The cylindrical container in which the melt was

The cylindrical container in which the melt was placed was made of Plexiglas. The container was surrounded by the inductor uniformly distributed over the height of the melt (Fig. 1).
Fig. 1 schematically shows the characteristic distribution of the Lorentz force (5), induced in the melt in an alternating magnetic field, and the flow lines indicating the main directions in which the fluid moves under the influence of the EM field force. As previously mentioned, the structure of the melt flow in the cylindrical volume at the above-noted density distribution of the Lorentz force is the circulation of two toroidal eddies. The density of the force applied to the melt oscillates at a frequency which is double the frequency of the EM field. At the same time, the change in the Lorentz force density can be neglected, as liquid metals are slow to respond (due to their inertness) to impacts with a frequency higher than 4Hz [10]. Varying the frequency of the EM field leads to a change in the depth to which the field can penetrate the conducting liquid (the skin depth) and in the area of the field\’s action. The density value of the EM field force decreases exponentially within the skin depth whose thickness also depends on the characteristics of the medium acted upon. The thickness of the skin depth can be described by the relation
where is the frequency with which the direction of the EM field changes, μ=1.256·10N/A2 is the magnetic constant, σ is the electric conductivity of the medium.
The δ value for Galinstan was 22mm; for the inside diameter D of the container equal to 62mm, this made up a substantial portion of the melt volume which the field affects by setting the fluid in motion.
The flow parameters were determined experimentally using the Doppler velocity meter. As the name implies, its operating principles are based on the Doppler effect, i.e., on changing the frequency of the ultrasonic wave emitted and received by the melatonin receptor agonist as the wave is reflected by the microparticles distributed in the fluid. The particle drift velocity within the ultrasonic beam is determined by the difference between the frequencies of the emitted and the reflected waves. This method is widely used for finding the flow parameters in low-melting liquid metals and model fluids [5,12,13].
In the experiment described, a DOP3010 ultrasonic Doppler velocity meter (Signal Processing SA, Switzerland) was used. During the measurements, the sensor (8mm in diameter) was placed directly into the fluid through the free surface to a depth of 1–2mm and fixed in that position throughout the whole experiment. The frequency of the signal transmitted by the sensor was 1428Hz at the speed of sound in the melt equal to 2730m/s (see Table 1). The sensor was secured on the cylindrical volume axis (point P (r = 0mm) in Fig. 1) for measuring the axial component of the velocity. This velocity component was predominant in the flow structure under investigation. At the same time, the velocity had a negative value if the flow parallel to the signal beam was directed toward the sensor and a positive one if the particles in the fluid were moving away from the sensor.
The experiment used two modes of applying the EM field force to the flow. In the first case, we measured the melt flow velocities under a continuous current in the inductor for the values ranging from 50 to 250Hz. Given that >> 4Hz, the effect on the melt was considered steady (from now on, the EM field parameters are denoted by the index st).
In the second case, we considered the motion of fluid under the action of the pulsed Lorentz force (parameters with the index p). As the force was modulated, the current in the inductor was interrupted to generate pulses with the frequency . If we denote the period of time during which the EM field force acts on the melt as , and the duration of the pause in which the Lorentz force is absent and the fluid moves under the influence of inertia as T0, the expression for determining the frequency and the period of the pulses can be written as follows:

