Identification and characterization of a novel peptide from rainbow trout (Oncorhynchus mykiss) with antimicrobial activity against Streptococcus iniae.

The overuse and misuse of antibiotics has led to the emergence of antibiotic-resistant bacterial species which remain a challenge to treat therapeutically. Novel and efficacious drugs are desperately needed to combat pathogens. One method to facilitate these discoveries is the use of in silico methods. Computational biology has the power to scan large data sets and screen for potential molecules with antibacterial function. In the current study, an in silico approach was used to identify an antimicrobial peptide (AMP) derived from rainbow trout von Willebrand Factor. The AMP was tested against a panel of aquatic bacterial pathogens and was found to possess antibacterial activity against Streptococcus iniae (S. iniae). Since S. iniae is a zoonotic pathogen, this may be useful in other species as well. The peptide was non-hemolytic and non-cytotoxic at the concentrations tested in rainbow trout cells. Pre-treatment of rainbow trout cells with the peptide did not result in an upregulation of immune genes but stimulating the rainbow trout macrophage/monocyte-like cell line, RTS11, with heat-killed S. iniae, did result in a significant upregulation of the tumor necrosis factor alpha (tnfa) gene. In this study, a new AMP has been identified but its expression, synthesis and role in vivo remains unknown. Nevertheless, the findings presented improve our understanding of fish gill and macrophage responses towards this important zoonotic pathogen.

Influence of genotype and perioral musculature on maxillary and mandibular development.

OBJECTIVES: To determine whether there is an association between skeletal jaw position and perioral musculature, and if genotypes can predict skeletal growth. MATERIALS AND METHODS: A prospective study on 42 patients over 1 year was performed. The study included 22 females and 20 males with and average age of 28.5 years. Lip strength was compared to radiographic cephalometric measurements. Allelic and genotypic frequencies from polymorphisms rs678397 and rs1815739 in ACTN3 and rs10850110 in MYO1H were compared to each variable. Chi-square and Fisher exact tests were used to determine if differences were statistically significant (alpha = 0.05). RESULTS: The data showed significant differences between rs678397 genotype and allele frequencies and SNA angle (P = .01; P = .003, respectively); between rs1815739 allele frequency and SNA angle (P = .01); between rs678397 allele frequency and ANB angle (P = .049); between rs678397 genotype and allele frequencies and lip strength in females (P = .045; P = .02); and between rs678397 allele frequency and overall lip strength (P = .049), after mean strength values used as cut off being customized by sex. CONCLUSIONS: Polymorphisms in ACTN3 are associated with weak lips and larger SNA and ANB angles.

Impact of analysis technique on our understanding of the natural history of labour: a simulation study.

OBJECTIVE: To evaluate the discrepancy between historical and more recent descriptions of the first stage of labour by testing whether the statistical techniques used recently (repeated-measures polynomial and interval-censored regression) were appropriate for detection of periods of rapid acceleration of cervical dilatation as might occur at the time of transition from a latent to an active phase of labour. DESIGN AND SETTING: A simulation study using regression techniques. SAMPLE: We created a simulated data set for 500 000 labours with clearly defined latent and active phases using the parameters described by Friedman. Additionally, we created a data set comprising 500 000 labours with a progressively increasing rate of cervical dilatation. METHODS: Repeated-measures polynomial regression was used to create summary labour curves based on simulated cervical examinations. Interval-censored regression was used to create centimetre-by-centimetre estimates of rates of cervical dilatation and their 95th centiles. MAIN OUTCOME MEASURES: Labour summary curves and rates of cervical dilatation. RESULTS: Repeated-measures polynomial regression did not detect the rapid acceleration in cervical dilatation (i.e. acceleration phase) and overestimated lengths of labour, especially at smaller cervical dilatations. There was a two-fold overestimation in the mean rate of cervical dilatation from 4 to 6 cm. Interval-censored regression overestimated median transit times, at 4- to 5-cm cervical dilatation or when cervical examinations occurred less frequently than 0.5- to 1.5-hourly. CONCLUSION: Repeated-measures polynomial regression and interval-censored regression should not be routinely used to define labour progress because they do not accurately reflect the underlying data. TWEETABLE ABSTRACT: Repeated-measures polynomial and interval-censored regression techniques are not appropriate to model first stage of labour.

