Methods: Japanese workers in Shanghai under treatment of as least one of diseases of HT, HL,

CKD or DM in outpatient clinic of Huashan Hospital World Wide Medical Center (HWMC) in Shanghai, China who stayed there for more than 6 months were enrolled. Medical Intervention were 1) medical treatment by collaboration PD0332991 of monthly visiting doctors from Kitano Hospital (KH) in Osaka, Japan and those of HWMC, 2) coaching of life style by KH nurses resident in Shanghai and 3) attending seasonal health care seminar were performed: Samples of disease status, life style status as behavior modification (BM) score calculated by division of number (N) of BM by N of interview minus 1 and health related QOL score by SF36 were obtained before and after intervention. Results: Within 28 enrolled patients, final 18 (17 male 1 female) were evaluated with full data of SF36. In 16 HT patients, systolic(s) and mean(m) selleck products blood pressure (BP) were significantly declined (P < 0.011, P < 0.023, respectively). Significant improvement of role-social QOL was observed (P < 0.046). Correlation between corrected BW and BM score and improvement of health related QOL were observed. Correlation between BM score and physical

and mental QOL improvement was observed. Multiple regression analysis indicated that role-social QOL improvement was independently affected by amelioration of mBP and BW (R-squared: 0.665 and 0.900, P-value: 0.002 and 0.001 respectively). Conclusion: International Joint medical Glutathione peroxidase intervention with intensive coaching of life style has brought about significant elevation of health related QOL of Japanese oversea worker patients

in Shanghai along with correction of BP and especially BW through BM. BUNANI EUNICE, DUMDUM1,2, BUNANI ARCHIE3 1Puerto Community Hospital; 2Cagayan de Oro Medical Center; 3Southwestern University College of Medicine Background: Literatures have emphasized that administration of anticoagulation in dialysis promotes minimal filter clotting and post dialysis bleeding, and improves patient quality of life through prolongation of the vascular access. Objective: This study evaluated the protocol plan designed to deliver both High and Low Molecular Weight Heparins (HMWH, LMWH) as bolus and cath-dwell and develop a relationship between filter clotting, post dialysis bleeding (PDB), blood flow rate (Qb), and activated Partial Thromboplastin Time (aPTT) among hemodialysis (HD) patients. Methods: 208 HD patients were included in an evaluative cross-over design; bolus-LMWH and HMWH as cath-dwell for the first 6 months and vis-à-vis on the next 6 months. Regression and ANOVA were used for analysis with R square as basis related to heparin adjustment and different filters in single-use basis. Results: Results indicated filter clotting among fistula (f = 8, spv = 0.742) and catheter (f = 17, spv = 0.

This indicates that ligation of a subset of TLRs generates proinflammatory cytokines that co-ordinate to potentiate human Th17 differentiation. In addition, the synergy between TLR-4 and TLR-7/8 in controlling the sequential production of regulatory and proinflammatory cytokines by naive CD4+ T cells was detected [78]. The observed polymorphism in DC responses to such TLR-mediated stimuli could explain differences in the susceptibility to infectious pathogens or autoimmune diseases within the human population. Furthermore, using agonists Tyrosine Kinase Inhibitor Library ic50 specific for TLR-7 (i.e. Imiquimod, Gardiquimod) or TLR-8 (ssPolyU), together with LPS, confirmed that a significant synergy in cytokine induction

is observed consistently after joint engagement of TLR-4 plus TLR-7 and/or TLR-8 [80,81]. However, the TLR-7, which is not present in DCs under normal conditions, is up-regulated dramatically in selected donors after stimulation Selleck Deforolimus by LPS, in agreement with a previous study [78,80]. Thus, the observed polymorphism between high and low DC responders is due probably to differences in TLR-7/8 up-regulation following TLR-4 stimulation, suggesting that a threshold stimulation of TLR-7 and/or TLR-8 is required to activate the joint secretion of multiple cytokines by DCs. Taken together, TLR-3, -4, -7 and -8 are required in the induction of Th17 cell differentiation and subsequent biological effects, but the role of TLR-9 is controversial,

which urgently needs to be illustrated (Fig. 3). In mice, coincidental activation of complement and several TLRs (TLR-3, -4, -7, -8 and -9) led to the synergistic production of serum factors that promote Th17 differentiation from anti-CD3/CD28 or antigen-stimulated T cells [82] (Fig. 3). Although multiple

