Challenges and Approaches to Measuring Repeat Fecal Immunochemical Test for Colorectal Cancer Screening.

BACKGROUND: Colorectal cancer screening with fecal immunochemical testing (FIT) can reduce colorectal cancer-related mortality. Effectiveness of FIT may be compromised when patients do not adhere to a regular schedule. However, having no standard measure of repeat FIT presents challenges for assessing effectiveness across populations and settings. We compared three measures of repeat FIT in a large, integrated health care system in Dallas, Texas. METHODS: We identified 18,257 patients age-eligible (50-60 years) for FIT in January 1-December 31, 2010 and followed over four rounds of screening. Measures included: (i) repeat FIT in prior screeners, or completion of FIT within 9-15 months of the previous; (ii) yes-no patterns, whereby patients were assigned yes or no in 9-15 month windows; and 3) proportion of time covered (PTC), or the amount of time patients were up-to-date with screening relative to time eligible. RESULTS: Repeat FIT varied by measure. Using a prior screeners measure, 15.8% of patients with a normal FIT in round 1 completed repeat FIT in round 2. Repeat FIT was notably higher (52.3%) using PTC. The most common yes-no pattern was YNNN or "one-and-done," and only 9.4% of patients completed two consecutive FITs across all rounds (YYNN). CONCLUSIONS: Different measures of repeat FIT yielded a range of estimates, making comparison across studies difficult. Researchers should weigh the advantages and disadvantages of each measure and select the most appropriate to their research question. IMPACT: Our study highlights the need for future research of repeat FIT measures that best approximate screening effectiveness.

Colonoscopy vs. Fecal Immunochemical Test in Reducing Mortality From Colorectal Cancer (CONFIRM): Rationale for Study Design.

RATIONALE: Colorectal cancer (CRC) is preventable through screening, with colonoscopy and fecal occult blood testing comprising the two most commonly used screening tests. Given the differences in complexity, risk, and cost, it is important to understand these tests' comparative effectiveness. STUDY DESIGN: The CONFIRM Study is a large, pragmatic, multicenter, randomized, parallel group trial to compare screening with colonoscopy vs. the annual fecal immunochemical test (FIT) in 50,000 average risk individuals. CONFIRM examines whether screening colonoscopy will be superior to a FIT-based screening program in the prevention of CRC mortality measured over 10 years. Eligible individuals 50-75 years of age and due for CRC screening are recruited from 46 Veterans Affairs (VA) medical centers. Participants are randomized to either colonoscopy or annual FIT. Results of colonoscopy are managed as per usual care and study participants are assessed for complications. Participants testing FIT positive are referred for colonoscopy. Participants are surveyed annually to determine if they have undergone colonoscopy or been diagnosed with CRC. The primary endpoint is CRC mortality. The secondary endpoints are (1) CRC incidence (2) complications of screening colonoscopy, and (3) the association between colonoscopists' characteristics and neoplasia detection, complications and post-colonoscopy CRC. CONFIRM leverages several key characteristics of the VA's integrated healthcare system, including a shared medical record with national databases, electronic CRC screening reminders, and a robust national research infrastructure with experience in conducting large-scale clinical trials. When completed, CONFIRM will be the largest intervention trial conducted within the VA (ClinicalTrials.gov identifier: NCT01239082).

Green Tea Induced Cellular Proliferation and the Expression of Transforming Growth Factor-ß1 in the Jejunal Mucosa of Fasting Rats.

OBJECTIVE: The aim of this study was to understand whether or not the protective effect of green tea after fasting-induced damage in the jejunal mucosa of rat is dependent on cell proliferation and the stimulation of specific growth factors. MATERIALS AND METHODS: Sixty adult male Wistar rats were used in this study. The animals were divided randomly into 5 groups, with 12 in each group (G1-5). The animals in G1 (control group) were fed a rat chow diet and water ad libitum. The animals in G2 (fasting group) were fasted for 3 days. The animals in the G3, G4, and G5 groups were fasted for 3 days as G2, but were given water (G3), green tea (G4), or a vitamin E (G5) solution, respectively, for another 7 days. The animals were euthanized, and the jejunum was removed and processed for histological and immunohistochemical analysis. RESULTS: Compared to the G3 group, the jejunal mucosa of G4 rats showed a 70.6% higher level (p < 0.001) of expression of proliferating cell nuclear antigen and 98% higher level (p = 0.0001) of the expression of transforming growth factor-ß1 (TGF-ß1), whereas the level of fibroblast growth factor-1 (FGF-1) and insulin-like growth factor-1 (IGF-1) expression was 22 and 11% lower, respectively, in G4 animals as compared to G3 rats. These differences in the expression of FGF-1 and IGF-1 in G4 animals were not statistically significant. CONCLUSION: In this study, green tea repaired the fasting-induced damage in the jejunal mucosa of rats, mainly by inducing a significant expression of TGF-ß1 in the jejunal mucosa.

