Sukces wprowadzenia szczepionek koniugowanych przeciw meningokokom serogrupy C i czterowalentnych przeciw serogrupom A, C, W-135, Y, pozwala mieć nadzieję, że wprowadzenie szczepionki białkowej, skutecznej również w stosunku do meningokoków serogrupy B, pozwoli znacznie ograniczyć liczbę

zakażeń meningokokowych. Polskie doświadczenia w opanowaniu ognisk epidemicznych, które wystąpiły na terenie woj. opolskiego w roku 2007, wskazują na wysoką skuteczność szczepień interwencyjnych [19]. A. Skoczyńska – zasadniczy wkład w koncepcję i projekt pracy, zebranie, analiza Talazoparib mouse i interpretacja danych, napisanie artykułu. A. Kuch – zasadniczy wkład w koncepcję i projekt pracy, zebranie find more i analiza danych. I. Waśko, A. Gołębiewska, P. Ronkiewicz, M. Markowska, K. Wasiak – zebranie i analiza danych, Waleria Hryniewicz – krytyczne zrecenzowanie artykułu pod kątem istotnej zawartości intelektualnej oraz akceptacja ostatecznej wersji do opublikowania. Badanie zostało częściowo sfinansowane

przez Ministerstwo Zdrowia w ramach programu polityki zdrowotnej pn. „Monitorowanie zakażeń szpitalnych oraz inwazyjnych zakażeń bakteryjnych dla celów epidemiologicznych, terapeutycznych i profilaktycznych na lata 2009–2013” jako Modułu I programu pt. „Narodowy program ochrony antybiotyków w Polsce”, przez Ministerstwo Nauki i Szkolnictwa Wyższego w ramach specjalnego urządzenia badawczego pn. Mikrobank 2 oraz w ramach grantu badawczego firmy GlaxoSmithKline. Pomoc umożliwiająca udział w spotkaniach naukowych oraz honoraria z tytułu wygłoszonych wykładów finansowane przez firmy Pfizer (AS, AK, WH), GlaxoSmithKline (AS, WH), Baxter i Novartis (AS). Pozostali autorzy: nie występuje. next Autorzy dziękują wszystkim Uczestnikom programu BINet oraz wszystkim pozostałym lekarzom i mikrobiologom biorącym udział w monitorowaniu inwazyjnej choroby meningokokowej w Polsce poprzez przekazywanie izolatów wraz z danymi. “
“Antibody production defects

are the most common primary immunodeficiencies. The hallmark of this pathophysiologically, clinically, and genetically heterogeneous group of immunodeficiencies is a defect in mounting the antigen-specific antibody response that is an indispensable condition for the effective adaptive immunity to pathogens. A broad spectrum of diseases represents this group of immune disorders, ranging from often asymptomatic selective IgA deficiency (sIgAD) and IgG subclass deficiencies (IgGsD) to severe agammaglobulinemias in which the production of all immunoglobulin isotypes is severely impaired [1]. The onset of clinical manifestation falls predominatingly on the second half of the first year of life due to the protective effect of transplacentally obtained maternal IgG antibodies over the first 3–6 months.

4E top). Upon activation, secretory lysosomes fuse with the plasma membrane and as a consequence of which, become accessible to FM4-64 (Fig. 4E, bottom). We found that recovery was reduced to 60 ± 1% (YFP-munc13-4) RG7422 in vitro and 59 ± 7% (munc13-4-YFP), which implies that an increased fraction of the munc13-4 molecules can no longer be exchanged from membranes (Fig. 4A, C, D). The t1/2 is increased to 60 ± 2 s (YFP-munc13-4) and 63 ± 5 s (munc13-4-YFP), suggesting tighter binding of munc13-4 with the secretory lysosome membrane. The difference in t1/2 of YFP-munc13-4 and munc13-4-YFP is diminished

