To analyze relative expression of different stage mRNAs, the amount of cDNA was normalized based on the signals from ubiquitously expressed β-actin mRNA (beta-actin5, 5′-GACCTGACAGACTACCTGAT-3′, and beta-actin3, 5′-AGACAGCACTGTGTTGGCAT-3′). To Selleckchem Obeticholic Acid provide negative controls and exclude

contamination by genomic DNA, the reverse transcriptase was omitted in the cDNA synthesis step, and the samples were subjected to the PCR reaction in the same manner with the same primer sets as indicated above for RT-PCR (−). PCR products were separated by electrophoresis in agarose or polyacrylamide gels, and the bands were visualized with ethidium bromide on an Alphaimager (Alpha Innotech). The identity of all the PCR products was confirmed by

sequencing. All obtained sequences were analyzed with the Genetyx software version 7.0 (GENETYX CORPORATION).The sequence was submitted to GenBank (Accession number; AB698464, Nivolumab cost AB698465, AB698466). In situ hybridization of zebrafish was performed as described previously ( Makino et al., 2005). Briefly, samples were fixed overnight at 4 °C in 4% paraformaldehyde in phosphate-buffered saline (PBS), washed briefly in two changes of PBS-0.1% Tween 20 (PBT), and transferred to 100% methanol for storage at − 20 °C. The samples were rehydrated stepwise through methanol in PBT and then washed Oxalosuccinic acid in four changes of PBT. Subsequently,

samples were incubated with 10 μg/ml proteinase K in PBT for 15–60 min and rinsed twice in PBT before a 20-min refixation. After washes in five changes of PBT, samples were prehybridized at 65 °C for 1 h in buffer consisting of 100% formamide, 20x SSC, 0.1% Tween 20, 10 mg/ml heparin, 9 mM citric acid, and 50 mg/ml yeast RNA and then hybridized overnight in hybridization buffer containing 0.5 mg/ml digoxigenin-labeled RNA probes with the sense or anti-sense sequence. Labeling of the RNA probe was performed with a labeling kit (Roche) using the pGEM-T-Easy vector cloned zMai1 sequence, which is common to all splice variants of zMsi1, and the probe was confirmed by sequencing. Samples were then washed at 65 °C for 10 min each in 75% hybridization buffer/25% 20x SSC, 50% hybridization buffer/50% 20 × SSC, and 25% hybridization buffer/75% 20 × SSC, followed by two washes for 30 min each in 0.2 × SSC at 65 °C. Further washes for 5 min each were conducted at room temperature in 75% 0.2 × SSC/25% PBT, 50% 0.2 × SSC/50% PBT, and 25% 0.2 × SSC/75% PBT. After a 1-h incubation period in PBT containing 2 mg/ml bovine serum albumin, samples were incubated for 2 h in the same solution with a 1:2000 dilution of sample-preabsorbed anti-digoxigenin antibody.

There find more are some limitations in the present study. The lack of inundation at the coastlines, coupled with the minimum depth requirement, means that the true free-surface variation at an arbitrary coastal location cannot yet be represented. Fluidity is capable of simulating inundation in a limited region (Funke

et al., 2011) and work is ongoing to link this technology to large-scale simulations. The virtual wave gauges must be contained within the mesh to record the free surface variations at a given location. As we varied coastlines and resolution, wave gauges were moved slightly between simulations to ensure they were not on land. Bondevik et al. (2005) used a similar methodology as the gauges specified there were not within their computational domain. They do not report the true location as the effect of this shift was thought to be small. The largest difference in the present study was less than 1 degree for the 50 km resolution simulation with the coarsest GSSHS coastline. All other simulations had differences of much less than 1 degree. The current model does not include inundation as the wave reaches the coastline. Therefore

www.selleckchem.com/products/BAY-73-4506.html comparisons are made between the estimated run-up height from sedimentary deposits and the maximum wave height in the vicinity of the deposit. The difference between the two estimates will depend on local factors, such as vegetation and small-scale (i.e. unresolved) bathymetric/topographic changes. We aim to include this in future work. Perhaps the most important simplifying assumption within this study is that the Storegga Slide moved as a single rigid block. This a priori   assumption is important because the way in which the original slide moves determines the initial dimensions of the resulting tsunami. Field observations ( Haflidason et al., 2005) suggest that much of the slide mass disintegrated, such that it was not a single rigid block. Moreover, there is evidence that GBA3 slope failure