Introduction Nowadays probiotics offer a promising

Nowadays, probiotics offer a promising alternative approach for controlling shrimp diseases and improving shrimp health through their potential to control pathogens, to stimulate the immune response, to improve water quality and to enhance nutrition through the production of digestive enzymes (Verschuere et al., 2000; Gullian et al., 2004; Wang, 2007). To achieve health benefits, probiotic bacteria must be viable and available at high concentration, typically 106–107 CFU/g of product (Kosin and Rakshit, 2006). Furthermore, incorporation into feed pellets is more effective in conveying probiotics into animals compared to direct application into rearing systems. It is also applicable for intensive aquaculture and requires no additional labor or shrimp handling (Gómez et al., 2007). The viability and stability of probiotics have been a technological challenge in feed manufacturing because probiotics, including Lactobacillus, Bacillus and yeast, are susceptible to the high temperature of the pelleting and drying process. According to Biourge et al. (1998), Bacillus CIP5832 spores in dog diet were found to have in excess of 99% loss after the extrusion, expansion and drying processes. Furthermore, the viable count of yeast in shrimp feed pellet decreased by 105 fold after extruding through a meat grinder at 72 °C for 31 s followed by drying at 65 °C for 6 h (Aguirre-Guzmán et al., 2002).
Fluidized bed drying is extensively used for drying wet particulate and granular materials. In a fluidized bed dryer, the probiotic cell suspension is mixed with a vibrating bed of absorbers or matrix molecules which helps to form capsules by adherence (Nag and Das, 2013). This process is comparatively economical. It involves low energy consumption, high throughput and imparts moderate heat stress to the bacterial cells (Beker and Rapoport, 1987; Nag and Das, 2013). Furthermore, this process was successfully used for the preparation of dried granules or powders containing lactic melatonin receptor agonist bacteria (Santivarangkna et al., 2007; Nag and Das, 2013). According to Nag and Das (2013), fluidized bed drying was able to retain viability of Lactobacillus casei CRL 431 of more than 7.7 log CFU/g during storage at 25 °C for 12 wk. Mille et al. (2004) revealed that the Lactobacillus plantarum viability in casein powder was up to 80% after fluidized bed drying at 35 °C for 30 min. Correspondingly, the survival of Lactobacillus brevis in fish feed was 99% (108–109 CFU/g) after fluidized bed drying at 40 °C for 40–60 min with a moisture content of 5% (Toledo et al., 2010). Several studies have reported that the viability of lactic acid bacteria during drying and storage was enhanced by the addition of protective agents such as trehalose, skim milk, whey protein, soy protein isolate, monosodium glutamate, sucrose, lactose, sorbitol and polymers such as carboxymethyl cellulose, dextran and acacia gum (Morgan et al., 2006; Santivarangkna et al., 2007; Golowczyc et al., 2011; Lapsiri et al., 2013). In the present study, an overnight culture of Lactobacillus lactis 1464 was incorporated into shrimp feed pellets prior to the pelleting process at ambient temperature and dried in a fluidized bed dryer to achieve a moisture content lower than 11%. The effect of the drying temperature, culture pH and protectants on the strain survival during drying was determined. Additionally, the storage stability of the strain in the pellets at 4 °C and 30 °C was also evaluated.

Materials and methods

Results and discussion

Conflict of interest

Dendrobium is one of the largest genera of Orchidaceae, with over 1000 species worldwide (Leitch et al., 2009). Many Dendrobium orchids are important horticultural crops and have been used in commercial trade due to their popular flowering abundance, wide range of flower colors, sizes, shapes, year-round availability and lengthy post-harvest life (Hossain, 2011). Most Dendrobium species are suspected of becoming extinct. Of these, Dendrobium friedericksianum Rchb.f. is a native epiphytic orchid found in tropical rain forests in Eastern Thailand according to the checklist of international species, (Convention on International Trade in Endangered Species of Wild Fauna and Flora, 2014). Due to their beautiful, bright golden-yellow flowers with diameters around 4–5 cm each, the orchid has attracted collectors and so specimens are in high demand for illegal trading. Consequently, this species is on the verge of being seriously endangered in the wild.