after activation (Fig. 4B), which agrees well with changes seen on β-hexosaminidase release. The FHL3 mutant YFP-Δ608-611 did not localize to membranes and recovery resembles that of a cytosolic molecule with free diffusion, resulting in diffusion like kinetics that approach effective diffusion models (Fig. 4A, B) (Sprague et al., 2004). FHL3 is a severe genetic disorder caused by mutations in UNC13D which codes for an essential factor in degranulation of cytotoxic lymphocytes ( Feldmann et al., 2003). A hallmark of the disease is the uncontrolled proliferation of activated lymphocytes and histiocytes

buy SB431542 that invade healthy organs as liver, spleen and brain. FHL3 usually presents before the age of 2 and is fatal unless treated. UNC13D is a large 4.4 kb gene with 32 exons, in which over 50 mutations have been reported ( Cetica et al., 2010). A number of which encode point mutants or truncated forms of munc13-4. Information on large scale phenotype–genotype correlations is not available, and we know little about the properties of the mutants that are expressed. This is in part caused by the difficulty in obtaining sufficient material from the young and oftentimes very sick patients for research. The RBL-2H3 cell line already showed its potential as a model for the molecular analysis of ectopically expressed perforin (FHL2) mutations (Shiver et

al., 1992, Risma et al., 2006, Voskoboinik and Trapani, 2006 and Voskoboinik et al., 2007). We extended its utility and Adenosine triphosphate here developed a simple complementation assay to study the requirements of munc13-4 in degranulation. The method relied on the stable expression of YFP-munc13-4 constructs using lentiviral transduction in combination with siRNA of endogenous munc13-4. We showed here that the assay can be used for the quantitative analysis of FHL3 mutants in secretory lysosome degranulation. Other fundamental aspects of munc13-4 function can also be investigated conveniently by using this model system examples including its role in maturation of secretory lysosomes (Menager et al., 2007) and the function of munc13-4 in complex with its upstream regulator rab27 in the degranulation process. The RBL-2H3 assay also provides incentives for the development of screens with small compound libraries to search for drugs that can overcome degranulation defects caused by expressed FHL3 mutants.

S1). By comparison of the amino acid sequences in the WRKY domain regions from Gossypium and Arabidopsis, 120 cotton WRKY candidate genes were classified

into three groups (groups I, II, and III), and group II genes were further classified into five subgroups (groups IIa–e; Fig. 2), based on the classification rules employed for the WRKY family genes in Arabidopsis [4]. Among the three groups, there were 20 members in group I, 88 in group II, and 12 in group III. Furthermore, in group II, subgroups IIa–e contained 7, 16, 37, 15, and 13 members, respectively. The types and chromosome distribution of these members are described in Table 1. It is noteworthy that WRKY108 in group I contained three WRKY domains (WRKY108N1, WRKY108N2, and WRKY108C). However, the three WRKY GDC-0941 chemical structure domains were not clustered in the N-terminal WRKY domain (NTWD) and the C-terminal WRKY domain (CTWD). The phylogenetic results showed that WRKY108N1, WRKY108N2, and WRKY108C were clustered into group IIc, group III, and group IId, respectively ( Fig. 2). According to D5 genomic sequence information, there was at least one intron insert in the WRKY candidate genes, with WRKY108 and WRKY109 having the most complex structures. The intron splices

in the conserved WRKY domain could be classified into learn more two major types, the R type and the V type. V-type introns were observed only in groups IIa and IIb ( Fig. S2). In addition to the WRKY domain, the WRKY family members were also predicted by MEME to contain other conserved motifs. However, six WRKY proteins,

