An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder tumor model. The mouse model is established by urethral catheterization under inhaled general anesthetic. Chemical burn is then introduced to the bladder mucosa using intravesical silver nitrate solution to disrupt the bladder glycosaminoglycan layer and allows cells to attach. Following several washes with sterile water, human bladder cancer KU-7-luc2-GFP cells are instilled through the catheter into the bladder to dwell for 2 hours. Subsequent growth of bladder tumors is confirmed and monitored by in vivo bladder ultrasound and bioluminescent imaging. The tumors are then treated intravesically with saRNA formulated in lipid nanoparticles (LNPs). Tumor growth is monitored with ultrasound and bioluminescence. All steps of this procedure are demonstrated in the accompanying video.     

Contrast-enhanced versus non-enhanced three-dimensional echocardiography of left ventricular volumes

Background In three-dimensional echocardiography (3DE), individual endocardial trabeculae are not clearly visible necessitating left ventricular (LV) volumes to be measured by tracing the innermost endocardial contour. Ultrasound contrast agents aim to improve endocardial definition, but may delineate the outermost endocardial contour by filling up intertrabecular space. Although measurement reproducibility may benefit, there may be a significant influence on absolute LV volume measurements. Methods Twenty patients with a recent myocardial infarction and good ultrasound image quality underwent 3DE using the TomTec Freehand method before and during continuous intravenous contrast infusion. LV volumes were measured offline using TomTec Echo-Scan software. Results The use of contrast enhancement increased end-diastolic (110±35 vs. 144±53 ml; p<0.01) and end-systolic volume measurements (68±31 vs. 87±45 ml; p<0.01) significantly compared with non-contrast; the ejection fraction remained unchanged (40±13 vs. 41±14%, p=NS). Measurement reproducibility did not improve significantly, however. Conclusion Volumes measured by 3DE are significantly larger when ultrasound contrast is used. Possibly, intertrabecular space comprises a substantial part of the LV cavity. In the presence of an adequate apical acoustic window, ultrasound contrast does not improve LV volume measurement reproducibility. (Neth Heart J 2008;16:47-52.)     

Novel pre-treatment processes to promote linen-containing fabrics properties

This study was undertaken to investigate the effect of plasma pre-treatment, followed by enzymatic treatment in the absence and presence of bleaching agent on the properties of linen and linen-containing fabrics. Different plasma gases (air, oxygen and nitrogen), enzymes (acid-cellulases, neutral-cellulase and alkaline-pectinase) as well as bleaching agents (peracetic acid and H₂O₂) were used. The changes in physico-mechanical properties, surface morphology and dyeing properties of the treated substrates have been investigated. The obtained results indicated that plasma pre-treatment followed by subsequent acid-cellulases/peracetic acid or alkaline-pectinase/H₂O₂ treatment result in: a dramatic improvement in hydrophilicity and wettability as well as in the degree of whiteness of the treated substrates, an improvement in reduction of surface roughness and extent of post-reactive dyeing, along with a weight loss and a drop in the tensile strength. The extent of surface modification as well as the changes in the above-mentioned properties are governed by the characteristics of the substrate, the plasma gas, the nature and dose of the used enzyme, as well as the type of bleaching agent and additive. The optimal treatment sequence for attaining better performance properties was O₂-plasma followed by alkaline-pectinase/H₂O₂ treatment in presence of PEG 400.     

一个适于最大似然法估计物种分化年代的进化速率平滑近似方法

众所周知 ,物种分化年代的估计对分子钟 (进化速率恒定 )假定很敏感。另一方面 ,在远缘物种 (例如哺乳纲不同目的动物 )的比较中 ,分子钟几乎总是不成立的。这样在估计分化时间时考虑不同进化区系的速率差异至为重要。最大似然法可以很自然地考虑这种速率差异 ,并且可以同时分析多个基因位点的资料以及同时利用多重化石校正数据。以前提出的似然法需要研究者将进化树的树枝按速率分组 ,本文提出一个近似方法以使这个过程自动化。本方法综合了以前的似然法、贝斯法及近似速率平滑法的一些特征。此外 ,还对算法加以改进 ,以适应综合数据分析时某些基因在某些物种中缺乏资料的情形。应用新提出的方法来分析马达加斯加的倭狐猴的分化年代 ,并与以前的似然法及贝斯法的分析进行了比较     

Behavioral and reproductive effects of ultrasound on the Indian meal moth, Plodia interpunctella

