Notably, loading of DCs with heat-stressed tumour cells has been shown to permit enhanced cross-priming, most likely due to concomitant upregulation of heat-shock proteins and tumour antigen expression [40]. Importantly, loading with heat-stressed tumour cells during DC maturation in our present study did not negatively affect the production of CXCR3 ligands or recruitment of NK and NKT cells, nor did it negatively affect production of CCL3, CCL4 or IL-12p70 upon subsequent CD40 ligation. We therefore believe that this could be of potential relevance in vaccination strategies for patients with CLL where tumour antigens still are poorly

identified. Despite enriching monocytes by a three-step protocol, selleckchem we were unable to achieve desired purity in some cases. With contaminating cells of up to 30%, most being CD19+ CLL cells, the risk of tumour cells affecting DC function and yield must be taken into account. Indeed, when autologous DCs are developed from monocytes in patients with progressive disease, DC dysfunction has been observed, most probably due to negative CHIR-99021 ic50 influence from circulating CLL cells [12, 41]. Yet, in the present study, the function of αDC1 was not seemingly affected by contaminating CLL cells, indicating that this maturation

cocktail could overcome the possible suppressive effect from such cells and thus be effective even in a clinical setting. On the other hand, as all patients in our study were non-progressive and previously untreated, GNE-0877 it is conceivable that the negative influence of the CLL cells is much less than in patients with progressive disease. Indeed, patients with advanced CLL have an upregulated expression of immunosuppressive cytokines, including IL-10 and TGF-β, which suppresses Th1 cell immune responses [42]. In support of this, we have previously

shown that patients with advanced disease had higher expression of inhibitory killer immunoglobulin-like receptors (KIR) on CD8+ cells than patients with non-progressive disease, indicating that also potential tumour-reactive CTL is inhibited by tumour-related causes in patients with progressive disease [43]. Accordingly, one possible interpretation could be that for DC-based immunotherapy to be successful, it should reasonably be administered to patients with non-progressive disease, before immunosuppression caused both by the disease itself and possibly by chemotherapy, makes this approach impossible. In conclusion, we found that tumour-loaded αDC1 derived from patients with CLL produced substantially higher levels of NK/NKT/CD8+ cell-recruiting chemokines and that they were superior to PGE2DC in the recruitment of NK and NKT cells. Instead, PGE2DCs produced higher levels of Th2- and Treg-attracting chemokines.

IgG concentrations prior to and at the time of rituximab

correlated with nadir IgG. IgG replacement was initiated because of recurrent infection in 12 (4.2%) patients and a lower IgG increased the odds ratio of receiving IgG replacement. IgG replacement therapy decreased the incidence and severity of infections, and recovery of IgG concentrations allowed cessation of IgG replacement in two patients after 4 and 7.5 years of replacement treatment. Conclusions: Monitoring of IgG is recommended for patients receiving rituximab. IgG replacement for sustained hypogammaglobulinaemia with recurrent selleckchem infections appears effective. The IgG treatment course is prolonged in most patients, but IgG recovery is reported. 175 PARENTAL PERSPECTIVES ON THE FINANCIAL IMPACT OF CARING FOR A CHILD WITH CHRONIC KIDNEY DISEASE M MEDWAY1,2, Nutlin-3a mw A TONG1,2, JC CRAIG1,2,

S KIM2, F MACKIE3, S MCTAGGART4, B BARTON5, K HOWARD1, G WILLIAMS1,2, A WALKER6, G WONG1,2 1Sydney School of Public Health, The University of Sydney, Sydney, NSW; 2Centre for Kidney Research, The Children’s Hospital at Westmead, Sydney, NSW; 3Department of Nephrology, Sydney Children’s Hospital, Sydney, NSW; 4Department of Nephrology, Royal Children’s Hospital, Brisbane, QLD; 5Children’s Hospital Education Research Institute, The Children’s Hospital at Westmead, Sydney, NSW; 6Department of Nephrology, Royal Children’s Hospital, Melbourne, Victoria, Australia Aim: This study aims to describe parental perspectives on the financial impact of caring for a child with CKD. Background: Chronic kidney disease (CKD) can impose a significant social

