Bacteriophages are bacterial viruses used to recognize

Bacteriophages are bacterial viruses used to recognize and infect host bacterial cells with high specificity in various environments (Weinbauer, 2004; Mahony et al., 2011). During phage-bacterial infection and replication, other neighboring bacteria should not be affected by phages; this suggests their evolution of host specificity (Chan et al., 2013; Lee et al., 2014). Although the development of phage resistance is when phage is very specific and only one phage is used more often, as already proposed in several literature reports (Labrie et al., 2010; Chan et al., 2013; Mateus et al., 2014). This can be achieved by a combination of more than one phage at the same time. The phage potential is to lyse bacteria and they also allow the treatment of multiple pathogens simultaneously (Gu et al., 2012; Sillankorva et al., 2012; Chan et al., 2013). Moreover, previous studies suggest that virulent bacteria that become resistant to phage infection are less fit or could lose their pathogenic properties (Capparelli et al., 2010; Filippov et al., 2011). The bacterial cell surface components, that act as receptors for phage adsorption can also act as virulent factors (Labrie et al., 2010; Mateus et al., 2014). This may undergo mutation when bacteria develop resistance to the phages and render them not virulent (Sorek et al., 2008).
In addition, a phage cocktail can increase the efficiency of phage therapy, approved as novel types of food preservatives that delay the development of drug resistance by the bacteria in aquaculture production (Flynn et al., 2004; Lee et al., 2014). Therefore, phage approach would be a suitable biocontrol to emerging V. harveyi in shrimp aquaculture, and it may also be able to control multiple antibiotic resistant (MAR) vibriosis in raw seafood for therapeutic purposes.
In this study, three V. harveyi specific phages that infect both shrimp pond and sea coastal environmental isolates were isolated and characterized by the determined specificity of host range, survival, one-step growth curve analysis, morphology, and phage-mediated biocontrol of aquatic animal model observation. Furthermore, the genomic level of information was provided by the nature of nucleic acids, randomly amplified polymorphic DNA (RAPD), and restriction fragment length polymorphism (RFLP) markers for evaluating the genetic similarity. The major structural proteins were detected by SDS-PAGE. This report will be helpful to extend our knowledge of V. harveyi infecting bacteriophages in future applications as potential biocontrol or therapeutic agents against multiple mps1 resistant V. harveyi from the aquaculture environment.

Materials and methods


The discovery of phages by Frederick Twort (1915) and the classic double layer agar plate method is extensively used in phage research to isolate, identify, and enumerate new phages and mutants. Numerous phages from large and well-defined plaques are easily observed so that they can be enumerated when plated by the plaque assay (Adams, 1959; Twort, 1915). However, the plaque development and plaque size seems to be directly proportional to continuous phage adsorption, burst size, and the diffusion of phages in the medium (Abedon and Culler, 2007).
The host range and efficiency of phage lytic potential confirmations indicate that members of the Siphoviridae family are ubiquitous in the aquaculture environment and might be effective to control the Harveyi clade of Vibrio sp. in the aquaculture system. The host range of the purified Siphoviridae had relatively wide host ranges, including some but not all tested isolates of the closely related species. The Siphoviridae phage VhCCS-02 and VhCCS-20 determined that the host range additionally included V. parahaemolyticus which were analyzed (Sawabe et al., 2007; Crothers-Stomps et al., 2010). Thus, their lytic potential against V, harveyi clade bacterial species needs to be further investigated. The best candidates for phage therapy should be viruses able to lyse the majority of the bacterial target’s isolates (Chan et al., 2013). Thus, VHM1, VHM2, and VHS1 are suggested as potential phage therapy candidates against infection caused by V. harveyi, since their lytic spectrum include all the V. harveyi isolates tested and even isolate from other species. The study is also unique in observation of Pasharawipas et al., (2005) in that V. harveyi is susceptible within the family Siphoviridae phages used (Shivu et al., 2007). Moreover, these phages were able to influence the development of several isolates isolated from the aquaculture environments. It is realized that several marine phages can infect different isolates of the same species or different closely related species (Comeau et al., 2006; Crothers-Stomps et al., 2010), however, specificity of the phage–host interaction in marine water is not completely elucidated and needs further investigation (Holmfeldt et al., 2007). Interestingly, there is a wide host range of Vibrio-phage isolated using V. alginolyticus host isolates able to infect several V. harveyi isolates (Lin et al., 2012). The conservative nature of the structure of Lipopolysaccharide (LPS) phage receptor on the outer membrane of many of the Gram-negative bacterial species could be a possible clarification for this wide lytic spectrum (Rakhuba et al., 2010). Further, the genetic similarity of these bacterial species which belong to the Harveyi clade may also have contributed to this observation (Hoffmann et al., 2012; Urbanczyk et al., 2013).

