To test this hypothesis, immunized mice were treated with an agonistic anti-GITR

mAb to disrupt the suppressive activity of Treg cells.48–50 Splenic GCs persist for at least 4 weeks so the GC response was monitored at days 8, 12, 18 and 24 post-challenge. Preliminary experiments tested the effects of continuous anti-GITR mAb injections on the GC response. When injected twice weekly for up to 4 weeks, however, anti-GITR mAb administration resulted in a high death rate in immunized mice, preventing an appropriate kinetic analysis (data not shown). Similar to previous studies18,22,23,26 a three-injection Ganetespib in vitro protocol was therefore used whereby 250 μg of either anti-GITR mAb or control rat IgG (rIgG) was injected on days −2, +1 and +5. Mice were immunized with SRBC on day 0 and splenic GCs were analysed during the ensuing 4 weeks. Naive mice kept in specific pathogen free conditions do not have detectable GC B cells in their spleens, as previously described1,5 and Dasatinib manufacturer shown in Fig. 1(a). Upon challenge with SRBC, a robust GC response is induced and easily detected as a B220+ PNAhi population (refs. 1,5 and Fig. 1a). Using a multi-colour approach, the IgM+ (non-switched)

B cells and switched GC B cells can be further delineated (Fig. 1a). When comparing the GC response from immunized mice injected with anti-GITR mAb or rIgG, it is clear that Treg-cell disruption resulted in a higher frequency and total number of splenic B220+ PNAhi GC B cells at all time-points Casein kinase 1 examined

(Fig. 1b). As expected, the ratio of IgM+ to switched GC B cells remained steady over the course of the response in control rIgG-treated mice, even as the reaction waned (Fig. 1c). However, immunized mice treated with anti-GITR mAb exhibited a higher frequency and total number of IgM− switched GC B cells at day 8, an imbalance which increased over time (Fig. 1c). When comparing the distribution of IgG isotypes expressed on switched GC B cells in anti-GITR mAb and rIgG treated mice, a significant increase in the percentage of IgG1+ GC B cells was observed at day 8 in the Treg-cell-disrupted group (data not shown). At all other time-points, IgG isotype expression within the switched GC pool did not differ between the two groups. Taken together, disruption of Treg cells led not only to a larger GC response, but to an inability to control the proportion of IgM+ to switched GC B cells. Given the marked changes observed in splenic GC B cells after Treg-cell disruption, the non-GC (B220+ PNAlo/neg) B-cell population was also monitored. As shown in Supplementary material, Fig. S1(A), a significant difference in the total number of non-GC B cells was observed after anti-GITR mAb treatment only at day 12 post-challenge. To assess which non-GC B-cell sub-sets were affected at day 12, a detailed analysis of follicular, pre-marginal zone, marginal zone, transitional 1 (T1), T2 and B1 B cell percentages was performed (see Supplementary material, Fig. S1B,C).

In the case of splenic macrophages, 10 units/mL IFN-γ was added to the culture medium to prime cells. In addition, 5 μg/mL polymyxin B was also added to avoid cell activation by LPS in the IFN-γ sample. Separately, RAW264.7 cells were incubated with pDNA/LA2000 complex for 2 h, then the cells were washed with RPMI-1640 Caspase inhibitor and incubated with fresh growth medium for an additional 6 h, and the supernatants were collected for ELISA and kept at −80°C until use. PMDC05 cells were incubated with ODNs for 24 h, then the supernatants were collected for ELISA and kept at −80°C until

use. The level of murine TNF-α and murine IL-6 in the media was determined by ELISA using the OptEIA set (BD Biosciences Pharmingen, San Diego, CA, USA). The level of human TNF-α in the media was determined by human TNF-α ELISA set (eBioscience, San Diego, CA, USA). RAW264.7 cells were incubated with Alexa488-labeled ODN1668 with or without ODN1720 or DNase-treated ODN1720 for 4 h and click here washed