and financial burden on patients and caregivers, however little is known about how caregivers experience and cope with the financial impact of CKD. Methods: Face-to-face semi-structured interviews were conducted with 27 parents of children with CKD across three centres in New South Wales and Queensland. Transcripts were thematically analysed. Results: We identified five themes: loss of freedom (prioritizing demands of care, limiting occupational opportunities, appreciating socio-economic advantage); burden of sole responsibility (inability to rely HAS1 on others, lack of respite, increased separation of family roles, self-reliance); adapting for survival (vigilant budgeting, redefining normality and expectations, rechanneling resources to basic needs, exploring new income sources, negotiating work flexibility); instability of circumstances (depleted capacity to work, unpredictability of child’s health, burden of travel-related costs, imposition of debt, domestic upheaval); and struggle in seeking support (‘falling through the cracks’, unmet information needs). Conclusions: Parents experienced meeting the complex needs of their child with CKD as an overwhelming focus, which they believed consumed much of their resources, time and energy.

Therefore, syk−/− DT40 B-cell mutants were reconstituted with a OneStrep-tagged version of human Syk and left untreated or stimulated through their BCR for six different time points. Cellular lysates were incubated with a streptactin affinity column and obtained proteins were size-separated by 1-D PAGE. Following in-gel digestion of Syk with endoproteinase trypsin, resulting Alpelisib phosphopeptide products were enriched by TiO2 microcolumns and identified by liquid chromatography (LC)-coupled tandem mass spectrometry (MS/MS) on an orbitrap mass

spectrometer. As shown in Fig. 1, we detected a total of 32 phosphoacceptor sites, 15 of which were on tyrosine, 11 on serine and six on threonine (see Supporting Information data 1 for annotated MS/MS spectra). Our analysis confirmed

all previously published phosphorylation selleck inhibitor events and revealed 19 novel acceptor sites. Notably, almost half of the Syk phosphosites mapped to interdomain B (see Fig. 1) previously implicated in the control of Syk functions 2, 3. Our data show that Syk is extensively modified by phosphorylation on a large number of acceptor sites, which might act individually or in concert to regulate Syk function. To monitor the phosphorylation kinetics of individual acceptor sites we used a quantitative SILAC-based mass-spectrometric approach 29–31. DT40 cells expressing OneStrep-tagged Syk were metabolically labeled in SILAC medium containing arginine and lysine residues with incorporated light or heavy isotopes of carbon and nitrogen.

Different combinations yielded three types of SILAC media. Using 12C6,14N2-Lys and 12C6,14N4-Arg resulted in “light medium” while the combination of 13C6,15N2-Lys and 13C6,15N4-Arg dipyridamole yielded “heavy medium”. “Intermediate medium” was obtained by using 2D4,12C6,14N2-Lys and 13C6,14N4-Arg. Cells cultured in “light medium” were left untreated and those cultured in “intermediate” or “heavy medium” were BCR-stimulated for different time points. This setup had important consequences. Proteins or peptides derived from the differentially labeled cells can be distinguished in the mass spectrometer by virtue of their distinct absolute molecular masses and hence can unambiguously be assigned to one of the three stimulation conditions. Proteins were purified from the three cell cultures via streptactin affinity chromatography, pooled at a 1:1:1 ratio and separated by 1-D PAGE. The gel slice containing the three pools of Syk was excised and subjected to trypsin digestion. TiO2-enriched phosphopeptides were analyzed by LC-MS/MS and individually quantified using MaxQuant software 32. This strategy allowed an unbiased relative quantification of Syk phosphorylation in resting and stimulated cells. Altogether, we monitored the phosphorylation kinetics of 16 phosphosites, which we grouped into three categories (Fig. 2).

One-ml fractions were collected and analysed by SDS–PAGE. The fractions that contained the desired proteins at the expected size were combined and desalted using PD-10 columns (Amersham-Pharmacia) equilibrated in PBS. Purification of recombinant Rv3619c protein.  The induction of expression and preparation of cell-free

extract from E. coli BL-21 cells carrying the plasmid pGES-TH/Rv3619c was carried out as described above for Rv3874 and Rv3875. The GST-Rv3619c fusion protein was recovered in the inclusion bodies as pellet, and therefore it was solubilized in successively higher concentrations XAV-939 of urea in phosphate-buffered saline (PBS), as described previously [20, 25]. Most of the fusion protein was recovered Selleckchem Y27632 in 4 m urea and was purified by affinity chromatography on glutathione-Sepharose column after proteolytic digestion of the column-bound