br Results br Discussion In the


In the present study, the prevalence of M. hyo-positive piglets around weaning (3–5 weeks of age) was in the same range as has been reported by many other recent studies (Calsamiglia and Pijoan, 2000; Fano et al., 2007; Sibila et al., 2007a; Nathues et al., 2010; Villarreal et al., 2010; Fablet et al., 2012). Nevertheless, when compared to Villarreal et al. (2010), on a herd level, a significantly lower percentage of M. hyo-positive herds was detected around weaning age (3–4 weeks of age), which may be due to differences in study design, principally the inclusion criteria and the sampling procedure. Villarreal et al. (2010) only included pig herds with a recent history of coughing in grower-finisher pigs, whereas our study herds had no indicative clinical signs of respiratory disease in peri-weaned or post-weaned piglets, or in fattening pigs.
In a recent French study which evaluated data from pigs from 125 herds (Fablet et al., 2012), it prostaglandin receptor was found that that 33.6% of the farms had at least one M. hyo PCR-positive piglet at 4 weeks of age increasing to 39.2% at 10 weeks of age, slightly higher than the prevalences reported in the present study (27.0 and 29.0% at 3–5 and 6–11 weeks of age, respectively). The within-herd prevalence was also higher, 14.1% and 16.1% at 4 and 10 weeks of age, respectively, compared to the 7.1% and 10.9% at 3–5 and 6–11 weeks of age in the present study. Again these differences are likely to be due to differences in prevalence of respiratory disease, with ~2/3 of the herds in the French study having respiratory disease detected at slaughter.
In our study, the difference in M. hyo prevalence between piglets at 3–5 weeks of age and the subsequent group of 6–11 weeks of age can be explained by an age effect and a batch-to-batch variation. Also, Fano et al. (2007) showed an important disparity between-batch prevalence (0 to 51.3%), which was observed when different batches were sampled over a year. However, in our study, different piglet batches in the same herd were sampled concurrently, in contrast to Fano et al. (2007) who sampled subsequent batches at weaning over the entire year. Nevertheless, during most seasons (winter, spring and autumn), an increase in M. hyo test-positive piglets between 3–5 weeks of age and 6–11 weeks of age was observed, which is in agreement with transmission results obtained in previous studies (Meyns et al., 2004, 2006; Villarreal et al., 2011), which show a clear increase in the percentage of M. hyo-positive piglets between young weanling piglets and those of older weaner pigs.
The effect of weather on M. hyo prevalence in peri- and post-weaned piglets was partly in accordance with the findings of Segalés et al. (2012), who showed a positive relation, at the piglet level, between M. hyo-positive nasal swabs and rainfall and between M. hyo-positive serology and weekly temperature. In our study, a significant negative association with rainfall was observed for piglets around weaning (3–5 weeks of age), whereas in older weaners (6–11 weeks of age), season of the year, outdoor relative humidity and minimum outdoor temperature were positively associated with the probability of being M. hyo-positive at the piglet level.
Overall, the odds of being M. hyo-positive at piglet level were highest during autumn (S4) and lowest during summer (S3), which, in Belgium and The Netherlands, coincides with the high rainfall season and the highest ambient temperatures with the lowest relative humidity, respectively. In previous studies, a higher frequency and transmission of respiratory problems occurred during cold wet seasons or lower environmental temperatures (Goodwin, 1985; Dee et al., 2010; Otake et al., 2010), with more favourable conditions for the survival of M. hyo. Mycoplasma hyopneumoniae is known to be very sensitive to UV-light and dry environments under summer conditions (Jorsal and Thomsen, 1988), whereas survival for at least 31 days in water at temperatures of 2–7 °C is possible (Goodwin, 1985). These associations with weather conditions explain the seasonal differences in M. hyo that we found.

