification (LOQs) ranged from 0.75 to 7.5 ng g^?1 and from 2.5 to 25 ng g^?1 lipid, respectively. The method was successfully applied to 20 samples of breast milk from different regions of the Rio Grande do Sul state, Brazil, of which 75% contained residues below the LOQs.     

Camptothecin@HMSNs/thermosensitive hydrogel composite for applications in preventing local breast cancer recurrence

The CPT was loaded into the HMSNs with the high loading capacity. Then the CPT@HMSNs were loaded into the PLEL thermosensitive hydrogels for local therapy to prevent the recurrence of breast cancer after the tumor was resected.