A novel inhibitory anti-invasive MAb isolated using phenotypic screening highlights AnxA6 as a functionally relevant target protein in pancreatic cancer.

BACKGROUND: Discovery and validation of new antibody tractable targets is critical for the development of new antibody therapeutics to address unmet needs in oncology. METHODS: A highly invasive clonal variant of the MDA-MB-435S cell line was used to generate monoclonal antibodies (MAbs), which were screened for anti-invasive activity against aggressive cancer cells in vitro. The molecular target of selected inhibitory MAb 9E1 was identified using immunoprecipitation/liquid chromatography-tandem mass spectrometry. The potential anti-tumour effects of MAb 9E1 were investigated in vitro together with immunohistochemical analysis of the 9E1 target antigen in normal and cancer tissues. RESULTS: MAb 9E1 significantly decreases invasion in pancreatic, lung squamous and breast cancer cells and silencing of its target antigen, which was revealed as AnxA6, leads to markedly reduced invasive capacity of pancreatic and lung squamous cancer in vitro. IHC using MAb 9E1 revealed that AnxA6 exhibits a high prevalence of membrane immunoreactivity across aggressive tumour types with restricted expression observed in the majority of normal tissues. In pancreatic ductal adenocarcinoma, high AnxA6 IHC score correlated with the presence of tumour budding at the invasive front of tumours (P=0.082), the presence of perineural invasion (P= <0.0001) and showed a weak correlation with reduced survival (P=0.2242). CONCLUSIONS: This study highlights the use of phenotypic hybridoma screening as an effective strategy to select a novel function-blocking MAb, 9E1 with anti-cancer activity in vitro. Moreover, through characterisation of the 9E1 target antigen, AnxA6, our findings support further investigation of AnxA6 as a potential candidate target for antibody-mediated inhibition of pancreatic cancer.

7B7: a novel antibody directed against the Ku70/Ku80 heterodimer blocks invasion in pancreatic and lung cancer cells.

Development of more effective therapeutic strategies for cancers of high unmet need requires the continued discovery of disease-specific protein targets for therapeutic antibody targeting. In order to identify novel proteins associated with cancer cell invasion/metastasis, we present here an alternative to antibody targeting of cell surface proteins with an established role in invasion; our functional antibody screening approach involves the isolation and selection of MAbs that are primarily screened for their ability to inhibit tumour invasion. A clonal population of the Mia PaCa-2, a pancreatic ductal adenocarcinoma (PDAC) cell line, which displays a highly invasive phenotype, was used to generate MAbs with the objective of identifying membrane targets directly involved in cancer invasion. Selected MAb 7B7 can significantly reduce invasion in a dose-responsive manner in Mia PaCa-2 clone 3 and DLKP-M squamous lung carcinoma cells. Using immunoprecipitation and liquid chromatography-tandem mass spectrometry (LC-MS-MS) analysis, the target antigen of anti-invasive antibody, 7B7, was determined to be the heterodimeric Ku antigen, Ku70/80, a core protein composed of the Ku70 and Ku80 subunits which is involved in non-homologous end-joining (NHEJ) DNA repair. RNA interference-mediated knockdown of Ku70 and Ku80 resulted in a marked decrease in the invasive capacity of Mia PaCa-2 clone 3 and DLKP-M cells, indicating that Ku70/Ku80 is functionally involved in pancreatic and lung cancer invasion. Immunohistochemical analysis demonstrated Ku70/Ku80 immunoreactivity in 37 PDAC tumours, indicating that this heterodimer is highly expressed in this aggressive cancer type. This study demonstrates that a functional MAb screening approach coupled with immunoprecipitation/proteomic analyses can be successfully applied to identify functional anti-invasive MAbs and potential novel targets for therapeutic antibody targeting.

Correlating transcriptional networks to breast cancer survival: a large-scale coexpression analysis.