The theme of the Special Issue of is veterinary oncology

The theme of the 2015 Special Issue of is veterinary oncology. The editorial team thought that this topic was timely, given the substantial recent progress in the understanding of the molecular pathobiology of cancer. This progress has thankfully been translated into new effective approaches to treat, prevent or detect cancer in humans, but also in animals. Likewise, improvement in the identification, design and delivery of experimental molecules with selective and potent activity against cancer AEBSF has led to the development of a new generation of chemotherapeutic drugs with improved safety profiles and enhanced therapeutic activity against some cancer types. While it would have been impossible to cover all these recent exciting developments in a single issue, the current Special Issue of contains multiple reviews and original articles authored by renowned colleagues from North America, Europe, Australia and Asia. This geographical diversity speaks to the global nature of veterinary science and is a good testament of the global stature that has acquired in recent years.
The advent of multiple novel technologies, especially the omics technologies, has helped generate and interpret a previously unfathomed volume of data. As a result, our understanding of the molecular pathobiology of cancer has improved dramatically, leading to significant changes in the approaches to cancer treatment and detection, but also in the directions of drug discovery efforts. This impact is well illustrated by the advances in personalized medicine reviewed by Professor Robert Klopfleisch from The Institute of Veterinary Pathology at The Freie Universität Berlin (). The objective of personalized medicine is to tailor treatments according to the molecular genotype and phenotype of the individual patient, and our ability to characterize these genotypes and phenotypes has been greatly enabled by the use of technologies such as next generation sequencing (NGS) and proteomics. Interestingly, as noted by , despite the wide diversity in molecular alterations observed among tumors, only a restricted number of driver genes or molecular pathways contribute to the development of most cancers, supporting the concept that personalized therapeutic approaches in oncology should result in enhanced efficacy and safety.
In the first part of a two-part review, Drs. from The University of Porto discuss the clinically relevant genetic and molecular pathways driving cell proliferation, apoptosis and DNA repair in canine mammary gland tumors (CMTs). In the second part of this review, these authors focus on pathways involved in metastasis, as well as the interactions between tumor cells and stromal components (inflammatory cells, cancer-associated fibroblasts, and endothelial cells) of the tumor microenvironment to put in context the clinical utility of different non-surgical therapeutic modalities for CMT treatment (). Since CMTs are the most common neoplasm in female dogs, it is of no surprise that this tumor type is a frequent focus of research in veterinary medicine. Hence, from Taiwan studied Kruppel-like factor 4 (KLF4) expression in CMTs and demonstrated that downregulation of this transcription factor was associated with decreased CMT cell proliferation and migration, and with induced cell death, suggesting that KLF4 is an oncogene in CMTs and may represent a suitable target for CMT therapy. Likewise, from The National Ilan University in Taiwan focus on the role of heat shock protein 27 (HSP27) in CMTs and provide evidence that canine HSP27 overexpression is associated with increased cell proliferation, clone formation, migration, and decreased sensitivity to doxorubicin, all supportive of an oncogene role.
Professor David Argyle and colleagues from The Royal (Dick) School of Veterinary Studies and The Roslin Institute at the University of Edinburgh also cover in two very comprehensive reviews some critical aspects of cancer biology in this Special Issue. The first review focuses on the cancer stem cell (CSC) model, a relatively recent area of cancer biology that has been a key focus of cutting-edge oncology research in the last few years (). In the second review, successfully took the daunting challenge of summarizing a large volume of recent data and studies to provide a comprehensive illustration of the link between chronic inflammation and cancer. This information provides further support that the tumor-promoting inflammatory environment is one of the major cancer hallmarks, as supported by the recent development of immuno-oncology therapies with exciting efficacy in human patients ().