br Discussion Knee cartilage is an

Knee cartilage is an important structure in the human body and plays a pivotal role in knee joint activity. It is known that cartilage morphology is influenced by many factors, such as age, gender, genetics, BMI, level of physical activity, osteoarthritis, pain and ACL or meniscal injuries. Among these factors, meniscal damage appears to be an important factor to induce cartilage morphological changes. Meniscal damage or loss alters the in vivo cartilage contact biomechanics by shifting the contact location to smaller regions of thinner cartilage, and by increasing the magnitude of the cartilage contact deformation. It is presumed that cartilage morphological alteration is accelerated in the knees with meniscal pathology; with altered knee joint kinematics, the mechanical demand eventually exceeds the ability of the joint to repair itself, setting the stage for OA development.
In this study the mechanism of injury was non-significant trauma in 50% of patients. This finding is in agreement with previous studies that reported that 52–92% of patients with symptomatic knee melatonin receptor agonist present with meniscal damage when assessed by MRI to the contralateral normal knees.
Quantitative MRI (qMRI), like 3D-WS-bSSFP sequence used in this study, provides non-invasive and reliable data on cartilage morphology in healthy subjects with different age groups and different genders. Comprehensive knowledge of the biochemical and biomechanical changes that occur with OA may require a combination of various qMRI techniques. The application of these qMRI techniques to the identification and monitoring of cartilage damage in individual patients for the early diagnosis and treatment of OA would be clinically significant. However, assessment of other joint structures is not possible. In fat-suppressed 3D-WS-bSSFP, the articular cartilage has very high signal intensity, joint fluid has an intermediate to low signal intensity, and subchondral bone and bone marrow are dark. Reported sensitivity and specificity for detection of cartilage loss are 75–85% and 95–97%, respectively.
The cartilage morphology (including cartilage volume by MRI and thickness by US) was investigated in the meniscectomized knees and the contralateral intact knees with a mean injury time of 6.4months. Using the 3D cartilage models (Fig. 1), the mean total cartilage volume was calculated for the femur and tibia. The cartilage thickness was measured within the weight-bearing area by US. In this study, the contralateral knees were used as the control side for analyzing the cartilage morphological changes in the post-meniscectomy knees. It was found that the meniscectomized side demonstrated a statistically significant lower total knee cartilage volume compared with the control side. The percentage of decrease in the total knee cartilage volume post-operatively from pre-operatively was >10% in 13 patients (65%), this value progressors of cartilage loss based on MRI measurements of the cartilage volume as a biomarker. However, this did not correlate with KOOS scores. As the articular cartilage is avascular and aneural, early changes could be asymptomatic or affecting the functional knee activity. Significant rates of cartilage loss are seen also in other studies; in subjects of post partial meniscectomy compared with healthy controls (difference 6.5% per year, 95% CI 3.7–9.3% per year; P<0.001). Even greater losses were observed at the central medial tibial cartilage and medial femoral condyle (15% and 12% respectively). Greater losses are observed in this cohort of Egyptian adults, this may have a genetic basis. The same genes that promote healing after cartilage damage also appear to protect against OA due to wear-and-tear processes. Osteoarthritis, like several other disorders, involves many genes that each contribute in a small way to the disease process. It is proved that there is a subtle genetic influence on OA risk, while other genes are protective.

Eosinophilia was reported in patients with a median eosinophil

Eosinophilia was reported in 30 patients (57.7%), with a median % eosinophil (IQR) of 18.1% (12.9–22.9) and a median absolute eosinophil count (IQR) of 1,482 (856.7–3,652.2) cells/μL. Both median % eosinophil and median absolute eosinophil were higher in patient whom allopurinol was the culprit drug (p = 0.009 and p = 0.003, respectively) (Table 4). Fourteen patients (26.9%) had atypical lymphocyte, with a median % atypical lymphocyte (IQR) of 10% (6–20.5). Thirteen patients (25%) had thrombocytopenia, with a median platelet count (IQR) of 84,000 (53,000–111,000) cells/μL. Lymphopenia was also reported in approximately a half the patients (51.9%). Blood test for human herpesvirus 6 (HHV-6) was performed in 6 patients. The results were positive for serum IgG antibody and negative for serum IgM antibody in 4 patients. No further quantitative test was done. The results for 2 remaining patients were unavailable.
The culprit drug was discontinued in every patient. Up to 85%, the culprit drug was discontinued in the first day of admission. The longest duration until discontinuation was 11 days (phenytoin was the culprit drug, this melatonin receptor agonist case was fortunately survived). Systemic corticosteroid, either intravenous dexamethasone or oral prednisolone, was administered to 30 patients (57.7%). Forty-nine patients (94.2%) received antihistamine, either chlorpheniramine or hydroxyzine (Table 5). However, when evaluated only in those who initial diagnosis were DRESS, systemic corticosteroid was administered in 23 out of 28 patients (82.1% versus 29.2% of whom initial diagnosis were DRESS versus other initial diagnosis, respectively, p < 0.001). There was no different in lag time of systemic corticosteroid administration between the 2 groups.
Drug reaction with eosinophilia and systemic symptoms (DRESS) is a rare but severe adverse drug reaction, with broad clinical features. The estimated incidence of DRESS ranges from 1 in 1,000 to 1 in 10,000 drug exposures. A recent study in Europe reported an overall prevalence of 37.6 cases per 100,000 inpatients. Multiple factors have been described to associate with DRESS development such as defect of drug detoxification, human leukocyte antigen-related gene, and HHV-6 reactivation. HHV-6 has been defined as a potential contributor to DRESS development.
Uneven yearly prevalence of DRESS is seen in Inverted repeats study (Fig. 1). During 11 years period, there were some trends regarding the culprit drugs. The number of nevirapine-induced DRESS patients contributed to approximately 50% of DRESS cases during 2004 and 2005. Interestingly, nevirapine was the second most common culprit drugs in our study (17.3%) (Fig. 2). The result is uncommon compared to previous reported worldwide (5%). Such finding could be explained by higher prevalence of underlying HIV infection in our study and more frequent prescription of nevirapine in Thailand in those years. Before 2010, nevirapine was the only preferred NNRTI in HAART in Thailand. The number of nevirapine-induced DRESS cases had declined since 2005, until the last case reported in 2010, the same year when the Thailand\’s national guideline for antiretroviral therapy recommended efavirenz as an additional preferred NNRTI to nevirapine.
The youngest patient in our study was a 2 years old girl diagnosed with carbamazepine-induced DRESS. Clinical presentations were high-grade fever, rash, lymphadenopathy, eosinophilia, and liver involvement. The onset time was 21 days and the patient stayed at the hospital for 5 days, which were both shorter than a case reported by EL omairi et al. on carbamazepine-induced DRESS in a 6 years old boy patient (4 weeks and 2 weeks, respectively). On discharged, levetiracetam was given as a substitute with no subsequent adverse reaction reported. There are few reports of DRESS in children; and to our knowledge, our patient was among the youngest patients diagnosed with DRESS.