TLR-triggered Methisazone cytokines were regulated by complement, Th17 cell-promoting activity in the serum was correlated with IL-6 induction, and antibody neutralization of IL-6 abrogated the complement effect [82]. These data establish a link between complement/TLR interaction and Th17 cell differentiation, and provide new insight into the mechanism of action of complement and TLR signalling in autoimmunity. Although CD4+ T cells are considered to be the major source of IL-17, especially in autoimmune diseases, recent studies have indicated that other T cell subpopulations such as CD8+ T cells, natural killer (NK) T cells and γδ T cells can also produce IL-17 [74,83]. It is reported that CCR6+ IL-17-producing γδ T cells, but not other γδ T cells, express TLR-1 and TLR-2, but not TLR-4 [84,85]. Ligands that target these pathogen recognition receptors can cause the selective expansion of IL-17+γδ T cells and functional consequences, such as neutrophil recruitment [86]. Studies have shown that γδ T cells activated by IL-1β and IL-23 are an important source of innate IL-17 and IL-21 and may act in an amplification loop for IL-17 production by Th17 cells [74,86].

The results revealed that the frequency of SIRT1 expression in medulloblastoma tissues was 64.17% (77/120), while only

one out of seven tumor-surrounding noncancerous cerebellar tissues showed restricted SIRT1 expression in the cells within the granule layer. Of the three morphological subtypes, the rates of SIRT1 detection in the large cell/anaplastic cell (79.07%; 34/43) and the classic medulloblastomas (60.29%; 41/68) are higher than that (22.22%; 2/9) in nodular/desmoplastic medulloblastomas Selleck GSK-3 inhibitor (P < 0.01 and P < 0.05, respectively). Heterogeneous SIRT1 expression was commonly observed in classic medulloblastoma. Inhibition of SIRT1 expression by siRNA arrested 64.96% of UW228-3 medulloblastoma cells in the gap 1 (G1) phase and induced 14.53% of cells to apoptosis at the 48-h time point. Similarly, inhibition of SIRT1 enzymatic activity with nicotinamide brought about G1 arrest and apoptosis in a dose-related fashion. Our data thus indicate: (i) that SIRT1 may

act as a G1-phase promoter and a survival factor in medulloblastoma cells; and (ii) that SIRT1 expression is correlated with the formation and prognosis of human medulloblastomas. In this context, SIRT1 would be a potential therapeutic target of medulloblastomas. “
“Both chordoma and Rathke’s cleft cyst are relatively rare diseases in the central nervous system. In this paper we report the first case of Dabrafenib nmr a chordoma coexisting with a Rathke’s cleft cyst. A 49-year-old man presented with a 19-month history of distending pain, movement dysfunction and diplopia of the left eye. The preoperative diagnosis was consistent with chordoma with cystic change. Final pathological diagnosis of chordoma coexisting

with Rathke’s cleft cyst was made according to histological and immunohistochemical studies and the clinical and radiological features are discussed. Considering the close relationship between the notochordal tissue and Rathke’s pouch during early embryogenic development, a possible mechanism is GNA12 also discussed with the literature review. “
“Optineurin is a gene associated with normal tension glaucoma and primary open-angle glaucoma, one of the major causes of irreversible bilateral blindness. Recently, mutations in the gene encoding optineurin were found in patients with amyotrophic lateral sclerosis (ALS). Immunohistochemical analysis showed aggregation of optineurin in skein-like inclusions and round hyaline inclusions in the spinal cord, suggesting that optineurin appears to be a more general marker for ALS.

A hallmark cytokine associated with tumor-induced immunosuppression is TGF-β1. Although we detected increased circulation of TGF-β1 in tumor-bearing animals in some experiments, it did not exert an apparent inhibition on the autoimmune Teff cells at a distal site in healthy tissues. At cellular levels, Treg cells and MDSCs have long been recognized as critical mediators of immunosuppression in cancer. Our studies with self-antigen-specific T cells highlighted an increased

potency of these regulatory mechanisms in tumors versus healthy tissues. The molecular mechanisms responsible for the local immunosuppression remain to be elucidated. Possibly, a suppressive cytokine milieu, directly or indirectly related to Treg cells and MDSCs, inactivates Teff cells at the tumor site, which could be reactivated by an agonistic cytokine stimulation [40] or a global alteration of tumor gene expression profiles [41]. This study implicates CTLA4. Buparlisib order Suggestive of the intertwining between autoimmunity and antitumor immunity, protection from cancer is often associated with the same polymorphisms of the CTLA4 locus that are linked to autoimmune susceptibility [15, 18-20]. A conditional knockout model