encoded by WRKY14, WRKY21, WRKY35, WRKY46, WRKY77, and WRKY90, contained only a WRKY domain ( Fig. S3). WRKYGQK residues are considered to be important regions of the WRKY transcription factor family. However, we found some genes with diverse amino acid residues Endonuclease in this region. Among the seven amino acid residues (WRKYGQK), mutations at the W and K sites were not observed; most variations involved Q to T, H, or K substitutions. For WRKY109 in group I, there were large variations in this seven residue regions in both NTWD and CTWD, with variations in three and four amino acid residues, respectively. In total, ten members showed divergence in the WRKY domain, of which seven belonged to group IIc (Table S3). In addition to the variations in amino acid residues in the WRKY DNA binding domain, some mutations were discovered in the zinc finger motif regions. Four members, including WRKY35 and WRKY114 in group I and WRKY108 and WRKY109 in group III, exhibited variations in amino acid residues in this motif (Table S4). By designing gene-specific primers (Table S5), we performed PCR cloning of WRKY genes and amplified the transcripts in given tissues of G. hirsutum acc. TM-1.

We therefore used a recently described method to identify specific intervention features likely to be associated successfully or unsuccessfully with the outcome of interest [31]. Interventions were analyzed based on their success in producing a significant change (p-value ≤ 0.05) in outcomes, in the hypothesized direction [31]. Outcome measures of interest were HbA1c levels, anthropometrics, physical activity, and diet outcomes. Studies that reported at least one of the four outcomes were included in the analysis. SCH772984 datasheet These four outcomes were selected based on what most studies investigated, although instruments measuring these outcomes varied across studies. For instance, anthropometrics

consisted of various measures including body mass index, thigh skinfold, body weight, tricep skinfold, waist-to-hip ratio, total body fat, percent body fat, trunk fat, and fat-free mass. Diet was assessed with a desirable change in any of the following: total kilocalorie intake, dietary risk score, mean vegetable consumption, fruit consumption, consumption of five fruits and vegetables per day, fried food consumption, healthy

eating plan adherence, fat-related selleck products dietary habits, dietary fat intake, dietary cholesterol intake, kilocalories from saturated fat, and percent kilocalories from fat. When a study used several instruments to measure an outcome (e.g., diet), at least 60% (an arbitrary cut-off) of the measures must have reported significant positive Flavopiridol (Alvocidib) results

to be considered a success for that outcome. Only post-test outcome data were used for all analysis. A rate difference determines which intervention feature has a positive or negative association with an outcome [31]. A rate difference was estimated for each intervention feature identified in the review using the following steps. First, a success rate was calculated for both the intervention with and without the feature. The success rate for the intervention feature (SRWF) is the number of studies reporting on the intervention having the feature of interest associated with a positive participant outcome, divided by all the studies reporting on intervention with the feature regardless of outcome; the specific formula used was: number of studies with feature with positive outcome/all studies with feature. Second, a success rate without a feature (SRWoF) is the number of studies reporting on the intervention without the feature of interest with a positive participant outcome, divided by all the studies without the feature regardless of outcome; the formula was: number of studies without feature with positive outcome/all studies without the feature. Third, rate differences were calculated for each intervention feature, by subtracting the success rate with feature (SRWF) from the success rate without the feature (SRWoF).

These mechanisms included changes in whole tissue buy PTC124 strain, hydrostatic pressure, and streaming potentials generated by bone fluid flow through a charged bone matrix. Streaming potentials were initially thought

to be generated by electrokinetic effects associated with a system of connected micropores associated with the collagen-apatite porosity [25]. Subsequently, Cowin et al. [26] and Zhang et al. [27] proposed that the pores were actually the canaliculi in the mineralized bone and these channels were the site of the strain generated potentials. Those electrokinetic effects might modulate the movement of ions such as calcium across the cell membrane [28] and [29].