br Conclusion br Conflict of interest statement br


Conflict of interest statement

This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MISP) (No. 2013R1A4A1069486).

Cyathostomin (small strongyle) parasites are ubiquitous in grazing horses world-wide and control programs are typically based on anthelmintic treatments applied routinely year-round with the objective being to avoid large burdens that can constitute a threat to equine health (Lloyd et al., 2000; O’Meara and Mulcahy, 2002; Earle et al., 2002).
Strongyle type parasites appear to have complex interaction with the immune system. In horses, experimental inoculation with cyathostomins and Strongylus vulgaris evoke a subtle acute phase inflammatory response (Andersen et al., 2014), and the number of mucosal larvae has been shown to correlate with large intestinal mucosal expression of the pro-inflammatory interleukin (IL-4) as well as the regulatory and anti-inflammatory cytokine IL-10 (Davidson et al., 2005). In human medicine it has become well-established that helminth infections can relieve the symptoms of inflammatory disorders such as melatonin receptor agonist (Araujo et al., 2004) and chronic inflammatory bowel syndrome (Crohn\’s disease) (Cooper, 2004; McKay, 2009), probably through parasitic stimulation of a release of the regulatory and anti-inflammatory IL-10 (McKay, 2009).
A number of studies have documented that anthelmintic treatment can evoke a systemic inflammatory response, and that the extent of this response is associated with the anthelmintic drug class used (Steinbach et al., 2006; Nielsen et al., 2013; Betancourt et al., 2014). When very pronounced, this inflammation can be part of the parasitic disease complex called larval cyathostominosis (Reid et al., 1995), which is characterized by mass emergence of large numbers of encysted larvae from the mucosal walls of the large intestine. To add to the complexity of this inflammatory response following anthelmintic treatment, a recent study with a murine asthma model suggested that ivermectin might have anti-inflammatory effects (Yan et al., 2011). To our knowledge this has not yet been investigated in any type of non-rodent mammals, but the observation is supported by equine studies reporting little or no signs of inflammatory reaction following treatment with moxidectin, which like ivermectin belongs to the macrocyclic lactone drug class (Steinbach et al., 2006; Nielsen et al., 2013; Betancourt et al., 2014).
Vaccination is another commonly applied practice in equine management, and the response to a vaccine is largely consists of both inflammatory and immunologic reactions. The acute phase inflammatory response was recently characterized in response to two commercially available equine influenza/tetanus vaccines: an inactivated influenza/tetanus toxoid Immune Stimulating COMplex (ISCOM) vaccine and an adjuvanted, live recombinant vector vaccine (Andersen et al., 2011). The study revealed a prominent acute phase response to both vaccines during the first 96h after vaccination, but with the response being more pronounced following administration of the ISCOM vaccine. Given the information presented above, there is reason to hypothesize that administration of an anthelmintic may interfere with (a) the inflammatory reaction to the vaccination and (b) the immune response to that vaccine. For convenience, horses are likely to often receive vaccinations and anthelmintics at the same time. A previous study suggested a possible interaction between gastrointestinal worm burdens and the immune response to antigen stimulation in ponies (Edmonds et al., 2001), and a recent study performed in cattle documented a beneficial effect of deworming calves two weeks prior or at the day of vaccination with a combination vaccine against respiratory viruses (Schutz et al., 2012).