three times with PBS. Then, the intensity of cell fluorescence was analyzed by flow cytometry (FACScan; BD Biosciences) using CellQuest software (version 3.1; BD Biosciences). Cellular uptake was estimated by subtracting the mean fluorescence intensity (MFI) at 4°C from that at 37°C (ΔMFI) and was plotted against the incubation time. ODN1668 was mixed with DNase I- or DNase II-treated ODN1720. The mixture containing 0.5 μg ODN1668 was incubated with DNase I (2 units/μg ODN1668) or DNase II (3 units/μg ODN1668) at 37°C. After 0, 5, 15, 30 min of incubation (DNase I treatment) or 0, 20, 40, 80 min of incubation (DNase II treatment), the mixture Clomifene was placed on ice and the reaction was terminated by the addition of 3 μL 0.2 M EDTA solution per 10 μL of samples. The ODN samples were run on a 21% PAGE and stained with ethidium bromide. The image of the gel was recorded using LAS 3000 (Fujifilm Life Science, Tokyo, Japan) and analyzed using Multi Gauge software (Fujifilm Life Science). Differences in the cytokine release were statistically evaluated by Student’s t-test. Differences in the thickness

of mouse footpad were statistically evaluated by one-way analysis of variance (ANOVA) followed by the Tukey–Kramer test for multiple comparisons. A p-value of less than 0.05 was considered to be statistically significant. We wish to thank Dr. Hiroyuki Yoshitomi (Graduate School of Medicine, Kyoto University) for providing technical assistance for subcutaneous injection of ODN into the footpad of mice. This work is partly supported by the 21st Century COE Program “Knowledge Information Infrastructure for Genome Science” and by a grant-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Sciences and Technology, Japan. 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”.

“
“The epidemiological and pathogenic relationship between Bordetella pertussis and Bordetella parapertussis, the two causes of whooping cough (pertussis), is unclear. We hypothesized that B. pertussis, due to its immunosuppressive activities, might enhance B. parapertussis infection when the two species were present in a coinfection of the respiratory tract. The dynamics of this

relationship were examined using the mouse intranasal inoculation model. Infection p38 MAPK inhibitor of the mouse respiratory tract by B. parapertussis was not only enhanced by the presence of B. pertussis, but B. parapertussis significantly outcompeted B. pertussis in this model. Staggered inoculation of the two organisms revealed that the advantage for B. parapertussis is established at an early stage of infection. Coadministration of PT enhanced B. parapertussis single infection, but had no

effect on mixed infections. Mixed infection with a PT-deficient B. pertussis strain did not enhance B. parapertussis infection. Interestingly, the depletion of airway macrophages reversed the competitive relationship between these two organisms, but the depletion of neutrophils had no effect on mixed infection or B. parapertussis infection. We conclude that B. pertussis, through the action selleck products of PT, can enhance a B. parapertussis infection, possibly by an inhibitory effect on innate immunity. The acute respiratory disease whooping cough (or pertussis) is caused by the gram-negative coccobacillus Bordetella pertussis, which binds to ciliated cells of the respiratory tract. However, a shorter and milder form of the disease is also caused by Bordetella Janus kinase (JAK) parapertussis (Heininger et al., 1994). Data suggest that B. parapertussis is the causative agent of a significant proportion of whooping cough cases in some global locations, including several European countries

(Watanabe & Nagai, 2004). In a clinical setting, it is hard to distinguish between these two pathogens, and involves costly laboratory tests such as PCR assays (Tatti et al., 2008). The treatment of infection is the same regardless of which of these two species of Bordetella is the infective agent, and therefore, tests to identify the causative pathogen are not always conducted. Mixed outbreaks and coinfection of patients with these two organisms have also been seen in clinical studies (Mertsola, 1985; Iwata et al., 1991), and there is little understanding of the epidemiological and pathogenic relationship between the two. Both of these pathogens are restricted to human hosts, although a distinct group of B. parapertussis strains that evolved independently (B. parapertussisov) infects sheep, causing a chronic nonprogressive pneumonia (Porter et al., 1994; Diavatopoulos et al., 2005). Bordetella pertussis and B.