Weighted gene coexpression network analysis (WGCNA) is a powerful 'guilt-by-association'-based method to extract coexpressed groups of genes from large heterogeneous messenger RNA expression data sets. We have utilized WGCNA to identify 11 coregulated gene clusters across 2342 breast cancer samples from 13 microarray-based gene expression studies. A number of these transcriptional modules were found to be correlated to clinicopathological variables (e.g. tumor grade), survival endpoints for breast cancer as a whole (disease-free survival, distant disease-free survival and overall survival) and also its molecular subtypes (luminal A, luminal B, HER2+ and basal-like). Examples of findings arising from this work include the identification of a cluster of proliferation-related genes that when upregulated correlated to increased tumor grade and were associated with poor survival in general. The prognostic potential of novel genes, for example, ubiquitin-conjugating enzyme E2S (UBE2S) within this group was confirmed in an independent data set. In addition, gene clusters were also associated with survival for breast cancer molecular subtypes including a cluster of genes that was found to correlate with prognosis exclusively for basal-like breast cancer. The upregulation of several single genes within this coexpression cluster, for example, the potassium channel, subfamily K, member 5 (KCNK5) was associated with poor outcome for the basal-like molecular subtype. We have developed an online database to allow user-friendly access to the coexpression patterns and the survival analysis outputs uncovered in this study (available at http://glados.ucd.ie/Coexpression/).

Olfactomedin III expression contributes to anoikis-resistance in clonal variants of a human lung squamous carcinoma cell line.

Three clonal subpopulations of DLKP, a poorly differentiated squamous lung carcinoma cell line, display striking differences in ability to survive in suspension (anoikis resistance). DLKP-SQ is anoikis resistant (7.5% anoikis at 24 h). In contrast, DLKP-M and DLKP-I are sensitive to anoikis (49.2% and 42.6% respectively). DLKP-I shows increased apoptosis consistently over all time points tested while DLKP-M appear to slow down metabolically and perhaps delays onset of anoikis by undergoing autophagy. Expression microarray analysis identified pronounced differential expression of Olfactomedin 3 (OLFM3) between the clones. High expression of OLFM3 was confirmed at the RNA level by qRT-PCR in DLKP-SQ and at the protein level by Western blotting (within the cell and secreted). Little or no OLFM3 was detected in the other two clones (DLKP-M and DLKP-I). Following siRNA knockdown of OLFM3 in DLKP-SQ, anoikis was increased 2.8-fold to 21% which was intermediate between the anoikis levels in DLKP-SQ and DLKP-M or DLKP-I. This knockdown correlated with increased apoptosis in suspension but not in attached culture conditions. Addition of recombinant OLFM3 reduced anoikis in DLKP-I. This is the first instance of OLFM3 being linked with anoikis resistance in a human cancer cell line.

Feasibility and relevance of global expression profiling of gene transcripts in serum from breast cancer patients using whole genome microarrays and quantitative RT-PCR.

BACKGROUND: Previous studies, by ourselves and others, have indicated that gene transcripts are detectable extracellularly. Advancing on this work, in order to investigate the feasibility of analysing global gene expression profiles and so the possibility in the future of identifying panels of circulating mRNA biomarkers that may be diagnostic, prognostic or predictive for cancer, here we performed the first whole genome microarray analysis of human serum. PATIENTS AND METHODS: RNA was isolated from pre-surgery serum and corresponding breast tumour and normal tissue biopsies, and from post-surgery and normal control serum. Specimens were examined using Affymetrix whole genome microarrays and quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). RESULTS: Of the 54,675 mRNAs/variants analysed, approximately 8% and 45% were called Present in serum and breast tissue specimens, respectively. Differentially expressed genes were identified for each group of specimens analysed. Analysis, by qRT-PCR, of 3 selected transcripts further indicated that the nucleic acids detected were mRNA, not DNA. mRNAs are apparently present in serum and their global detection and identification can be successfully achieved using microarray technologies. CONCLUSION: The potential implication of this novel finding is that using microarrays it may be possible to identify a panel of extracellular mRNAs that are diagnostic, prognostic and/or predictive of outcome for cancer patients.

Osteogenesis imperfecta.