started in deep water and moved retrogressively upslope ( Masson et al., 2010). This modelling also assumes a priori   that the slide accelerated to a speed of ∼∼35 m/s over 3365 s. The acceleration trajectory of the slide is unknown, although previous modelling suggests that such fast speeds are needed to generate a large far field tsunami. We have based our model on the work of ( Harbitz, 1992). This was later refined in terms of both the slide shape and initiation by Bondevik et al. (2005) but no comparison to Harbitz (1992) was carried out and hence it is difficult to ascertain what effect these modifications had on the model results. Bondevik et al. (2005) do not give an analytical expression for the modified slide and hence it could not be used in this study. In addition, Bondevik et al. (2005) also increased resolution of the mesh from 12.5 km to 2.08 km, possibly confounding any comparison.

24 Digital images were analysed using Sigma-Scan 2.0 software. The distance between the cemento–enamel junctions up to the height of alveolar bone of the first mandibular molar on the mesial side of the rat was recorded. Samples were homogenised in Trizol reagent (Invitrogen) for 1 min using a tissue homogenizer (Polytron-Agrgregate, Kinematica, Littau/Luzern, Switzerland) at maximum speed. Total RNA was isolated according to the manufacturer’s guidelines and quantified by a spectrophotometer. Fluorouracil The integrity of RNA was verified by agarose gel electrophoresis. Complementary DNA was prepared using 2 μg of total RNA and a reverse transcriptase. The primers used

in the experiments were the standard TaqMan (Applied Biosystems, Foster City, CA, USA) brand. The gene analysed were TNF-α

(primers: sense 5′ GGC ATG GAT CTC AAA GAC AAC C-3′ and antisense 5′-CAA ATC GGC TGA CGG TGT G-3′). Glyceraldehyde-3-phosphate dehydrogenase (GenBank NM_017008) was used as a housekeeping gene. Real-time polymerase chain reaction was carried out in the StepOne polymerase chain reaction cycler (Applied Biosystems, Foster City, CA, USA). The polymerase chain reaction conditions were 95 °C for 10 min, followed by 40 cycles at 95° for 10 s and 60 °C for 45 s. Real-time data were analysed using the Sequence Detector System 1.7 (Applied Biosystems, Foster City, CA, USA). Results are expressed as fold inductions compared with controls. Results FXR agonist are presented as means ± SEM for the number of rats (n) indicated. The data were analysed by the unpaired Student’s t-test for two mean comparisons and one-way ANOVA (with Bonferroni post hoc test) for bone reabsorption and TNF-α expression. The level of significance was set at P < 0.05. Table 1 shows that the body weight and naso-anal length were 29% and 15% respectively, O-methylated flavonoid lower in MSG groups when compared with CTL (P < 0.05), however, the Lee Index was 8% higher in the MSG rats (P < 0.003). The retroperitoneal and perigonadal fat pads weight doubled in

MSG rats when compared with CTL rats (P < 0.0001, Fig. 1A and B). The neonatal MSG treatment did not influence the plasma concentration of glucose, NEFA and total CHOL (P > 0.05). However, in the MSG group plasma and TG concentrations were 3.0 and 4.0 times higher (P < 0.0001 and P < 0.0002), respectively than, CTL group ( Table 2). According to Fig. 2, alveolar bone resorption was 44% lower in obese-MSG group compared with CTL group (P < 0.01). In the presence of ligature, there was a significant increase in alveolar bone resorption in both groups CTL L and MSG L compared with CTL and MSG group respectively (P < 0.001). However, alveolar bone resorption in the MSG L animals was similar to that occurring in the CTL group (P > 0.05) ( Fig. 3A–D). The TNF-α gene expression in periodontal tissue was similar in MSG and CTL animals in the absence of ligature (P > 0.

COTS were placed in individual 68-l plastic containers with flow-through seawater at ambient conditions. Injections of 10 ml of each solution (initially, 4 g l−1 of Bile Salts No. 3 and 6 g l−1 of Oxgall) were administered using a plastic

syringe with an 18-gauge needle. Sea stars were injected in (1) the distal portion of the arm, (2) the middle of the Ganetespib manufacturer arm, (3) the proximal portion of the arm, and (4) the central disk ( Fig. 1). A. planci used in the double dose treatments were all injected in the central disk. Two separate measures of the effectiveness are considered in this study: i) the time until death (in hours), recorded as the time from injection until all podia (tube feet) were completely immobile ( Rivera-Posada et al., 2011), and ii) the proportion of sea stars that actually died with 2–3 days. A total of 12 A. planci distributed in three groups of 4 sea stars were used for this experiment. Each A. planci was injected with 10 mL of 8 g l−1 Bile Salts No. 3 and time to death was estimated.

