Thus, it would be unreasonable to expect a stronger effect (in other words, after onset of diabetes) in humans. Secondly, no preclinical study ever tested the clinical GAD-Alum preparation, and no efficacy was noted in our recent studies in NOD and B6 diabetes models (Pagni, Boettler and von Herrath, unpublished). mTOR inhibitor Again, it is probably unreasonable to expect an antigenic formulation to work in humans when it does

not even prevent diabetes in otherwise permissive animal models. Several other theories have been proposed to account for the failure of GAD-Alum in humans, including the lack of GAD expression in β cells; this is a controversial area, as many studies have demonstrated expression of GAD-65 and 67 proteins in murine and human β cells [36]. Lastly, one could ask whether the dose of GAD-Alum was sufficient – as most patients mounted a clearly detectable immune response, this appears less likely. However, alum might have been a suboptimal adjuvant for an ASI, as the resulting mixed but T helper type 2 (Th2)-dominated cytokine response of induced GAD-reactive T cells (Arif, Sotrastaurin in vitro Roep and Peakman, unpublished) did not result in protective cell populations. In the absence of a functional mouse model of GAD-Alum preventing diabetes, it will be difficult at this point to clarify these issues. The question of the antigenic dose might have more bearing on the

issue of efficacy with oral insulin [15]. As predicted from animal models [37], prophylactic oral not insulin given at a daily dose of 7·5 mg had a very marginal effect in preventing diabetes in individuals at high risk (exhibiting multiple autoantibodies [38-41]), but not in any other patient groups. However, as has been evident from multiple studies in different mouse models, oral insulin dosages have to be comparatively

much higher to induce optimal disease preventive effects, which are seen at a dose of 1 mg given twice per week [42]. This dose would equate to approximately 1 g of oral insulin twice per week in humans. In addition, it is likely to be necessary to provide the drug in enteric-coated capsules, without which > 99·99% of the insulin is lost through digestion in the stomach and only minimal amounts of intact antigen or some peptides will reach the lower gut and the Peyer’s patches, the location at which oral insulin has been shown to induce its desired immune-regulatory response. Therefore, more precise dose calculations should have probably preceded the oral insulin trial and its current follow-up study. A further human/mouse mismatch relates to the overall management of expectations when devising trials for ASI. In rodent studies most, if not all, ASI is effective only for early and, at best, late prevention of disease, but never after onset of hyperglycaemia. Thus, we should not expect antigens to reverse human diabetes or even preserve C-peptide after onset (at least with effects detectable in reasonably sized studies); and this has indeed been the case.

The mice received a four-collagen–antibodies cocktail intravenously (i.v.) 10 days later (20 days after surgery, day 0). One week later, they received an intraperitoneal (i.p.) injection of LPS to enhance arthritis incidence and severity, and the experiment was terminated on day 14. Control

mice were injected with phosphate-buffered saline (PBS) i.v. and LPS i.p. Male transgenic ERE-luciferase mice were castrated and 11 days later immunized with chicken CII and adjuvant. After 9 days they received one subcutaneous injection of raloxifene, oestradiol or vehicle, and were then terminated 10 h later (day 10 after immunization). Mice were given subcutaneous injections 5 days per week of the raloxifene analogue LY117018 (generous gift from Eli Lilly, Indianapolis, selleck compound IN, USA) (60 µg/mouse/day) or 17β-oestradiol-3-benzoate (E2) (Sigma, St Louis, MO, USA) (1·0 µg/mouse/day)

dissolved in Miglyol812 (OmyaPeralta GmbH, Hamburg, Germany). Control mice received Miglyol812 (100 µl/mouse/day). The dosages of Ral and E2 have been shown previously to prevent osteoporosis equally well in mice [19–21]. LY117018 differs from raloxifene at only one site on the molecule, with a pyrrolidine ring on the basic side chain instead of a piperidine ring. This small difference does not affect its biological properties. Thus, Cabozantinib nmr Ral and LY117018 can be regarded as replaceable with respect to their biological properties. Experiment 1.  Two weeks after ovariectomy DBA/1 mice were immunized with 100 µg of chicken CII (Sigma, St Louis, MO, USA) dissolved in 0·1 m acetic acid and emulsified with an equal volume of incomplete Freund’s adjuvant (Sigma) supplemented with 0·5 mg/ml Mycobacterium tuberculosis (Sigma). A total volume of 100 µl was injected intradermally at the base of the tail. After 21 days, mice received enough a booster injection with CII emulsified in incomplete Freund’s adjuvant. Arthritis developed shortly thereafter, and was evaluated continuously for frequency and