"Fragile bones" have been described in medical literature for centuries. Cases dating from antiquity include dental and skeletal details eerily similar to those found among modern patients whose bones fracture easily and whose bodies show signs of muscular and other weakness. Osteogenesis imperfecta--whose name implies "imperfect birth of bone"--is one of these inherited fragile bone syndromes. A generalized disorder of the body's connective tissues, it is most obvious in its effect on bone, but also involves the body's ligaments, tendons, fascia, eyes, skin, teeth and ears. Radiographs, bone scans and other imaging tools are essential in the initial diagnosis, assessment of fracture risk, and planning and tracking of treatment.

Radiation protection.

One of radiologic technologists' most important professional obligations is protecting patients, other members of the health care team, the public and themselves from as much radiation-related harm as possible while also maximizing the screening, diagnostic and therapeutic potential of ionizing radiation. This article reviews the different types of radiation dose and how radiation affects the body. Patient shielding, personnel dosimeters and area monitors are discussed, along with beam collimation and filtration. The author also describes protocols to protect pregnant patients and pregnant technologists.

Investigation of the molecular profile of basal cell carcinoma using whole genome microarrays.

BACKGROUND: Skin cancer accounts for 1/3 of all newly diagnosed cancer. Although seldom fatal, basal cell carcinoma (BCC) is associated with severe disfigurement and morbidity. BCC has a unique interest for researchers, as although it is often locally invasive, it rarely metastasises. This paper, reporting the first whole genome expression microarray analysis of skin cancer, aimed to investigate the molecular profile of BCC in comparison to non-cancerous skin biopsies. RNA from BCC and normal skin specimens was analysed using Affymetrix whole genome microarrays. A Welch t-test was applied to data normalised using dCHIP to identify significant differentially-expressed genes between BCC and normal specimens. Principal component analysis and support vector machine analysis were performed on resulting genelists, Genmapp was used to identify pathways affected, and GOstat aided identification of areas of gene ontology more highly represented on these lists than would be expected by chance. RESULTS: Following normalisation, specimens clustered into groups of BCC specimens and of normal skin specimens. Of the 54,675 gene transcripts/variants analysed, 3,921 were differentially expressed between BCC and normal skin specimens. Of these, 2,108 were significantly up-regulated and 1,813 were statistically significantly down-regulated in BCCs. CONCLUSION: Functional gene sets differentially expressed include those involved in transcription, proliferation, cell motility, apoptosis and metabolism. As expected, members of the Wnt and hedgehog pathways were found to be significantly different between BCC and normal specimens, as were many previously undescribed changes in gene expression between normal and BCC specimens, including basonuclin2 and mrp9. Quantitative-PCR analysis confirmed our microarray results, identifying novel potential biomarkers for BCC.

Influence of multidrug resistance and drug transport proteins on chemotherapy drug metabolism.

INTRODUCTION: Chemotherapy involving the use of anticancer drugs remains an important strategy in the overall management of patients with metastatic cancer. Acquisition of multidrug resistance remains a major impediment to successful chemotherapy. Drug transporters in cell membranes and intracellular drug metabolizing enzymes contribute to the resistance phenotype and determine the pharmacokinetics of anticancer drugs in the body. AREAS COVERED: ATP-binding cassette (ABC) transporters mediate the transport of endogenous metabolites and xenobiotics including cytotoxic drugs out of cells. Solute carrier (SLC) transporters mediate the influx of cytotoxic drugs into cells. This review focuses on the substrate interaction of these transporters, on their biology and what role they play together with drug metabolizing enzymes in eliminating therapeutic drugs from cells. EXPERT OPINION: The majority of anticancer drugs are substrates for the ABC transporter and SLC transporter families. Together, these proteins have the ability to control the influx and the efflux of structurally unrelated chemotherapeutic drugs, thereby modulating the intracellular drug concentration. These interactions have important clinical implications for chemotherapy because ultimately they determine therapeutic efficacy, disease progression/relapse and the success or failure of patient treatment.

Galectin-3 expression alters adhesion, motility and invasion in a lung cell line (DLKP), in vitro.