Schisandrin B displays a protective role against primary pulmonary hypertension by targeting transforming growth factor ß1.

Pulmonary arterial smooth muscle cells (PASMCs) in the medial layer of the vessel wall are involved in vessel homeostasis, but also for pathologic vascular remodeling in diverse diseases, such as pulmonary arterial hypertension (PAH). Pulmonary vascular remodeling in PAH results in vascular disorders, but its underlying molecular mechanisms are still not to be fully disclosed. In this study, we investigated the expression and function of the transforming growth factor (TGF)-ß1 in human PASMC cultured under the condition of hypoxia and elucidated the effect of schisandra chinensis and its active ingredients on proliferation, migration, and apoptosis in human PASMCs. We demonstrated that schisandrin B (Sch.B) alleviated the severity of PAH in PASMCs cultured under the condition of hypoxia. Significant upregulation of TGF-ß1 was observed in hypoxia-induced human PASMCs. Interestingly, administration of Sch.B substantially attenuated TGF-ß1 level in these PASMCs. In order to elucidate Sch.B function, the hypoxia-induced human PASMC was stimulated with Sch.B or cotreatment with TGF-ß1 in vitro. In agreement with its TGF-ß1-reducing effect, Sch B relieved human PASMCs migration and promoted the apoptosis of human PASMCs, by activation of TGF-ß1 downstream signal pathways in PASMCs. In contrast, co-treatment with TGF-ß1 promoted human PASMC proliferation and migration and inhibited the apoptosis of human PASMC, which can attenuate the protective role of Sch.B in human PASMC. Taken collectively, these findings suggest that the vascular relaxation evoked by Sch.B was mediated by direct effect on vascular smooth muscle cell via TGF-ß1 downstream signal pathways.

Protective and curative role of Citrus sinensis peel on cadmium-induced testicular and spermatic damage: a morphometric and immunohistochemical evaluation using monoclonal antibodies against Ki-67 and proliferating cell nuclear antigen

This study examined the protective and curative effects of aqueous zest extract of Citrus sinensis on Cadmium-induced testicular tumor in animal models. Twenty four male Wistar rats (10 to 12 weeks old) weighing 165-275 g were divided into group A (treated orally with 2.5 ml/kg body weight/daily of normal saline), Group B (treated intraperitoneally with a single dose of 5mg/kg of cadmium), group C (Treated intraperitoneally with 5 mg/kg of cadmium before 10 mg/kg aqueous zest extract of Citrus sinensis orally), group D (treated with 5mg/kg of cadmium before 40 mg/kg extract), group E (treated with 10 mg/kg extract before 5 mg/kg of cadmium) and group F (treated with 40 mg/kg extract before 5mg/kg of cadmium). The procedure lasted for 8 weeks. Group B rats showed a significant (p< 0.05) decrease in testis weight, testis volume, sperm count (p > 0.001), sperm motility (p > 0.001), abnormal sperm morphology (p<0.001) and a significant decrease in tubular diameter, length (p <0.05), cross sectional area, width, germinal epithelia height, numerical density (p <0.01), perimeter, number (p < 0.001) and a significant increase in tubular lumen of the seminiferous tubules. Rats that were treated with cadmium without pre-treatment or post-treatment with extract showed marked degeneration and atrophied seminiferous tubules with absence of late stage germ cells. There was also a reduction in proliferative cell nuclear antigen (PCNA) materials and Ki67 positive cells in these rats. Interestingly, all these parameters were however attenuated in the groups that were pre-treated and post-treated with the extract. Taken together therefore, it was concluded that aqueous zest extract of Citrus sinensis have protective and curative roles in the abatement of cadmium-induced testicular tumor and that these effects might be as a result of the antioxidant and free radical scavenging potentials of these neutraceuticals