severity. Experiment 2.  Twenty days after OVX or sham-operation, DBA/1 mice received an intravenous shot of a four-antibody cocktail [monoclonal immunoglobulin (Ig)G antibodies specific for the C1, J1, D3 and U1 epitopes on the collagen type II molecule], according to the protocol of Nandakumar and Holmdahl [10]. Non-arthritic controls received equal volumes of PBS. One week later, all mice received an intraperitoneal injection of 25 µg LPS (Escherichia coli 055 : B5; Difco Laboratories, Detroit, MI, USA). Experiment 3.  ERE-luciferase mice were immunized with 100 µg of chicken CII (Sigma) dissolved in 0·1 m acetic acid and emulsified with an equal volume of incomplete Freund’s adjuvant (Sigma) supplemented with 0·5 mg/ml M. tuberculosis (Sigma). A total volume of 100 µl was injected intradermally at the base of the tail.

While the objectives of the review by Strippoli et al.17 Tamoxifen solubility dmso were to evaluate the benefits and harms of ACEi and ARBs in preventing the progression of CKD. Both reviews included studies of both type 1 and type 2 diabetes and Strippoli et al.17 people with either microalbuminuria or macroalbuminuria. While the reviews included both type 1 and type 2 diabetes the majority of selected trials enrolled only people with type 2 diabetes. The overall conclusions of the two systematic reviews are summarized below: A significant reduction in the risk of developing microalbuminuria

in normoalbuminuric patients has been demonstrated for ACEi only. This effect appears to be independent of BP and, kidney BGB324 datasheet function and type of diabetes. However, there is insufficient data

to be confident that these factors are not important effects modifiers.16 In relation to type 2 diabetes the following outcomes are of note:16,17 All-cause mortality The relevant trials comparing ACEi treatment with ARB treatment all included people with type 2 diabetes and no significant differences on all cause mortality, progression of microalbuminuria to macroalbuminuria or regression from microalbuminuria to normoalbuminuria were noted.17 However, as noted in the overall conclusion by the authors the trials were limited and provide insufficient evidence for comparison of effects. The objectives of the systematic review was to assess the RCT evidence for the effects of different therapeutic BP goals and interventions in the normotensive range on the decline of glomerular function.64 The search strategy was limited to studies of people with

2 years duration of type 1 or type 2 diabetes with incipient or overt nephropathy with or without elevated BP. The intervention was required to be treatment with one or more hypertensive agents. The review identified 5 RCTs meeting the search criteria. All of these studies have been identified and assessed.4,16,17 Only two studies that considered www.selleck.co.jp/products/cobimetinib-gdc-0973-rg7420.html the effect of BP targets within the normotensive range in people with type 2 diabetes were identified.70,73 Kaiser et al.64 considered GFR as surrogate endpoint in the absence of a renal failure endpoint such as need for dialysis and/or transplantation. The authors noted that no trial demonstrated any beneficial effect of lower target BP values on the progression of kidney failure. In short decreases in albuminuria were not accompanied by a decrease in the rate of decline in GFR. They conclude that the available evidence does not support a beneficial effect of BP lowering within the normotensive range on progression of diabetic nephropathy as assessed by the change in GFR. The systematic review and meta analysis pooled analyses from the number of small studies comparing combination treatment of ACEi + ARB with ACEi alone.77 A total of ten studies covering both type 1 and type 2 diabetes were included in the meta-analysis.

Reaction products were diluted five times upon addition of 10 mmol l−1 Tris-HCl (pH 8.3) buffer. Adapters were produced by mixing equimolar amounts of complementary oligonucleotides (Eurogentec, Seraing, Belgium): (5′-CTCGTAGACTGCGTACC-3′ and 5′-CGGGTACGCAGTC-3′ for HpyCH4IV; 5′-GACGATGAGTCCTGAC-3′ and 5′-TAGTCAGGACTCAT-3′ for MseI) and heating the mixture to 95 °C followed by slow cooling to ambient temperature. One microlitre of the diluted restriction-ligation mixture was used for amplification in a reaction volume of 25 μl containing