BACKGROUND: Galectin-3, a beta-galactosidase-binding protein, is involved in regulating many physiological and pathological cellular processes. The significance of galectin-3 in human lung and nasal carcinoma cells has not yet been elucidated. MATERIALS AND METHODS: Using RT-PCR and Western blotting techniques, the constitutive level of galectin-3 in the human non-small cell lung carcinoma cell line, DLKP, was investigated. Following galectin-3 cDNA transfection into these cells, growth, toxicity, adhesion, motility and invasion assays were used to investigate the relevance of galectin-3 over-expression. RESULTS: Galectin-3 over-expression did not induce a multi-drug resistance phenotype or significantly affect cell growth rate, but it did result in enhanced (i) adhesion to extracellular matrix components; (ii) cell motility; and (iii) in vitro invasiveness. Furthermore, studies of RPMI-2650 variants suggest that galectin-3 expression correlates with nasal carcinoma cell invasiveness. CONCLUSION: Our results suggest that galectin-3 expression levels in both lung and nasal tumour cells may play a role in cell motility, invasion, and metastasis.

Reducing errors in radiology.

Medical errors, even those that are relatively minor, can have serious consequences, such as misdiagnosis and longer and costlier hospital stays. Reducing errors requires all members of the health care clinical and administrative team to commit to the effort, and effective risk management addresses system-wide causes of errors. Errors often result from poor communication, inadequate training, chronic fatigue, and entrenched workplace hierarchies. Error reduction strategies support high-quality patient care, even in the most stressful and complex situations.

Use of plant growth regulators in fig tree seedling s 'roxo de valinhos' / Uso de reguladores vegetais em mudas de figueira 'Roxo de Valinhos'

The application of plant growth regulators can promote better development of the seedlings, essential for the success of fruit growing. The current study aims to evaluate the use of plant growth regulators in fig tree seedlings 'Roxo de Valinhos'. The experiment was carried out in a suspended nursery covered with plastic, using rooted cuttings of the cultivar Roxo de Valinhos. Four applications were made with a commercial product, known as Stimulate®, at doses of 0, 50, 100, 150, and 200 mL L-1. Using a randomized block experimental design, the study was subdivided into plots (doses X days of the last application [after 0, 7, 14 and 21]). The following traits were evaluated: stem and root length; stem diameter; root volume; leaf number; leaf area; stem, leaf and root dry mass; specific leaf area, leaf weight ratio; leaf area ratio; and chlorophyll total. Results indicated that the product promoted greater seedlings development at a dose range from 100 to 150 mL L-1. Different dose levels did not vary according to the days after product application. Furthermore, using such technique enabled seedlings of higher quality and vigour.

A aplicação de reguladores vegetais pode configurar uma técnica viável para promoção do melhor desenvolvimento das mudas, imprescindível para o sucesso da fruticultura. Deste modo, objetivou-se avaliar o uso de reguladores vegetais na produção de mudas da figueira 'Roxo de Valinhos'. O experimento foi realizado em viveiro suspenso com cobertura plástica, utilizando-se estacas enraizadas da cultivar Roxo de Valinhos. Foram realizadas quatro aplicações com o produto comercial Stimulate® nas concentrações de 0, 50, 100, 150, e 200 mL L-1. Utilizou-se o delineamento experimental em blocos casualizados com parcelas subdivididas [concentrações X épocas de avaliação (após 0, 7, 14 e 21 dias da última aplicação)]. Avaliou-se: comprimento do ramo e da maior raiz; diâmetro do ramo; volume de raízes; número de folhas e área foliar; massa seca de folhas, ramos e raízes; área específica foliar, razão de peso foliar e razão da área foliar; e teores de clorofila total. O uso do produto, nas concentrações de 100 a 150 mL L-1, promoveu maior desenvolvimento das mudas. O efeito das diferentes doses não variou em função dos dias após aplicação do produto. O uso de tal técnica permite a obtenção de mudas de maior qualidade e vigor.