fusion protein with thrombin protease, as described previously [16, 25]. The fractions that contained the desired protein at the expected size were combined and analysed for purity by 15% SDS–PAGE gels, as described previously [24]. Raising polyclonal antibodies against recombinant proteins in rabbits.  Polyclonal antibodies were raised in rabbits against the purified and GST-free Rv3874, Rv3875 and Rv3619c recombinant proteins according to standard procedures [26]. In brief, purified proteins (50 μg/ml) were emulsified with an equal volume of incomplete Freund’s TCL adjuvant (Sigma) and injected intramuscularly in the right and left thigh. The rabbits were boosted twice with the same amount of protein at 2 weeks intervals. The animals were bled from the ear before the immunization and 2 weeks after the last immunization. The sera were tested for antigen-specific antibodies using 15% SDS–PAGE gels, as described previously [26]. Enzyme-linked immunosorbent assay (ELISA).  ELISA was performed to detect antibodies in rabbit

sera against full-length purified recombinant proteins and overlapping synthetic peptides corresponding to each protein using standard procedures [32]. In brief, wells of 96-well PolySorb plates (Nunc, Rochester, NY, USA) were coated with antigens/peptides (10 μg/ml), blocked with the blocking buffer, incubated with the primary antibody (rabbit sera at 1:100) followed by secondary antibody (alkaline phosphatase-conjugated anti-rabbit immunoglobulin G) and addition of substrate for colour development, as described previously [33]. The colour intensity was measured by determining the optical density (OD) at 405 nm. Antigen-/peptide-coated wells in the presence of secondary antibody alone, i.e. without adding primary antibody, were used as negative controls. The results were expressed as E/C, which is defined as: E/C = OD in antigen-coated wells having primary and secondary antibodies/OD in antigen-/peptide-coated wells having secondary antibody alone. The values of E/C > 2 were considered positive. PCR using gene-specific primers and genomic DNA from M.

05), while antagonistic cytokines like IFN-γ were increased in acute phase of KD (P < 0.05) and reduced after therapy with IVIG (P < 0.05). These results suggest that aberrantly decreased levels of NKG2D expression on NK cells and CD8+T cells might be one of the factors

led to disturbed immunological function in patients with KD. PF-01367338 ic50 Cytokines milieu could be important factors causing reduced expression of NKG2D. Kawasaki disease (KD) is an acute systemic vasculitis that affects infants and children. At present, the pathogenesis of KD remains to be further investigated. However, there is a large body of evidence that immunological disturbances play a key role in the pathogenesis of KD. A great many studies have found that the levels of many proinflammatory cytokines such as tumour necrosis factor (TNF)-α and interleukin (IL)-6 are elevated in acute KD, but the mechanisms resulting in aberrant immune function or overexpression of proinflammatory cytokines are not completely clear [1-3]. NKG2D is a C-type lectin-like type II transmembrane glycoprotein. It expressed on immunocompetent cells, such as natural-killer (NK) cells, CD8+ cytotoxic lymphocytes (CD8+T), NKT cells and γδT cells and participates in the regulation of innate and adaptive immune response through enhancing their killing activity. It has been

demonstrated that NKG2D expression is induced Proteasome inhibition assay on NK cells and CD8+T cells by their activation [4-6]. Accumulated evidences suggest that peripheral CD8+T cells may be functionally suppressed in acute phase of KD. Previous studies have shown a reduction in the total number of CD8+T cells in the peripheral blood of KD patients [7]. However, the expression of NKG2D on NK cells and CD8+T cells in the acute phase of KD is still required to be investigated. In this study, flow cytometry (FCM) was used to detect the expression of NKG2A/NKG2D on CD8+T cells and CD3−CD56+ NK cells in patients with KD, both in the acute phase and after IVIG therapy. The cytokines regulating expression of NKG2D such as IL-1β, IL-6, TNF-α, IL-7, IL-12, IL-15, interferon (IFN)-γ selleck chemicals llc and transforming growth factor

(TGF)-β were also evaluated in this study. Aberrantly, decreased levels of NKG2D expression were found in acute phase of KD patients, suggesting that downregulation expression of NKG2D might be one of the factors led to disturbed immunological function in KD. Forty-six children with KD admitted to the Shenzhen Children Hospital between June 2011 and April 2012 were included in the study. The patients comprised 26 males and 20 females (mean age: 26.33 ± 23.82 months; age range: 2 months–5 years). The diagnosis was carried out according to the clinical criteria of the Kawasaki Disease Research Committee of Japan. Blood samples were obtained before treatment with 2 g/kg/day intravenous immunoglobulin (IVIG, mean duration of illness, 6.3 days; range, 3–12 days) and after IVIG treatment (mean duration of illness, 12.0 days; range, 8–20 days).