The fungal aggregation ratio is defined as We extended the analysis of fungal aggregation by computing the cluster distributions for both strains. These are plotted in Fig. 7 for resting, swollen and opsonised spores, respectively. Interestingly, a statistical analysis using the Wilcoxon signed-rank

test revealed that these distributions were not significantly different from each other. This implies that – even selleck inhibitor in cases where af was found to be significantly different – clusters of a given spore number occurred with roughly the same frequencies, independent of the considered strains and spore conditions. In this comparative study of phagocytosis assays for L. corymbifera, we established a workflow for the automated analysis of fluorescence microscopy images suitable for high-throughput screening. We focused on two strains that deviate in virulence: JMRC:FSU:9682 (virulent strain) and JMRC:FSU:10164 Ceritinib (attenuated strain). The most striking finding was an increased phagocytosis ratio for the virulent strain compared to the attenuated strain. This result is counterintuitive given that alveolar macrophages represent the first line of innate immune defence. We speculate that the virulent strain could survive in alveolar macrophages and use these phagocytes as vehicles for dissemination via the blood stream causing systemic infections. As a prerequisite spores of the virulent strain would have to be efficiently recognised by phagocytes,

which is consistent with the observed difference in the effect of opsonisation between the virulent and the attenuated strain. A similar phenomenon was described for the encapsulated basidiomycete yeast Cryptococcus neoformans that causes disseminating infections in immunocompromised hosts.[21, 22] These cells survive in macrophages and are readily phagocytised allowing the pathogen to remain concealed from the immune system and protecting

it from exposure to antifungal agents.[21] The expulsion of Cryptococcus was reported to be blocked by a novel actin-dependent process (Arp2/3 complex-mediated actin polymerisation) on infected phagosomes, which may have significant implications for the dissemination of an invasion by C. neoformans.[22] Whether or not actin polymerisation is involved in the inhibition of the escape of L. corymbifera from macrophages Vorinostat in vitro is a subject of ongoing investigations. In passing we note that we applied a definition of the phagocytosis ratio pr that is relative to the number of adherent spores. This is motivated by the fact that about the non-adherent spores in the images we cannot be sure that they ever were in contact with macrophages. Thus, a definition of pr relative to the total number of spores, would likely underestimate the phagocytosis ratio. The possibility to distinguish between adherent and non-adherent spores is a clear advantage of image-based analyses compared with, for example, flow cytometry analyses of phagocytosis assays.

Interactions between naive T cells and DCs are believed to control both primary T cell activation and subsequent T cell fate, and thus the outcome of the adaptive immune response. How DCs perform such a complex feat remains unclear. The currently see more accepted view is that immune outcomes are determined primarily by factors external to both DCs and T cells, such as the microbe-derived signals that radically alter the activation state of DCs [1]. An alternative view is that the DC lineage is comprised of distinct DC subpopulations committed to predetermined functions [2, 3]. These functions, including generation of T cell tolerance or immunity,

are then amplified by exposure to microbial signals. In this model, the outcome C59 wnt mouse of an immune response depends upon how T cells integrate signals derived from the mix of preprogrammed DCs to which they are exposed during priming. The DC lineage in the mouse has been subdivided into populations on the basis of surface phenotypes that correlate with differences in ontogeny, microanatomical location and requirements for specific cytokines and transcription factors. In the currently

accepted schema, expression of high levels of CD11c and MHC II defines conventional DCs (cDCs), which are generated from precursors residing Interleukin-2 receptor in secondary lymphoid organs such as LN and spleen [1]. cDCs are then subdivided into CD8+ (Xcr1+Clec9a+) and CD11b+ (Sirpa+) subsets that correlate with the human CD141+ (Xcr1+Clec9a+) and CD1c+ (Sirpa+) DC subsets (reviewed in [4, 5]). In addition to cDCs, LNs contain migratory DCs (mDCs) that have entered the LN via afferent lymphatic vessels.

In murine LNs draining the skin, mDCs are defined as CD11cintMHC IIhigh, and comprise four distinct subsets: radioresistant migratory epidermal Langerhans cells (mLCs) and three subsets of radiosensitive migratory dermal DCs (mDDCs) that differ in expression of CD11b and CD207/Langerin [6] and/or CD103 (reviewed in [1, 7]). Migration of antigen-bearing DCs into the LN is essential for generating both peripheral adaptive immune responses and tolerance to antigens present within non-lymphoid tissues such as the skin [6, 8]. Migratory DC subset equivalents in humans have not been established fully, but recent reports have identified multiple distinct DC populations in human skin and LNs [9-11]. Attributing specific functions to individual DC subsets has proven far more difficult than the analysis of phenotype. DC subsets capable of driving CD4 and CD8 responses, regulating T helper type 1 (Th1)/Th2/Th17 bias, generating inducible regulatory T cells (Tregs) and/or inducing tolerance are highly model-dependent (see Table 1).