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Lesiones detectadas en seis programas poblacionales de cribado de cáncer colorrectal en España. Proyecto CRIBEA / CRIBEA Project: Lesions Detected in Six Spanish Colorectal Cancer Screening Population Based Programmes. Spain

Fundamentos: En este estudio se presentan los resultados de seis programas poblacionales de cribado de cáncer colorrectal desde 2005 a 2012 (Cataluña, Valencia, Murcia, Cantabria, País Vasco y Canarias) que utilizan diferentes tipos de test de cribado de sangre oculta en heces (SOH) bienal. El objetivo fue describir y comparar los resultados en cuanto a lesiones detectadas tanto por programa, participación, sexo, edad, tipo de test y comunidad autónoma. Metodos: Estudio de cohorte retrospectivo de las personas participantes en al menos una ronda completa cuya edad estaba comprendida entre los 50 y los 74 años. Lesiones consideradas: adenomas avanzados (AA), cáncer aolorrectal invasivo (CCR) y la suma de ambos, neoplasia avanzada (NA). Se realizó un nálisis de regresión logística y estudio de tendencias temporales. Resultados: Se obtuvieron 1.995.719 participaciones, lo que supuso el 46,7% de las invitaciones a participar. Se detectaron 21.228 neoplasias avanzadas (2.813 CCR y 18.415 AA). Se observaron diferencias en la detección de neoplasia avanzada (NA) entre los programas variando entre 15,1‰ y 35,8‰ participantes. La participación se relacionó con las tasas de detección (OR: 1,25 en 40-60% de participación). El test inmunoquímico cualitativo obtuvo una OR de 4,79 y el cuantitativo de 7,30 sobre guayaco. Los hombres tuvieron una OR de 2,73 sobre las mujeres, observándose en el 2012 una tasa de detección de neoplasia avanzada en hombres y mujeres de 33,1 y 14,2 x 1.000 respectivamente. Conclusiones: El tipo de test resultó el factor más determinante en la detección de lesiones. Las tendencias temporales mostraron un aumento de la tasa de detección por el cambio de test a partir del 2010 (AU)

Background: In this study, the results of six Colorectal Cancer Screening Population Programmes are shown (Catalonia, Valence, Murcia, Cantabria, the Basque Country and the Canary Islands collected between 2005 and 2012. These programmes use the faeces occult blood test (FOBt) biennial. Objective: To determine and compare the results of lesions detected by the programmes, participation, sex, age and test used. Methods: Retrospective cohort study based on people invited, aged between 50-74 years, in at least a complete round. Lesions considered: Advanced Adenomas (AA), Colorectal Invasive Cancer (CRC) and both of them, known as Advanced Neoplasia (AN). Logistic Regression and time trends are used. Results: 1,995,719 of invitations registered, with an average participation- rate of 46.7%. 21,228 Advanced Neoplasias (2,813 CRC and 18,415 AA). Differences in detection rates observed between programmes (varying from 15.1‰ to 35.8‰ between participants). Participation rates were related to lesions’ detection rates (OR 1.25 in 40-60% of participation). Inmunochemical qualitative test showed an OR of 4.79 and quantitative test an OR of 7.30 over the guaiac test. Men showed an OR of 2.73 with respect to women. In 2012 the Advanced Neoplasia rate for women and men was 33.1 and 14.2 by 1,000 participants. Conclusions: The test used was the most important factor for detecting lesions. Time trends showed an increase in detected lesions caused by the change of the type of test in 2010 (AU)

Neuronal activity and the expression of hypothalamic oxytocin and vasopressin in social versus cocaine conditioning.

Although drug rewards and natural rewards share neural substrates, the neuronal activation patterns and mechanisms behind the interaction between cocaine and social reward are poorly understood. Here, we investigated the conditioned place preference (CPP) in social (conspecific) vs cocaine conditioning, and the expression of central c-Fos, hypothalamic oxytocin (OT) and vasopressin (AVP) in ICR mice. We found that the mice produced CPP when conditioned with unfamiliar conspecific or cocaine alone. However, the mice failed to produce CPP when the two stimuli were concurrently conditioned. Compared to conditioning with conspecific alone, the mice decreased preference for conspecific when conditioning with social vs cocaine. We observed differential expression of c-Fos-immunoreactive neurons in the ventral anterior cingulate cortex, posterior cingulate cortex, accumbens (shell and core), medial nucleus of the amygdale and the ventral pallidum when comparing the control (CK), social (SC) or cocaine conditioning (CC) group, and social vs cocaine conditioning (SCC) group. Compared to the CK group, the SC or CC group had higher OT expression in the paraventricular nucleus (PVN) and lower AVP expression in the PVN and supraoptic nucleus. The SCC group showed lower OT expression compared to the SC group, and higher OT and AVP expression in the PVN compared to the CC group. These results indicate that cocaine impairs social preference through competing with social reward. The differential activations of neurons within specific reward areas, and differential expression of OT and AVP are likely to play an important role in mediating the interaction between social and cocaine rewards.