established an essential role for CTLA4 in Treg cells Epacadostat [8]. Its intrinsic role in Teff cells has also been well-documented [9, 10]. Our study with a CTLA4 shRNA model indicates a distinction between quantitative variation in CTLA4 and the “all-or-nothing” model of CTLA4 knockouts. A subtle reduction of CTLA4 did not impair Treg-cell function, but substantially promoted Teff-cell capacity in tumor settings. An expansion of immunotherapy trials has generated a plethora of novel ideas in cancer immunology. The entangling of auto-immunity toxicity with antitumor benefit has provoked a shift of perspective whereby autoimmune side effects are considered

not only a welcome marker but actual effectors for antitumor immunity [7]. A direct comparison of about cancerous versus healthy tissue in interaction with self-antigen-specific Teff cells revealed their intrinsic potential in tumor eradication. However, they were subjected to regulatory mechanisms that have been evolved to induce tolerance to nonmalignant self-tissue, even more so in the tumor microenvironment. Therefore, self-antigen can be effectively targeted for antitumor immunity, but harnessing the tumor-destruction capacity of self-antigen-specific T cells requires effective strategies to overcome the suppressive microenvironment at the tumor site. CTLA4 blockade therapies can abrogate suppressive tumor milieu by reverting the local predominance of Treg cells over self-antigen-specific Teff cells. On the other hand, a subtle reduction of CTLA4 reflecting genetic variations may substantially alter an immunoprivileged environment evolved in a solid tumor through an intrinsic impact on Teff cells.

These composite www.selleckchem.com/products/Liproxstatin-1.html findings support the hypothesis that specific CXCL12 analogues with ancillary antibiotic treatment are beneficial in experimental sepsis, in part, by augmenting PMN recruitment and function. This article is protected by copyright. All rights reserved. “
“Filoviral hemorrhagic fever (FHF) is caused by ebolaviruses and marburgviruses, which both belong to the family Filoviridae. Egyptian fruit bats (Rousettus aegyptiacus) are the most likely natural reservoir for marburgviruses and entry into caves and mines that they stay in has often been associated with outbreaks of MVD. On the other hand, the natural reservoir for ebola viruses remains elusive;

however, handling of wild animal carcasses has been associated with some outbreaks of EVD. In the last two decades, there has been an increase in the incidence of FHF outbreaks in Africa, some find more being caused by a newly found virus and some occurring in previously unaffected areas such as Guinea, Liberia and Sierra Leone, in which the most recent EVD outbreak occurred in 2014. Indeed, the predicted geographic

distribution of filoviruses and their potential reservoirs in Africa includes many countries in which FHF has not been reported. To minimize the risk of virus dissemination in previously unaffected areas, there is a need for increased investment in health infrastructure in African countries, policies to facilitate

collaboration between health authorities from different countries, implementation of outbreak control measures by relevant multi-disciplinary teams and education of the populations at risk. Ebolaviruses and marburgviruses are single-stranded, negative-sense, non-segmented RNA viruses belonging to the family Filoviridae, order Mononegavirales (Table 1). These filoviruses are known to cause hemorrhagic fever in humans and nonhuman primates [1]. Most of the known filoviruses are endemic to Africa: several different virus species belonging to the genus Ebolavirus have been found in central and western African rain forests, within approximately 10° north and south of the equator [2], and single species belonging Oxaprozin to the genus Marburgvirus in open dry areas of eastern and south central Africa [3] (Fig. 1). The first case of MVD in Africa was reported in 1975, when a tourist who had visited Zimbabwe developed hemorrhagic fever in South Africa [4, 5]. There were a few subsequent outbreaks of this disease, but after 1987 there was a period of quiescence until the DRC outbreak in 1998. The first outbreak of EVD was reported in Zaire (now the DRC) in 1976, subsequently outbreaks occurred in Sudan (now South Sudan) in 1976 and 1979 [4]. These were followed by 15 years of no reported outbreaks in Africa.