Hyperactivity shortly after injection was used as an indicator that the sea star was correctly injected. Three different types of injection guns were tested ( Fig. 2): (1) DuPont™ Velpar® Spotgun®, (2) Simcro™ STV 12-ml Plastic Syringe, and (3) prototype metal injection gun. The DuPont™ Velpar® Spotgun® was fitted with a 50-cm needle, 4-mm tip, and 5-L plastic bladder, which is currently used in the field to inject sodium bisulfate (dry acid) solution. Although this gun provides good reach to cryptic A. planci, the INCB024360 research buy width of the tip creates large holes, which raises concerns that chemicals injected could easily leak out of these openings without any effect or without killing the sea star. It is important to note that this gun was originally designed to spray herbicides and not to inject A. planci. Simcro™ STV 12-ml Plastic Syringe is cheap, lightweight, requires minimal maintenance, and offers the possibility to attach Montelukast Sodium any size and length of syringe needle. This gun has been successfully used in A. planci control efforts around Japan ( Kuroshio Biological Research

Foundation, 2011). A prototype metal injection gun with a 50-cm spear and Luer-lock to attach a 16 Ga × 1/2″ needle was designed for more accurate injections of small amounts of solution ( Fig. 2, inset). A thinner and shorter needle was used to minimize the puncture size and leakage after injection and to avoid overshooting (tip of needle exits the sea star arm and solution is not injected internally) during injection, as what usually happens with longer needles. Fish, corals, and other echinoderms (Table 1) were collected from back reef habitats around Lizard Island. Smaller fishes (i.e. Pomacentridae, Chaetodontidae) were collected using clove oil, which is noteworthy because clove of its hepatotoxic properties (Javahery et al.

Total serum creatine

kinase (CK) and its isoenzyme MB (CK–MB) are well established and widely accepted markers for the diagnosis and follow-up of heart injury or myocardial infarction (Bachmaier et al., 1995). Biochemical analyses showed an increase in CK and CK-MB levels (Fig. 3A and B, respectively). Increased CK concentrations were observed click here in envenomed animals (4850 UI/mL) compared to the control group (injection of PBS) (1293 UI/mL). Levels of CK–MB were also higher in the envenomed group (1980 UI/mL) than in the control group (413 UI/mL). We have demonstrated previously, in dogs envenomed with Tityus fasciolatus scorpion venom ( Pinto et al., 2010), that the occurrence of myocardium damage is correlated with high serum levels of CK and CK–MB. As we

also observed higher levels of these two markers of heart injury of the envenomed rats, we suggest in this study that H. lunatus venom has cardiotoxic effects, possibly through the action of neurotoxins acting on voltage gated ion channels present in the heart ( Chen and Heinemann, 2001; Korolkova et al., 2004). The soluble venom of H. lunatus was fractionated by HPLC and showed more than 20 components ( Fig. 4A). As with other chromatographic profiles of scorpion venoms ( Batista et al., 2004), the separation in C18 reverse column is completed at approximately 60 min of the buy Gefitinib gradient, at a flow rate of 1 mL/min. According to the authors mentioned above, the fractionated components during

the first 20–40 min of the gradient would be the minor peptides corresponding to K+- and Na+- channel neurotoxins. It is known that most scorpion oxyclozanide toxic peptides have molecular masses lower than 10 kDa. These basic peptides are neurotoxins of low molecular mass that bind to ion channels with high affinity, exerting their noxious effect ( Catterall et al., 2007). These small proteins may be responsible for the typical symptoms of neurotoxic envenoming observed in inoculated mice. Several components purified after RT (retention time) 30 min showed PLA2 activities (peaks 10 to 19, except the fraction 13). Fraction 15 (from 35 to 40 min RT), which showed highest PLA2 activity, was further analyzed by MALDI–TOF and the individual components clearly identified. The molar masses ( Fig. 4B) found were 11,914.5 Da and 13,650.6 Da. In the final part of our study and as a preliminary step in the production of an anti-H. lunatus anti-venom with therapeutic properties, we have attempted to study by ELISA and immunoblotting the antigenic/immunogenic potential and cross-reactivity of rabbit anti-H. lunatus serum. Immune sera anti-H. lunatus and anti-T. serrulatus (for comparative purposes), were raised in rabbits and their reactivities against H. lunatus, T. serrulatus, C. sculpturatus and Androctonus australis hector venoms evaluated. Fig. 5 shows the ELISA (absorbance at 490 nm) at different serum dilutions (1:100 to 1:12,800).

