Analytical variables influencing the HCV RNA determination by TaqMan real-time PCR in routine clinical laboratory practice

Hepatitis C virus (HCV) quantification is used as a prognostic marker for treatment success. In a routine clinical laboratory some infinitesimal sample handling factors can contribute to variability and loss of precision in HCV quantification. This may include blood collection tubes, blood drawing procedure, sample processing and storage temperatures. In current study blood was collected in tubes with different anticoagulant type (spray vs. liquid), group 1, blood was drawn with possible suck of methylated spirit through needle (experimental group) while avoiding the methylated spirit suck (control group) group 2, plasma separation was delayed from 0 to 60 min for group 3, plasma storage at different temperatures group 4. All samples were analyzed using Corbett research real time PCR system using AJ Roboscreen Kit. Mean viral load difference between spray vs. liquid was found 3.6 × 10(5) IU/ml (p < 0.001). Methylated spirit inhibited the viral load quantification with a value of 4.8 × 10(5) IU/ml (p < 0.001). Mean viral load difference was found 1.2 × 10(5) IU/ml (p < 0.05). Delay in centrifugation from 0 to 60 min and plasma placement at 25 °C for 15 min before freezing had no effect (p = 0.5996). Plasma storage temperature at -80 and -20 °C did not affect significantly on RNA levels (p > 0.05). In conclusion blood collection tubes and procedures can be a key factor in variability of results, that might affect the treatment response decision.     

Denoising of ECG signals based on noise reduction algorithms in EMD and wavelet domains

This paper presents a new ECG denoising approach based on noise reduction algorithms in empirical mode decomposition (EMD) and discrete wavelet transform (DWT) domains. Unlike the conventional EMD based ECG denoising approaches that neglect a number of initial intrinsic mode functions (IMFs) containing the QRS complex as well as noise, we propose to perform windowing in the EMD domain in order to reduce the noise from the initial IMFs instead of discarding them completely thus preserving the QRS complex and yielding a relatively cleaner ECG signal. The signal thus obtained is transformed in the DWT domain, where an adaptive soft thresholding based noise reduction algorithm is employed considering the advantageous properties of the DWT compared to that of the EMD in preserving the energy in the presence of noise and in reconstructing the original ECG signal with a better time resolution. Extensive simulations are carried out using the MIT-BIH arrythmia database and the performance of the proposed method is evaluated in terms of several standard metrics. The simulation results show that the proposed method is able to reduce noise from the noisy ECG signals more accurately and consistently in comparison to some of the stateof-the-art methods.     

Breast cancer suppressor candidate-1 (BCSC-1) is a melanoma tumor suppressor that down regulates MITF

Understanding the molecular aberrations involved in the development and progression of metastatic melanoma (MM) is essential for a better diagnosis and targeted therapy. We identified breast cancer suppressor candidate-1 (BCSC-1) as a novel tumor suppressor in melanoma. BCSC-1 expression is decreased in human MM, and its ectopic expression in MM-derived cell lines blocks tumor formation in vivo and melanoma cell proliferation in vitro while increasing cell migration. We demonstrate that BCSC-1 binds to Sox10, which down regulates MITF, and results in a switch of melanoma cells from a proliferative to a migratory phenotype. In conclusion, we have identified BCSC-1 as a tumor suppressor in melanoma and as a novel regulator of the MITF pathway.     

Development and evaluation of Leishmania infantum rK26 ELISA for serodiagnosis of visceral leishmaniasis in Iran

The purpose of this study was to prepare recombinant K26 antigen from Leishmania infantum and evaluate its performance by enzyme-linked immunosorbent assay (ELISA) test for serodiagnosis of visceral leishmaniasis (VL) in endemic regions of Iran. The results were compared with those obtained by direct agglutination test (DAT) and whole cell ELISA using crude parasite antigen. Of 93 sera from patients with confirmed VL, 90 sera were positive with rK26 ELISA (sensitivity=96.8%), whereas 85 sera were positive with DAT (sensitivity=91.4%) and 89 sera were positive with whole cell ELISA (sensitivity=95.7%). Of 130 subjects who either had other infectious diseases (n=30) or were healthy (n=100), rK26 ELISA were negative in all cases (specificity=100%), whereas DAT were negative in 116 cases (specificity=89.2%) and whole cell ELISA was negative in 114 cases (specificity=87.7%). The results of this study indicate that the rK26 ELISA is more sensitive and specific than conventional methods and could be used for reliable diagnosis of VL caused by Leishmania infantum.     