3A) drastically decreased the ratio of Treg and effector T cells. To see whether these relative differences in Treg were the result of MOG-immunization or already established in the steady state,

we analyzed Treg in the spleen of nonimmunized LFA-1+/+ and LFA-1−/− mice. Although around 14% of CD4+ T cells in WT mice were FoxP3+, only approximately 5.5% Treg were found in LFA-1 KO mice (Fig. 6A). In contrast to the situation in the spinal cord, the absolute numbers of Treg were also diminished, whereas the numbers of CD44high CD62Llow effector-memory phenotype T cells were unaltered in steady state (Fig. 6A). To get more information about the phenotype of Treg in LFA-1 KO mice, we analyzed several markers ICG-001 concentration defining subsets or which are known to be important for the function of Treg, such as CTLA-4, GITR, OX40, and 4-1BB (Supporting Information Fig. S1). However, we could not find any differences. Generally, a diminished population of Treg can be explained

by either reduced generation in the thymus or altered survival and homeostasis in the periphery. To discriminate these two possibilities, we directly examined Treg in the thymus of LFA-1−/− and LFA-1+/+ mice. As reported earlier 14, there were neither obvious differences in the size and cellularity of the thymus nor in the distribution of CD4/CD8 thymic subsets (Fig. 6B and data not shown). Also histologically, the thymus did not display any abnormalities. PD0325901 nmr However, when we analyzed the frequency

of FoxP3+ Chloroambucil T cells in the different thymic subsets, we found a significant reduction of Treg in the CD4+ single-positive subset of LFA-1−/− mice (Fig. 6B). Taken together, our results clearly show that a reduced generation of naturally occurring Treg in the thymus of LFA-1 KO mice results in a substantial lower frequency of Treg in secondary lymphoid organs. To test whether the reduced number of Treg in LFA-1 KO mice alone would be sufficient to explain the aggravated course of EAE, we suboptimally depleted Treg from WT mice. This was achieved by a single injection of the anti-CD25 mAb PC61. As shown in Supporting Information Fig. 2, this treatment resulted in a Treg frequency resembling LFA-1−/− mice. WT mice, PC61-depleted WT mice, and LFA-1 KO mice were immunized with MOG peptide. Figure 7 shows that the P61-depleted animals developed EAE scores absolutely comparable to LFA-1−/− mice. Interestingly, they even showed accelerated disease development. Until now, the exact role of LFA-1 in the pathogenesis of EAE is still elusive. There are several early studies using blocking mAb against LFA-1 which provided conflicting results. In one case, this treatment resulted in a clear amelioration 4, whereas Welsh et al. 5 reported an augmentation of EAE. In a third study 15, the animals simply died from the injection of the Ab.

Statistical analyses were performed using GraphPad Prism statistical analysis software. Group differences were analyzed by unpaired, two-tailed Student’s t-test, while a two-way ANOVA with repeated measures was applied when comparing different experimental groups over time. p-values of 0.05 or less were considered significant. The authors thank Dr. Thomas Lane for providing antisera to CXCR3 and CXCL10.

We also thank Dr. Julie Rumble and Dr. Nick Lukacs for critical reading of the manuscript. This work was supported by the National Multiple Sclerosis Society Grant CA 1037A (B.M.S) and by the National Multiple CHIR-99021 clinical trial Sclerosis Society Grant FG 1985-A-1 (S.J.L.). The authors declare no financial or commercial conflict of interest. As a service

to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information Selleck Doxorubicin (other than missing files) should be addressed to the authors. “
“Antiviral RNA silencing has been recognized as an important defense mechanism in arthropods against RNA viruses. However, the role of this pathway in DNA virus infection remains largely unexplored. A report in this issue of the European Journal of Immunology provides new insight into the role of RNA silencing in antiviral defense against DNA viruses. Huang and Zhang [Eur. J. Immunol. 2013. 137–146] found that the dsDNA virus white spot syndrome clonidine virus, an agriculturally important pathogen of shrimp, is targeted by the shrimp RNA-silencing machinery via