But most assays require various components, two to three substrates, cofactors, activators, and reagents for stabilization or prevention from deactivating processes, like oxidation or proteolysis. These components can

be added step by step to the assay until, with the last addition, the reaction PD0325901 ic50 starts. Such a procedure is not only laborious and time consuming, especially for extensive test series; it is also not very accurate. Pipetting is usually the severest source of error and, therefore, pipetting steps should be reduced as far as possible. Especially pipetting of small volumes proceeds with higher uncertainty than of larger volumes. Therefore it is advantageous to prepare a larger quantity, an assay mixture for the whole test series instead of executing each assay sample separately. The assay mixture should contain all necessary components in their final concentrations, with the exception of one, which is added finally to the individual assay sample to start the reaction. If, for example, 5 components of 2 µl must be added step by step to

an assay sample of 1 ml, 500 pipetting steps are required for 100 tests, while only 5 pipetting steps of 0.2 ml are required to prepare 100 ml assay mixture. Besides time saving the accuracy increases significantly, as the scatter of the data will considerably be reduced, because all samples (with the sole exception of the science last component FK228 mw to be added to start the reaction) possess exactly the same composition. This opens, however, the risk, that an error of one single step, e.g. wrong pipetting, obligatorily affects all assays, while by direct pipetting only the one sample, where the error happens, will be concerned. Nevertheless, the risk is minor, since preparation of a large quantity with few single steps can (and should) be done with great care, while such care cannot be given to any of the separate assays. The required components are preferentially added to the assay mixture

from concentrated stock solutions. They can be prepared in a larger quantity and frozen for storage. Immediately before usage they will be thawed and the portions not consumed can be frozen again. Since sensitive substances, like NADH, do not stand repeated freezing and thawing, such solutions may be divided into small portions, each sufficient for one test series, and frozen separately. Reagents which are not stable in solution at all must be prepared directly before usage. Some solutions, like buffers and inorganic salts, are principally stable at room temperature, but for long-term storage to avoid microbial contamination they should also be frozen. Care must be taken that all components of the assay mixture are compatible with one another. Any reaction, like oxidation, reduction, precipitation or complexing (e.g.

In addition

to the diagnostics of intracranial vascular disease, this technique is valuable in intensive care and stroke units for follow-up examinations in vasospasm after subarachnoid hemorrhage and for intraoperative monitoring. In difficult anatomical conditions, the application of echo contrast agents can improve the diagnostic reliability of the examination. Based on advances in computer and transducer technology http://www.selleckchem.com/products/obeticholic-acid.html TCCS as a noninvasive method has a great potential in further innovative imaging and therapeutic solutions such as cerebral perfusion imaging, sonothrombolysis, and site targeted ultrasound contrast agents for drug delivery to the brain. “
“The National Institute of Neurological Disorders and Stroke trial of recombinant tissue plasminogen activator (tPA) showed that JQ1 intravenous thrombolysis with acute ischemic stroke within 3 h from onset had favorable clinical recovery compared with placebo-treated patients [1]. However, a thrombolytic effect was not evaluated with monitoring of occlusion artery in this study. Cerebrovascular ultrasonography was useful clinically for evaluating cerebral hemodynamics rapidly and in real-time for the patients with acute ischemic stroke compared with magnetic resonance angiography (MRA). The timing and speed of recanalization after (tPA) therapy monitoring by transcranial Doppler (TCD) correlates with clinical recovery [2] and [3]. These real-time flow informations are

useful in developing next therapies and in selection for interventional treatment. The aim of this study was to analyze if the patients had early recanalization or not using transcranial color-coded sonography (TCCS) in order to evaluate the usefulness of real-time monitoring in systemic thrombolysis. Dipeptidyl peptidase Consecutive patients who had acute ischemic stroke with intravenous tPA within 3 h from onset between April 2010 and January 2011 were included in this study.