1 μmol l−1 Alvelestat solubility dmso M HpyCH4IV primer with one selective residue (underlined) (5′-Flu-GTAGACTGCGTACCCGTT-3′), 1 μmol l−1 MseI primer with four selective residues (underlined) (5′-GATGAGTCCTGACTAATGAA-3′), 0.2 mmol l−1 of each deoxynucleoside triphosphate, PF-02341066 concentration and 1 U of Taq DNA polymerase (Roche Diagnostics) in 1× reaction buffer containing 1.5 mmol l−1 MgCl2. Amplification was performed as follows. After an initial denaturation step at 94 °C for 4 min in the first 20 cycles, a touchdown procedure was applied: 15 s denaturation at 94 °C; 15 s annealing at 66 °C with the temperature for each successive cycle lowered by 0.5 °C and 1 min

of extension at 72 °C. Cycling was then continued for further 30 cycles with an annealing temperature of 56 °C. After completion of the cycles, incubation at 72 °C for 10 min was included before the reactions were cooled to room temperature. Products were diluted 1 : 10 with distilled water. To 1 μl of diluted product, 0.25 μl of ET400-R size marker (GE Healthcare, Osimertinib clinical trial Diegem, Belgium) and 8.75 μl of distilled water were added. Following 1-min denaturation step at 94 °C, the samples were

quickly cooled to room temperature and analysed on a MegaBACE 500 automated DNA analysis platform equipped with a 48-capillary array according to the instructions of the manufacturer (GE Healthcare). AFLP data were imported into BioNumerics v. 6.0 Software (Applied Maths, Sint-Martens-Latem, Belgium) and analysed by UPGMA clustering using the Pearson correlation coefficient. The analysis was restricted to DNA fragments in the range from 60 to 300 bp. Clinical isolates were identified to the species level based on the similarity of their AFLP profile to those of the type strains. To assign specific AFLP genotypes, the obtained profiles were also inspected visually and scored for presence/absence of clearly recognisable DNA fragments. Two profiles differing by at least one clearly recognisable DNA band were considered to represent different genotypes.

Our results revealed that CML-specific CTL crucially contribute to disease control and are characterized by high IL-7Rα expression. Interestingly, CML cells produced IL-7 that was crucial for the

maintenance of specific CTL. Therefore, CML maintains Vismodegib mouse leukemia-specific CD8+ T-cell-mediated immunosurveillance by IL-7 signaling. Bone marrow was cotransduced with retroviral particles encoding for BCR/ABL and NUP98/HOXA9 and injected into irradiated syngeneic recipient mice. As shown previously, coexpression of BCR/ABL and NUP98/HOXA9 led to the development of CML and progression to blast crisis within several weeks 17. Granulocyte counts rose up to 9×107/mL (C57BL/6 control mice:<2×106 granulocytes/mL blood). Phenotypically, the leukemic cells consist of a population MAPK Inhibitor Library nmr of immature myeloid blasts (MAC-1+,

GR-1+ and c-kit+) of up to 10% and a majority of mature granulocytes 17. This cotransduction was chosen to model the transition from chronic phase to blast crisis. To study antigen-specific immune responses, H8 transgenic mice were chosen as bone marrow donors. In this experimental setup, all leukemia cells expressed the immunodominant CTL epitope gp33 of lymphocytic choriomeningitis virus (LCMV) on MHC class I molecules as a model leukemia antigen (H8-CML mice). To analyze the impact of CD8+ T cells on disease progression, H8-CML mice with high granulocyte counts (>5×107 granulocytes/mL blood) were depleted of CD8+ Progesterone T cells by monoclonal antibody or were left untreated. Depletion of CD8+ T cells

led to disease progression and death of 83% of the animals within 4–19 days (Fig. 1A). CML progression was significantly slower in untreated control mice and 50% of the mice survived up to 75 days. On the contrary, treatment with IgG from rat serum (as control for αCD8 antibody YTS169.4) did not prolong survival when compared with untreated mice (Supporting Information Fig. 1). These results suggest that CD8+ T cells are crucially involved in the control of CML disease progression. The fact that a minority of the CD8+ T-cell-depleted animals still could control CML suggests that other effector mechanisms may contribute to the immunosurveillance of CML. In agreement with our earlier results, in CML mice no gp33-specific CTL response was detectable in the blood by tetramer staining 17. Naïve C57BL/6 mice and LCMV-immune mice which had been infected 8 wk previously with 200 pfu LCMV-WE were used as controls (Fig. 1B and D). The absence of specific CD8+ T cells in blood of CML mice by tetramer staining was in contrast to the rapid leukemia progression in CD8+ T-cell-depleted animals. Using a dextramer enrichment approach, we could detect gp33-specific CTL in pooled spleens and lymph nodes of H8-CML animals above the background of naïve C57BL/6 mice (Fig. 1C and D and Supporting Information Fig. 2A and B) 18.