Correlation of micro vessel density and c-Myc expression in breast tumor of mice following mesenchymal stem cell therapy

Stem cell therapy for degenerative diseases has been established; however there are controversies over the treatment of solid tumors with stem cell transplantation. In the present study, the anti-tumor action of mesenchymal stem cells (MSCs) has been examined in a mouse model of breast cancer with emphasize on tumor growth, angiogenesis and c-Myc expression in breast tumors. For this purpose, MSCs were isolated from bone marrow of Balb/c mice and characterized. A Balb/c mouse model of breast cancer was developed and subjected to cell therapy intra venous (I.V) or intra tumor (I.T) with MSCs. Tumor growth was measured using a digital caliber for until the end of experiment (30 days). Then the mice were sacrificed and their tumors were removed and processed for histopathological examination, immunohistochemical assay of CD31 and measuring of c-Myc expression using quantitative PCR. Detection of the labeled-MSCs in tumors following injection of the cells (I.V or I.T) clearly showed the homing of MSCs into tumors. Tumor growth in case of MSC-treated mice by I.V and I.T routes was inhibited by approximately 28% and 34% respectively compared to controls. The suppression of angiogenesis was reflected in Micro Vessel Density (MVD) following I.V or I.T delivery of the MSCs. c-Myc gene expression in tumor tissues of mice treated I.V or IT with MSCs was down-regulated to 28.0% and 16.0% respectively compare to control groups. In conclusion, growth inhibition of breast tumors in mice due to MSC therapy is associated with modulation of c-Myc activation and angiogenesis markers.     

Effects of bio fertilizer and nano Zn-Fe oxide on physiological traits,antioxidant enzymes activity and yield of wheat (Triticum aestivum L.) under salinity stress

In order to evaluate the effects of nano Zn-Fe oxide and bio fertilizer on physiological traits, antioxidant activity and yield of wheat under salinity stress, a factorial experiment was conducted based on RCBD with three replications.Treatments were included salinity in three levels (no-salt, salinity 25 and 50 mM NaCl), four bio fertilizers levels (no bio fertilizer, seed inoculation by Azotobacter, Azosperilium, Pseudomonas) and nano oxide (without nano, application of nano Zn oxide, nano Fe oxide and nano Fe-Zn oxide 1.5 g/lit). Salinty stress decreased the chlorophyll-a, chlorophyll-b, total chlorophyll, photochemical efficiency of PSII and yield of wheat, whearas electrical conductivity, soluble sugars, proline content, and the activities of Catalase (CAT), Peroxidase (POD) and Polyphenol Oxidase (PPO) enzymes increased. Similar results were observed in CAT, POD and PPO activities due to inoculation by bio fertilizers and nano oxide. Maximum of soluble sugars and proline content were observed in the highest salinity level and application of Pseudomonas. Application of nano Zn-Fe oxide increased about 17.40% from grain yield in comparision with no application of nano oxide in the highest salinity level. Generally, it was conducted that bio fertilizer and nano oxide can be used as a proper tool for increasing wheat yield under salinity condition.     

Differential allelopathic expression of bark and seed of Tamarindus indica L.

Allelopathic performance of the bark and seed of Tamarindus indica L. tree was evaluated through bioassay-guided studies using seven common agronomic crops (asparagus, cucumber, lettuce, radish, sesame, tomato and welsh onion) and seven weed species (barnyard grass, Chinese milk vetch, perennial ryegrass, phacelia, timothy grass, white clover and wild ginger) under laboratory conditions. As demonstrated by a sandwich method, the bark of the tamarind tree caused strong growth inhibition (compared to the corresponding controls) in both radicles and hypocotyls of the species tested, and the inhibitory effect was highest in barnyard grass (52–65%) and lowest in welsh onion (19–13%). The crude-water soluble extracts of bark at different concentrations (1, 5 and 10%) (w/v) exhibited a strong growth inhibition in all the plant species tested, and a proportional increase in the percentage of growth inhibition was observed with an increase in the concentrations of the extracts. The magnitude of inhibition in weed species was higher (5–60%) than those of agronomic crop species (3–40%). The growth of all the weed species tested was strongly inhibited (17–56%), while the agronomic crop species showed both inhibited (5–21%) and stimulated (5–27%) growth due to the effect of crude-water soluble exudates of tamarind seed. Among the agronomic crop species tested, lettuce (22–27%) followed by radish (20–25%) and sesame (5–8%) showed stimulatory growth with the crude-water soluble exudates of seed. In the pot culture experiments using four agronomic crops (lettuce, radish, tomato and cucumber) and two weed species (barnyard grass and white clover), spraying of crude-water soluble extracts of tamarind seed-coat at three different concentrations (1, 5 and 10%) (w/v) showed that the growth of lettuce (35–62%) and radish (32–56%) was stimulated, while all other species tested showed growth inhibition (29–61%). When the spraying of crude extracts of seed-coat was turned off, the growth of both lettuce and radish continued to be stimulated (4–7%) and all other previously inhibited species recovered well, the recovery percentage ranging between 78 and 82%. However, when spraying of crude extracts of seed-coat was continued, growth increased (10–14%) in lettuce and radish, and reduced (37–76%) in four other species tested. The inhibitory or stimulatory effects of the crude extracts on agronomic crop and weed species were higher in the radicle than the hypocotyl and reached a peak with 10% (w/v) concentrations. These results clearly demonstrated the differential allelopathic effects (inhibitory and excitatory) of bark and seed of tamarind tree in the species tested. Thus, it is evident that these two organs contain certain biologically active true growth regulator(s) and are either additively or synergistically involved in the plant-specific expression, particularly by the seed-coat.     