(Z)-3,4,3',5'-tetramethoxystilbene, a natural product, induces apoptosis and reduces viability of paclitaxel-and cisplatin-resistant osteosarcoma cells.

AIM OF STUDY: Osteosarcoma is a common bone tumor and the development of drug resistance in therapy of osteosarcoma is a general rule. The natural compounds isolated from medicinal plants represent a valuable resource for anticancer therapeutics. (Z)-3,4,3', '-tetramethoxystilbene is one of them. In this work, we investigated the potential anti-cancer activities of (Z)-3,4,3' ,'5-tetramethoxystilbene in paclitaxel- and cisplatin-resistant osteosarcoma cells. MATERIALS AND METHODS: ATP assay was used to examine cell viability. Cell nuclei staining assays with Hoechst or propidium iodide (PI) were used to evaluate cell apoptosis. Xenograft tumor model was used to evaluate the in vivo anti-cancer activities of (Z )-3,4,3',5'-tetramethoxystilbene. TUNEL staining assay was used to evaluate the apoptosis of tumor cells. RESULTS: We found that (Z)-3,4,3',5'-tetramethoxystilbene could effectively reduce viability of both paclitaxel-and cisplatin-resistant osteosarcoma cells. Moreover, (Z)-3,4,3',5'-tetramethoxystilbene induced dramatic apoptosis in resistant cells. Importantly, (Z)-3,4,3',5'-tetramethoxystilbene significantly suppressed in vivo tumor growth of cisplatin-resistant osteosarcoma. CONCLUSION: This is the first report on anti-cancer activity of (Z)-3,4,3','5- tetramethoxystilbene in resistant osteosarcoma cells. Our studies suggest that (Z)-3,4,3',5'-tetramethoxystilbene is a promising therapeutic drug for overcoming drug resistance in osteosarcoma.

[Maternal iron overload results in liver damage in rat offspring].

OBJECTIVE: To investigate the effects of iron overload on the liver tissue and function of maternal rats' offspring. METHODS: Forty pregnant rats were randomly divided into four groups, high-dose group, middle-dose group, low-dose group (120, 60 and 30 mg/kg BW) and control group. Iron detran was administered by intraperitoneal injection every other day and the entire trial lasted for 6 weeks. After 6 weeks, the iron serum levels, GOT and GPT of rats' offspring were determined. Histological changes of the liver injury were measured. The expressions of Bcl-2 and Bax in liver were measured by immunohistochemistry method. In addition, MDA, SOD and GSH-Px in liver were assessed by spectrophotometry. RESULTS: After 6 weeks, the level of serum iron in control group was significantly lower than in middle-dose and high-dose group (P < 0.05), and the result of level of liver iron was similar with that. According to the HE staining, it showed that liver cell gradually was damaged with the increasing of accumulation of hepatic iron. The expression of Bcl-2/Bax was significantly higher in control group than in middle-dose and high-dose groups (P < 0.05). The levels of serum GPT and GOT in high-dose group were significantly higher in control group (P < 0.05). The levels of Liver MDA in middle-dose and high-dose groups were both significantly higher than in control group (P < 0.05). Moreover, the levels of liver GSH-Px and SOD in high-dose group were significantly lower than in control group (P < 0.05). CONCLUSION: Iron overload in maternal rats may result in excessive iron deposition and oxidative damage of liver tissue and cell function in maternal rats' offspring.

High-density self-contained microfluidic KOALA kits for use by everyone.