The result shows that the expression of relevant cytokines decreased after deactivation. In addition, the expression of IL-12p40 and IL-6 was higher in GM-BMMs from Klf10-deficient mice than that from WT mice after deactivation (Fig. 4B). Moreover, the downregulation of Klf10 was abolished to some

extent (Fig. 4C), which may enhance its inhibitive function on the cytokines. These data may indicate that Klf10 alone is insufficient to inhibit the inflammatory factors in GM-BMMs; other factors are possibly involved in the suppression of inflammation in deactivated GM-BMMs. Klf11, another member of the KLF family, was also identified as a downregulated selleck screening library gene in LPS-stimulated M-BMMs. Klf11 is described as a TGF-β inducible early gene 2 and shares a highly conserved C-terminal DNA-binding domain with Klf10 [18]. In addition, Klf10 and Klf11 GPCR Compound Library ic50 have three repression domains in the

N-terminal, which define them as a subfamily of repressor. We supposed that Klf11 may have a similar role in the regulation of inflammatory factors in M-BMMs. First, we found that overexpression of both Klf10 and Klf11 can inhibit the production of IL-12p40 and IL-6 in M-BMMs from WT mice and can rescue their overproduction in M-BMMs from Klf10-deficient mice (Fig. 5A). Therefore, we knocked down Klf11 through RNA-mediated interference. The efficiency of RNAi was confirmed by qPCR (Supporting Information Fig. 6A). The inhibition of Klf11 resulted in an increased production of IL-12p40 in WT cells, and this phenomenon was more notable in Klf10-deficient cells (Fig. 5B). Therefore, Klf10 Selleck C225 and Klf11 may have a similar function in the regulation of IL-12p40. However, interference of Klf11 in the same conditions did not result in a significant change of IL-6 as that in the overexpression assay. Moreover, we

used another SiRNA, as previously reported [42], to confirm the inhibitory function of Klf11 in the regulation of IL-12p40 (Supporting Information Fig. 6B and C). These data indicated that Klf11 can act similarly to Klf10 in the inhibition of IL-12p40 production. The KLF family members are characterized by a DNA-binding domain capable of binding to target genes to regulate their transcriptional activities and gene expressions. IL-12p40 promoter was sequenced to determine whether Klf10 can regulate the expression of IL-12p40 in a direct manner and a CACCC site was found therein (Fig. 6A). The binding site was highly conserved in mammals (Fig. 6B), similar to the CACCC-binding site of erythroid Krüppel-like factor in human macrophages. Subsequently, a series of luciferase reporter construct that can encode a WT IL-12p40 promoter (−283 to +99 bp), a mutant with 2-bp mutations in the CACCC site (at –233 bp), or a 5′ deletion promoter construct (−223 bp) were constructed to investigate whether Klf10 can repress the transcription of IL-12p40.

None. Table S1. Differentially expressed gene-sets [from gene-set enrichment analysis (GSEA)] in the distal colon of appendicitis–appendectomy (AA) mice compared to the distal colon of

sham–sham (SS) mice. “
“Citation Ozornek H, Ergin E, Jeyendran RS, Ozay AT, Pillai D, Coulam C. Is Apolipoprotien E codon 112 polymorphisms associated with recurrent pregnancy loss? Am J Reprod Immunol 2010; 64: 87–92 Problem  To compare the prevalence of 112T>C point mutations among women experiencing RPL with fertile control women. Method of Study  Buccal swabs were obtained from 232 individuals: 136 with a history of ≥2 abortions, 37 with at least 2 live Gemcitabine births and 59 with a history of deep vein thrombosis (DVT). DNA was extracted and PCR amplification

of Apo E codons was performed. Results  The allelic frequency of a cytosine at position 112 was 11.4% (31/272) among patients experiencing RPL, compared with a frequency of 5.4% (4/74) among the fertile controls (P = 0.19) and 19.5% (23/118) among individuals with a history of DVT. However, significantly more E3/E4 and E4/E4 genotypes were seen among individuals experiencing RPL and DVT than fertile controls (P < 0.05). Conclusion  Apo E4 codon 112C point mutation is, by itself, not associated with an elevated risk of recurrent pregnancy loss, but rather codon 112C in association with codon 158C is a risk see more factor for RPL. “
“Type I diabetes is a disease caused by autoimmune destruction of the beta cells in the pancreas that leads to a deficiency in insulin production. The aim of this study was to evaluate the prophylactic potential of a prime-boost strategy involving bacille Calmette–Guérin (BCG) and the pVAXhsp65 vaccine (BCG/DNAhsp65) in diabetes induced by streptozotocin (STZ) in C57BL/6 mice and also in spontaneous