Performance of mango cultivars under subtropical conditions in the state of São Paulo / Desempenho de cultivares de mangueira em condições subtropicas do estado de São Paulo

Mango is quite popular among tropical fruits, and has traits of a fast-expanding market. There is a huge variety of mango cultivars and their potential should be studied in order to be introduced in new producing regions with favourable edaphoclimatic characteristics for planting. The current study aimed to evaluate mango productive performance under subtropical conditions in the state of São Paulo, Brazil. The experiment was conducted with 'Espada Vermelha', 'Keitt' and 'Palmer' cultivars grafted onto rootstock 'Espada' over three crop cycles 2012-2013, 2013-2014 and 2014-2015. The experimental design was a randomized complete block, consisting of 3x3 factorial design, in which the first factor corresponds to the cultivars and the second factor to the evaluated crop cycles, with 10 replications and one plant per plot. The following variables were evaluated: number of fruits per panicle, number of fruits per plant, panicles percentage to the full aborted fruits, fruit fresh weight, production, yield, canopy volume, production efficiency and crop seasonality. Data analysis by Tukey test and Pearson´s correlation coefficients. Among the evaluated cultivars, Palmer has stood out with the best productive performance under the experiment conditions in the three evaluated crop cycles. The seasonal availability of cultivars should be explored so that there are fruit productions over a longer period of time. Correlations among productive traits varies according to the cultivar.

A manga é uma das frutas tropicais mais apreciadas e seu mercado está em expansão. Existe uma grande variedade de cultivares de mangueira e seu potencial deve ser estudado, sendo importante a introdução do cultivo desta frutífera em novas regiões produtoras, principalmente em locais que apresentem características edafoclimáticas favoráveis para seu cultivo. Desta forma, objetivou-se com este trabalho, avaliar o desempenho produtivo de três cultivares de mangueira nas condições subtropicais do estado de São Paulo, Brasil. O experimento foi conduzido em três ciclos agrícolas, (2012-2013, 2013-2014 e 2014-2015) com as cultivares Espada Vermelha, Keitt e Palmer enxertadas sobre a cultivar Espada. O delineamento experimental foi em blocos casualizados com esquema fatorial 3 x 3, em que o primeiro fator corresponde às cultivares e o segundo fator aos ciclos agrícolas avaliados. Utilizaram-se 10 repetições e uma planta por parcela. As seguintes variáveis foram avaliadas: número de frutos por panícula, número de frutos por planta, porcentagem de panículas com abortamento total de frutos, massa fresca do fruto, produção, produtividade, volume da copa, eficiência produtiva e sazonalidade de colheita. Os dados obtidos foram analisados por meio do teste Tukey e correlação de Pearson. Dentre as cultivares avaliadas a Palmer destaca-se com o melhor desempenho produtivo independente do ciclo agrícola avaliado. As correlações entre as características produtivas avaliadas variam conforme a cultivar.

Evaluation of papaya genotypes using agronomic descriptors and estimation of genetic parameters / Descritores agronômicos e parâmetros genéticos na avaliação de genótipos de mamoeiros

In Brazil, the improvement of papaya agribusiness is correlated to the estimation of genetic parameters and evaluation of agronomic descriptors; thus enabling less favourable areas for the development and implementation of papaya crops. Therefore, the current study aimed to evaluate quantitative descriptors and to estimate genetic parameters in genotypes of papaya grown under subtropical climate in the state of São Paulo. The experimental design was randomized in block. The experimental units were grouped into seven blocks of five plants each and three treatments, which consisted of three different varieties: Sunrise Solo, Tainung nº1 and local. The psychochemical properties were analysed for plant height; intersection height of first fruits; stem diameter; male, female and hermaphroditic flower percentage; fruit number per plant; production; yield; normal fruits percentage, carpeloid and pentandric fruits; pulp matter; pulp length; pulp diameter; pulp shape; pulp cavity; pulp thickness; peel performance; pulp performance; seeds performance; seeds number; soluble solids; pH; titratable acidity; ratio; technological index; ascorbic acid and reducing sugars. Furthermore, most agronomic descriptors showed high heritability; besides that, the varieties Sunrise Solo and Tainung nº1 performed the best results in Botucatu.