They are positive for B-cell markers and CD38, negative for CD138 (a marker for plasma cells; [45]) and exhibit low PNA-binding activity. Histology revealed the average number of IgG1+ memory B cells in splenic follicles to be comparable between wild-type and mutant mice with conditional deletion of Bcl6 in B cells, consistent with the results of the flow cytometric analysis. These results suggest that in the spleen both unmutated and mutated memory B cells mostly localize to B-cell follicles. In wild-type mice, large numbers of GC B cells accumulate mutations in their Ig

VH genes by day 7 after immunization Palbociclib [2]. In parallel with GC development, studies of the frequency of mutated VH genes among IgG1 memory B cells showed an increase from ≤5% at day 7 to 50–60% at day 40 after immunization. There was no significant increase in the frequency of mutated cells in the memory B-cell population from day 40 to day 100 after immunization [2]. This observation supports the notion that memory B cells that develop independently of GCs within the first week of the response are maintained for a long period, and

then are joined by mutated GC B-cell progeny as the immune response progresses. Despite the recruitment of substantial numbers of GC B-cell progeny into the memory compartment over time, the absolute numbers of memory cells were similar between normal wild type mice and mice in which the GC response was ablated by Bcl6 U0126 deletion. These results indicate that the splenic environment has a limited capacity to sustain memory B cells. We speculate that GC-dependent and -independent memory B cells compete for hypothetical “niches” in the spleen for survival. It seems unlikely that competition for mafosfamide antigen is the determining factor in this process, since memory B cells persist over a long period in the apparent absence of immunizing antigen,

under competitive conditions [44]. The postulated niches for memory B cell maintenance in peripheral lymphoid organs may thus serve some trophic function. For over two decades, GCs have been considered to be the sole site for memory B-cell generation. Recent studies challenge this dogma and demonstrate that memory B cells can also develop before the onset and independently of the GC reaction (Fig. 2). How important are such low-affinity memory B cells and their immune response for protective immunity? We have recently shown that IgG1 memory B cells proliferate in response to antigen re-exposure and accumulate somatic mutations in their rearranged VH genes [10], regardless of whether they express unmutated or mutated VH genes. In this process, unmutated memory B cells generate large numbers of progeny expressing somatically mutated antibodies with high affinity for the antigen to which they responded.

After 6 months treatment the ARB treatment group had a reduced albumin excretion rate and ACR, while the ACEi was higher.94 However, the baseline conditions differed between treatment groups and the majority of individuals were normoalbuminuric thus the relevance of the outcomes for individuals with microalbuminuria is questionable. The GEMINI trial involved 1235 Selleck RAD001 people with type 2 diabetes with elevated BP under either an ACEi or ARB hypertension

treatment randomized for treatment with two different β-blockers (carvedilol and metoprolol).95 A post hoc analysis of differential effects of the β-blockers on the progression of albuminuria indicated a greater reduction in microalbuminuria for carvedilol compared with metoprolol. In those with normoalbuminuria fewer progressed to microalbuminuria on carvedilol. These 5-Fluoracil concentration effects were not related to BP. Multivariate analysis demonstrated only baseline urine ACR and treatment were significant predictors of changes in albuminuria. In a separate analysis the presence of metabolic syndrome at baseline corresponded with an OR of 2.68 (95% CI: 1.36–5.30) over the duration of the study. The DETAIL study involved 250 people with type 2

diabetes with mild to moderate hypertension and eGFR ≥ 70 mL/min per 1.73 m2 from 6 European countries.96 The study compared an ARB and an ACEi treatment over 5-years. After 5 years the difference in eGFR between the ARB and the ACEi was −3.1 mL/min per 1.73 m2 and was insignificant. The mean annual declines in eGFR were 3.7 mL/min per 1.73 m2 for the ARB and 3.3 mL/min per 1.73 m2 for the ACEi. These results were considered by the authors to be similar to eGFR decline reported in the IRMA 2, IDNT, and RENAAL studies and compare to an expected untreated type 2 diabetes the annual decline in the order of 10 mL/min per 1.73 m2. Telmisartan was