The results of our study show that

The results of our study show that after multi-level upper airway surgery, all clinical signs associated with brachycephalic airway syndrome improved substantially. The most encouraging result was the striking 90% urotensin ii in the frequency of life-threatening events. Dogs with severe brachycephalic airway syndrome frequently experience choking fits, cyanosis and even collapse (Wykes, 1991; Roedler et al., 2013). Air hunger is a most unpleasant sensation and could have the greatest potential to compromise animal welfare (Beausoleil and Mellor, 2015).
Marked improvement in the frequency of reported sleeping problems was also observed after surgery. Preoperatively, 34/62 (55%) owners reported that their dogs showed numerous sleeping problems. Postoperatively, sleeping difficulties were almost completely resolved, except for two dogs that had sleep apnoea. Upper airway resistance in brachycephalic dogs is increased by multiple anatomic abnormalities and can be aggravated during sleep by relaxation of muscles that dilate the upper airways (Hobson, 1995). Thus, some brachycephalic dogs attempt to relieve respiratory difficulties by assuming particular body positions during rest to reduce upper airway resistance (Roedler et al., 2013). Widdicombe and Davies (1988) demonstrated that nasal obstruction significantly increases upper airway resistance, which might explain why dogs with severe nasal obstruction can only sleep with their mouths open. Approximately half of the dogs in our study experienced apnoeic episodes. Interestingly, brachycephalic dogs have been used as animal models for sleep disordered breathing in humans (Hendricks et al., 1987). The results of our study indicate that successful surgical treatment of upper airway obstruction can eliminate sleep disordered breathing and demonstrated that multi-level airway surgery can reduce the incidence of canine obstructive sleep disorders markedly.
Breathing sounds in brachycephalic dogs are comprised of a combination of different stridors caused by narrowing in various areas of the upper airways. Distinguishing between these stridors appears to be impossible for dog owners, but they can recognise the occurrence and frequency of breathing sounds (Packer et al., 2012). In our survey, dog owners were required to report the occurrence of breathing noises while their dogs slept, rested, and exercised. Following multi-level surgery, the occurrence of breathing sounds improved in all circumstances as follows: at rest 17/41 (41%), during exercise 34/62 (55%) and while sleeping 32/59 (54%). Snoring is thought to result from vibration of the soft palate and adjacent tissues (Liistro et al., 1991). Oechtering (2010) summarised the factors affecting the width and collapsibility of the nasopharynx in brachycephalic dogs. The increased volume of soft tissue in the nasopharynx (hypertrophic soft palate, hypertrophic tonsils and obesity) and negative pressure during inspiration due to nasal stenosis, both result in pathological narrowing of the nasopharynx. Additionally, when the mouth is closed and the ventral head or neck is subjected to pressure (i.e. prone position), the relatively large tongue further increases airway narrowing. While nasal stenosis can be treated satisfactorily by surgery (Schuenemann and Oechtering, 2014b; Oechtering et al., 2016b), surgical interventions have limited impact on the nasopharyngeal area. Weight loss can improve nasal stenosis, but is rarely curative, as it shortens and thins the enlarged soft palate, thereby affecting only the ventral margin of the nasopharynx (Oechtering, 2010). As a result, persistence of some breathing sounds should be expected despite upper airway surgery.
Brachycephalic dogs must generate strong negative pressure in the thorax to overcome the resistance produced by multifactorial upper airway obstruction. Typically, this inspiratory effort is evident as laboured breathing (Carvalho et al., 2010; Oechtering, 2010). Preoperatively, two-thirds of owners observed inspiratory effort in their dogs during physical exercise. Improvement was reported in 80% of all dogs postoperatively, which is further evidence that surgery can reduce upper airway resistance. Interestingly, the questions concerning inspiratory effort in the preoperative survey revealed that a high number of dog owners felt unable to answer these questions correctly. This suggests that recognising inspiratory effort can be challenging for dog owners.

igf1r inhibitor br Material and methods br Results and discussion br Conclusion

Material and methods

Results and discussion

With regard to all the studied traits, Djenah Khetifa was selected as the most water-stress tolerant genotype under in vitro conditions, followed by Oued Zenati. Waha appeared to be the most sensitive, but also the most embryogenic, under non-stress conditions. The results verified a remarkable variation for callus induction ability in genetic materials under water stress condition that can be used in durum wheat breeding program. These genotypes should be tested in a field trial and then looking for the association or correlation between in vitro and in vivo conditions. The selected igf1r inhibitor and plants will provide a tool for determining the mechanisms involved in tolerance to water stress. We suggest that in vitro screening method for drought tolerance would be an appropriate route to develop drought-tolerant lines in durum wheat.

This work was supported by the Ministry of Higher Education and Scientific Research of Algeria and Walloon Agricultural Research Centre in Belgium. We thank the both institutions for their assistance.

Morus indica L., commonly known as Indian mulberry, is a woody, perennial plant belonging to the family Moraceae. The cultivated varieties are mostly diploid and monoecious, which flower during August and the fruit set starts in early September. M. indica is considered as an economically important plant, which is cultivated commercially for its edible fruit and its foliage as the primary feed of silkworm (Bombyx mori L.). Anthocyanins derived from this plant are a natural food colorant and fabric-tanning agent [1]. Moreover, different clinical trials have established the presence of therapeutic agents showing hypoglycemic, hypolipidemic, diuretic and hypotensive properties [2,3]. The presences of different bioactive compounds such as quercetin, rutin, and isoquercetin were reported in the mulberry plant [4]. This plant is currently under various pharmacognostic examinations to explore the mechanism of action in different traditional medicines in which various parts of the plant are the major component.
Conventionally, the propagation of Indian mulberry is mainly limited to cuttings, as the seeds are mostly heterozygous due to cross pollination. However, there is a major concern over the difficulty in rooting through cuttings [5]. Furthermore, the successful rooting through conventional cutting depends on the genotype, environmental conditions and physiological state of the donor plant [6]. Under the intensive care and standardizing the limiting factor, only less than 40% success rate is obtained in the final transplantation stage [7]. This shows the exigency for an alternative more efficient method for the clonal propagation of this species.
Plant tissue culture technique has been successfully utilized to propagate uniform Indian mulberry platelets in mass scale while maintaining the genetic fidelity. In vitro propagation of M. indica has been reported by various workers utilizing different protocols. In the majority of the work reported, 6-benzylaminopurine (BAP) was considered effective in inducing shoots from axillary bud culture [8–10]. Lalitha et al. [11] obtained a high frequency of multiple shoots in excised nodal explants supplemented with thidiazuron (TDZ, 9.08μM). The effectiveness of 2,4-dichlorophenoxyacetic acid (2,4-D) was found to act both as a shooting and as a rooting agent at varying concentrations in M. indica. Chitra and Padmaja [12] reported that 0.3mgL−1 2,4-D induced healthy shoots which subsequently induced rooting in the presence of 1mgL−1 2,4-D.
Despite various successful micropropagation reports, in vitro contamination by the microorganisms trapped in lenticels and scale leaves is one of the most challenging problems particularly for the field-grown explants of M. indica[13,14]. Another challenge is the long multistep acclimatization process [15–17] which causes economically unviable cost of production. The quality of roots of in vitro plantlets along with the number and status of stomata greatly influences the successful acclimatization [18].

