In the 1970s and 1980s, early days of the Green Revolution, plant

In the 1970s and 1980s, early days of the Green Revolution, planthoppers became major threats and today the same pests have Navitoclax supplier returned with a vengeance, causing even more destruction

and misery throughout South and Southeast Asia. Since 2008 Thailand’s rice bowl has suffered continuous outbreaks for 14 consecutive seasons. From 2010 rice farmers in Thailand have been losing a million tons of paddy a year due to the planthoppers. Similarly, Indonesia is suffering the same threats and lost about a million tons in 2011. Smaller patches of outbreaks occur in Malaysia, India, Myanmar, Bangladesh, Philippines and India while China continues to lose about 1 million tons a year. In 2012 the southern provinces of China suffered the worst planthopper outbreaks in the last 20 years. Besides economic loss, thousands hundreds of farmers have suffered crop failures, pesticide poisoning and severe debt problems which have forced them into poverty and hunger and even suicides. Planthoppers are secondary pests that are normally under natural control. Outbreaks are symptoms of unsustainable practices that destroy vital biodiversity and ecosystem services triggering exponential population growth resulting in outbreaks. Although abnormal weather like droughts and floods

can also trigger outbreaks, the most consistent factor in Asia is insecticide misuse. Insecticide misuse in Asia is due to weak marketing regulations that permit pesticides to be sold as FMCGs (fast moving consumer goods), like tooth paste (Heong et al., 2013; ADB Sustainable Development Working Paper # 27. Asian Development Bank, Manila Philippines). Pesticide retailers are uncertified and EX 527 in vitro often adopt multi-level Fenbendazole marketing systems and provide incentives to promote sales. Insecticides are packaged in hundreds of trade names, and mixed into cocktails,

further confusing farmers. At the village level retailers often serve as local pest control advisors to farmers as the government extension services are inadequate. When pesticides are marketed to encourage prophylactic applications and overuse it is difficult to sustain attempts to implement IPM. There is an urgent need to prioritize the strengthening of pesticide marketing regulations and their enforcement. Plant protection services in Asia were designed more than 50 years ago for “hunt and kill” operations. Today with increased evidence of the value of ecosystem services, plant protection systems need to be reformed to focus on information, diagnostics and accreditation that can provide reliable information and recommendations to farmers. To strengthen natural control mechanisms ecological engineering approaches that involve biodiversity restoration and conservation may be promoted to enable change (Gurr et al., 2012; In Biodiversity and Insect Pests: Key issues for sustainable management. John Wiley & Sons. pp. 214–229). Heong, K.L., Wong, L. and Delos Reyes, J.H. 2013.

However, further analysis revealed that, although the vast majori

However, further analysis revealed that, although the vast majority of TCR/pMHC complexes crystallized within the remit of these conditions, a number of structures crystallized in conditions outside of this range (Fig. 4). Thus, although it could be tempting to limit the number of conditions in a protein crystal screen to improve efficiency and reduce protein consumption, PLX4032 concentration broader screens are required to ensure that crystallization conditions are not missed for important proteins. The ability of T cells to respond to antigen depends on the productive

interaction between the TCR and pMHC. The crystal structures of a number of TCR/pMHC complexes have been solved and show that the TCR has a relatively conserved mode of binding to pMHC in which the GSK458 supplier TCR lines up approximately diagonally to the MHC peptide binding groove, with the TCR α

chain contacting the MHC α2 domain and the TCR β chain contacting the MHC α1 domain. The antigen specific portion of the TCR/pMHC interaction occurs between the pMHC surface and the TCR complementarity determining region loops (CDR-loops) (Rudolph et al., 2006). These CDR-loops serve different roles during TCR binding to pMHC: the variable (V)-gene encoded CDR2-loops contact mainly the conserved helical region of the MHC surface, the V-gene encoded CDR1-loops can contact both the MHC and the peptide and the more variable somatically rearranged CDR3-loops contact mainly the antigenic peptide. Although the general features of TCR/pMHC binding have been defined, there remains a number of conflicting models that describe the structural basis of T cell MHC-restriction, cross-reactivity, autoimmunity and alloreactivity. Furthermore, each previous TCR/pMHC complex has been governed by a unique set of contacts that enable T cell antigen recognition. Thus, there is still a pressing need to increase the number of TCR/pMHC complex structures in the literature in order to: (1) determine an accepted set of rules

