A Discussion of Some of the Factors Causing Variable Results with Flagella Stains

Most flagella stains would probably give more consistent results if they were of stable composition and more was known of the factors influencing the mechanism of their action. Many of the mordants that have been recommended change rapidly, both chemically and physically, after preparation.

The reaction of some mordants, such as that of Loeffler, seems to be an important factor in their use. It is complicated, however, by temperature effects, oxidizing and reducing agents, the age of the bacterial cultures and variations in different species.

Users of flagella stains will probably always have to select or alter methods to suit the cultures to be stained. Any method which makes use of a minimum of complicated solutions and operations is most likely to give consistently good results.

No clearing or decolorizing agent tried changed poor preparations to good ones, but sometimes good ones can be improved by this treatment.     

Co-Expression of p16, Ki67 and COX-2 Is Associated with Basal Phenotype in High-Grade Ductal Carcinoma In Situ of the Breast

We assessed the co-expression of cell cycle-related biomarkers in a series of 121 consecutive cases of high-grade ductal carcinoma in situ (DCIS), pure or associated with invasive carcinoma, and their associations with the different immunoprofiles of DCIS. Cases were identified from the histopathology files of the Breast Pathology Laboratory, Federal University of Minas Gerais, Brazil, from 2003 to 2008. The expression of estrogen receptor, progesterone receptor, HER2 overexpression, cytokeratin 5, epidermal growth factor receptor 1, cyclooxygenase-2, p16 and Ki67 were assessed. Tumors were placed into five subgroups according to their immunohistochemical profile: luminal A, luminal B, HER2, basal-like and “not classified”. We found that the basal phenotype was associated with a higher frequency of p16-positive cases (83%) and the luminal A phenotype showed a higher frequency of p16-negative cases (93%; p=0.000). The association of biomarkers p16⁺/Ki67⁺/COX2⁺ was expressed in 02/06 cases (33.3%) of the basal phenotype but in only 01/70 cases (1.4%) of the luminal A phenotype (p=0.01). The co-expression of p16⁺/Ki67⁺/COX2^- was associated with a basal phenotype (p=0.004). P16 expression, p16⁺/Ki67⁺/COX2⁺ and p16⁺/Ki67⁺/COX2^- co-expression showed significant associations with the basal phenotype and these profiles could be used to guide more aggressive treatment strategies in patients with high-grade DCIS.     

A pharmacogenomic method for individualized prediction of drug sensitivity

Identifying the best drug for each cancer patient requires an efficient individualized strategy. We present MATCH (M erging genomic and pharmacologic A nalyses for T herapy CH oice), an approach using public genomic resources and drug testing of fresh tumor samples to link drugs to patients. Valproic acid (VPA) is highlighted as a proof‐of‐principle. In order to predict specific tumor types with high probability of drug sensitivity, we create drug response signatures using publically available gene expression data and assess sensitivity in a data set of >40 cancer types. Next, we evaluate drug sensitivity in matched tumor and normal tissue and exclude cancer types that are no more sensitive than normal tissue. From these analyses, breast tumors are predicted to be sensitive to VPA. A meta‐analysis across breast cancer data sets shows that aggressive subtypes are most likely to be sensitive to VPA, but all subtypes have sensitive tumors. MATCH predictions correlate significantly with growth inhibition in cancer cell lines and three‐dimensional cultures of fresh tumor samples. MATCH accurately predicts reduction in tumor growth rate following VPA treatment in patient tumor xenografts. MATCH uses genomic analysis with in vitro testing of patient tumors to select optimal drug regimens before clinical trial initiation.     

Inhibitors of phosphoprotein phosphatases 1 and 2A cause activation of a 53 kDa protein kinase accompanying the apoptotic response of breast cancer cells

Treatment of MCF-7 breast cancer cells with 50 nM okadaic acid triggers an apoptotic response which is accompanied by a 7-fold increase in the activity of a protein kinase with a relative molecular mass of 53 kDa. The activity of the kinase was stimulated by cell treatment with inhibitors of phosphoprotein phosphatase 1 and 2A, but not by stressing conditions. Okadaic acid-induced stimulation of the 53 kDa protein kinase was not abolished by coincubation of cells with cycloheximide. We conclude that stimulation of the 53 kDa protein kinase by inhibitors of phosphoprotein phosphatases involves pre-existing molecular components whose activity depends on the phosphorylation state of serine/threonine residues.     