br Introduction The field of nanotechnology is considered

The field of nanotechnology is considered as one of the most important and active areas of research. Its fundamental building blocks, nanoparticles, are distinguished by some exclusive physicochemical properties such as optical, electronic, catalytic and magnetic ones. These unique properties of nanoparticles could be attributed to high surface area in comparison to volume ratio, which potentially results in high reactivity [6].
Silver nanoparticles (AgNPs) have been reported to be applied in agricultural, medical and environmental fields [11]. Additionally, this type of nanoparticles is the best choice for potential applications in the field of living organisms and biological systems.
The increased usage of nanoparticles especially for medical purposes created a new challenge of synthesizing them by untraditional methods to overcome the undeniable disadvantages of other physical and chemical methods [21]. Plant mediated nanoparticles synthesis (green method) is preferred as it is clean, cost effective, nontoxic and safe for human therapeutic use [4].
Onion (Allium cepa) is a widely cultivated plant all over the world. It is rich in carbohydrates, proteins, sodium, potassium and phosphorus. It has been reported to have numerous important properties such as antimicrobial, antioxidant, antiparasitic and anti-inflammatory activities [7]. Recently, it is found that onion can be used as a potential candidate for synthesis of silver nanoparticles.

Materials and methods

Results and discussion
In this study, silver nanoparticle (AgNP) synthesis by reducing Ag+ ions present in the aqueous solution of silver nitrate with the help of onion (Allium cepa) extract was investigated. The color change from colorless to dark brown indicated the formation of AgNPs [19]. Change in the state of a Nutlin-3a from the molecular scale to nano scale is accompanied with color change due to excitation of surface plasmon vibrations in nanoparticles [18].

Among the most important biotechnologically important industrial enzymes, amylases (especially α-Amylases, E.C. play a great role as it has multiple applications in various industries such as dairy, soft drinks, chocolates, pharmaceuticals, food processing, leather, textile, paper, wine, meat, fish processing and many others [1]. α-Amylases degrade starch by the hydrolysis of internal α-1,4 and α-1,6-glycosidic linkages to produce smaller carbohydrates such as glucose, maltose and maltotriose units [2]. In starch processing industry, a shift from the acid hydrolysis of starch to enzymatic digestion was notable in the last century [3]. Amylases are found in microbes, plants and animals but industrially important commercial enzymes are produced from microbes [3]. Among all sources of amylase, bacteria are the most significant because of the limited space and short period of time required for their cultivation and their feasibility to genetic manipulation [4,5]. Several different microorganisms are reported to produce extracellular amylases [6–9]. Commercially α-amylases are mainly produced from the genus Bacillus. Among many species, Bacillus subtilis, B. stearothermophilus, B. licheniformis and B. amyloliquefaciens are reported extensively as industrial amylase producers [6,7,10]. However, there are very few reports on Chryseobacterium sp. as amylase producers. Recently it has been shown that this bacterial species has the capacity to utilize raw organic materials, mung bean, to produce extracellular amylases [11].
Reports on using waste organic matter for the production of different industrially important enzymes had shown increasing trends lately [7–9,12]. In general, commercial carbon and nitrogen sources are used for the production of amylase by bacteria. To cut the cost, the use of organic waste matter might be a potential alternative to commercial carbon and nitrogen sources. A decade ago, about 7690 tons of municipal solid wastes (MSW), to which kitchen wastes are the chief source as organic matters, were generated per day in the six major cities of Bangladesh [13]. Based on these data, it is predicted that ∼47,000 tons of wastes will be produced per day by 2025 due to the rapid urbanization and economic growth in Bangladesh. About 75% constituent of the MSW in Bangladesh are organic that comprises mainly carbohydrates, proteins, peptides, fatty acids and their esters. Thus the organic materials of kitchen wastes can be used as raw materials for fermentation to produce many pure and fermented products [12,14,15]. The use of waste materials has potentials as the Bangladesh government has taken steps to implement national 3R strategy for waste management including “reducing waste”, “reusing”, and “recycling resources”.