Fenbendazole that describe the generalities of T cell specificity, and (2) understand the unique features of individual TCR/pMHC interactions that allow T cells to target different disease epitopes. The study of TCR/pMHC complexes has been limited by the challenges in expression, purification and successful crystallization of these soluble proteins. Here, we report a new systematic and directed approach for the design of a TCR/pMHC Optimized Protein crystallization Screen (TOPS) that has proved to be useful for the crystallization of this family of immuno-proteins. With this novel crystallization screen, we have successfully generated the majority of our current portfolio of structures that includes 21 TCR/pMHC complexes (13 derived from a common parent complex), 3 TCRs and 8 pMHCs. We found that TCR/pMHC complex crystals most commonly formed at a neutral pH, with 15%–20% of PEG 4000 and 0.2 M ammonium sulfate.

Another important mechanism appears to be the turbulent mixing ta

Another important mechanism appears to be the turbulent mixing taking place along the so-called Turkish Straits (TS) conduit (consisting of the Sea of Marmara, the Straits of Istanbul and the Dardanelles), thus increasing the total salt content of BSW outflow in the North Aegean Sea. Indeed, during the late May–early June 2001 period, strong south-westerly gales prevailed along

the TS, rapidly changing to vigorous north-easterly Etesians. Under south-westerly winds, the denser North Aegean Sea water increases its thickness along the Dardanelles, supporting vertical mixing and promoting salt diffusion to the upper layer, thus returning salt back to the Mediterranean (Yüce, 1996, Özsoy and Ünlüata, 1997 and Stashchuk SP600125 price and Hutter, 2001).

In contrast, north-easterly winds, dominant during the 1998, 1999 and 2000 summer sampling periods, cause southward surface BMN-673 currents to increase and northward bottom currents to decrease (Yüce 1996). Under these conditions, the thickness of Mediterranean water decreases and vertical mixing is limited as a result. At the sub-basin scale field of gyres and flows, the BSW-LIW frontal zone and the Samothraki Anticyclone appear as the most prominent surface features of the North Aegean Sea. Horizontal density gradients across the frontal interface appear stronger during the 1998 conditions Δσt = 0.11 per km), reducing to 0.05 per km in 2001, due to horizontal CYTH4 and vertical mixing induced by southerly winds. A significant cross-frontal horizontal geopotential anomaly gradient (ΔФ5/40 = 0.012–0.018 m2 s−2 per km) remains almost constant throughout the samplings. The Samothraki Anticyclone appears as a permanent feature in the area, containing a low density core (supplied by the less saline BSW) that produces both an upward doming of the sea surface, detectable by satellite altimeters ( Larnicol et al. 2002), and a strong clockwise geostrophic circulation ( Theocharis & Georgopoulos 1993). The horizontal

distribution of the geopotential anomaly (contour of ΔФ0/40 > 0.8 m2 s−2) was used to identify the anticyclone’s core water. It occurred that in summers 1998 and 2000, under northerly winds, the anticyclone was located to the north-west of Lemnos Island ( Figure 4d) and to the south-west of Samothraki Island ( Figure 7d) respectively, while in summer 2001, under the influence of strong south to south-westerly winds, it moved to the north-west of Samothraki Island ( Figure 9d). Figure 12 illustrates the eastward/westward baroclinic transport in the 0/40 m layer along the 25°E meridian. It turns out that in summers 1998–2000, under the influence of northerly winds, the Samothraki Anticyclone achieved almost symmetrical forms in terms of eastward/westward surface layer transport. Moreover, westward baroclinic transport induced by the BSW outflow was observed in deep water.

Putative target genes were manually selected from these candidate

Putative target genes were manually selected from these candidates based on their location in the maize genome. Functions of the predicted target genes were assigned manually according to the functions of the best hits from the BLAST search [41] and [43] against the NCBI database ( For the predicted novel miRNA sequences, conservation in other plant species was examined by searching

the nucleotide databases with BLASTn [41] to identify their homologs and surrounding sequences. These germination-related miRNAs were also aligned with the maize genome using PatScan [42]. To analyze whether the matched sequence could form a suitable hairpin, the sequences of candidate precursors were analyzed using BAY 73-4506 purchase RepeatMasker ( to eliminate repetitive sequences. Sequences surrounding the matched sequence (100–200 nt to either side) were extracted and run through RNAfold ( Most targets of miRNAs in plants have one miRNA-complementary site located in the coding region and occasionally