Glycoengineered cell models for the characterization of cancer O-glycoproteome: an innovative strategy for biomarker discovery

Glycosylation is one of the most abundant forms of protein posttranslational modification. O-glycosylation is a major type of protein glycosylation, comprising different types and structures expressed in several physiologic and pathologic conditions. The understanding of protein attachment site and glycan structure is of the utmost importance for the clarification of the role glycosylation plays in normal cells and in pathological conditions. Neoplastic transformation frequently shows the expression of immature truncated O-glycans. These aberrantly expressed O-glycans have been shown to induce oncogenic properties and can be detected in premalignant lesions, meaning that they are an important source of biomarkers. This article addresses the recent application of genetically engineered cancer cell models to produce simplified homogenous O-glycans allowing the characterization of cancer cells O-glycoproteomes, using advanced mass spectrometry methods and the identification of potential cancer-specific O-glycosylation sites. This article will also discuss possible applications of these biomarkers in the cancer field.     

Changes in the fucose content of haptoglobin in breast and ovarian cancer: Association with disease progression

There is increasing evidence for changes in fucosylation in cancer. Previously, we showed that the fucose-specific lectin,Lotus tetragonolobus, extracts an abnormal form of haptoglobin (Hp) from cancer sera. This study investigates the monosaccharide content of Hp obtained from women with ovarian and breast cancer at different stages of their disease. In both cancers, Hp fucose was low when the disease was benign or in remission and much higher when the disease was progressive. This occurred whether the data was expressed per mole of protein or per three mannose residues. Changes in other monosaccharides were minor compared with fucose. There were small increases in theN-acetylglucosamine and galactose content (per three mannoses) in ovarian cancer, suggesting that some glycan chains have increased branching. The latter was independent of disease activity which may be due to some indirect cause such as cytotoxic therapy or an inflammatory response. When ovarian cancer patients were in remission, the number of glycosylation sites on Hp was reduced. Hp isolated from patients with early, but not advanced breast cancer also appeared to have increased glycan branching. The increased fucosylated Hp may interfere with fucose-mediated adhesion reactions of cancer cells.Part of this work was published in abstract form,Glycoconjugate J 1993;10; 318.     

MICB基因多态性与乳腺癌的易感关联性研究

目的:研究海南汉族人群MICB等位基因的多态性与乳腺癌易感性之间的关联性。方法:采用PCRSSP(PCR sequence-specific primers)和PCR-SBT(PCR sequence-based typing)方法对样本MICB等位基因的多态性进行检测。结果:乳腺癌患者中检出14种MICB等位基因;和对照组相比较,MICB*002和MICB*014等位基因在乳腺癌患者组分布频率较少,MICB*002和MICB*014等位基因可能对乳腺癌不易感(MICB*002:OR=0.31,95%CI:0.19-0.51,Pc0.05;MICB*014:OR=0.32,95%CI:0.17-0.60,Pc0.05)。MICB*016和MICB*003等位基因在乳腺癌患者组分布较多;MICB*016和MICB*003等位基因可能对乳腺癌易感(MICB*016:OR=10.68,95%CI:2.52-45.28,Pc0.05;MICB*003:OR=3.57,95%CI:1.34-9.49,Pc0.05);MICB*002/002和MICB*014/014基因型可能对乳腺癌不易感(MICB*002/002:OR=0.12,95%CI:0.04-0.36,Pc0.05;MICB*014/014:OR=0.30,95%CI:0.10-0.89,Pc0.05)。结论:MICB等位基因的多态性与乳腺癌的易感性之间存在关联性。     

乳腺癌腋窝淋巴结清扫术后上肢淋巴水肿的危险因素分析

研究背景：乳腺癌是女性常见的恶性肿瘤之一,同时也是治愈率最高的癌症,文献报道早期乳腺癌术后5年生存率达85．16％。上肢淋巴水肿是腋窝淋巴结清扫术后常见并发症,术后10—35％患者出现上肢水肿,就上肢淋巴水肿的风险因素,国内外做了许多研究,但结果不一,分歧较大。方法：抽取60例腋窝淋巴结清扫术后患者,调查统计可能与腋窝淋巴结清扫术后上肢淋巴水肿相关的8个因素（变量）：年龄、临床分期、是否放疗、是否出现延迟愈合／感染／积液等术后其它并发症、术后上肢功能锻炼、是否是优势侧,平时是否参加体育锻炼,是否有高血压等合并症。数据采用SPSS13．0分析软件以logistic回归方法进行分析。结果：四项关联因素分别为：1、是否放疗（OR=8．966）2、延迟愈合／感染／积液等其它术后并发症（0R=8．493）3、术后上肢功能锻炼（OR=0．194）4、高血压（0R=5．609）。结论：放疗、其它术后并发症、高血压为腋窝淋巴结清扫术后上肢淋巴水肿的风险因素,而术后上肢功能锻炼为术后水肿的保护因素。     