The daily activity patterns of adult movement, female calling, and mating of the Indian meal moth, Plodia interpunctella (Hübner) (Lepidoptera: Pyralidae), were examined both in the absence and presence of ultrasound. Moths were exposed to ultrasound from a commercial ultrasonic device (Cix 0600) that produces constant sound patterns, and from a device developed at Kansas State University (KSU device) that produces random sound patterns. Daily activity patterns of adult movement, female calling, and mating followed a similar trend in the absence or presence of ultrasound. Female calling and mating, both in the absence and presence of ultrasound, primarily occurred during scotophase (21.00–07.00 hours). Ultrasound from the two devices significantly reduced the frequency of female calling and mating relative to unexposed moths. Consequently, the number of spermatophores transferred by males to females and egg production were lower in females exposed to ultrasound compared with unexposed females. In the absence of ultrasound, female P. interpunctella mated 2.9 times, resulting in 2.8 spermatophores/female. In the presence of ultrasound from the Cix 0600 device, a female mated 2.1 times and had 1.7 spermatophores. Corresponding values for the KSU device were 1.9 and 1.4, respectively. In the absence of ultrasound, 78% of the matings lasted 30–90 min, whereas in the presence of ultrasound 45–58% of the matings lasted either less than 30 min or more than 90 min. Moths exposed to ultrasound laid 96–130 eggs female^?1 compared with 229 eggs female^?1 for unexposed moths. Ultrasound did not affect the pre‐oviposition period and adult longevity of P. interpunctella.     

Combining Information from Similar Experiments: II. Quantifying Qualitative Results

Suppose that independent experiments each indicate general qualitative results, such as higher than normal incidence rates of tumors for exposed populations. This paper suggests methods for amalgamating the qualitative results from several such experiments into a more quantitative form, such as a dose-response relationship. The methods are designed to be robust both to systematic bias in one of the experiments and also to procedural variability across experiments. Data from four rodent experiments with tolazamide are used to illustrate the methods.     

Evaluation of secondary infertility in the gorilla

Reproductive failure in two female gorillas in the Los Angeles Zoo led to evaluation for secondary infertility. Male factor was excluded by noting routine conceptions in other individuals in the same group, and by behavioral observations of coital activity with the infertile females. Biochemical evidence of ovarian function was obtained by urinary sex steroid analysis. Continuity of the reproductive tract of the infertile apes was then studied by hysterosalpingography and laparoscopy while their general health was evaluated by ultrasonography and physical examination. Progressive, multiorgan involvement with Echinococcus vogeli was documented and mebendazole therapy begun. The investigation of secondary infertility in great apes by combined methods is safe and accurate, but requires special equipment and experienced personnel.     

Comparison of accuracy of intramuscular fat prediction in live pigs using five different ultrasound intensity levels

The objective of this study was to evaluate the possibility of prediction of intramuscular fat (IMF) in live pigs using ultrasound method. Moreover, the accuracy of prediction at five different ultrasound intensity levels was investigated. Cross-sectional images of longissimus dorsi muscle (LD) at right last rib area, from hybrid pigs, were taken. Each pig was scanned at the same frequency (3.5 MHz) and at the five different ultrasound intensity levels 70%, 75%, 80%, 85% and 90% of total amplifying of sonograph, using the device ALOKA SSD-500. The video image analysis was used to predict IMF content (ultrasound intramuscular fat (UIMF) 70 to UIMF90). The second day after slaughter, the dissection of right half carcass was done. A sample of LD at the last rib was taken for laboratory analysis of IMF content (LAIMF). Scatter plots with UIMF on the x-axis and LAIMF on the y-axis were constructed to account for individual variability within and between intensity levels. Correlations between LAIMF and UIMF were significantly different from zero (r = 0.40–0.52), except for correlation between LAIMF and UIMF90 (r = 0.14). Statistical model with LAIMF (the dependent variable), UIMF (the same model for each intensity level), live weight (the covariates) and sex (the fixed effect) was developed. Coefficients of determination (R²) were 0.33, 0.38, 0.34, 0.25 and 0.17 with UIMF at the intensity level 70%, 75%, 80%, 85% and 90%. Root mean square errors ranged from 0.516% to 0.639%. Standard errors of individual prediction ranged from 0.523% to 0.649%. Goodness-of-fit of the model was also justified by testing the residuals for normality. Although the results are not quite unequivocal in favour of the one intensity level, it seems that intensity levels 75% and 80% are the most suitable to predict IMF in live pigs. Further research is needed, mainly to increase accuracy of collecting, processing and evaluating the sonograms using video image analysis.