tPA was administered in a dose of 0.6 mg/kg (10% bolus, 90% continuous infusion during 1 h) according to Japanese standard protocol [4]. The patients with insufficient acoustic window were excluded. An experienced neuro-sonographer performed all TCCS studies using a EUB-7500 or 8500 with a 2 MHz sector transducer (S50A, HITACHI Medical Corporation, Japan). We evaluated occlusion of intracranial arteries from transtemporal or suboccipital window by TCCS with Thrombolysis in Brain Ischemia (TIBI) flow-grading system [5] and monitored residual flow in real-time every 15 min until 120 min after the t-PA bolus. An insonation time with TCCS was not longer than 5 min in each examination. No head frame was used during insonation. Complete recanalization was defined as TIBI 0–3 to 5, and partial recanalization was defined as TIBI 0–2 to 3. National Institutes of Health Stroke Scale (NIHSS) scores were obtained before tPA treatment, every 15 min until 1 h and every 30 min after 1 h by a neurologist.

This work was supported by a European Commission Marie Curie Intra-European Fellowship (011457) and a Brunel Research Initiative (BRIEF) Award to CR and a Wellcome Trust Senior Fellowship to MH. We are indebted to all our participants. “
“Goal-directed action requires the ability to identify the consequences Selleck Crizotinib of our behaviour in the external world. We use the term ‘agency’ to refer to

this fundamental aspect of human self-consciousness (Pacherie, 2008). Recent theoretical work distinguishes between two important aspects of agency (Synofzik et al., 2008a, 2008b). First, people can make explicit judgements about their agency (“I did that”). Second, there is a subjective feeling of control that accompanies one’s own actions, even in the absence

of any conscious awareness or reflective thought, known as sense of agency. The dominant experimental model for studying agency involves explicit judgements of whether a sensory event is caused by one’s action, or by that of another agent. Several studies have used self-recognition paradigms to investigate this explicit sense AZD2281 nmr of agency ( Daprati et al., 2007; Tsakiris et al., 2005). In the typical design, the participant makes a manual action, and sees video feedback which may either show their own action or the action of another person. Participants judge whether they are viewing their own hand action or not. Other studies have extended this paradigm from recognition of one’s own hand action to judging whether arbitrary effects, such as appearance of a symbol on a computer screen, are caused by one’s own key press actions or another person’s ( Sato and Yasuda, 2005; Wegner and Wheatley, 1999). Spatial ( Daprati et al., 2007) and temporal ( Farrer et al., 2008; Wegner and Wheatley, 1999) congruence of one’s own action and sensory feedback are key cues for self-attributing agency. Another prominent approach to investigate agency has been to manipulate agency as an independent variable by either giving participants control or not giving them

control over some external event, and contrasting different levels of control ( Metcalfe and Greene, 2007). Level of control is often manipulated by introducing a feedback delay. Interestingly, Interleukin-3 receptor recent neuroimaging studies failed to find any clear neural correlates for positive judgements of agency, but showed that the right angular gyrus plays a key role in rejecting agency based on lack of temporal congruence ( Farrer et al., 2003, 2008). The importance of the parietal areas in general, and the angular gyrus in particular, in processing of agency was confirmed by patient studies. Lesions including this area were reported to produce a deficit in recognising visual feedback of one’s own action ( Sirigu et al., 1999). It remains unclear whether angular gyrus activation is linked to explicit judgement of agency, or whether automatic monitoring of action outcomes is sufficient. For example, Miele et al.

nimh.nih.gov/lcmr/snge/Protocols/ISHH/ISHH.html). Slides were exposed to Amersham Hyperfilm MP film for 2 months at room temperature with appropriate 14C-labeled standards (Amersham, Little Chalfont,

UK). No specific hybridization was detected with sense probes and no APJ mRNA signal above background was detected in tissues from APJ KO mice. Some slides were subsequently dipped in Ilford K5 nuclear emulsion and stored desiccated at 4 °C for 4–6 months before development using Kodak D19 at room temperature. Tissue sections were counterstained with toluidine blue. Mouse cryostat sections (20 μm) were cut and thaw mounted onto subbed (gelatin, vanadium oxide) slides. APJ receptor autoradiography was performed with check details modifications of the procedure described by Katugampola et al. [21]. Brain sections were fixed in 0.1% PFA in PBS for 5 min and rinsed in 10 mM Hepes pH 7.5.