乳腺癌与MICA基因多态性的相关性研究

目的:研究海南汉族人群MICA等位基因的多态性与乳腺癌的相关性。方法:采用PCR-SSP和PCRSBT方法对样本MICA等位基因的多态性进行检测分析。结果:乳腺癌患者中有检测出10种MICA等位基因,其中MICA*002/019基因型频率较对照组显著偏低(OR=0.32,Pc0.05)。结论:MICA*002/019基因型可能与乳腺癌的保护相关。     

乳腺癌患者循环肿瘤细胞检测的研究进展

血行播散是乳腺癌转移的重要途径,检测腋窝淋巴结已经不能完全准确判断乳腺癌患者是否存在转移。肿瘤细胞进入外周血是肿瘤远处转移的前提,对乳腺癌患者循环肿瘤细胞的检测将有助于判断预后,指导治疗,监测治疗效果。简要综述了乳腺癌循环肿瘤细胞及其检测方法的研究进展。     

MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis

To investigate the global expression profile of miRNAs in primary breast cancer (BC) and normal adjacent tumor tissues (NATs) and its potential relevance to clinicopathological characteristics and patient survival, the genome-wide expression profiling of miRNAs in BC was investigated using a microarray containing 435 mature human miRNA oligonucleotide probes. Nine miRNAs of hsa-miR-21, hsa-miR-365, hsa-miR-181b, hsa-let-7f, hsa-miR-155, hsa-miR-29b, hsa-miR-181d, hsa-miR-98, and hsa-miR-29c were observed to be up-regulated greater than twofold in BC compared with NAT, whereas seven miRNAs of hsa-miR-497, hsa-miR-31, hsa-miR-355, hsa-miR-320, rno-mir-140, hsa-miR-127 and hsa-miR-30a-3p were observed to be down-regulated greater than twofold. The most significantly up-regulated miRNAs, hsa-mir-21 (miR-21), was quantitatively analyzed by TaqMan real-time PCR in 113 BC tumors. Interestingly, among the 113 BC cases, high level expression of miR-21 was significantly correlated with advanced clinical stage (P = 0.006, Fisher's exact text), lymph node metastasis (P = 0.007, Fisher's exact text), and shortened survival of the patients (hazard ratio [HR]=5.476, P < 0.001). Multivariate Cox regression analysis revealed this prognostic impact (HR=4.133, P = 0.001) to be independent of disease stage (HR=2.226, P = 0.013) and histological grade (HR=3.681, P = 0.033). This study could identify the differentiated miRNAs expression profile in BC and reveal that miR-21 overexpression was correlated with specific breast cancer biopathologic features, such as advanced tumor stage, lymph node metastasis, and poor survival of the patients, indicating that miR-21 may serve as a molecular prognostic marker for BC and disease progression.     

Cytological and immunocytochemical evaluation of thyroid and breast masses in patients with a previous neoplasm: case reports

The diagnosis of secondary tumours represents one of the most important fields in the application of fine needle aspiration cytology (FNAC). We studied two patients, one with a history of breast cancer and one with a previous tumour of the thyroid, who showed a second mass, in the thyroid and in the breast, respectively, during follow up. The aim of our study was to evaluate if cytology, performed on FNAC smears, may distinguish a metastatic lesion from a second primary tumour, or if further immunocytochemistry should be performed. Our data demonstrate that, while cytology may be indicative of a second primary tumour, the histotype should be confirmed by immunocytochemical staining.     