Cell-based assays are essential tools used by research labs in a wide range of fields, including cell biology, toxicology, and natural product discovery labs. However, in some situations, the need for cell-based assays does not justify the costs of maintaining cell culture facilities and retaining skilled staff. The kit-on-a-lid assay (KOALA) technology enables accessible low-cost and prepackageable microfluidic platforms that can be operated with minimal infrastructure or training. Here, we demonstrate and characterize high-density KOALA methods for high-throughput applications, achieving an assay density comparable to that of a 384-well plate and usability by hand with no liquid-handling equipment. We show the potential for high-content screening and complex assays such as quantitative immunochemistry assays requiring multiple steps and reagents.

Identification and monitoring of host cell proteins by mass spectrometry combined with high performance immunochemistry testing.

Biotherapeutics are often produced in non-human host cells like Escherichia coli, yeast, and various mammalian cell lines. A major focus of any therapeutic protein purification process is to reduce host cell proteins to an acceptable low level. In this study, various E. coli host cell proteins were identified at different purifications steps by HPLC fractionation, SDS-PAGE analysis, and tryptic peptide mapping combined with online liquid chromatography mass spectrometry (LC-MS). However, no host cell proteins could be verified by direct LC-MS analysis of final drug substance material. In contrast, the application of affinity enrichment chromatography prior to comprehensive LC-MS was adequate to identify several low abundant host cell proteins at the final drug substance level. Bacterial alkaline phosphatase (BAP) was identified as being the most abundant host cell protein at several purification steps. Thus, we firstly established two different assays for enzymatic and immunological BAP monitoring using the cobas® technology. By using this strategy we were able to demonstrate an almost complete removal of BAP enzymatic activity by the established therapeutic protein purification process. In summary, the impact of fermentation, purification, and formulation conditions on host cell protein removal and biological activity can be conducted by monitoring process-specific host cell proteins in a GMP-compatible and high-throughput (> 1000 samples/day) manner.

Long-term ethanol effects on acute stress responses: modulation by dynorphin.

The brain stress-response system is critically involved in the addiction process, stimulating drug consumption and the relapse to drug taking in abstinent addicts. At the same time, its functioning is affected by chronic drug exposure. Here, we have investigated the role of the endogenous opioid peptide dynorphin as a modulator of effects of long-term ethanol consumption on the brain stress-response system. Using the two-bottle choice paradigm, we demonstrate an enhanced ethanol preference in male dynorphin knockout mice. Exposure to mild foot shock increased ethanol consumption in wild-type control littermates, but not in dynorphin-deficient animals. Blood adrenocorticotropic hormone levels determined 5 minutes after the shock were not affected by the genotype. We also determined the neuronal reactivity after foot shock exposure using c-Fos immunoreactivity in limbic structures. This was strongly influenced by both genotype and chronic ethanol consumption. Long-term alcohol exposure elevated the foot shock-induced c-Fos expression in the basolateral amygdala in wild-type animals, but had the opposite effect in dynorphin-deficient mice. An altered c-Fos reactivity was also found in the periventricular nucleus, the thalamus and the hippocampus of dynorphin knockouts. Together these data suggest that dynorphin plays an important role in the modulation of the brain stress-response systems after chronic ethanol exposure.

Determination of aflatoxins B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil using immunoaffinity column cleanup, postcolumn derivatization, and liquid chromatography/fluorescence detection: collaborative study.

The accuracy, repeatability, and reproducibility characteristics of a method using immunoaffinity column (IAC) cleanup with postcolumn derivatization and LC with a fluorescence detector (FLD) for determination of aflatoxins (AFs; sum of AFs B1, B2, G1, and G2) in olive oil, peanut oil, and sesame oil have been established in a collaborative study involving 15 laboratories from six countries. Blind duplicate samples of blank, spiked at levels ranging from 0.25 to 20.0 microg/kg for AF, were analyzed. A naturally contaminated peanut oil sample was also included. Test samples were extracted with methanol-water (55 + 45, v/v). After shaking and centrifuging, the lower layer was filtered, diluted with water, and filtered through glass microfiber filter paper. The filtrate was then passed through an IAC, and the toxins were eluted with methanol. The toxins were then subjected to RPLC-FLD analysis after postcolumn derivatization. Average recoveries of AFs from olive oil, peanut oil, and sesame oil ranged from 84 to 92% (at spiking levels ranging from 2.0 to 20.0 microg/kg); of AFB1 from 86 to 93% (at spiking levels ranging from 1.0 to 10.0 microg/kg); of AFB2 from 89 to 95% (at spiking levels ranging from 0.25 to 2.5 microg/kg); of AFG1 from 85 to 97% (at spiking levels ranging from 0.5 to 5.0 microg/kg); and of AFG2 from 76 to 85% (at spiking levels ranging from 0.25 to 2.5 microg/kg). RSDs for within-laboratory repeatability (RSD(r)) ranged from 3.4 to 10.2% for AF, from 3.5 to 10.9% for AFB1, from 3.2 to 9.5% for AFB2, from 6.5 to 14.9% for AFG1, and from 4.8 to 14.2% for AFG2. RSDs for between-laboratory reproducibility (RSDR) ranged from 6.1 to 14.5% for AF, from 7.5 to 15.4% for AFB1, from 7.1 to 14.6% for AFB2, from 10.8 to 18.1% for AFG1, and from 7.6 to 23.7% for AFG2. Horwitz ratio values were < or = 2 for the analytes in the three matrixes.