Materials and methods

melatonin receptor agonist Introduction Piezoelectric ultrasonic motors are fascinating

Piezoelectric ultrasonic motors are fascinating actuators. They combine fast dynamics, high force and offer many advantages in comparison to electromagnetic motors. Ultrasonic motors worked via the vibration of the piezoelectric comprise body (stator) in the ultrasonic frequency band and the converse piezoelectric effect of the PZT elements [1]. The mechanical movement and torque are obtained by means of the frictional force between the stator and rotor or slider. Ultrasonic motor, nowadays, is a good candidate for application in fields of aerospace, robots and medical instruments for its good characteristics, such as simple size, light weight, high torque at low speed, quick response, no electromagnetic interference and higher position accuracy, etc. [2].
Ultrasonic motors are also named vibrating motors and can be divided into traveling wave type ones [3–6], standing wave ones [7,8] and composite vibration modes type ones [9–11] from the viewpoint of vibration characteristics. Up to date, ultrasonic motors are mainly used for driving and/or positioning in fields of aerospace mechanism, robot and medical instrument, and so on. For example, Egashira et al. [12,13] developed a nonresonant ultrasonic motor to drive a lithography precision stage fed back by a Sony melatonin receptor agonist BS76 linear scale (resolution to 10nm). Tanaka et al. [14] utilized neural network (NN) –PID controller to overcome the time-variant characteristics of the ultrasonic motor for precision motion control. Fan and Lai [15] presented a linear positioning system driven by an ultrasonic motor and its control algorithm design with nano resolution. Cheng et al. [16] proposed an improved BPNN-PID controller for the long-stroke and nanopositioning control of a new co-planar linear stage driven by an ultrasonic motor. In some special occasions, however, such as toxic sealing containers, dangerous locations, etc., it requires that ultrasonic motors not only have the function of driving and/or positioning, but also have the function of melatonin receptor agonist harvesting to power small electronic devices without external electric sources or internal batteries. This is because the use of batteries naturally entails several shortcomings in these special occasions, such as short lifetime, contribution to environment damage, and a requirement of frequent maintenance. Energy harvesting can be a good alternative for eliminating or reducing the needs for batteries in these special occasions.
Based on the direct piezoelectric effect of PZT elements, an energy harvesting type ultrasonic motor is presented in this study. The proposed motor has similar working principle with the traveling wave type ones [17], and the traveling wave in the elastic body (stator) is still generated by superposition of two mode responses with equal amplitude and phase difference π/2 both in time and space, which are excited by the converse piezoelectric effect of two groups PZT elements applied with two alternating voltages with equal amplitude, the same frequency and phase shift π/2 in ultrasonic range.
The most notable difference between the present ultrasonic motor and other traveling wave type ones previously reported [3–6,17] may be in the stator structure, as well as the adjustment mechanism of the pre-pressure between the stator and rotor. In the stator of the new motor, there are two pieces of PZT rings, one is bonded on the bottom surface of the stator metal body, and is excited to generate a traveling wave in the stator, and the other one is boned on the outside top surface of the stator metal body, and is used to harvest the vibration-induced energy of the stator based on the direct piezoelectric effect. In previously published traveling wave ultrasonic motors, however, there exists only one piece of PZT ring bonded on the bottom surface of the stator metal body to excite to generate a traveling wave. Furthermore, in the previous development of traveling wave motors, a disk spring was usually used between the stator and rotor to produce the pre-pressure and the contact pressure. By contrast, the disk spring is deleted in the new motor and the pre-pressure between the stator and rotor is obtained by the deformation of the rotor. The rotor with approximate flexibility is designed to improve the contact performance and reduce the power loss from the radial friction between the stator and rotor.