Load that is rapidly placed on bone first pressurizes the interstitial fluid around the osteocytes, before the fluid is driven to flow. Zhang et al. [27] estimated that the fluid component could carry as much as 12% of the applied mechanical load and produce peak pressures of 2–3 MPa. More recently, Gardinier et al. [30] have predicted that the magnitude of the pressure experienced by osteocytes in vivo could reach up to Histone Methyltransferase inhibitor 5 MPa. Klein-Nulend et al. [5] subjected osteocytes, osteoblasts, and periosteal fibroblasts from chicken calvarial bone to two different mechanical stimuli, i.e. hydrostatic

compression (IHC) and pulsatile fluid flow (PFF). Osteocytes were particularly sensitive to fluid shear stress, more so than to hydrostatic stress, although one Urease can argue that the hydrostatic pressure applied, i.e. 13 kPa, is much lower than the 5 MPa predicted to occur in vivo [30]. More recent research has shown that cyclic hydraulic pressures of 68 kPa can modulate signaling molecule production in cells of the mouse MLO-Y4 osteocyte cell line [31]. Over the past decade a number of theoretical and experimental studies have appeared that have put forth evidence strongly favoring interstitial fluid flow and direct cell strain as opposed to streaming potentials or hydrostatic pressure as the most likely mechanism for mechanosensation. Osteocytes form a ‘network’ throughout the bone matrix by connecting with each other and with surrounding lining cells on the bone surface. These anatomical characteristics of osteocytes make them ideally placed in bone to sense external mechanical loads imparted on bone. Osteocytes are directly connected with each other via gap junction-coupled long slender cell processes which run along the central axis of the canaliculi except where there are ridges created by transverse collagen fibrils.

We compared data taken at 1 and 10 m perpendicular to shoreline using a linear regression analysis to determine coherence with distance. We tested for differences in the concentration of PAHs

in wetland soils categorized in the SCAT surveys using a one-way ANOVA, and tested for differences between oil and un-oiled sites using a Student’s t-test and a Tukey’s HSD post hoc test for significant differences, with an alpha = 0.05. We created box and whisker plots (minimum to maximum; 25th to 75th percentile) of the concentration of alkanes and aromatics for the three estuaries (Breton Sound, Barataria Bay and Terrebonne Bay) selleck products that were sampled before the oil reached the marsh in May 2010. We divided Barataria Bay into east and west components using Grand Isle as the border and compared the concentrations of alkanes and aromatics in September 2010. We then used a Kruskal–Wallis non-parametric analysis to test for differences in the concentration of total alkanes and total aromatics among estuaries for all data in May 2010 and September 2010, and amongst sampling at the four Bay Batiste sampling trips to the same 30 sites. The total target alkane and PAH concentrations in the 405 samples ranged

from 0.4 to 8,640 mg kg−1, and from below detection limits (0.1 μg kg−1) to 355,744 μg kg−1, respectively. Samples with the lowest concentrations were collected during the pre-impact sampling in Enzalutamide in vitro May 2010, when the concentration of target alkanes and PAHs averaged 0.98 ± 0.005 mg kg−1 and 23.9 ± 1.61 μg kg−1, respectively. Some samples from May 2010 had measurable traces of petroleum in them, but no identifiable MC252 oil. We consider these May 2010 data to

be a baseline against which we compared oiling amounts from after the MC252 spill in 2010 and subsequent re-distributions. MC252 oil was detected in 34 of the 94 samples collected in September 2010 and February 2011. The average concentration of target alkanes and PAHs in these 34 samples was 991 ± 377 mg kg−1 and 29,977 ± 11,410 μg kg−1, respectively (Table 3). The average target alkane and PAH concentrations Dapagliflozin in the MC252 oiled wetlands was, therefore, over 1,015 and 1,255 times, respectively, the concentration of these alkanes and aromatics in the relatively un-oiled wetland sediments sampled in May 2010. All samples contained numerous alkanes, with some samples having obvious odd carbon preferences and others not. Samples with significant oiling contained normal alkanes with the typical pattern of alkanes, as well as the isoprenoid alkanes pristane and phytane seen in crude oils. Except for samples with highly elevated amounts of oil, many alkane patterns had biogenic and petrogenic source signatures. In general, the samples with low levels of alkanes exhibited a pattern associated with the various biogenic sources, with only some having odd carbon preferences. The average concentration of target alkanes within 1 m of the water’s edge for 91 paired samples was 37.3 ± 26.