concluded to be not inferior to enalapril in providing long-term renoprotection. However, the results do not necessarily apply to more advanced nephropathy but support clinical equivalence of ARB and ACEi in persons with conditions that place them at high risk for CV events. The large ONTARGET trial comparing ARB and ACEi of in excess of 25 000 participants included a large proportion with diabetes and microalbuminuria.97 Relevant secondary outcomes are kidney impairment and kidney failure requiring dialysis. The only significant differences between treatments (ACEi, ARB and ACEi + ARB) were for increased kidney impairment in the combination therapy compared with the ACEi. Further analysis of renal outcomes,98 indicated a significantly higher increase in ACR in the ACEi treatment group compared with the ARB and ACEi + ARB (31% vs 24% and 21%). The risk of developing new microalbuminuria was not different between ACEi and ARB treatment groups, but was significantly lower in the combination treatment group.

This may result in their aberrant activation or prevent their survival

if their endogenous ligands were no longer present. Therefore identifying such adipose-resident lipid antigens would provide immense insight into the physiological basis of iNKT cell accumulation in adipose tissue and a potential pathway that could be manipulated to prevent their loss in obesity. Whether or not targeting iNKT cells in the clinic would result in meaningful clinical effects on diabetes and weight loss remains to be seen, but pursuing this avenue seems well justified. Lipid antigens that target iNKT cells, as well as other bioactive lipids, have been used clinically to treat patients with cancer. They are also the subject of many clinical trials for various BMS-777607 concentration cancers, including melanoma and prostate cancer, as well as autoimmune diseases. Lipid-based drugs for therapeutics for other purposes are also available. Furthermore, studies have shown that lipids given parenterally can activate iNKT cells, making the idea of targeting adipose iNKT cells in obesity a promising

and viable strategy. AZD1208 in vitro Adipose iNKT cells represent a unique iNKT cell population, which appear to be poised towards anti-inflammatory cytokine production. Whether anti-inflammatory iNKT cells are destined to migrate to adipose tissue from the thymus, or whether adipose tissue influences their phenotype and function remains to be seen. Nevertheless, the recent surge of reports on adipose iNKT cells have revealed one of

the clearest examples of the regulatory function of an iNKT cell population, indicating that they maintain healthy adipose tissue under normal conditions and correct obesity and metabolic disorder when stimulated under high fat diet conditions.[3, 39, 57, 58, 7] In keeping with their role as a bridge between the innate and adaptive immune systems, iNKT cells seem to be one of the first cells Liothyronine Sodium that are affected by obesity, even as early as a few days after commencing an HFD. Therefore, analogous to their key role in autoimmune diseases including type 1 diabetes, multiple sclerosis and systemic lupus erythematosus, and in various cancers, iNKT cells are also early and key players in the immune regulation of metabolism. It is likely that future studies will reveal the mechanism by which iNKT cells are lost in obesity, which may provide insight into how to prevent this loss and a greater understanding of the basis of their accumulation in adipose tissue. It is hoped that adipose lipid antigen(s), if any, will be identified, which would no doubt be very beneficial to answering some of these outstanding questions. We are very grateful to Professors Michael Brenner, Mark Exley, Donal O’Shea and Cliona O’Farrelly for insight and helpful discussions. The author has nothing to disclose.

, 2005). Whilst reductions in bacterial

susceptibility have been observed following repeated exposure (Perron et al., 2003), HDPs are reportedly less likely to induce bacterial resistance, in comparison with conventional antibiotics (Steinberg et al., 1993; Ge et al., 1999; Mosca et al., 2000). Human salivary HDPs comprise various short-chain peptides that are commonly associated with mucosal surfaces and which exhibit broad-spectrum antimicrobial activity. They have been classified into three subcategories: defensins, histatins and cathelicidin LL37 (Boman, 2000); defensins and cathelicidins constitute < 1% of total salivary proteins, whilst histatins constitute c. 5% (van Nieuw Amerongen et al., 2009). find protocol Oral HDPs are derived from various sources (Fig. 1) including neutrophils, which produce human neutrophil proteins (HNPs) (Selsted et al., 2009, 1992); gingival epithelia, which produce β defensins (Krisanaprakornkit et al., 1998); and salivary glands that secrete Everolimus in vivo histatins (Imamura et al., 2009) (Fig. 1). HDP production levels may vary in a stimulus-dependent manner (reviewed by Dale & Fredericks, 2005; Gorr & Abdolhosseini, 2011) and their