the production of virus-derived siRNAs. Furthermore, the authors show that the RNA-silencing pathway, and crucially, Dicer-2, is important for restricting viral infection. This study provides novel insights not only into shrimp antiviral defenses but also potentially into antiviral immunity against DNA viruses in a larger spectrum of hosts, as discussed in this Commentary. Furthermore, this study may contribute to the future development of immune-based therapeutics to combat viral pathogens, not only in aquaculture, but also in insect vectors of human diseases. RNA silencing is an innate antiviral pathway used to target foreign RNA for degradation. This mode of recognition is based on the fact that all RNA viruses produce double-stranded (ds)RNA during their life cycle. Since dsRNAs are not naturally produced in higher organisms, the development of dsRNA-based recognition systems provides a simple strategy for the selective targeting of RNA viruses. Organisms including plants and arthropods use RNA silencing to control both endogenous gene expression and foreign RNAs derived from viruses.

Mitochondria are motile organelles, utilizing tracks of microtubules to distribute themselves evenly along axons, and travel to areas of

metabolic demand. Mitochondria form branched networks throughout the cytoplasm, via the dynamic processes of fusion and fission. Mitochondrial fusion is a two-stage process of outer membrane fusion mediated by the proteins mitofusin 1 and 2, and inner membrane fusion involving OPA1 [7–9]. Conversely, Fis1 PKC inhibitor and Drp1 mediate mitochondrial fission events [10]. Through this physically interconnected network, mitochondria are able to create an efficient system for the delivery of ATP throughout the cell [11], buffer calcium levels [12], facilitate the exchange of lipid membranes buy EPZ-6438 [13] and allow complementation of mitochondrial DNA. All of these are crucial for the maintenance of healthy mitochondria [14]. Indeed, investigation of mitochondrial morphology in fibroblasts revealed that the cell responds to an increase in superoxide production with an increase in mitochondrial branching [11]. This observation supports the notion that networking of the mitochondria represents an adaptive mechanism, allowing the mitochondria to function more efficiently, thus coping with cellular stress [11,13]. Accordingly, it has been noted that a loss of connectivity, concomitant with the formation of punctate mitochondria, is seen under conditions

of mitochondrial dysfunction [15,16]. Furthermore, fragmentation, by either an inhibition of fusion or an increase in fission, facilitates the induction of the intrinsic apoptotic cascade by aiding release of mitochondrial pro-apoptotic factors into the cytoplasm [17]. Thus, the morphology of mitochondria may have a significant impact on the ability of the organelle

Bay 11-7085 to function efficiently, and as discussed later, aberrant mitochondrial function influences mitochondrial morphology, which may lead to further deleterious effects [11]. Consequent to these diverse functions and alterations in morphological state, mitochondrial pathology is now implicated as causal or contributory to several neurodegenerative diseases [18]. This review will be focused on a common motor neurone disorder, amyotrophic lateral sclerosis (ALS). Axonal transport is required for the correct distribution of organelles, synaptic vesicles and products of protein synthesis, as well as the transport of signalling factors endocytosed at the cell membrane to the cell body. Axonal transport, including mitochondrial axonal transport, is facilitated by the cytoskeleton and molecular motor proteins. Microtubules, made of tubulin, are arranged longitudinally and are polarized in axons, with the minus end originating from the microtubule organizing centre in the cell body, and the plus end extending to the growth cone.

The unique receptor repertoire of dNK cells further includes the expression of several Ly49 receptors, the expression of activation markers such as CD69 and KLRG1 (which is considered as a marker for active NK cell proliferation37) and the expression of CD117 (the c-kit receptor). Another study, by Mallidi et al.17 described the phenotype of NK1.1+ dNK cells as DX5+ NKp46+ CD27+ CD11b+ CD11c+ CD69+. Interestingly, the NK1.1+ dNK cells expressed more B220 and CD69 than NK1.1+ eNK cells and also expressed ICOS (which is expressed on activated NK cells38), whereas eNK cells did not express ICOS at all. In the fetal-maternal interface, the maternal uterine tissue is

in close contact with the fetal-derived trophoblast cells. This interface contains immune cells, which constitute Dabrafenib concentration 40% of the cells in the human decidua.39 Analysis of this immune population has revealed that, unlike any other tissues or mucosal surfaces, 50–70% of the human decidual lymphocytes are NK cells, while the remainder are CD14+ macrophages, dendritic cells,