Constatou-se evolução clínica favorável, com remissão espontânea da hemorragia digestiva e sem recorrência das perdas hemáticas. Com o intuito de identificar uma etiologia subjacente à amiloidose realizou estudo

complementar. Efetuou medulograma que revelou a presença de 20% de plasmócitos de origem monoclonal, compatível com o diagnóstico de mieloma múltiplo, confirmado posteriormente pela imunofenotipagem medular. Diminuição das imunoglobulinas séricas, nomeadamente G 2,5 g/L (7,0-15,0), A < 0,24 g/L (0,6-4,0), M < 0,16 g/L (0,6-3,0). Cadeias GSK-3 inhibitor leves livres no soro Kappa 0,18 g/L (0,33-1,90), Lambda 0,62 g/L (0,57-2,63). Eletroforese das proteínas séricas sem alterações e urinárias com vestígios de proteinúria tipo tubular. Imunofixação sérica com acentuada hipogamaglobulinémia. Sem alterações na imunofixação urinária. Cadeias leves livres na urina Kappa 2,6 mg/dL (0,135-2,42) e Lambda 0,8 mg/dL (0,024-0,666).

Clearance da creatinina 46,3 ml/min e proteinúria das 24 horas de 103 mg (42,0-255,0). Realizou ressonância magnética à coluna que revelou vários focos hipointensos sugestivos de infiltração mielomatosa a nível cervical, torácico e lombar. Além das alterações referidas, identificaram-se alterações degenerativas da coluna com unco-discartroses e protusões disco-osteofitárias, motivando Akt inhibitor ligeira compressão da medula a nível cervical. A TAC do tórax, abdómen e pélvis não mostrou alterações de relevo. O ecocardiograma transtorácico identificou hipertrofia

moderada do septo basal anterior e acentuada do septo interventricular, com ligeiro aumento da refringência e padrão de disfunção diastólica do tipo pseudonormal. Pelo diagnóstico de mieloma múltiplo, não secretor, sintomático, iniciou quimioterapia com melfalan e prednisolona. Foi orientado para reabilitação e pelo elevado risco de fraturas ósseas colocado colete dorso-lombostato. A reavaliação por colonoscopia esquerda, realizada 4 meses depois de diagnóstico de amiloidose gastrointestinal, e ainda durante o tratamento Ureohydrolase com quimioterapia, revelou a nível do sigmoide a persistência das lesões descritas previamente. O doente não apresentou recidiva da hemorragia digestiva. Contudo, registaram-se 2 internamentos posteriores por intercorrências infeciosas, nomeadamente infeções respiratórias. A amiloidose não é uma doença única, mas sim um grupo de doenças que partilham a característica comum de depósito de proteínas na matriz extracelular1. Pode ser adquirida ou hereditária, sistémica ou localizada a um único órgão, como o trato gastrointestinal3 and 6. A verdadeira incidência da amiloidose é desconhecida pois apenas os doentes sintomáticos são investigados8. A nomenclatura atual da doença consiste na primeira letra – A (de amiloide) – seguida pela descrição da natureza da proteína precursora que forma os respetivos depósitos1 and 2. Existem 6 tipos diferentes de amiloidose. A AL, com deposição de cadeias leves, é a forma mais comum.