A estimativa de parâmetros genéticos e a avaliação de descritores agronômicos do mamoeiro pode contribuir com o desenvolvimento do agronegócio do mamão no Brasil, permitindo a expansão de novos cultivos em regiões menos favoráveis para sua exploração comercial. Frente ao exposto, objetivou-se com este trabalho avaliar descritores quantitativos e estimar parâmetros genéticos em genótipos de mamoeiro, cultivados em clima subtropical do estado de São Paulo. O delineamento experimental foi em blocos casualizados, com três tratamentos (variedade Sunrise Solo, híbrido Tainung nº1 e variedade local) e sete blocos, utilizando cinco plantas por parcela experimental. Foram mensuradas as seguintes caraterísticas: altura das plantas e da interseção dos primeiros frutos; diâmetro do caule; porcentagens de plantas masculinas, femininas e hermafroditas; número de frutos por planta; produção; produtividade; porcentagem de frutos normais, carpelóides e pentândricos; massa, comprimento, diâmetro, formato, diâmetro da cavidade interna e espessura da polpa dos frutos; rendimento de casca, de polpa e de sementes; número de sementes; sólidos solúveis; pH; acidez titulável; ratio; índice tecnológico; ácido ascórbico e açúcares redutores. Foi possível inferir com o estudo que há alta herdabilidade quanto à maioria dos descritores agronômicos avaliados e que os bons resultados para a variedade Sunrise Solo e o híbrido Tainung nº 1, possibilitam a indicação desses genótipos para cultivo, na região de Botucatu ­ SP.

How cultural competency can help reduce health disparities.

Radiologic technologists encounter patients from a variety of racial, ethnic, and socioeconomic backgrounds. Reduced income and access to health care among minority and immigrant populations accounts for many health disparities. Enhancing cultural competency among health care providers and providing culturally appropriate services can improve communication, access to health care, and eventually health outcomes. Although awareness of a patient's ethnic or racial background can aid in screening and diagnosis, patient-centered care requires that everyone be seen as individuals first and members of a racial or ethnic minority second.

Ergonomics in radiology.

Radiologic technologists and radiologist assistants are at risk for developing a variety of work-related musculoskeletal injuries, especially those caused by repeated strain on the upper extremities and lower back. This article examines the types of activities most likely to cause on-the-job injuries in technologists, such as transferring, positioning, and transporting patients, as well as the types of equipment that tend to be associated with injury, including ultrasound probes, heavy protective garments, and picture archiving and communications system workstations. Psychosocial factors also are explored, along with management's role in minimizing work-related injuries. Practical techniques for preventing injury are emphasized throughout.

Caracterização física e química dos frutos nos diferentes quadrantes da planta e germinação de sementes do portaenxerto cítrico tangerineira 'Sunki' / Physical and chemical characterization of fruit in different plant quadrants and germination rootstock of citrus tangerine 'Sunki'

A tangerineira 'Sunki' apresenta grande relevância para a citricultura brasileira por apresentar importantes características para o melhoramento genético e por ser considerada um ótimo portaenxento. Neste contexto, objetivou-se com o presente trabalho estudar a caracterização dos frutos do portaenxerto cítrico 'Sunki', em função da posição dos mesmos na planta e também, avaliar a germinação de suas sementes. As características físicas avaliadas foram massa do fruto, diâmetros longitudinal e equatorial dos frutos, relação diâmetro longitudinal/diâmetro equatorial, número de gomos por frutos, massa de gomos e de casca e número de sementes viáveis por fruto. Quanto à caracterização química, foi avaliado acidez titulável, sólidos solúveis, "Ratio", pH e vitamina C. Para a avaliação dessas variáveis, os frutos foram obtidos dos quadrantes nordeste, sudoeste, noroeste e sudeste. Com relação à avaliação da germinação das sementes, foram utilizadas diferentes doses de ácido giberélico e Stimulate. As sementes, extraídas de frutos maduros, foram lavadas, secadas à sombra e armazenadas em BOD. Logo após, estas foram tratadas com biorreguladores por 24 horas. Depois da embebição, as sementes foram colocadas no germinador em papel germitest. Após o inicio do processo germinativo, as avaliações foram realizadas diariamente até o trigésimo quarto dia. Frutos da tangerineira 'Sunki' apresentam boa uniformidade quanto à posição dos frutos na copa; o uso de ácido giberélico e de bioestimulante é promissor para uso comercial na germinação de sementes desse portaenxerto.