SIGNR1 resides in the spleen marginal zone 28 and lymph node medulla 34 captures antigens from distal infection sites via blood and lymph, respectively. Therefore, SIGNR1 in confined parts of the body in vivo plays a role as the first sensing machinery against infection. For instance,

it is known that SIGNR1 in the spleen marginal zone is involved in systemic complement activation by sensing blood-borne CPS of S. pneumoniae35. Likewise, rpMϕ are also the first interceptors for peritoneal infection and a major source of oxidative burst in peritoneal cells, as shown Fig. 4D, possibly leading to subsequent inflammatory responses in the cavity. The host innate immune system simultaneously recognizes various types of ligands on microbes via a variety of receptors on the various types CHIR-99021 chemical structure of cells. Recently, Dectin-2 36, 37 has been shown to also be important for host response to C. albicans. Nevertheless, our finding sheds light on the cooperation this website of different and/or similar types of PRRs in innate responses. Like the intracellular crosstalk of distinct PRR-mediated signaling pathways, PRRs also collaborate to recognize and capture

microbes and to transduce signals for enhancing cellular responses. Collectively, although the cooperative action pathway between SIGNR1 and Dectin-1 in the oxidative response is not entirely definitive, our results suggest that the anti-microbial activity/oxidative burst induction is due to efficient recognition of cell wall mannoproteins via SIGNR1 and their subsequent internalization, possibly along with the association with Dectin-1, allowing Dectin-1 to access the limited β-glucans and leading to the activation of Syk-mediated signaling. Female else BALB/c mice were purchased from Japan SLC (Hamamatsu, Shizuoka, Japan). The mice were maintained under specific pathogen-free conditions, and used at 8–12 wk of age. All experiments were conducted according to our institutional guidelines. HEK293T cells, the mouse monocytic cell line RAW264.7 cells and RAW-transfectants (RAW-SIGNR1, RAW-control and RAW-SIGNR1Δcyto

cells) were maintained as described previously 26. Expression levels of SIGNR1 and Dectin-1 of these transfectants were analyzed with biotinylated anti-SIGNR1 clone 22D1 28 with PE-streptavidine and anti-Dectin-1 clone 2A11 (AbD Serotec, Oxford, UK) with PE-anti-rat IgG, respectively. Substitutions of glutamic acid 285 with glutamine (E285Q) in SIGNR1 were introduced by overlapping PCR. cDNA fragments of SIGNR1ΔCRD (192–325) was PCR amplified using forward primer 5′-GATCGAATTCATGAGTGACTCCACAGAAGCC-3′ in combination with reverse primer 5′-GATCCTCGAGCTACAGGCGGAAGAGTTCAGTCTTC-3′. pcDNA4/HisMax-SIGNR1 23 was used as a template, and the resulting PCR products were cloned into the EcoRI-XhoI site of pcDNA4/HisMax (Invitrogen, Carlsbad, CA). Surface expression of these mutant proteins was confirmed by flow cytometry with polyclonal anti-SIGNR1 (R&D Systems, Minneapolis, MN).

Tetanus toxoid is a protein antigen and elicits a strong specific antibody response. In our experience, impaired response to tetanus toxoid is observed only in severe immune deficiency; even patients with common variable immunodeficiency who have impaired specific antibody response to pneumococci do not display impaired specific antibody response to tetanus toxoid. Only two patients in this study had impaired protective levels to most of the 14 polysaccharide antigens; the majority of patients had impaired responses to serotypes

3, 8, 9N and 12F. Oxelius et al.[3] reported normal responses to polysaccharide antigens in their mixed sample of 10 adults and children (although they had data only for pneumococcal serotypes 3, 6A, 19F and 23F). This is in contrast to a report by Popa et al.[8], who observed decreased response Tamoxifen to tetanus and Haemophilus influenza vaccines in IgG3-deficient adults. Soderstrom et al.[11] reported that 75% of Barasertib chemical structure adults with selective IgG3 deficiency had low B cell function, as defined by EBV- or PWM-stimulated protein