Only in the younger age group and in

Only 12% in the younger age group and 7% in the older age group did not have any detectable islet cell autoantibodies (Table 1). The frequency of islet cell autoantibodies detected in the younger age group was 62% in GADA and 69% in IA-2A. These were not significantly different from those detected in the older age group. However, 37% of patients in the younger age group had IAA detected. This was significantly higher than the 10% of the older age group.
In the younger age group, remission occurred in 21 patients (40%), being complete in two (4%). However, 55 children (59%) in the older age group achieved remission, being completed in 11 (12%; Table 2). The remission rate of the younger age group was significantly lower.
In the younger age group, the proportion of boys (57% vs. 46%) and girls (43% vs. 54%) were comparable between the remitter and the nonremitter (p = 0.397). Most patients, 90% in the younger age group and 89% in the older age group, had an onset of remission during the 1st 3 months (Table 2), which was not statistically significant. Twelve (57%) in the younger age group achieved remission < 9 months, whereas 59% in the older age group achieved remission < 6 months. However, six out of 21 (19%) in the younger age group and 13 out of 55 (24%) in older age group achieved remission longer than 1 year. The difference was not statistically significant.
In literature, 9% of young Caucasian children with T1DM have a first degree family member with T1DM, with a predominance of fathers over mothers. However, in this study, only one patient had a mother with T1DM. This may be partially due to ethnic differences.
In this study, serum C-peptide levels, both basal and glucagon-stimulated, of patients younger than 6 years were lower than those of older patients, implying that the younger age group had less insulin reserve than older age group. This is consistent with findings of our previous report and other studies. The present study also showed that the frequency of previous respiratory tract infection in children younger than 6 years was higher than that of the older age group, which is similar to the results of another study.
In this study, ∼87% of patients younger than 6 years had ketoacidosis upon diagnosis. They also had higher blood chemokine receptor levels, lower pCO2 levels, and lower bicarbonate levels upon diagnosis than the older patients. Taken together, these suggest that patients younger than 6 years not only have a higher frequency, but also more severe degrees of ketoacidosis than older patients. Thus, less insulin reserve with stress related to viral infection may play an important role in unstable situations of patients aged < 6 years upon diagnosis. The frequency of islet cell autoantibodies has been reported to be significantly less in early-onset diabetes. However, in this study, 88% of diabetic patients aged younger than 6 years had at least one autoantibody detected. Their frequency of GADA and IA-2A positivity was similar to those of the older age group, but their frequency of IAA positivity was significantly higher, similar to findings of our previous report and other studies. This may be explained by the fact that IAA is the first detectable β-cell autoantibody among high-risk prediabetic children and that it disappears more rapidly than other autoantibodies. The present study confirms that autoimmune destruction of pancreatic β-cell is an important cause of T1DM in Taiwanese children younger than 6 years old. This study also demonstrates a higher rate and severity of DKA with lower HbA1c levels upon diagnosis of diabetic children aged < 6 years. The results may chemokine receptor suggest more rapid and extensive pancreatic β-cell damage by an autoimmune process, resulting in a more severe metabolic decompensation at diagnosis, consistent with results of previous reports.
In newly-diagnosed type 1 diabetic children, there is often a period of remission during the natural course of the disease, characterized by a striking fall in insulin doses to maintain good metabolic control. Different criteria have been used to define remission. Daily insulin requirement alone or HbA1c alone has been used to define remission in some studies, whereas other studies include daily insulin requirements and HbA1c in defining remission. As remission defined by daily insulin requirement alone or HbA1c tends to overestimate remission rates, insulin dose-adjusted A1c, proposed by Mortensen et al to define remission, was used in the present study. The reported remission rates vary between 25% and 100%, which partially reflects the different definitions used and the difference in patients\’ ages.