in the 3′ or 5′ un-translated regions (UTRs) [21], [36], [38], [44] and [45], and plant miRNAs exhibit perfect or near-perfect complementarity with their target mRNAs [46]. We adopted a set of previously reported rules to predict miRNA targets [36] and [47]. These rules allow one mismatch in the region complementary to nucleotide positions 2 to 12 of the miRNA, do not allow a mismatch at position 10/11, which is a predicted cleavage site, and allow three additional mismatches between positions 12 and 22, but with no more than two continuous find more mismatches. Therefore, candidate miRNA

target genes were determined using publicly available prediction algorithms, including miRU [48], the target search in WMD web [49], and the prediction tool in the UEA plant sRNA toolkit. These programs were used with their default settings. The microarrays used in this study were obtained from GSE9386, entitled “Genome-wide analysis of gene expression profiles during the kernel development of maize (Z. mays L.)”. The raw data from microarray hybridization was exported from GenePix suites Selleck Venetoclax 6.0 (Axon, USA) and imported into LIMMA with annotation and spot types [50]. Spots with a negative flag value were assigned a weighting of 0.1 in the subsequent analysis. Background-subtracted signal intensities were normalized using two-step normalization, consisting of print-tip group loss (within-array normalization) and between-array scale normalization. The adjusted p value was then assessed using the false discovery rate. To identify a statistically significant differential expression of genes, p = 0.01 was used as a criterion. To obtain probe annotations, the consensus representative sequences of all probes were searched using BLAST against the TIGR rice protein database (http://www.tigr.

Such reliable SCAR marker has been achieved in Mercurialis annua,

Such reliable SCAR marker has been achieved in Mercurialis annua, Carica papaya, and Cannabis sativa [14], [15] and [24]. The availability of markers linked to sex-associated genes would allow cloning the gene/s involved in this process and this information will help in the development of gene specific markers. It is possible to differentiate male, female, and hermaphrodite plants of Simarouba precisely and rapidly using the RAPD markers. Authors are thankful to the Gulbarga University for providing work facility and University of Agricultural Sciences

Dharwad and Bangalore for research material. “
“Lactic acid is widely used in the food processing, cosmetics, pharmaceutical and chemical learn more industry. Increasing prices of fossil fuels lead to increasing interests in lactic acid as a component for the production of biodegradable polymer polylactic acid [24]. There have been various attempts to produce lactic acid efficiently in bio-refineries from inexpensive feedstock such as lignocellulosic raw

materials, e.g. wheat straw or hard- and soft-wood [4] and [16]. Lignocellulose as part of the secondary cell wall of rooted plants is one of the most abundant natural materials. Ku-0059436 supplier It contains cellulose, hemicellulose and lignin [8]. Cellulose and hemicellulose represents polymeric carbohydrates formed from glucose, xylose, and arabinose amongst other sugars [22] and [16]. Therefore, lignocellulose is also the most abundant carbonate storage. After a hydrolysation

process, lignocellulose can serve as a potential substrate in a biotechnological microbial fermentation for the formation of valuable products such as lactic acid [11], [12] and [23]. Unfortunately, a non-specific chemical hydrolysis treatment, e.g. high temperature acid or alkali pre-treatment, leads to solvation of lignin and to the formation of complex sugars and inhibitory compounds such as furfural [18], [19], [20] and [21]. One way of reducing the inhibitory effect of lignin for oxyclozanide process optimization is the reduction of the lignin concentration in the fermentation medium [7]. Another option is the use of microorganisms inhibited by lignin only to a low level, or those that can transform lignin into another compound like vanillate [10] and [13]. In order to improve the screening of microorganisms usable in complex and inhibitory media like lignocellulosic hydrolysates, it is necessary to characterize their growth behaviour. High throughput methods for kinetic analysis of the lignin inhibition are useful to achieve information about the lag time (λ) and the maximum growth rate (μm). These screening methods provide the chance to investigate the growth behaviour under different working conditions. In order to get access to lignin stable natural microorganisms (MOs) it is crucial to screen interesting bacteria in an inhibitory environment.