A plaque-forming cells lower than 50% of healthy controls. Data on T cell function in selective IgG3 deficiency are limited. We observed that 30–40% of patients display impaired T cell proliferative response to mitogens and recall antigens. Soderstrom et al.[11] reported decreased T cell function (defined as PHA or ConA stimulation indices of <0·8) in 40% of IgG3-deficient adult subjects. In their study, data were presented as stimulation index, Montelukast Sodium which may be skewed due to differences in background counts. In our study, we analysed data as net counts per minute after subtracting the background. T helper-1 (IFN-γ) and T helper-2 (IL-5) cytokine production was analysed in seven subjects; abnormal IFN-γ production was observed in one patient and abnormal IL-5 production in two patients. It is not possible to suggest the significance of these cytokine results in IgG3 subclass deficiency, as the number of samples tested is small. Finally, NK cell cytotoxicity

and neutrophil oxidative burst (reactive oxygen species generation) were relatively normal. In two patients oxidative burst was modestly reduced; however, it was not to a level observed in chronic granulomatous disease. Furthermore, patients did not have diabetes mellitus. In general, IgG1 or IgG2 deficiencies are reported to cause more severe infections, and there is greater acceptance of the use of immunoglobulin prophylaxis in such cases [7]. In our study, clinical response to IVIG was observed in the majority of patients with IgG3 deficiency. Six of 13 patients who received IVIG had dramatic relief from their recurrent infections, five patients experienced moderate clinical improvement and two patients had no response. We did not observe any correlation between response to IVIG and immunological parameters. However, our sample size is too small to reach a definitive conclusion. Olinder-Nielsen et al.

For CD137, there was a significant increase in the percentage of both CD28null/CD4+ and CD28null/CD8+ buy Anti-infection Compound Library T cells expressing this co-stimulatory receptor in patients with BOS compared with stable transplant patients and controls (Fig. 5a). The increase was significantly greater for the CD8+ subset compared with the CD4+ cells for all groups (Fig. 5a). For CD28+ cells (both CD8 and CD4 subsets) there was decreased expression of CD137 in stable transplant patients and patients with BOS

compared with controls [75 ± 22·2%, 33·6 ± 18·6% and 37·1 ± 18·7%; and 60·1 ± 21·4%, 31·5 ± 16·7% and 28·3 ± 18·2% (mean ± s.d.) CD28+/CD137+/CD4+ and CD28+/CD137+/CD8+ for controls, stable patients and patients with BOS, respectively] (all P < 0·05). For CD152, there was a significant increase in the percentage of both CD28null/CD4+ and CD28null/CD8+ T cells expressing this co-stimulatory receptor in patients with BOS

compared with stable transplant patients and controls (Fig. 5b). There were no significant changes in expression of CD152 by CD28+ (either CD4+ or CD8+) for any group BVD-523 [50·5 ± 18·9%, 42·8 ± 18·9% and 39·4 ± 20·1%; and 19·0 ± 10·6%, 14·7 ± 12·3% and 12·8 ± 11·9% (mean ± s.d.) CD28+/CD152+/CD4+ and CD28+/CD152+/CD8+ for controls, stable patients and patients with BOS, respectively] (all P > 0·05). For CD154, there was a significant increase in the percentage of CD28null/CD4+ (note: unchanged in the CD8+ subset) expressing this co-stimulatory receptor in patients with BOS compared with stable transplant patients and controls (Fig. 5c). There was decreased expression in the CD28+/CD4+ subset compared to the

CD8+ subset in stable transplant Carbohydrate patients and patients with BOS compared with controls [81 ± 19·9%, 63·1 ± 17·1% and 48·9 ± 24·2%; and 16·9 ± 4·6%, 6·0 ± 3·1% and 6·4 ± 4·7% (mean ± s.d.) CD28+/CD4+/CD152+ and CD28+/CD8+/CD152+ for controls, stable patients and patients with BOS, respectively] (all P > 0·05). For CD134, there was significantly increased expression by CD28null/CD4+ T cells (note: unchanged in the CD8+ subset) in patients with BOS compared with stable transplant patients and controls (Fig. 5d). There were no differences in the percentage of CD28+/CD4+ and CD28+/CD8+ cells expressing CD134 in stable transplant patients and patients with BOS compared with control subjects [52·0 ± 21·3%, −51·3·1 ± 22·7% and 35·5 ± 28·2%; and 28·1 ± 16·4%, 18·6 ± 17·8% and 13·8 ± 12·9% (mean ± s.d.) CD28+/CD134+/CD4+ and CD28+/CD134+/CD8+ for controls, stable patients and patients with BOS, respectively] (all P > 0·05).