Evaluation of the Dimension Vista 1500 Chemical Autoanalyzer / 임상검사와정도관리

BACKGROUND: Laboratory tasks involve the analysis of a large number of specimens and generation of accurate results in a short period of time. The Dimension Vista 1500 (Siemens, Germany), an automated chemistry analyzer, has been introduced in our hospital to improve the efficiency of laboratory analysis. In order to assess the performance and usability of the analyzer, we evaluated its analytical performance and clinical usefulness, and compared these factors to those of the formerly used Modular DP analyzer (Roche Diagnostics, USA), Vitros 5.1 FS analyzer (Ortho Clinical Diagnostics, USA), and ADVIA Centaur analyzer (Siemens). METHODS: The accuracy, linearity, recovery factor, and sample carryover of the Dimension Vista 1500 analyzer were determined for 19 routine, immunochemistry, and cardiac marker tests (blood urea nitrogen, creatinine, glucose, calcium, phosphorus, total protein, albumin, AST, ALT, gamma-glutamyl transferase, creatine kinase [CK], lactate dehydrogenase, alkaline phosphatase, total bilirubin, high-sensitivity C-reactive protein, CK-MB, cardiac troponin I [cTnI], myoglobin, B-type natriuretic peptide), and values obtained for Modular DP, ADVIA centaur, and Vitros 5.1 FS analyzers were used to make comparisons. RESULTS: The coefficient of variation (CV) showed excellent values of or =0.996 with excellent linearity between 0.99 and 1.02. In addition, the recovery factor values of the tests were 88-107%, and percentage sample carryover values of the tests were or =0.96, except for cTnI (0.935), and showed good correlation (P<0.001). CONCLUSIONS: The Dimension Vista 1500 analyzer showed good analytical performance (linearity, precision, and accuracy) for 15 routine chemistry and 4 cardiac marker tests. Furthermore, results from the tests performed on the Dimension Vista 1500 analyzer correlated well with those obtained from similar tests performed on the Modular DP, ADVIA centaur, and Vitros 5.1 FS analyzers. Therefore, the Dimension Vista 1500 analyzer is appropriate for a tertiary care hospital where large volumes of tests have to be processed within a short period of time.

Tumor topoisomerase II alpha status and response to anthracycline-based neoadjuvant chemotherapy in breast cancer.

OBJECTIVES: Individualized chemotherapy for breast cancer improves the outcome. Anthracyclines target the enzyme topoisomerase IIα (TOP2A). We set out to perform a retrospective study of the presence of gene abnormalities and the expression of TOP2A in a cohort of breast cancer patients treated with neoadjuvant anthracycline-based chemotherapy. METHODS: Forty-three patients with 45 breast cancers were treated with neoadjuvant docetaxel-epirubicin with/without capecitabine chemotherapy. The TOP2A status of the cancers, determined retrospectively by fluorescent in situ hybridization and immunohistochemistry, was analyzed in relation to the standard clinical and pathological data. RESULTS: Clinically and pathologically complete remission (pCR) was achieved in 15 (33.3%) and 9 (20%) cases, respectively. The TOP2A gene was amplified in 2 human epidermal growth factor receptor 2 (HER2)-positive cancers (8%), and 32 (84.2%) overall exhibited TOP2A expression in >15% of the cells. The expression of TOP2A exhibited a strong correlation with the expression of Ki67 (R = 0.743, p < 0.001), and was negatively correlated with estrogen receptors (ER; R = 0.404, p = 0.012) and progesterone receptors (R = 0.430, p = 0.007). The expression of TOP2A was not related to the amplification of the TOP2A gene or the HER2 status of the tumor. The proportions of Ki67- and TOP2A-positive tumor cells were significantly reduced after chemotherapy (56.1 ± 23.6 vs. 19.0 ± 27.7%, p = 0.004, and 41.0 ± 27.9 vs. 12.7 ± 24.8%, p < 0.001, respectively). The development of pCR was related to a high grade (p = 0.054), ER negativity (p = 0.027) and high TOP2A expression (p = 0.037). The expression of TOP2A was an independent predictor of pCR (OR = 1.460, for every 10% increase, 95% CI: 1.016-2.096, p = 0.041). After a median follow-up time of 31.0 months, neither relapse-free survival nor overall survival was related to the tumor response. CONCLUSIONS: TOP2A expression is a marker of the tumor's proliferation rate and sensitivity to anthracycline-based chemotherapy, and does not depend on the amplification of its gene.

