Retrospective analysis of double-strand break rejoining data collected using warm-lysis PFGE protocols

Sample preparation procedures for the pulsed-field gel electrophoresis (PFGE) assay usually involve a lysis step at temperatures as high as 50°C. During this warm-lysis procedure, multiply damaged sites containing heat-labile sites (HLS) can be converted into double-strand breaks (DSB). Once formed, these DSB cannot be distinguished from the DSB formed directly by ionizing radiation. This paper develops a method to correct DSB estimates for the effects of HLS in warm-lysis protocols. A first-order repair model is used to predict the number of HLS available for conversion into DSB as a function of the time available for repair before initiating warm-lysis. A mathematical expression is derived to separate prompt DSB from those formed through the artefactual conversion of HLS into DSB. The proposed formalism only requires the specification of two adjustable parameters, both of which can be estimated from measured data. Estimates of prompt DSB yields obtained by correcting warm-lysis data are in good agreement with estimates obtained using cold-lysis protocols, which do not include the effect of HLS. The retrospective analyses of two published datasets suggest that corrections for HLS have a substantial impact on DSB yields within the first 20?–?30 min after irradiation. Bi-exponential fits to the DSB data for Chinese hamster ovary cells suggest that corrections for HLS reduce the half-time for fast DSB rejoining by about 15%, whereas the half-time for the slow DSB rejoining only decreases by 4%. The total DSB yield and the fraction of fast-rejoining DSB decrease by 24 and 38%, respectively, when the correction is applied. The proposed formalism can be used to characterize trends and uncertainties in DSB rejoining kinetics associated with the artefactual conversion of HLS into DSB. The retrospective application of the methodology to warm-lysis data enhances their relevance and usefulness for studies of DSB rejoining kinetics.     

Rapid bioassay-guided screening of toxic substances in vegetable oils that shorten the life of SHRSP rats

It has been consistently reported that vegetable oils including canola oil have a life shortening effect in Stroke-Prone Spontaneously Hypertensive Rats (SHRSP) and this toxic effect is not due to the fatty acid composition of the oil. Although it is possible that the phytosterol content or type of phytosterol present in vegetable oils may play some role in the life shortening effect observed in SHRSP rats this is still not completely resolved. Furthermore supercritical CO₂ fractionation of canola oil with subsequent testing in SHRSP rats identified safe and toxic fractions however, the compounds responsible for life shortening effect were not characterised. The conventional approach to screen toxic substances in oils using rats takes more than six months and involves large number of animals. In this article we describe how rapid bioassay-guided screening could be used to identify toxic substances derived from vegetable oils and/or processed foods fortified with vegetable oils. The technique incorporates sequential fractionation of oils/processed foods and subsequent treatment of human cell lines that can be used in place of animal studies to determine cytotoxicity of the fractions with structural elucidation of compounds of interest determined via HPLC-MS and GC-MS. The rapid bioassay-guided screening proposed would require two weeks to test multiple fractions from oils, compared with six months if animal experiments were used to screen toxic effects. Fractionation of oil before bio-assay enhances the effectiveness of the detection of active compounds as fractionation increases the relative concentration of minor components.     

Salicylic acid reduces the production of several potential virulence factors of Pseudomonas aeruginosa associated with microbial keratitis

PURPOSE: Pseudomonas aeruginosa is a common cause of contact-lens-related microbial keratitis. This bacterium is becoming increasingly resistant to antibiotics, and even if the infection can be treated with antibiotics, damage to the cornea resulting from the combined effect of bacteria and host factors can lead to loss of vision. The purpose of this study was to test the effect of salicylic acid on the production of potential virulence factors during the growth of P. aeruginosa. METHODS: Bacterial cells were grown in a subinhibitory concentration of salicylic acid, and supernatants were collected and analyzed for presence of proteases by using zymography and hydrolysis of chromogenic substrates. The supernatants were also analyzed for the amount of acetylated homoserine lactones by using bacterial reporter strains. Pseudomonas cells from salicylic acid cultures were analyzed for their twitching and swimming motility as well as their ability to invade or cause the death of corneal epithelial cells. RESULTS: Growth in a subinhibitory concentration of salicylic acid resulted in a significant reduction in the number of bacterial cells and a reduction in the rate of the number of bacteria increasing during logarithmic growth, but the time to reach the stationary phase of growth was unchanged. These changes in growth pattern affected the amount of acylated homoserine lactones produced by P. aeruginosa 6294. Also affected by growth in salicylic acid was the ability of strain 6294 to show twitching or swimming motility. Salicylic acid also reduced the invasion of strain 6294 into corneal epithelial cells and the epithelial cell death caused by strain 6206. Furthermore, production of proteases by P. aeruginosa was significantly reduced by growth in salicylic acid. CONCLUSIONS: The results of this study clearly demonstrate that salicylic acid has a significant impact on several potential virulence factors of P. aeruginosa that may be involved in the production of microbial keratitis. These effects were probably mediated by reduction in the cell density and concomitant reduction in the quorum-sensing signaling molecules, the acylated homoserine lactones, produced by P. aeruginosa.     