br Methods br Results Overall consecutive


Overall, 145 consecutive patients with a mean age of 41.9 years were investigated. AKI was diagnosed in 52 patients (35.8%). Thirty-five patients died during hospitalization (mortality rate: non-AKI, 7/86=8.1%; Stage 1, 11/35=31.4%; Stage 2, 7/12=58.3%; Stage 3, 10/12=83.3%; p<0.001). Patient data, including age, sex, laboratory parameters, and risk model scores, are listed in Tables 1 and 2 according to mortality and AKI, respectively. Patients who died during the hospital course were older and had larger total burn surface area (TBSA), more ventilator use, poor conscious levels, worse renal function, and higher phosphate levels at admission. These patients also exhibited lower c-kit inhibitor (Hb), albumin, and urine output levels. In the admission period, they had longer ICU stays and renal replacement therapy. Patients with AKI were older and had larger TBSA, more ventilator use, lower Glasgow Coma Scale scores, and lower Hb. Patients with Stevens–Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) exhibited a significantly higher mortality rate compared to patients with other etiologies. The etiologies on admission are listed in Table 3.
The predictive ability of APACHE II, APACHE III, SOFA, and AKI staging was compared, and the calibration and discrimination of the models are listed in Table 4. Among these risk models, APACHE III exhibited the highest prediction ability (AUROC, 0.889±0.029), followed by APACHE II (AUROC, 0.841±0.037) and AKI staging (AUROC, 0.819±0.045). Both the APACHE III and APACHE II were significantly superior to SOFA and OSF in evaluating the mortality (APACHE II vs. SOFA p=0.004; APACHE II vs. OSF p<0.001; APACHE III vs. SOFA p=0.033; APACHE III vs. OSF p = 0.002, respectively). Severity of AKI was also superior to OSF (p=0.022). To determine the selected cutoff points for predicting in-hospital mortality, the sensitivity, specificity, and overall accuracy of the prediction were determined (Table 5). Among the models, APACHE III exhibited the highest Youden index score and overall correctness at the 48-point cutoff. The AKI system exhibited the highest sensitivity for prognostic prediction. Multivariate logistic regression analysis with backward elimination was conducted to identify demographic and clinical characteristics associated with mortality in patients with burns (Table 6). Age, TBSA, ventilator usage, Glasgow Coma Scale scores, Hb, creatinine, albumin, phosphate, and SJS/TEN that were statistically significant were included in the multivariate analysis by applying a multiple logistic regression based on forward stepwise regression to obtain independent predictors. Only TBSA, ventilator usage, AKI, and SJS/TEN were associated with mortality.
Figure 1 shows the cumulative survival rates according to the AKI stage. The cumulative survival rates revealed that mortality increased with the AKI stage.

AKI is not uncommon in ICUs and has been associated with increased mortality, ICU length of stay, and medical costs for critically ill patients. Patients admitted to burn units typically exhibit complex syndromes with numerous pathways that affect renal function and involve c-kit inhibitor hemodynamic changes, tissue breakdown products, inflammatory cytokines, septicemia, and drug toxicity. AKI is also a key factor in mortality. Recent investigations using APACHE III examined acute physiology scores, age, and chronic health problem scores, which are currently widely used to predict clinical outcomes in ICU settings and exhibit the accuracy of APACHE III in estimating mortality in patients with burns. Moore et al reported an AUROC of 0.833 while using APACHE III, which is similar to our results. Further investigation supported the efficacy of SOFA scores in assessing the extent of organ dysfunction in various groups, including critically ill patients with burn injuries, but did not provide related AUROC information. Coca et al reported the association between different RIFLE stages of AKI with morbidity and mortality in burn patients, but did not compare the complex scoring systems with AKI stage. Chung et al compared the outcome between early intensive renal replacement therapy and conventional therapy in which AKI was defined by AKIN criteria. Patients with either Stage 2 AKI in combination with shock or Stage 3 AKI received continuous venovenous hemofiltration. Patients with early treatment had less 28-day mortality compared with those who had conventional treatment (38% vs. 71% p=0.011). To our knowledge, integument study is the first to compare the usefulness of these complex scoring systems with the 2012 AKI definition for outcome prediction. Although the discriminative power of APACHE III is higher, the KDIGO AKI staging approach is a considerably simpler method that requires fewer parameters to assess the prognosis in patients with burns. For the further interventional trial, our result also provided an overview of the definition.

High correlations between tick initial

High correlations between tick initial weight and egg mass weight (Table 1) suggest that both traits were affected by similar host immunological factors, agreeing with results presented by Barriga et al. (1995). Nearly perfect association (0.97) between those traits in susceptible heifers indicates that egg mass produced by ticks of this group were almost entirely dependent on blood intake capacity during tick development on the cattle. Although also positive and high, the correlation of 0.80 observed between initial weight and egg mass weight for resistant heifers indicates that a small portion of the variation observed in oviposition weight was due to factors other than tick initial weight in this group. The lower correlation between tick initial weight and egg mass weight observed in the resistant group, as well as the negative association between initial weight and egg production index of ticks from resistant heifers, may be an indicative of occurrence of a defense mechanism that disturbs conversion of ingested blood to egg mass in animals of the resistant group.
In conclusion, genetic variability for tick resistance observed in Braford heifers affected both the number of ticks carried by the animals as well as egg mass weight, egg production index and nutrient index of engorged females of R. (B.) microplus. Thus, in addition to economic losses related to prostanoid receptors of cattle productivity or to higher demand for treatments, maintaining animals with high tick susceptibility in the herd implies higher environmental infestation levels, perpetuating high prevalence of ticks on farm. Genetic evaluation of livestock for tick resistance and inclusion of tick counts as selection criteria in breeding programs can be implemented as an auxiliary tool for the strategic control of R. (B.) microplus in production systems. Finally, is important to note that further studies to uncover genetic factors responsible for immunological mechanisms involved in the expression of resistant phenotypes could enable genetic evaluations for tick resistance based on genomic information (e.g., molecular markers); thereby, increasing prediction accuracy, accelerate identification of superior genotypes, and avoid cattle exposure to ticks required by current genetic evaluation methods.