All sections were pre-incubated for 20 min in 20 mM Hepes pH 7.5 containing 1 mM EDTA, 0.3% BSA and Sigma Protease Inhibitor Complex (Sigma, Dorset, UK). Slides were then incubated in 20 mM Hepes, pH 7.5, 100 mM NaCl, 5 mM Alpelisib price MgCl2, 10 mM KCl, 1 mM EDTA, Sigma protease inhibitor complex and 0.3% BSA containing radiolabeled (Glp65, Nle75, Tyr77) (125I)-Apelin-13 [0.5 nM] (Perkin Elmer, Cambridgeshire, UK) in the absence or presence of unlabeled (Pyr1)-apelin-13 [1 μM] (Bachem, Germany) as a displacer. Binding specificity was assessed by comparison of the distribution of [125I]-(Pyr1)apelin-13 binding sites in wildtype tissue to that in APJ KO tissue. Incubation lasted 1 h at RT in a humid chamber and was followed by 2 × 10 min washes in ice-cold 20 mM Hepes pH 7.5, 0.3% BSA with stirring

and 2 × 15 min washes in ice-cold 10 mM Hepes pH 7.5. Slides were then rinsed in ice-cold dH2O and air-dried Levetiracetam at 4 °C before being exposed to X-ray film (Amersham Hyperfilm MP) for 2 weeks. Following this some slides were re-exposed to emulsion-coated film (Amersham Hyperfilm 3H) for 1 month to obtain better macroscopic resolution. Films were developed as described for ISHH, except emulsion-coated films, which were developed manually as per manufacturer’s instructions. ISHH with antisense APJ riboprobes was used to map the distribution of APJ mRNA in the male and female mouse brain and peripheral tissues. Sections from all tissues were also hybridized with sense APJ riboprobes as controls and showed only background level of labeling. A number of tissues, including the pituitary, lung, heart, ovary and uterus, showed high levels of hybridization, with representative photographs shown in Fig. 1, Fig. 2 and Fig. 3. Within the brain APJ mRNA had a very restricted distribution where the PVN and SON hypothalamic regions showed high levels of gene expression (Fig. 1A and B). No labeling of other structures throughout the brain was observed.

4C). Differential loading of the antigen on the resin could result

in different OAg chain conformations and a different exposure of the epitopes and/or determine a different interaction with the corresponding antibodies. The present study describes the successful coupling of OAg from S. Typhimurium to NHS-Sepharose in order to produce an affinity matrix that is capable of purifying specific antibodies from polyclonal human serum. The method can be applied to OAg from different bacteria and it does not modify OAg chain epitopes. Columns were filled with different OAg–ADH preparations with consistent results and columns were used at least 10 times with no deleterious effect on recovery of antibodies. This process could potentially be adapted for the purification of antibodies against other bacterial polysaccharides, as well as for the large scale purification of antibodies against Salmonella OAg. GSK2126458 datasheet As such, the method will be useful for the investigation of the protective capacity of OAg antibodies with different fine specificities in the context of

immunity to NTS, both in HIV-infected African adults and in individuals immunised with OAg-based vaccines. We selleck thank Simona Rondini for the extraction of OAg from S. Typhimurium D23580. We are grateful to Robert Heyderman and the Malawi-Liverpool-Wellcome Trust Clinical Research Programme for S. Typhimurium D23580. We thank Simona Rondini, Yunshan Goh and Adam Cunningham for helpful discussions. This work was supported by a PhD studentship from the Medical Research Council, UK (to CMO’S) and a Clinical Research Fellowship from GlaxoSmithKline (to CAM). The authors Molecular motor declare that they have no conflict of interest. “
“Assessing cytokine profiles in small

tissue biopsies presents a significant technical challenge, particularly the quantification of multiple cytokines when some are present at low concentrations. Multiplex methods using Luminex technology may offer an attractive solution. However these are often developed using soluble materials such as sera or cell culture supernatants spiked with recombinant cytokines and standard protocols are not available for tissue samples. Luminex assays use multiple sets of polystyrene or paramagnetic beads or ‘microspheres’ — see Vignali (2000) and Houser (2012). Each set is fluorescently colour-coded to be identifiable on a dedicated flow cytometer or other platform and pre-coated with antibody to capture a specific cytokine or other analyte, around which a sandwich immunoassay is built. Different bead sets can be combined to enable simultaneous measurement of multiple cytokine concentrations in a single sample against standard curve preparations. These assays require substantially less sample than traditional enzyme-linked immunosorbent assays (ELISAs) – typically 25–50 μL for multiple analytes compared with 200 μL for a single analyte – yet may offer similar sensitivity to Luminex (Vignali, 2000, Biagini et al.