Antimicrobial Peptides of Salmonid Fish: From Form to Function.

Antimicrobial peptides (AMPs) are small, usually cationic, and amphiphilic molecules that play a crucial role in molecular and cellular host defense against pathogens, tissue damage, and infection. AMPs are present in all metazoans and several have been discovered in teleosts. Some teleosts, such as salmonids, have undergone whole genome duplication events and retained a diverse AMP repertoire. Salmonid AMPs have also been shown to possess diverse and potent antibacterial, antiviral, and antiparasitic activity and are induced by a variety of factors, including dietary components and specific molecules also known as pathogen-associated molecular patterns (PAMPs), which may activate downstream signals to initiate transcription of AMP genes. Moreover, a multitude of cell lines have been established from various salmonid species, making it possible to study host-pathogen interactions in vitro, and several of these cell lines have been shown to express various AMPs. In this review, the structure, function, transcriptional regulation, and immunomodulatory role of salmonid AMPs are highlighted in health and disease. It is important to characterize and understand how salmonid AMPs function as this may lead to a better understanding of host-pathogen interactions with implications for aquaculture and medicine.

Xela DS2 and Xela VS2: Two novel skin epithelial-like cell lines from adult African clawed frog (Xenopus laevis) and their response to an extracellular viral dsRNA analogue.

The skin epithelial layer acts as an important immunological barrier against pathogens and is capable of recognizing and responding to pathogen-associated molecular patterns (PAMPs) in human and mouse models. Although presumed, it is unknown whether amphibian skin epithelial cells exhibit the ability to respond to PAMPs such as viral double-stranded RNA (dsRNA). To address this, two cell lines from the dorsal skin (Xela DS2) and ventral skin (Xela VS2) of the African clawed frog (Xenopus laevis) were established. Xela DS2 and Xela VS2 cells have an epithelial-like morphology, express genes associated with epithelial cells, and lack senescence-associated beta-galactosidase activity. Cells grow optimally in 70% Leibovitz's L-15 medium supplemented with 15% fetal bovine serum at 26 °C. Upon treatment with poly(I:C), a synthetic analogue of viral dsRNA and known type I interferon inducer, Xela DS2 and Xela VS2 exhibit marked upregulation of key antiviral and pro-inflammatory transcripts suggesting frog epithelial cells participate in the recognition of extracellular viral dsRNA and production of local inflammatory signals; similar to human and mouse models. Currently, these are the only known Xenopus laevis skin epithelial-like cell lines and will be important for future research in amphibian epithelial cell biology, initial host-pathogen interactions, and rapid screening of the effects of environmental stressors, including contaminants, on frog skin epithelial cells.

Frog Skin Innate Immune Defences: Sensing and Surviving Pathogens.

Amphibian skin is a mucosal surface in direct and continuous contact with a microbially diverse and laden aquatic and/or terrestrial environment. As such, frog skin is an important innate immune organ and first line of defence against pathogens in the environment. Critical to the innate immune functions of frog skin are the maintenance of physical, chemical, cellular, and microbiological barriers and the complex network of interactions that occur across all the barriers. Despite the global decline in amphibian populations, largely as a result of emerging infectious diseases, we understand little regarding the cellular and molecular mechanisms that underlie the innate immune function of amphibian skin and defence against pathogens. In this review, we discuss the structure, cell composition and cellular junctions that contribute to the skin physical barrier, the antimicrobial peptide arsenal that, in part, comprises the chemical barrier, the pattern recognition receptors involved in recognizing pathogens and initiating innate immune responses in the skin, and the contribution of commensal microbes on the skin to pathogen defence. We briefly discuss the influence of environmental abiotic factors (natural and anthropogenic) and pathogens on the immunocompetency of frog skin defences. Although some aspects of frog innate immunity, such as antimicrobial peptides are well-studied; other components and how they contribute to the skin innate immune barrier, are lacking. Elucidating the complex network of interactions occurring at the interface of the frog's external and internal environments will yield insight into the crucial role amphibian skin plays in host defence and the environmental factors leading to compromised barrier integrity, disease, and host mortality.