L., L.A., M.H. and J.P. analyzed data and M.L., L.A. and G.G. wrote the paper. Conflict of interest: The authors declare no financial or commercial conflict of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. “
“To discriminate between viable and non-viable Enterococcus faecalis, the predominant pathogen in apical periodontitis, a real-time PCR method combined with propidium monoazide (PMA) was developed and

evaluated. Selleck Doxorubicin PMA had no antimicrobial effect on E. faecalis cells and permitted enumeration of both viable and non-viable cells. Therefore, E. faecalis cells from the root canals of nine patients with apical periodontitis were analyzed to evaluate the diagnostic usefulness of this approach. Viable and non-viable

E. faecalis cells were successfully discriminated in these clinical specimens. A real-time PCR assay combined with PMA will contribute to the precise diagnosis of apical periodontitis. Enterococci are present in small numbers in the oral find protocol cavities of healthy individuals; however, they dominate the oral cavity in patients with apical periodontitis, which is primarily caused by anaerobic oral bacteria surviving on the teeth in apical biofilms post-treatment. The enterococci recovered from biofilms in the root canals of patients with apical periodontitis are often antimicrobial-resistant (1, 2). E. faecalis is a major pathogen in apical periodontitis (3); thus, monitoring

this organism in periapical biofilms during the treatment of apical periodontitis is crucial. Quantitative PCR-based methods have been developed for enumerating bacteria (4, 5); however, DNA-based detection methods cannot differentiate between signals originating from live and dead bacteria. Such differentiation is diagnostically important, especially for antimicrobial-resistant organisms. Therefore, a PCR-based method that can discriminate between DNA derived from viable and dead bacterial cells is needed. Recently, the DNA-binding Nutlin-3 ic50 dyes EMA and PMA were used for PCR-based differentiation of viable and dead bacterial cells (6–8). These dyes exclusively penetrate dead cells following membrane damage and cross-link the DNA via photo-activation, thereby inhibiting amplification (9). However, recent data has shown that EMA cross-linking during genomic DNA extraction renders the DNA insoluble and causes its loss in concert with cellular debris (7). EMA can also penetrate live cells of some bacterial species (6); however, it is toxic to viable cells (8, 10). In this study, we evaluated a PMA-based quantitative detection method that distinguished viable from non-viable E. faecalis cells in root canals. The bacteria used in this study are listed in Table 1. Enterococcus faecalis was grown anaerobically in trypticase soy broth (Becton-Dickinson, Sparks, MD, USA).

One important implementation mTOR inhibitor of Rep-Seq is in estimating the number of unique receptors, i.e. the size of the expressed repertoire in an individual at any given moment.14,17,19,20,33 Estimates of the number of non-sampled receptors

are key for an accurate quantification of the total diversity. A solution for an analogous problem was identified > 60 years ago by the legendary statistician Fisher. The problem, termed the ‘unseen species problem’, refers to the attempt to estimate the total number of species in a given large population, based on random samples of species.35–37 Fisher et al.37 developed an analytic solution, assuming a Poisson distribution, which was later extended by Efron and Thisted.35 This analytical solution is mainly a capture–recapture method, associated with statistical analysis of these repeatedly sampled collections of sequences. Various estimation attempts were made, by estimating the number of unique V(D)J combinations. Since receptor diversity is also created by nucleotide insertions and deletions (indels) and somatic hypermutations in B cells, these estimations are only lower boundaries to the actual number

of possible combinations. Most studies focused on a single chain of the immune receptor and therefore resulted in describing only a portion of the total diversity obtained Ruxolitinib by the combination of the two chains constructing the heterodimer. For example, Wang et al.20 estimated 0·47 × 106 TCR-α unique nucleotide sequences and 0·35 × 106 TCR-β sequences. Robins et al.19 suggested that CD8+ T cells express < 0·1% of the combinatorial landscape of the β chain (5 × 1011). Weinstein et al. showed a lower limit of 5000–6000 unique antibodies Rho in the zebrafish.33 Although these are only lower limits to the actual size of the repertoire, it is clear that any individual expresses only a small fraction of the potential diversity (Figs 2 and 3). In spite of substantial advances in repertoire size estimates, there remain three important issues with the capture–recapture approach that

require further attention: First, the common assumption is that the number of unique clones is distributed according to a Poisson distribution. However, recent studies show evidence of a power law distribution.33 Moreover, Fisher et al. demonstrated that several estimation approaches conflict; in terms of receptor sequences, they determined a ratio of the number of new and unique sequences discovered in a new sample divided by the total size of the data (i.e. the whole repertoire expressed in an individual). When this ratio is < 1, i.e. only a portion of the sample contains new sequences, all estimations agree. However, when the ratio is > 1, some approaches converge and stabilize while others completely diverge.