A fundamental problem in wavelet analysis is the selection of the

A fundamental problem in wavelet analysis is the selection of the mother wavelet function. For analysing the echo envelope of the acoustic signal, Ostrovsky & Tęgowski (2010) applied six differently defined mother functions. The use of so many different functions did not yield a larger amount of information, however. In the present case, the number of wavelet mother functions was reduced to two: one symmetric and the other asymmetric. The Mexican Hat (mexh) buy AZD4547 was selected as the symmetric wavelet mother function, while the family of Daubechies wavelets exemplifies the asymmetric mother functions. A wavelet

of the order of 7 (db7) was selected from this family. In order to account for wavelet asymmetry, profiles were analysed in both directions, in the same direction as the measurements according to (db7 +) and in the opposite direction (db7 −). The following parameters were determined for each of the transforms: – wavelet energies for a given scaling parameter (EMVj, wav, ELTj, wav, ESTj, wav): The use of a fractal dimension in the analysis of bottom bathymetry should result from the following assumptions (Herzfeld et al. 1995): – bathymetry has a non-trivial structure at every scale;

It is evident that the bathymetry of a water body formed by numerous geological processes has a non-trivial structure and that it cannot be described by simple geometric figures. The work involving the analysis of bathymetric profiles from the eastern Pacific (Herzfeld et al. 1995) indicates that bathymetry can be treated as a fractal because the assumption that DH > DT is fulfilled; selleck inhibitor however, the assumptions of self-similarity are not satisfied when the image scale is being changed. The fractal dimension is considered to be an appropriate parameter for describing the morphological diversification of bottom surfaces ( Wilson et al. 2007).

In the case of a flat bottom, the fractal dimension calculated for the bathymetric CYTH4 profile should take a value equal to unity; as irregularities in the seafloor appear and their magnitudes change, its value will rise. In this work, the fractal dimension was determined using indirect methods, such as the box dimension, semivariogram analysis of spectral parameters and wavelet analysis. For determining the box fractal dimension of the deviations from the bathymetric profile segments (DMVbox, DLTbox, DSTbox), the definition given by Hastings & Sugihara (1994) was used: equation(14) Dbox=limΔs→0log10NΔs−log10Δs, where N(Δs) determines the number of squares covering a depth profile of a side length Δs. In case of the bathymetric profiles, both the length and depth have the same dimension. The proposed procedure for determining this parameter consists of four consecutive steps: – normalisation of the distance, taking the unit profile length to be 256 m; Application of a uniform standardisation is valid, taking a standard distance and depth of 256 m, equal to the length of the analysed section.

791 g and 0 828 g SFA per 120 g (data not shown) In Brazil, at p

791 g and 0.828 g SFA per 120 g (data not shown). In Brazil, at present, both MF–I and MF–WPC could not receive the comparative “reduced saturated fat” claim (Table 7), once a reduction of at least 25% less saturated fat and a difference higher

than 1.5 g/100 g in buy Dasatinib this nutrient compared to the control MF are required (Brasil, 1998). Standards for “reduced saturated fat” products are planned to be at least 30% SFA of the control product in Brazil, besides the conditions that the decreased saturated fat content must not result in an increased trans-FA, the reference product is not able to fill the requirements for a “low saturated fat” product, and

the energy given by SFA must not be above 10% of the total energy of the product ( ANVISA, 2011). According to these requisites, mousses I, WPC, I–WPC, and MF–I–WPC could receive the “low saturated fat” claim, oppositely to mousses MF–I and MF–WPC ( Table 7). For this kind of product, the U.S. and the E.U. legislation showed to be less restrictive regarding the comparative “reduced saturated fat” claim. This claim can be applied in the U.S. for all modified mousses, with the exception of mousse MF–WPC, once they all presented at least 25% less SFA than control mousse MF ( Table 6 and Table 7) ( US CFR, 2010f). Similarly, only mousse MF–WPC, with less than 30% SFA than control MF, also did not fill the requisite to receive this claim in the E.U. ( Table 6 and Table 7) ( EC, Talazoparib supplier 2007). In Brazil, the current nutritional information and claims for specific nutrients such as trans-FA ( ANVISA, 2003b) already consider their amounts per serving portion, which is equivalent to ½ cup (120 g) for milk-derived desserts ( ANVISA, 2003a). For all mousses studied, trans-FA content is lower than 0.2 g per serving portion of ½ cup (data not shown) and might be declared in Brazil as “zero” ( Table 7) ( ANVISA, 2003b). The acceptable upper limit for a “zero” trans-FA product is proposed