Impact of biting midges on residential property values in Hervey Bay, Queensland, Australia

Biting midges (Culicoides spp.) are an important environmental health issue in Hervey Bay, an area of rapid population growth in Australia. It is also the gateway to a World Heritage area (Great Sandy Strait) and a destination for tourists. The spread of housing developments into suburbs close to midge breeding habitats has led to a problem for the local government responsible for managing biting insects in its area. Suburbs with a severe biting midge problem were found to have significantly lower residential property values than less affected suburbs. The gross reduction in value in due to the midge problem was estimated to range from more than AUS dollar 25 million, based on actual sale price, to more than AUS dollar 55 million, based on the perceptions of the most severely affected residents.     

Aurantosides G, H, and I: three new tetramic acid glycosides from a Papua New Guinea Theonella swinhoei

Aurantosides G-I (1-3) have been isolated from the lithistid sponge Theonella swinhoei from Papua New Guinea. Their structures were established by spectroscopic and chemical methods. Compounds 1-3 represent new monochloropentaenoyl tetramic acids with mono-, di-, and tri-N-saccharide substituents, respectively. Aurantosides G-I (1-3) failed to show any significant cytotoxicity against the human colon tumor cell line HCT-116.     

Eeffects of pyrethroid insecticide ICON（lambda cyhalothrin） on reproductive competence of male rats

AIM: To assess the effect of ICON (trade name of lambda-cyhalothrin) on sexual competence and fertility of male rats. METHODS: Male rats were gavaged daily for 7 consecutive days with different doses of ICON (63 mg/kg and 100 mg/kg) or vehicle (distilled water). Their sexual behaviour and fertility were evaluated at different time points during treatment and post-treatment using receptive females. RESULTS: Treatment had no effect on fertility, but sexual competence was seriously impaired: libido (assessed in terms of pre-coital sexual behaviour, and numbers of mounting, intromission and ejaculation), sexual arousability/motivation (in terms of latencies for mounting, intromission and ejaculation), sexual vigour (judged by frequencies of mounting and intromission or copulatory efficiency). In addition, ICON suppressed intromission ratio, indicating erectile dysfunction. These effects on sexual function had a rapid onset and was reversible. ICON-induced sexual dysfunction was mediated by multiple mechanisms, mainly toxicity, stress, sedation and possibly via GABA and dopaminergic systems. CONCLUSION: Exposure to ICON may cause sexual dysfunction in male rats.     

Ablative Carbon Dioxide Laser Treatment for Papular Scars of Nose and Chin Due to Acne: A Case Series

Introduction:Papular scarring of the nose and chin due to acne is an under-reported,under-recognized,and under-treated condition.Ablative carbon dioxide laser t...     

Sch?tzung des Lebensalters kindlicher Opfer auf Bilddokumenten

In recent years more and more digital images with suspected pedopornographic content are being subjected to analysis by the law enforcement authorities. In this procedure the age of the victims plays a central role. The conventional age estimation from photographs is typically based on the evaluation of physical signs of sexual maturity, dental status (if observable) and body proportions. The analysis of facial proportions is a new approach which supplements the former criteria. As the development of the face is in general not fully completed until the age of 18 years, this approach provides an additional benefit due to the recently established age threshold of 18 years for juvenile pornography in Europe. The development of software for an automated age estimation based on facial proportions will contribute to a reduction of invested time and effort during the investigations into cases of suspected child abuse.