Research supported by CNPq – National Council for Scientific and Technological Development grant 478992/2012-2, Embrapa – Brazilian Agricultural Research Corporation grants and, and CAPES – Coordination for the Improvement of Higher Level Personnel grant PNPD 02645/09-2. Authors acknowledge the Delta G Connection for providing animals and data for this research and Dr. Concepta M. Pimentel for reviewing the manuscript and providing a constructive contribution to its final version.

, an important protozoan belonging to the Apicomplexa family, causes the fatal disease East Coast fever in cattle, restricted to the East Coast and the central part of Southern Africa. This disease, together with tropical theileriosis (), causes major economic losses to the farming community in developing countries.

The use of Specific Pathogen Free SPF pigs as

The use of Specific Pathogen Free (SPF) pigs as a model to test parasite populations has been extensively studied (Eriksen et al., 1992; Kringel and Roepstorff, 2006; Nejsum et al., 2009; Petersen et al., 2014; Roepstorff et al., 1997; Taira et al., 2003). Increasingly, minipigs are used in biomedical research and in pharmaceutical safety studies due to their similarity to human physiology, their small size, and high degree of uniformity as a result of melatonin receptor agonist and rearing under controlled stable conditions (Swindle et al., 2012). Whereas production pigs (including SPF pigs) are only free of a limited number of pathogens, Göttingen minipigs selected and bred for laboratory use offer a much higher level of absence of parasites, bacteria and viruses. In the present study, Göttingen minipigs ( were inoculated with increasing doses of TSO to validate the dose-dependent establishment of T. suis larvae in the minipig infectivity model by a standardized method. The assessment of TSO biological potency is needed for evaluation of the stability (quality) of each egg batch prior to their use as an active pharmaceutical ingredient (API) in clinical trials against several immune-mediated diseases.
The present study is the first to validate a dose-dependent establishment of T. suis larvae under GMP conditions.

Materials and methods


The present study demonstrates a clear positive linear relationship between inoculated embryonated eggs and larval establishment in the minipig infectivity model within the dose range 1000–10,000 TSO. Thereby, the minipig model appears to be a reliable and accurate (larval establishment in relation to the TSOee) model for the assessment of biological potency of TSO used as trial parasite-based medication. In addition, the validation of dose-dependent establishment of T. suis larvae was performed under controlled environmental conditions and guidelines of Good Manufacturing Practices (GMP), which is a requirement for medicinal products.
Earlier T. suis studies in which pigs were inoculated with very high doses of 50,000 (Batte et al., 1977; Beer and Lean, 1973) and 100,000 eggs (Batte et al., 1977) have reported severe symptoms of diarrhoea, bloody faeces, retardation of growth or weight loss, and even death. Clinical manifestations and gross pathological lesions in the large intestine have also been shown in outbred Yorkshire pigs at a lower egg dose (2500 eggs), but the authors suggested that concurrent bacterial infection (including Campylobacter) may have caused the severe pathological lesions and disease outcome at 45dpi (Mansfield and Urban, 1996). Apart from such observations, T. suis is generally described to be well tolerated in pigs, and the present validation study confirmed that inoculation doses up to 10,000 eggs (89.1% embryonation) are not associated with any clinical symptoms during intestinal establishment of the larvae (up to 21dpi).
Previously, it was observed that by increasing T. suis inoculation doses up to 42,405 eggs (1650eggskg−1 body weight) there is a reduction in the average weight gain of conventional pigs (Hale and Stewart, 1979). Even though not statistically significant, there was a tendency of lower weight gains in pigs inoculated with 5000 embryonated T. suis eggs compared to uninfected control pigs (average weight of all pigs was 26.8kg±6kg at the start of the experiment) (Kringel and Roepstorff, 2006). In our study, we did not observe any (negative) relationship between infection dose and daily weight gain in minipigs (received average 122 to 1333 TSOeekg−1 body weight per group). Similarly, no significant difference in weight gains between larger pigs (average 77kg at the start of the experiment) was observed in a study with low (400 eggs), medium (4000 eggs), and high (40,000 eggs) inoculation doses with T. suis (Pedersen and Saeed, 2000).