Moreover, data suggest that BIL fails to induce apoptosis in cultured human nontransformed cells. These results suggest that BlL has a promising potential for application in the therapy and/or diagnosis of cancer. Future studies are needed to elucidate the details of BlL induced-apoptosis mechanism in several tumor cell lines. The authors declare that there are no conflicts of interest. The authors express their gratitude to the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for research grants and fellowship (LCBBC and MTSC) and to the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

(CAPES) and Fundação de www.selleckchem.com/erk.html Amaparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE) for research grants. Authors are deeply grateful to Maria Barbosa Reis da Silva, Maria D. Rodrigues and João Antônio Virgínio for their technical assistance. “
“Loxoscelism is a set of signs and symptoms caused by the bite of spiders of the genus Loxosceles ( Da Silva et al., 2004). Loxosceles (Araneae, Sicariidae) can be found in temperate and tropical regions of America, Oceania, Asia, Africa and Europe ( Swanson and Vetter, 2006, Hogan et al., Enzalutamide molecular weight 2004 and Souza et al., 2008). This genus represents a public health problem in Brazil, mainly in South and Southeast regions, with more than 3000 cases reported annually by

the Ministry of Health ( Hogan et al., 2004). Usually, the clinical manifestations of loxoscelism are characterized by necroulcerative dermatitis Nintedanib (BIBF 1120) at the site of the bite (83.3% of the cases). However the envenoming can also cause systemic effects (16% of the victims) leading to acute renal failure, which may be lethal ( Málaque et al., 2002, Hogan et al., 2004 and Abdulkader et al., 2008). Locally, lesions caused by Loxosceles venom present edema, hemorrhage, inflammation with predominance of neutrophils, rhabdomyolisis, damage to the vessels wall, thrombosis, and dermonecrosis ( Futrell, 1992, Ospedal et al., 2002 and Pereira et al., 2010). In addition, according to some studies, Loxosceles venom causes cytoplasmic vacuolization, loss of adhesion ( Hogan et al., 2004, Veiga et al., 2000 and Veiga et al.,

2001) and apoptosis of endothelial cells ( Pereira et al., 2010). The family of Loxtox proteins ( Kalapothakis et al., 2007), such as: sphingomyelinase-d, SMA protein, phospholipase-d dermonecrotic protein (DP) and dermonecrotic factors (DNF) were found and characterized in the venom of Loxosceles and were associated with local and systemic loxoscelism ( Barbaro et al., 2005, Felicori et al., 2006 and Da Silveira et al., 2007). The systemic and local effects of the venom are well described in human, rabbit, and guinea pig cutaneous tissue. The use of the murine model in loxoscelism study is restrained to inflammatory events analysis, since the dermonecrotic lesion does not develop in mouse following intradermal injection of the venom ( Sunderkötter et al., 2001 and Barbaro et al., 2010).

Furthermore it is necessary to identify all novel gene Selleck GSI-IX isoforms from PSCs. Based on the SGS short reads (75 bp), ENCODE

project predicted novel transcripts from 15 cell lines, including hESCs (H1 cell line) [3••]. Although 73,325 transcripts from 31,204 genes in intergenic and antisense regions were reported, the detailed description of novel transcripts from hESCs is lacking. More importantly, the validation rate by overlapping targets 454 Life Sciences (Roche) of these novel transcripts (from 15 cell lines) were only 70–90%. In 2013, two research groups sequenced (by SGS) single-cell human embryo transcriptomes from oocytes to late blastocyst [7• and 8•]. With the SGS prediction tools (Cufflinks, Trinity and PASA), Yan et al. predicted 7420 novel transcripts from 3866 potential transcription units, including 253 possible protein-coding genes and 7167 possible novel long non-coding RNAs (lncRNAs) [ 1, 2 and 36]. However, the accuracy of transcript prediction by SGS was not

reported. Moreover, Yan et al. imposed a strong arbitrary constraint on novel transcript unit definition: >10 kb apart from two transcripts, which narrowed the novel transcript identification. Au et al. filled the gap of reliable novel transcript identification by using long reads of TGS. In Au and colleagues’ experiments, multiple long reads covered the full lengths of novel or annotated gene isoforms or their significant fragments, which resulted in a very reliable direct detection or prediction Selleckchem Galunisertib under certain FPR control (<5%). 2103 novel transcripts were identified which were not annotated by RefSeq, Ensembl, UCSC Genes or Gencode. Au et al. also predicted 111 lncRNAs from these novel transcripts by very high stringency modes for two ncRNA prediction methods (P ≥ 0.9 for RNAz; MFE ≤ −15 and Z score ≤ −4 for alifoldz), 50 of which SDHB have specific expression in hESCs. These novel lncRNAs are much longer and contain more junctions than the annotated lncRNAs predicted from SGS, such as Gencode library. Among the novel lncRNAs identified in Au et al., only 4 of the Gencode-annotated