A hundred-year-old question: is the moss calyptra covered by a cuticle? A case study of Funaria hygrometrica.

BACKGROUND AND AIMS: The maternal gametophytic calyptra is critical for moss sporophyte development and ultimately sporogenesis. The calyptra has been predicted to protect the sporophyte apex, including the undifferentiated sporogenous region and seta meristem, from desiccation. We investigate the hypothesis that this waterproofing ability is due to a waxy cuticle. The idea that moss calyptrae are covered by a cuticle has been present in the literature for over a century, but, until now, neither the presence nor the absence of a cuticle has been documented for any calyptra. METHODS: The epidermis of the calyptra, leafy gametophyte and sporophyte sporangia of the moss Funaria hygrometrica were examined using scanning and transmission electron microscopy. Thicknesses of individual cuticle layers were quantified and compared statistically. The immunochemistry antibody (LM19) specific for pectins was used to locate cell wall material within the cuticle. KEY RESULTS: A multi-layered cuticle is present on the calyptra of F. hygrometrica, including layers analogous to the cuticular layer, cell wall projections, electron-lucent and electron-dense cuticle proper observed in vascular plants. The calyptra rostrum has a cuticle that is significantly thicker than the other tissues examined and differs by specialized thickenings of the cuticular layer (cuticular pegs) at the regions of the anticlinal cell walls. This is the first documentation of cuticular pegs in a moss. CONCLUSIONS: The calyptra and its associated cuticle represent a unique form of maternal care in embryophytes. This organ has the potential to play a critical role in preventing desiccation of immature sporophytes and thereby may have been essential for the evolution of the moss sporophyte.

Fish oil concentrate delays sensitivity to thermal nociception in mice.

Fish oil has been used to alleviate pain associated with inflammatory conditions such as rheumatoid arthritis. The anti-inflammatory property of fish oil is attributed to the n-3 fatty acids docosahexaenoic acid and eicosapentaenoic acid. Contrarily, vegetable oils such as safflower oil are rich in n-6 fatty acids which are considered to be mediators of inflammation. This study investigates the effect of n-3 and n-6 fatty acids rich oils as dietary supplements on the thermally induced pain sensitivity in healthy mice. C57Bl/6J mice were fed diet containing regular fish oil, concentrated fish oil formulation (CFO) and safflower oil (SO) for 6 months. Pain sensitivity was measured by Plantar test and was correlated to the expression of acid sensing ion channels (ASICs), transient receptor potential vanilloid 1 (TRPV1) and c-fos in dorsal root ganglion cells. Significant delay in sensitivity to thermal nociception was observed in mice fed CFO compared to mice fed SO (p<0.05). A significant diminution in expression of ion channels such as ASIC1a (64%), ASIC13 (37%) and TRPV1 (56%) coupled with reduced expression of c-fos, a marker of neuronal activation, was observed in the dorsal root ganglion cells of mice fed CFO compared to that fed SO. In conclusion, we describe here the potential of fish oil supplement in reducing sensitivity to thermal nociception in normal mice.

Estimation of ochratoxin A in some Turkish populations: an analysis in urine as a simple, sensitive and reliable biomarker.