type 1 diabetes in non-obese diabetic (NOD) mice. BCG/DNAhsp65 vaccination in NOD mice determined weight gain, protection against hyperglycaemia, decreased islet inflammation, higher levels of cytokine production by the spleen and a reduced number of regulatory T cells in the spleen compared with non-immunized NOD mice. In the STZ model, however, there was no significant difference in the clinical parameters. for Although this vaccination strategy did not protect mice in the STZ model, it was very effective in NOD mice. This is the first report demonstrating that a prime-boost strategy could be explored as an immunomodulatory procedure in autoimmune diseases. Type 1 diabetes (T1D) is an autoimmune disease characterized by T cell-mediated destruction of the β cells in pancreatic islets. It affects the insulin production and leads to hyperglycaemia, polyuria and hypoinsulinaemia [1]. As a chronic condition, it may cause blindness, cardiovascular injury and harm in other systems at later stages [2].

It is important that only studies matching the inclusion criteria are included in the systematic review, so that the systematic review answers a specific clinical question. Prospective criteria for study inclusion and exclusion should be explicitly ABT888 stated in the review to minimize selectivity by authors. These criteria are a requirement before commencing Cochrane reviews, when a study protocol is developed, peer reviewed and published before initiating the review. The decision regarding which studies to include in a systematic review may have an important effect on a conclusion, say regarding the overall utility

of a healthcare intervention.13 Therefore, study inclusion assessment should be completed independently by at least two authors and generally is arbitrated by a third. Readers of systematic reviews can look for a flow chart (usually presented as a Fig. 1) describing the details of studies identified, studies excluded, reasons for exclusion and numbers of studies included in the final review. If the outcome of interest is dichotomous (the outcome

is one of two possibilities – example, death or survival) the treatment effect is calculated for each trial as a risk ratio, an odds ratio or a risk difference together with the 95% confidence interval (95% CI; the range Z-IETD-FMK molecular weight within which we are 95% confident that the effect calculated is likely to exist). While full discussion of all methods Tenoxicam is beyond the scope of this review, dichotomous outcomes are frequently evaluated as a relative risk (RR), which deserves a brief explanation. A RR divides the event rate in the intervention group (number of events divided by the total number of individuals randomized in that group) by the event rate in the comparison group. For example, if 20 of 100 patients in the active intervention group who are randomized to

erythropoietin to normalize haemoglobin levels experienced an event and 10 of 100 patients in the control group (those randomized to a lower haemoglobin target), experienced the event, then the RR is 2 (20/100 divided by 10/100), indicating that the intervention is twice more likely than the comparison treatment to result in the outcome. Interpretation of this risk for the specific patient is possible when the actual risk of the outcome for that patient without treatment is known (e.g. when RR = 2, a doubling of risk from 2% to 4% is quite different from the doubling of risk from 10% to 20% in the present example). If the outcome of interest is a continuous variable (an example is systolic blood pressure, mmHg), then the effect size of the intervention is summarized as a mean difference (MD; and its 95% CI). The MD for the outcome in each trial is the amount by which an intervention changes the outcome on average compared with the control.

In addition to standard microbiological culture, we examined explanted suture and tissue specimens using CM to determine whether bacterial selleck compound biofilms were present. Specimens were prepared as described previously (Kathju et al., 2009a, b). Briefly, suture

and tissue recovered at surgery were placed in Hanks balanced salt solution (HBSS) and placed on wet ice, directly after removal. After rinsing in the HBSS (to remove unattached bacteria) and blotting on sterile paper, specimens were mounted on the bottom of a 35-mm Petri plate on partially solidified agar (Kathju et al., 2009a, b). Specimens were stained for viability assessment using Molecular Probes BacLight Live/Dead kit (Molecular Probes, Eugene, OR). The BacLight kit consists of two nucleic acid stains, Syto9 (green), which enters all bacteria, and propidium iodide (red), which can only enter bacteria with porous cell walls. Once inside the bacteria, the propidium iodide suppresses the Syto9 fluorescence so that live bacteria appear green, whereas dead or damaged cells appear red. In some cases, bacteria stain with both dyes

and appear yellow – these have been interpreted as live, but nonculturable. The nuclei of human cells also take up these nucleic acid stains, but rapidly turn red. They are readily distinguished from bacteria on the basis of size and morphology. In addition, these stains have been used to stain extracellular bacterial DNA Phospholipase D1 (eDNA), which is commonly found in the EPS and appears as a diffuse staining MLN0128 nmr surrounding the bacterial cells (Böckelmann et al., 2006; Thomas et al.,