The tangerine ' Sunki' are highly relevant for the Brazilian citrus industry by presenting important traits for genetic improvement and to be considered a great portaenxento. In this context, the objective of the present work to study the characterization of the fruits of citrus rootstock ' Sunki', depending on the position on the plan and also the germination of their seeds . The physical characteristics evaluated were fruit weight, longitudinal and equatorial diameters of the fruit longitudinal diameter / equatorial diameter, number of buds per fruit , weight of buds and bark and number of viable seeds per fruit. As for the chemical characterization was evaluated titratable, acidity, soluble solid, "Ratio ", pH and vitamin C. For the evaluation of these variables, the fruits were obtained from the northeast, southwest, northwest and southeast quadrants. Regarding the evaluation of seed germination, different doses of gibberellic acid and Stimulate were used. The seeds extracted from ripe fruit, washed, dried in the shade and stored in BOD. Immediately after, they were treated for 24 hours with plant growth regulators. After soaking, the seeds were placed in a germinator in germitest paper. After initiation of the germination process, the evaluations were performed daily until the thirty- fourth day. Fruits of 'Sunki' mandarin with uneven as the fruit position within the canopy, the use of gibberellic acid and growth promoter is promising for commercial use on seed germination of this rootstock.

Update on bone densitometry.

As the U.S. population ages, osteoporosis will become more prevalent and its detection and treatment more critical. This article provides an update on various techniques used to measure bone density and discusses the indications for bone densitometry and special patient populations that may undergo the exam. In addition, the article discusses the role of the radiologic technologist in bone densitometry, operator safety and the need for precision testing of densitometry equipment. This article is a Directed Reading. Your access to Directed Reading quizzes for continuing education credit is determined by your area of interest. For access to other quizzes, go to www.asrt.org/store.

Drug metabolism-related genes as potential biomarkers: analysis of expression in normal and tumour breast tissue.

The complex role of drug metabolism-related enzymes and their possible influence in cancer development, treatment and outcome has not yet been completely elucidated. There is evidence that these enzymes can activate certain environmental procarcinogens to more toxic derivatives and thus a role has been proposed for them in carcinogenesis. The fact that they can also inactivate a number of chemotherapeutic drugs has raised the possibility of these enzymes influencing the sensitivity of tumour cells to anticancer agents. In this report, we analyse the expression of drug metabolism-related genes within a whole genome microarray study of 104 breast cancer and 17 normal breast specimens. Kaplan-Meier survival curves, Chi-squared, and Cox Regression analyses were used to identify associations between expression of gene transcripts and patients' clinicopathological and survival data. Our results show that several of these genes are significantly expressed in both normal and tumour tissue; in many cases, expression is altered in the tumour specimens as compared to normal breast tissue. Moreover, expression of ARNT2 and GST A1 was correlated with prognosis. Kaplan-Meier analysis showed expression of ARNT2 mRNA to correlate significantly with favourable disease outcome for patients, in terms of both their disease-free survival (P = 0.0094) and overall survival (P = 0.0018) times from diagnosis, while detection of GST A1 mRNA correlated with shortened disease-free survival (P = 0.0131) and overall survival (P = 0.0028). Multivariate analysis indicated GST A1 expression to be an independent prognostic factor for overall survival (P = 0.045). Our results suggest a possible use of ARNT2 and GST A1 as prognostic breast cancer biomarkers.

RNA Interference with siRNA.

Over the last decade, RNA interference has emerged as an effective mechanism for silencing gene expression. This ancient cellular antiviral response can be used to allow specific inhibition of the function of any chosen target gene, including those involved in diseases such as cancer, AIDS and hepatitis. It has become an invaluable research tool to aid in the identification of novel genes involved in disease processes. It has advanced from the use of synthetic RNA for the endogenous production of small hairpin RNA by plasmid and viral vectors, and from transient inhibition in vitro to longer-lasting effects in vivo. However, as with antisense and ribozymes, the efficient delivery of siRNA into cells is currently the limiting factor to successful gene expression inhibition in vivo. This review gives an overview of the mechanism of action of siRNA and its use in cancer research. It also discusses the successes and shortcomings of this new gene knockdown tool.