CD4+ T cells, a few CD8+ T cells, γδ T cells, and NKT cells.35 dNK cell numbers are the highest in the first trimester of pregnancy and their numbers decline during the second trimester. As in mice, only few dNK cells are present in the human decidua at term.36 The majority of dNK cells are CD56bright CD16− (as opposed to mouse dNK cells which express high levels of CD1618). Indeed, dNK cells Torin 1 price resemble peripheral blood CD56bright CD16− NK cells also in the high expression levels of CD94/NKG2.40 However, similar to eNK cells, dNK cells resemble CD56dim CD16+ NK cells in the expression of KIRs41 and in their granules cell content. In fact, dNK cells differ from peripheral Carbohydrate blood NK cells both in phenotype and in function. Comparison analysis of the gene expression in dNK cells versus peripheral blood NK cells showed that dNK cells

should be considered as a unique NK subset.27 dNK cells over-expressed several genes, compared with the two peripheral blood NK subsets and several genes were exclusively expressed in dNK cells. For example, granzyme A was significantly over-expressed in dNK cells, as were the C-type lectin-like receptors NKG2C and NKG2E. dNK cells have been shown to express several activating receptors, including NKp46, NKp30, NKp44 (in contrast to human eNK cells which lack NKp30 and NKp44 expression, as discussed above), NKG2D, and 2B4.42–44 The expression of NKp44 (which is not expressed on non-activated peripheral blood NK cells) and the expression of the activation marker CD6945 (which is also expressed on mouse dNK cells) suggest that dNK cells might already be activated in the local environment of the decidua.

24 Median follow up was for 37.8 months. This survival advantage persisted when late referral and observation for <1 year were excluded. Riegel et al.,

in a prospective study of 551 patients from Germany, showed that only 38.7% of patients selleck kinase inhibitor with CKD stage 4 were under nephrological care.25 These patients had a higher incidence of planned initiation of dialysis (81.0% compared with 48.0%), less hospitalization (54.5% vs 83.7%) and a shorter duration of hospital stay (11.4 vs 17.4 days). Roderick et al. studied 250 patients referred for renal replacement therapy over a 12-month period.26 Ninety-six patients (38%) were referred late (<4 months), which were further defined as avoidable and unavoidable late referrals. These patients were less likely to receive standard CKD therapies, were in a poorer clinical state and more frequently commenced dialysis emergently. Mortality at 6 months was 16% in the early referral group compared with 28% in the avoidable late referral group and 35% in the unavoidable late referral group, respectively. Starck in 2001 studied a prospective cohort of 2264 patients in the Dialysis Morbidity and Mortality (DMM) Study Wave 2.27 Late referral

(within 4 months of initiation of dialysis) was associated with higher mortality at 1 and 2 years with RR 1.68 (95% CI: 1.31–2.15) and 1.23 (95% CI: 1.02–1.47), respectively. Patients who were seen by a nephrologist at least twice in the year before dialysis commencement had a lower risk of death with Nutlin-3 mw RR 0.8 (95% CI: 0.62–1.03). Late referral patients were less

likely to have a fistula, to be on erythropoietin and to have had two or more predialysis nephrologist visits. Stehnan-Breen et al. also used data from the DMM Study.28 Only 34.4% of patients had permanent access at the initiation of dialysis; 67% of patients had an AV graft rather than a fistula. Early referral was an important predictor of permanent access with OR 0.33, along with serum albumin (OR 1.55), erythropoietin use (OR 1.79) and fewer predialysis nephrologist visits (OR 0.1) – all surrogate markers of timely referral. Wauters et al., in a prospective Suplatast tosilate study of 279 patients in three countries (France, Italy and Switzerland), found 71.6% were referred early (>6 months), 15.1% intermediate (1–6 months) and 13.3% late (<1 month).29 Late referral was associated with an active cancer, rapid progression of CKD, the structure of the dialysis centre (city worse than private or regional centres) and the nature of the referring physician (nephrologists and general practitioners better). Sesso and Belasco in 1996 reported the outcomes of 205 consecutive patients with non-diabetic nephropathy who were commenced on dialysis between October 1992 and March 1995 in the Nephrology Division of Hospital São Paolo, Brazil.