2008). Fully hydrated specimens were then imaged by CM using a Leica DM RXE upright microscope attached to a TCS SP2 AOBS confocal system (Leica Microsystems, Exton, PA) using either a × 20 air objective or a × 63 long working distance water immersion objective. Live (green) and ‘dead’ (red) bacteria were imaged using 488 and 594 nm lasers; the suture and xenograft were imaged using reflected CM (blue) and bright-field microscopy (gray). Examination of one of the pieces of explanted Surgisis xenograft by CM showed heterogeneously distributed patches of live and dead bacteria and evidence of associated eDNA attached to the xenograft material (Fig. 2a). These organisms had a primarily coccal appearance, consistent with the solitary finding by culture of staphylococci. Interestingly, only one of the four specimens yielded a positive culture result, illustrating the inherent difficulty in detecting biofilm infections and making the case for multiple specimens to be sent for clinical culture, as well as the utility of using independent culture-free methods. Biofilms are commonly patchy on surfaces and this heterogeneity might partly explain the inconsistency in culture data. Examination of explanted suture material also showed evidence of attached and viable biofilm bacteria (Fig. 2c and d).

Our study is the first to evaluate the percentage of blood monocytes in CRPS patients. Although the percentage of total monocytes MDV3100 in vitro (CD14+ peripheral blood mononuclear cells) remained unchanged in CRPS, the percentage of the CD14+CD16+ monocyte subgroup was elevated significantly (P < 0·01) in individuals afflicted with CRPS compared to healthy controls. Previous studies have

determined that these cells represent a potent antigen-presenting and proinflammatory subpopulation of monocytes [28] that has been shown to be expanded in inflammatory conditions [34]. Although there was no correlation between the increased number of CD14+CD16+ monocytes in the CRPS group and the patients’ overall pain level, there was a correlation between increased numbers of CD14+CD16+ monocytes in CRPS patients demonstrating cold allodynia. This finding

suggests that the increased percentage of CD14+CD16+ monocytes may be associated with central sensitization. As reported previously, there was no difference in plasma levels of TNF-α, IL-10, IL-8, IL-6 and IL-1β between CRPS patients and controls [35,36]. However, individuals with high levels of CD14+CD16+ monocytes demonstrated a significantly Abiraterone purchase lower (P < 0·05) plasma level of IL-10 compared to individuals with low levels of CD14+CD16+. This is consistent with a study showing that CD14+CD16+ monocytes produce similar levels of the proinflammatory cytokines TNF-α, IL-6 and IL-1β and lower levels of the anti-inflammatory cytokine IL-10 [26]. This study also showed that the percentage of lymphocytes (T helper cells, T cytotoxic cells, NK cells or B cells) did not differ between CRPS patients and healthy control individuals. These results are in agreement with the study of Ribbers and colleagues that reported no association between lymphocyte subpopulations and patients with reflex sympathetic dystrophy (currently referred to as CRPS-type 1) [37]. A subsequent study by Kaufmann and colleagues also found no changes in the percentage of T cytotoxic cells, NK cells and B cells in CRPS patients [38]. However, they reported a reduction

of T helper cells (CD8+ lymphocytes) as well as an increase in the CD4/CD8 ratio [38] in CRPS patients compared to healthy controls. Although our study also Demeclocycline found a small reduction of CD8+ lymphocytes and an increase in the CD4/CD8 ratio, these changes were not statistically significant (P > 0·05). The elevation in the percentage of CD14+CD16+ monocytes seen in CRPS patients in this study could be due to the syndrome itself or may result from other factors. Factors such as physical inactivity, morbid obesity and sleep have been shown to alter the percentage of CD14+CD16+ monocytes [39–41]. Morbidly obese individuals have been reported to show elevated levels of the CD14+CD16+ monocyte subset [39]. The percentage of obese individuals (BMI > 30) in both the CRPS and control groups was approximately 20%.