biological functions include chemotaxis, where HNPs 1 and 2 for example attract monocytes (Territo et al., 2000); stimulation of epithelial cell turnover during wound healing; neutralization of bacterial lipopolysaccharides; antiviral activity, and direct antibacterial effects (Dimond et al., 2009). Antibacterial activity occurs by interaction with the cell envelope, causing progressive leakage of cell contents (Zasloff, 2002).

Previous investigations into the antimicrobial activity of human oral HDPs suggest that they exhibit varying potency against oral bacteria when pure cultures are exposed in endpoint susceptibility tests (Hancock & Devine, 2004; Dimond et al., 2009). The role of HDPs in influencing the microbiological composition Carnitine dehydrogenase of the oral microbiota has been suggested by investigations of human subjects. For example, a single nucleotide polymorphism in Type I diabetics which reduces the efficacy of β defensin 1 has been associated with increased carriage of the pathogenic yeasts Candida glabrata and Candida tropicalis (Jurevic et al., 2003), whilst salivary proteomics of diabetic children has revealed a lack of histatins, which has been correlated with an increased periodontitis incidence (Cabras et al., 2010). In Chediak–Higashi syndrome, where individuals lack neutrophil azurophilic granules (a major source of HDPs), elevated susceptibility to bacterial and fungal infections has been reported (Ganz et al., 1988). Finally, levels of LL37 increase in response to the progression of periodontitis, and this HDP may therefore act as an inducible protective factor (Turkoglu et al., 1989).

The aim of preoperative urodynamic examination for POP surgery patients is to estimate LUT function. Videourodynamic examination for POP patients provides accurate information about morphological findings of the bladder and urethra, and LUT function. Morphological finding is informative and impressive for the physician and patient. Chain cystogram can precisely evaluate the anatomical relationship of the bladder and urethra. The advantage of videourodynamic examination is that it can simultaneously evaluate morphological and functional findings.

Preoperative urodynamic evaluation of SUI and detrusor function was useful for predicting postoperative urinary conditions in POP patients.3 Preoperative impaired detrusor contractility seems to be related to postoperative voiding difficulties.2 In our study, four patients needed CIC due to failure to empty after TVM with TOT placement. Cisplatin mw In three of these patients PFS was not applicable due to inability to void during urodynamic examination, and in one patient the evaluation of PFS was weak- detrusor. Four patients developed SUI after TVM without TOT placement. Three patients had UDS SUI, while the other patient had no UDS SUI. All 4 patients required postoperative additional TOT placement. Preoperative UDS SUI seems not to be

an absolute indication for combined TVM and TOT placement. UDS SUI was detected in the majority of 22 patients at cough maneuver in the standing position among

four conditions. LPP ACP-196 at cough maneuver in the standing position had a highest value of 91.7 cm H2O among LPP in four conditions. LPP measurement at cough maneuver in the standing position is important to detect UDS SUI. Observation of SUI during urodynamic examination with prolapse reduction by gauze pack or ring pessary was not positive in all 19 patients. SUI was not observed at prolapse reduction by gauze pack in four patients. Prolapse reduction C1GALT1 procedure is not perfect for the detection of SUI. To detect unmasked SUI due to POP, absolute value of LPP is not important, but specialized physical examination including cough test in the standing position with reduction by gauze packing or pessary in the vagina is recommended. Videourodynamic examination for POP patients provides accurate information about morphological findings of the bladder and urethra, and LUT function. LPP measurement at cough maneuver in the standing position is important to detect UDS SUI. Prolapse reduction procedure is not perfect for the detection of SUI. The authors declare no conflict of interest. “
“Objectives: Low power diode Iaser (830 nm) irradiation is a useful analgesic tool in superficial pain. Pulse laser irradiation allows us to increase the laser power because the non-irradiation time reduces heating effects and/or direct tissue damage at the irradiation area.