Introduction T lymphocytes encompass functionally specialized populations that can differently

T lymphocytes encompass functionally specialized populations that can differently contribute to host defence against a broad variety of pathogens. Beside well characterised αβ T cells, all jawed vertebrate species possess a numerically smaller proportion of γδ T melatonin receptor agonist (Holderness et al., 2013). Their numbers in blood range between 0.5-10% for human and mice (Hayday, 2000) and up to 20–50 % in domesticated animal species, such as cattle (Mackay and Hein, 1989), pigs (Hirt et al., 1993; Yang and Parkhouse, 1996) or chickens (Sowder et al., 1988; Arstila and Lassila, 1993). Despite emerging knowledge about their functions, γδ T cells are still an enigmatic cell population with lots of features being characteristic for innate and adaptive immune responses.
Subpopulations of γδ T cells have been defined according to special functions, specific phenotypic features, TCR gene segment usage or tissue localization (Turchinovich and Pennington, 2011; Pang et al., 2012; Chien et al., 2014). Unfortunately, a standard system for classification of γδ T-cell subsets in distinct species is still lacking. Avian γδ T cells have been grouped with regard to their CD8α-co-receptor expression (Tregaskes et al., 1995; Berndt et al., 2006).
The influence that γδ T cells exert on the immunological defence against pathogenic agents is not only dependent on their sufficient number but also on their activation and differentiation into competent effector, regulator or memory cells. Up to now, there is no information on avian γδ T cells and their requirements for activation and subset differentiation as well as for an adequate maintenance and expansion of these cells in in-vitro cultures for further functional studies. To maintain, activate and multiply lymphocytes in cultures or simply for use as positive control in basic research, different non-specific mitogens have been employed for a longer time. Commonly applied mitogens to trigger proliferation are Phorbol-myristate-acetate (PMA)/Ionomycin, or Concanavalin A (ConA), but also IL-2, which have been repeatedly used for studies with bovine γδ T cells (Sathiyaseelan and Baldwin, 2000; Baldwin et al., 2002; Fikri et al., 2002). For chickens, it has been published that ConA is sufficient to induce proliferation of splenocytes (Hanieh et al., 2012; Ren et al., 2015; Yu et al., 2015) and also γδ T cells (Kasahara et al., 1993). Beyond, Phytohaemagglutinin (PHA) has often been used to induce avian T-cell proliferation as positive control in infection or vaccination studies (Theis et al., 1975; Yu et al., 2013).
The prevailing readout for T-cell activation is proliferation or expression of typical surface markers, as CD25. CD25, the α-subunit of the Interleukin-2-receptor, has already been described as a marker for T-cell activation in chickens (Hála et al., 1986). Recently, numbers of CD25+ γδ T cells have been found in whole-blood cultures of white leghorn chickens (Braukmann et al., 2015). However, there is no data available that compare CD25 expression and proliferation after non-specific in-vitro stimulation of avian γδ T-cell subsets in PBL cultures.
In the present study, the commonly used mitogens PMA, Ionomycin, PHA, ConA as well as different recombinant interleukins, as IL-2, IL-12 and IL-15, were analysed in terms of their capacity to maintain, multiply and activate avian γδ T cells in cell cultures. For that reason, peripheral blood lymphocytes (PBLs) of four different chicken lines varying in their egg laying performance and phylogeny (Lieboldt et al., 2015a) were co-cultured with the commonly used stimulants, and the CD25-antigen expression was scrutinized in comparison to proliferation of the following CD8α-characterised γδ T-cell subpopulations: CD8α−, CD8αhiβ+, CD8ααhi+ γδ T cells.

Material and methods

Results and discussion
In the present study, untreated PBL controls showed spontaneously increased percentages of CD25+ cells in all γδ T-cell subsets at 72 and 96h compared to 48h, but most pronounced in the CD8α− γδ T-cell population (p≤0.05) (Fig. 1a–c). A likewise heightened amount of CD25+ γδ T lymphocytes has recently been reported for avian whole blood cultured without any addition (Braukmann et al., 2015). The reason for this phenomenon is not yet clear, but might has been the result of a de-novo production of IL-2 by the cultured cells themselves, which could have enhanced the IL-2R expression or even cell proliferation. Notwithstanding, the CD25-expression intensity was only marginally increased (Fig. 2a–c), and proliferation of γδ T cells was not elicited in our untreated PBL samples (Fig. 3a–f). A stress-induced response, perhaps in consequence of the isolation method, the culture conditions and/or the material of the culture dishes cannot be excluded as possible cause for the slight increase of the CD25 antigen expression on the cells. Other authors reported on augmented non-specific proliferation of CD4+ and CD8+ cells in avian PBMC cultures, which was reduced after substitution of FBS with serum from an immune chicken (Norup et al., 2011). In any case, the use of FBS and chicken serum, as applied in our experiments, seems to be a suitable combination for the study of avian γδ T cells in PBL cultures, at least because of the unchanged proliferation and only slightly enhanced CD25-expression intensity.