The Relationship of Body Mass Index to Reproductive Factors in Pre- and Postmenopausal African-American Women With and Without Breast Cancer

To date, there are virtually no existing data on the relationship between obesity, menopausal status, and breast cancer in African-Americans. Therefore, the present study was designed to test the following hypotheses in an African-American population: (1) there exists a positive association between BMI and breast cancer among postmenopausal women; (2) there exists an inverse association between BMI and breast cancer among premenopausal women; and (3) similar associations between BMI and reproductive factors exist for both pre- and postmenopausal breast cancer cases. The study population comprised 357 African-American women (n=193 breast cancer cases; n=164 controls). No significant differences were observed between premenopausal cases and controls for BMI, obesity categories, and reproductive factors. Among the postmenopausal women, the cases had significantly lower weight and BMI levels than the controls. Age at first pregnancy and parity were significantly lower among postmenopausal cases than their controls. No significant associations were revealed between body mass index and breast cancer for pre- and postmenopausal women. In the present study, early age at menarche was the only reproductive factor that was an independent predictor of BMI for both pre- and postmenopausal women, irrespective of breast cancer status. Also, these findings strongly suggest the need to consider reproductive factors, particularly age at menarche, as a covariate of BMI and other obesity-related diseases.     

DNA methylation changes associated with risk factors in tumors of the upper aerodigestive tract

Cancers of the upper aerodigestive tract (UADT) are common forms of malignancy associated with tobacco and alcohol exposures, although human papillomavirus and nutritional deficiency are also important risk factors. While somatically acquired DNA methylation changes have been associated with UADT cancers, what triggers these events and precise epigenetic targets are poorly understood. In this study, we applied quantitative profiling of DNA methylation states in a panel of cancer-associated genes to a case-control study of UADT cancers. Our analyses revealed a high frequency of aberrant hypermethylation of several genes, including MYOD1, CHRNA3 and MTHFR in UADT tumors, whereas CDKN2A was moderately hypermethylated. Among differentially methylated genes, we identified a new gene (the nicotinic acetycholine receptor gene) as target of aberrant hypermethylation in UADT cancers, suggesting that epigenetic deregulation of nicotinic acetycholine receptors in non-neuronal tissues may promote the development of UADT cancers. Importantly, we found that sex and age is strongly associated with the methylation states, whereas tobacco smoking and alcohol intake may also influence the methylation levels in specific genes. This study identifies aberrant DNA methylation patterns in UADT cancers and suggests a potential mechanism by which environmental factors may deregulate key cellular genes involved in tumor suppression and contribute to UADT cancers.     

A Standardized Test of Renal Concentrating Capacity in Adults with Some Results in Essential Hypertension

Three groups of patients: A with normal glomerular filtration rate, B with moderate and C with advanced renal damage, were dehydrated and fasted for 30 hours. At regular intervals measurements were taken of urine osmolality, urine specific gravity and serum osmolality. The time required to reach maximum urine osmolality varies with the degree of dehydration and inversely with the severity of kidney damage. In patients with normal glomerular filtration rate, maximum urine osmolality is not attained by 30 hours of dehydration. Thus, for shorter periods, all “normal ranges” of concentrating capacity must be related to specific durations of dehydration. Carefully measured urine specific gravities parallel urine osmolalities closely, especially when proteinuria and glucosuria are absent. The measurement of U/P osmolality ratio offers no clinical advantage in the assessment of renal concentration capacity over the measurement of urine osmolality alone. In Group A, hypertensives achieved higher urine concentrations than did the nonhypertensives under identical test conditions. A normal range for renal concentrating capacity has been presented.     

Comparative analysis of the cytotoxic activity of extracts from different parts of A. absinthium L. on breast cancer cell lines and correlation with active compounds concentration

The aim of the present study was to compare the cytotoxicity of different extracts of the plant Artemisia absinthium on breast cancer cell lines and to establish the correlation between the cytotoxicity and the active constituent’s level in these extracts. The cytotoxicity of the extracts was evaluated on the breast cancer cell lines, MCF-7 and MDA MB-231 by MTT assay and LDH release assay. An HPTLC method was developed for the simultaneous estimation of active constituents, that is, artemisinin, artemisinic acid, and alpha-thujone in different parts of A. absinthium. The whole extract was best among all the extracts tested with least IC₅₀ value and high LDH release that is, 491.19?µg/µL with 27.92% for MCF-7 and 459.97?µg/µL with 29.43% for MDA MB-231 cell lines respectively. Although, the concentration of all three quantified active compounds was higher in the extract from aerial part; however, the whole extract showed the best cytotoxicity among all extracts evaluated on the breast cancer cell lines. Surprisingly, our results demonstrate that the quantified active compounds were not solely responsible for the cytotoxic activity of the plant parts and further studies may be conducted to identify the compounds with synergistic, allosteric or antagonistic effects.