to be more severe, Mirabegron reducing to 0.1 g of this component per serving portion ( ANVISA, 2011), which implies that control mousse MF (0.109 g trans-FA/120 g) could not be declared as “zero trans-FA” following this standard ( Table 7). In the U.S., on the other hand, the legislation is more flexible in this situation: products that contain less than 0.5 g of trans-FA per serving portion, as in case of all mousses studied, are considered as “zero” or the statement “not a significant source of trans fat” is placed at the bottom of the table of nutrient values ( US CFR, 2010a). Such specific claims for trans-FA in food products are not contemplated by the E.U. legislation ( EC, 2007).

The Zhoushan region is located in eastern China in the northeaste

The Zhoushan region is located in eastern China in the northeastern region of the province of Zhejiang. The study was carried out in the obstetrical wards of Zhoushan Women’s & Children’s Health Hospital, a major maternal and child health hospital in Zhoushan city. From July 2005 to October 2006, every month approximately 30 pregnant women were recruited at their first or second prenatal medical examination, and the total study cohort consisted of 440 women. Neurobehavioral development of the neonates was assessed at 3 days of age using the Neonatal Behavioral Neurological Assessment (NBNA). Mothers and neonates with disorders highly associated with adverse neurodevelopment such as traumatic brain

injury, meningitis, epilepsy, and severe neonatal illnesses were excluded from the study after delivery. Sixteen infants were excluded because of preexisting or acquired medical problems that may seriously affect development. Six mothers were excluded because of incomplete PI3K Inhibitor Library manufacturer questionnaires. In total, 418 mother-neonate pairs were included in the study after written consent

was obtained, and they completed questionnaires. The study protocol was approved by the Medical Ethics Committee of Zhoushan Women’s and Children’s Health Hospital. A detailed questionnaire was administered to collect information on fish consumption and the general nutritional status of the mothers during the third trimesters of pregnancy. All mothers were asked to estimate the quantity and type of fish consumption in a week. In addition, the questionnaire included questions regarding potential confounding factors such as demographic data, maternal education, abortion history, use of skin whitening cosmetics, dental amalgam treatment, occupational exposures, monthly household income per capita and/or month, and some paternal-related information. All questionnaires were administered by trained interviewers. Prenatal mercury exposure was assessed by measuring mercury concentrations in maternal blood, hair, urine, and neonate cord blood. Maternal hair samples were collected about 1-3 days

after delivery. The samples in the proximal 3 cm length to the scalp and weighing 0.5-0.75 g were collected for mercury concentration. selleckchem All samples were handled in a class 100 clean hood. Approximately 0.3 g of hair weighed in a quartz boat. We precleaned plastic and glassware by soaking them in 10% HNO3 for 24 hours and then rinsing them several times with deionized water. Hair samples were sonicated for 15 minutes in approximately 10 mL of 1% Triton X-100 solution in precleaned 50-mL Pyrex beakers. After sonication, samples were rinsed several times with distilled deionized water and dried at 60°C for 24 hours. Urine specimens from the first morning voided urine samples (approximately 50 mL) were collected over a period of 24 hours in polypropylene vessels in the third trimester of pregnancy.

The risks of exposure to, and severe disease from RSV, should be

The risks of exposure to, and severe disease from RSV, should be carefully assessed when administering Palivizumab. Down’s syndrome itself has been shown to be a risk factor for severe RSV infection, even in the absence of congenital

heart disease. For infants and children with Down’s syndrome ≤24 months of age at the beginning of the RSV season, Doxorubicin ic50 the prevention of severe RSV disease using Palivizumab may be considered when the patient suffered any of following past or present complications, or has abnormal laboratory test results: • Anatomical, physiological or functional abnormalities of the respiratory system: Airway obstruction and/or associated apnea due to marked megaloglossia, Bioactive Compound Library glossoptosis, respiratory tract malacia, or other airway abnormalities, pulmonary hypertension,