Mid Infrared MIR spectroscopy could be a suitable candidate in

Mid-Infrared (MIR) spectroscopy could be a suitable candidate in this regard. It has become a common spectroscopic technique in the food industry. It is used for characterizing molecules, but it could also be applied for evaluating the quality of food matrices (e.g.: determining the composition of milk [11], the quality of oils [12], the authenticity of honey [13]. Literature shows that only a few studies have determined the MIR spectral profile of phenolic compounds. MIR spectroscopy has been used as a new tool for determining in situ rosmarinic melatonin receptor agonist (caffeic acid ester) in Lavandula officinalis culture suspensions [14]; for better identification in the melatonin receptor agonist culture suspension, the MIR spectrum of a pure commercial rosmarinic acid standard was recorded, interpreted then compared with dried cells of L. officinalis. Similarly, Pei et al. used MIR spectroscopy to analyze Herba epimedii (traditional Chinese medicine); the correlation value, representing the similarity of two spectra of an herbal sample and the standard phenolic substance, icariin (flavonol glycoside), in the 1280–1200cm−1 region, has been found to be a good indicator for fast and effective quality control and could be used as a screening criterion for the herbs [15]. MIR spectroscopy was also used in the framework of a study on the chelation of isoflavones like biochanin A (O-methylated isoflavone) and genistein (isoflavone) with metal [16]. In the same period, the technique was used by Gorinstein et al. to determine the bioactivity of exotic fruits [17]. Spectra of fruits (Avocado, durian and mango) were compared to pure phenolic standards (catechin (flavan-3-ol) and gallic acid (phenolic acid)) to detect the presence of those phenolic compounds in the fruits. Bands of these two compounds were pointed and interpreted. Another study has been realized with the objective to develop a quantitative method for the determination of anthocyanin content in sweet cherries by using MIR spectroscopy, the spectrum of keracyanin (anthocyanin) as chloride salt was recorded and interpreted. A linear relationship between anthocyanin content and the area of ​​the band between 1640 and 1630cm−1 was established (coefficient of correlation 0.99) [18]. Mangolim et al. have characterized, by MIR spectroscopy, curcumin before and after its complexation with ß-cyclodextrin. They studied the stability, the solubility and the food application of the inclusion complex [19].
In recent times, two phenolic acids; trans-ferulic acid and gallic acid were analyzed and characterized by MIR spectroscopy in the field of a possible interaction that they can occur in a co-encapsulated complex [20]. More recently, MIR technique associated multivariate analysis was effective in characterizing and distinguishing hydrolysable and condensed tannins [21].
Identification of phenolic compounds in natural products is of a great interest. Several papers attempted to investigate this topic [22–24]. It seems clear that MIR spectroscopy has a good potential for the analysis of phenolic compounds in addition to its usefulness to determine the bioactivity and/or the antioxidant potential of plant materials. It is then important to set a spectral database to easily do identification and characterization. The aim of this study is to perform the analysis of series of pure phenolic standards (powders) in order to identify the main spectral bands and to explore the possibility to establish rules allowing the detection of specific or a family of phenolic compounds by MIR spectroscopy.

Material and methods
A total of 61 standards corresponding to 36 phenolic compounds (HPLC grade purity) belonging to phenolic acid and flavonoid families have been analyzed. Table 1 presents the phenolic standards purchased; their name, families (phenolic acids/flavonoids) and the provider company (Vwr via merck, Sigma Aldrich, Extrasynthese, Vwr via alpha aesar, Molekula, Vwr via Molekula, Vwr via Cayman, Vwr, Cayman) as well. Sixteen phenolic compounds were purchased from more than one supplier, giving a total of respectively 32 and 29 samples of phenolic acids and flavonoids.

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).