In this study, ochratoxin A (OTA) occurrence and level in human urine samples, collected from four different regions of Turkey was analyzed by NaHCO(3) dilution, immunoaffinity column clean-up and high-performance liquid chromatography with fluorescent detection. For the repeatability of the method, RSD (%) values were found between 3.83 and 8.86, for the accuracy, the recovery values were found between 85.7% and 110.5% and limit of detection and limit of quantification of the method were measured as 0.006 and 0.018 ng mL(-1) respectively. For the analysis, first morning urine samples were collected and the results were adjusted with creatinine levels. From the total collected samples of 233 larger amounts of 83% was contaminated with OTA. Among the calculated to be OTA levels, positive sample rate, average OTA amount and the highest contamination was found in Ankara. (Positive sample rate; 90.1%, average OTA concentration; 14.34 ng g(-1) creatinine and highest OTA value; 75.60 ng g(-1) creatinine). In order to define the exposure profile of OTA in human a questionnaire was conducted among the voluntaries as well. But related to the gender, age, dietary habits, coffee consumption, smoking and alcohol habits of the volunteers, no correlation was found with the OTA exposure.

Screening and diagnosis for colorectal cancer: present and future.

AIMS AND BACKGROUND: Colon cancer is the third most common cause of cancer diagnosed in the United States, and the second leading cause of cancer death. Although rates of the disease have been going down in recent years, results can be further improved. About 90% of people whose colon cancer is caught before it has spread to nearby lymph nodes or organs survive more than 5 years after diagnosis. However, only 10% of patients whose cancer has spread to distant parts of the body survive 5 years. Diagnosis at an early stage aims to reduce the incidence of tumors in an advanced stage and hence mortality. METHODS AND STUDY DESIGN: We analyzed the literature to understand what new tests to use and new directions to take. RESULTS: There is evidence to support the screening of average-risk individuals over the age of 50 years to detect and prevent colon cancer. Screening of these people can only reduce mortality rates, not incidence, identifying cancer at an early stage and through the removal of clinically significant adenomas. Patient preferences and availability of resources play an important role in the selection of screening tests, because each test presents specific risks and specific benefits. CONCLUSIONS: The American Cancer Society has added two new screening methods, CT colonography, also known as virtual colonoscopy, and stool DNA tests to the list of options. A small revolution in screening for colon cancer is in the making. The availability of these less invasive tests should increase the number of people who undergo regular screening.

Phosphorylated ERK is a potential predictor of sensitivity to sorafenib when treating hepatocellular carcinoma: evidence from an in vitro study.

BACKGROUND: Sorafenib is the first agent that has demonstrated an improved overall survival benefit in advanced hepatocellular carcinoma (HCC), setting a new standard for first-line treatment. However, no one has yet been able to predict sensitivity to sorafenib. Pre-treatment pERK level has been shown to be associated with favorable response to such therapy in a phase II clinical study, indicating that pERK may be a potential biomarker for treatment of HCC with sorafenib. METHODS: The effects of sorafenib and 5-fluorouracil (5-FU) on cell proliferation were evaluated by cell viability assays in four HCC cell lines (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) with different metastatic potential and basal pERK expression levels. Expression levels of pERK were determined by immunocytochemical quantification together with western blot analysis, and pERK density values were also calculated. Correlation analyses were then carried out between the IC50 values of drugs and pERK density values. After basal ERK phosphorylation was down-regulated with U0126 in MHCC97-H cells, cellular responsiveness to sorafenib was assessed by cell viability assay. RESULTS: Basal pERK levels increased stepwise in cell lines in accordance with their metastatic potential. Sorafenib inhibited ERK phosphorylation in a dose-dependent manner in all four cell lines at a concentration between 5 and 20 microM, but the degree of inhibition was significantly different according to their basal pERK expression level (P < 0.0001). In contrast, no significant change was observed after 5-FU treatment. Correlation analyses between the IC50 values and pERK densities revealed that the effects of sorafenib on cell proliferation were significantly correlated with basal pERK levels (Spearman r = -0.8671, P = 0.0003). Resistance to 5-FU was also significantly associated with basal pERK expression in these HCC cell lines (Spearman r = 0.7832, P = 0.0026). After the basal ERK phosphorylation level in MHCC97-H cells was reduced with U0126, they were significantly less sensitive to sorafenib-mediated growth inhibition, with an IC50 of 17.31 +/- 1.62 microM versus 10.81 +/- 1.24 microM (P = 0.0281). CONCLUSION: In this in vitro study, pERK was confirmed to be a potential biomarker predictive of sensitivity to sorafenib in treating HCC. The RAF/MEK/ERK pathway may be involved in drug resistance to traditional chemotherapy in HCC.