lncRNAs are longer than 2000 bp, while 72 other novel lncRNAs (65%) are longer than 2000 bp with the averaged lengths around 2300 bp; only 6 of Gencode-annotated lncRNAs contains more than 5 exons, while 78 novel lncRNAs (70%) contain more than 5 exons. All together, the study conducted by Au and colleagues, in combination with other studies of RNA-Seq and sequencing of the human genome, resulted in the identification of novel genes and provided a complete exon structure complexity. This is particularly important for investigating the functional role of the unique human transcriptome, including lincRNAs/lncRNAs, and regulative secondary structures in maintaining pluripotency. Overall, a comprehensive profiling of hPSC transcriptome is critical for addressing their pluripotency.

Our research concerns the characterisation of immune responses to the pathogen Helicobacter pylori (Hp) which are linked to peptic ulceration and gastric cancer development ( Atherton, 2006 and Robinson et al., 2008). The challenges are broadly similar in other fields, particularly for gastrointestinal mucosal researchers: how to study immune responses using methodology that better reflects cytokine levels in the mucosa in vivo. Endoscopic mucosal biopsies are small (typically around 5–10 mg) and concentrations of many of the cytokines of interest are low,

so assay Venetoclax in vivo sensitivity and sample volume requirements are critical. Other investigators have used semi-quantitative methods including immunohistochemistry ( Lindholm et al., 1998, Lehmann et al., 2002 and Holck et al., 2003) and western blotting ( Luzza et al., 2000 and Tomita et al., 2001), or PCR-based methods to quantify cytokine mRNA which are not always fully reflected at the protein level ( Luzza et ATM/ATR inhibitor drugs al., 2001, Robinson et al., 2008 and Serelli-Lee et al., 2012). Cytokines have been measured in gastric biopsy homogenates using ELISA ( Yamaoka et al., 2001, Shimizu et al., 2004, Caruso et al., 2008, Robinson et al., 2008 and Serelli-Lee et al., 2012), but additional volume is needed for each analyte assayed

which may require sample dilution. Therefore the number of cytokines, particularly those present at low concentrations, that can be assayed using this method is limited. Another common approach is to culture gastric biopsies in vitro, with or without stimulation, and measure cytokine concentrations in culture supernatants ( Crabtree et al., Unoprostone 1991, Bodger et al., 1997 and Mizuno et al., 2005). However, these methods may alter the cytokine profile ( Veldhoen et al., 2009). The cytokine concentrations in homogenates of gastric biopsies should more closely reflect those found in the gastric mucosa in vivo. Luminex-based methods have been used to assess murine immune responses to Hp infection ( Taylor et al., 2008) and vaccination ( Taylor et al., 2007) in splenocyte culture supernatant and recently to quantify gastric

cytokine concentrations in Hp-infected mice ( Schumacher et al., 2012). A method to measure Hp-specific IgG in human saliva samples has also been developed, using Luminex beads conjugated with antigens including Hp whole cell sonicate and recombinant urease ( Griffin et al., 2011). However, to our knowledge, Luminex assays have not been optimised for human gastrointestinal mucosal tissue samples, though were recently used to quantify interleukin-1β, interleukin-1 receptor antagonist, interleukin-6 and tumour necrosis factor-α in gastric tissue samples ( Serelli-Lee et al., 2012). Careful kit selection and optimisation of tissue sample preparation in a limited volume of extraction buffer will theoretically facilitate cytokine detection in these samples.