pulmonary hypoplasia/dysplasia, or emphysematous lung. * Although the normal values vary depending on the months of age, one suggestion would be 2000/mm3 or lower and 1000/mm3 or lower for lymphocyte counts and T-lymphocytes, respectively. (1) If patients have a tendency to bleed due to thrombocytopenia (such as because of Wiskott–Aldrich syndrome and myeloablation) or other coagulopathy, or they are receiving anticoagulants and/or antiplatelet drugs, bleeding resulting from an intramuscular injection of Palivizumab may be serious. It is recommended that Palivizumab be carefully given to such patients, for example, with application of pressure to

the injection site for an appropriate length of time to ensure hemostasis. It is important to employ strict infection control measures even when using Palivizumab. It is particularly important to educate guardians, since their cooperation is essential in managing high-risk children. It is also important to provide instructions not only for RSV infection, but also for other pathogens causing respiratory tract infections. In addition, guardians should understand that adhering to the administration schedule is critical to maximize the effectiveness. Recent medical advances have improved the lives of immunocompromised patients, but as a result, the chance of exposure to and infection by RSV among these high risk patients has increased. Severe RSV infections in immunodeficiency disorders such as SCID have long been recognized, at least since GNAT2 the 1980s 2, 4 and 5. Hall et al. examined the immunological status of 608 infants under the age of five who were hospitalized with an RSV infection over a ten year period [2]. They identified 47 patients with immunologic abnormalities, including those receiving chemotherapy (20 cases) or steroids (22 cases) and those with primary immunodeficiency syndrome (5 cases). The frequency of nosocomial infections, as well as the rates of infection in the lower respiratory tract, admissions to the ICU and mortality was compared with immunologically normal children.

, 1995) After incubation, cells were washed twice with FACS buff

, 1995). After incubation, cells were washed twice with FACS buffer and were either used for intracellular staining or fixed with a solution of 2% paraformaldehyde in PBS. Incubation with primary antibodies to MHC I (Salomonsen et al., 1987) and MHC II (Kaufman et al., 1990) was followed by Alexa-647 conjugated goat anti-mouse antibody (Life Technologies). Secondary antibody alone or unconjugated goat anti-mouse antibody (Life Technologies) was used as an unstained control for surface MHC staining. Intracellular staining JQ1 chemical structure was carried out as described previously (Ariaans et al., 2008). Briefly, splenocytes from challenged birds or non-infected controls were seeded in a 96-well round-bottom plates (Nunc)

at 106 cells/well in a final volume of 200 μl of culture media or culture media supplemented with the different stimuli at the concentration described in the ELISpot technique (except PMA which was used at 50 ng/ml). Cells

were cultured using the conditions described above for ELISpot assays (24 h culture). Selleckchem Epacadostat For intracellular staining, during the last 2 h of culture, cells were treated with Brefeldin A according to the manufacturer’s instructions (Cytofix/Cytoperm™ Plus Fixation/Permeabilization kit, BD Biosciences). To avoid non-specific binding signal, we preincubated cells with Cytofix/Cytoperm™ buffer containing 2% normal mouse serum and further staining steps involved Cytofix/Cytoperm™ washing buffer containing 1% normal mouse serum (Biosource). To confirm the specificity of the anti-IFNγ antibody EH9 we also employed a validated anti-IFNγ [mAb80 (Ariaans et al., 2008)]. Purified fractions of both antibodies were conjugated using Alexa Fluor® 647 monoclonal antibody labeling kit (Molecular Probes) according to the manufacturer’s selleck screening library instructions. A mouse isotype matched control antibody IgG1 Alexa Fluor® 647 (Life Technologies) was employed at the same concentration as EH9 and Mab80. For analysis, a gate on the FSC/SSC region of lymphocytes was selected and a minimum of 10,000 events were acquired on a FACSCalibur instrument using Cell Quest software (BD Becton Dickinson). Flow cytometry

analysis indicates that non-adherent CKC were not present at significant levels (data not shown). FlowJo software (TreeStar) was used to analyze flow cytometry data. A paired or unpaired t-student test or one-way ANOVA was performed using GraphPad Prism (version 6.0 for Windows, GraphPad Software, San Diego, California, USA). Screening identified two anti-chicken IFNγ antibodies (clones EH9 and AF10) which were shown by ELISA to bind recombinant chicken IFNγ and to work effectively as an antibody pair in capture ELISA (Supplementary Fig. 2A–C). We subsequently compared this antibody pair with commercially available antibodies [from Life Technologies (Ariaans et al., 2008 and Reemers et al., 2012)] in ELISpot assays.