Early expression of vimenti in human mammary cultures

Summary The intermediate filaments of most epithelial cells in vivo consist solely of cytokeratins. Using monoclonal antibodies to vimentin or keratin, we have examined the expression of vimentin in homologous specimens of frozen tissue sections and primary cultures of normal human mammary epithelium. In frozen sections, only epithelial cells reacted with the antikeratin antibody, whereas antivimentin reactivity was associated with stromal cells. All epithelial cultures were positive for cytokeratin and in addition coexpressed vimentin as strongly as cultured fibroblasts and as early as the 4th d after initiation of the culture. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis of cytoskeletal preparations of secondary cultures of normal mammary epithelium have also demonstrated the appearance of a moiety identical to the vimentin found in cultured fibroblasts. Our observations are consistent with the hypothesis that vimentin expression is induced, possibly as a result of changes in cell shape or growth rate, when cells are freed from three-dimensional restirctions imposed by the tissue of origin     

Circadian resonance in the development of two sympatric species of Camponotus ants

Circadian clocks provide adaptive advantage to their owners by timing their behavioural and physiological processes in accordance with the external environment. Here we report the results of our study aimed at investigating the effect of the interaction between circadian timing system and environmental light/dark (LD) cycles on pre-adult development time of two sympatric species of Componotus ants, the night active Componotus compressus, and the day active C. paria—both species develop in dark underground nests, under fairly constant conditions of humidity and temperature. We estimated pre-adult developmental durations in these ants under three different LD cycles (T20 = 10 h of light and 10 h of darkness, T24 = 12 h of light and 12 h of darkness, and T28 = 14 h of light and 14 h of darkness). We find that both species exhibit significantly faster pre-adult development under T24 compared to T20 and T28. Given that faster development in insects is considered as an adaptive strategy these results can be taken to suggest that Camponotus ants accrue greater fitness advantage under T24 compared to T20 and T28 LD cycles, possibly due to “circadian resonance” between circadian timing system and environmental LD cycle. Thus our study reveals that boreal species of ants could serve as a case for the study of adaptive significance of circadian organization.     

The role of albumin and PPAR‐α in differentiation‐dependent change of fatty acid profile during differentiation of mesenchymal stem cells to hepatocyte‐like cells

Differentiation of mesenchymal stem cells (MSCs) to hepatocyte‐like cells is associated with morphological and biological changes. In this study, the effect of hepatogenic differentiation on fatty acid profile and the expression of proliferator‐activated receptors‐α (PPAR‐α) have been studied. For this purpose, MSCs isolated from human umbilical cord were differentiated into hepatocyte‐like cells on selective culture media. The morphological and biochemical changes, PPAR‐α expression and reactive oxygen species (ROS) levels were studied during the differentiation process. Besides, the cells were processed to determine changes in fatty acid profile using gas chromatography analysis. The results showed that hepatic differentiation of the MSCs is associated with a decrease in major polyunsaturated fatty acids in mature hepatocytes, whereas there was an increase in the saturated fatty acid (SFA) levels during hepatocyte maturation. The differentiation‐dependent shift in the ratio of SFA/USFA was associated with changes in albumin and PPAR‐α expression, whereas changes in fatty acid profile were independent of ROS production and lipid peroxidation in differentiating cells. In conclusion, these data may suggest that hepatocyte formation during the stem cell differentiation is associated with a shift in the fatty acid profile that is probably a normal phenomenon in hepatogenic differentiation of the MSCs. Copyright © 2014 John Wiley & Sons, Ltd.