Zinc transporter 8 (ZnT8) autoantibody epitope specificity and affinity examined with recombinant ZnT8 variant proteins in specific ZnT8R and ZnT8W autoantibody-positive type 1 diabetes patients.

Variant-specific zinc transporter 8 autoantibodies (ZnT8A) against either arginine (R) or tryptophan (W) at amino acid (aa) position 325 of the zinc transporter 8 (ZnT8) has been identified in type 1 diabetes (T1D) patients. Reciprocal cross-over tests revealed differences in half-maximal binding to indicate variable affinity of patient ZnT8 autoantibodies. Insufficient recombinant ZnT8 variant proteins have precluded detailed analyses of ZnT8 autoantibody affinity. The aims in the present study were to (i) generate recombinant ZnT8R- and ZnT8W-aa275-369 proteins; (ii) test the ZnT8R- and ZnT8W-aa275-369 proteins in reciprocal competitive radiobinding assays (RBA) against ZnT8R- and ZnT8W-aa268-369 labelled with (35) S-methionine; and (iii) determine the specificity and affinity of sera specific for either ZnT8 arginine (R) or ZnT8 tryptophan (W) autoantibodies in newly diagnosed T1D patients. The results demonstrate, first, that it was possible to produce recombinant human MBP-ZnT8-aa275-369 protein purified to homogeneity for RBA reciprocal competition experiments. Secondly, high-titre ZnT8WA sera diluted to half maximal binding showed significant specificity for respective variants of either ZnT8R or ZnT8W. Thirdly, ZnT8WA-positive sera showed high affinity for ZnT8W compared to ZnT8RA for ZnT8R. These data demonstrate that T1D patients may have single amino acid-specific autoantibodies directed against either ZnT8R or ZnT8W and that the autoantibody affinity to the respective variant may be different. Further studies are needed to assess the mechanisms by which variant-specific ZnT8A of variable affinity develop and their possible role in the pathogenic process leading to the clinical onset of T1D.

Serological evaluation of possible exposure to Ljungan virus and related parechovirus in autoimmune (type 1) diabetes in children.

Exposure to Ljungan virus (LV) is implicated in the risk of autoimmune (type 1) diabetes but possible contribution by other parechoviruses is not ruled out. The aim was to compare children diagnosed with type 1 diabetes in 2005-2011 (n = 69) with healthy controls (n = 294), all from the Jämtland County in Sweden, using an exploratory suspension multiplex immunoassay for IgM and IgG against 26 peptides of LV, human parechoviruses (HPeV), Aichi virus and poliovirus in relation to a radiobinding assay (RBA) for antibodies against LV and InfluenzaA/H1N1pdm09. Islet autoantibodies and HLA-DQ genotypes were also determined. 1) All five LV-peptide antibodies correlated to each other (P < 0.001) in the suspension multiplex IgM- and IgG-antibody assay; 2) The LV-VP1_31-60-IgG correlated with insulin autoantibodies alone (P = 0.007) and in combination with HLA-DQ8 overall (P = 0.022) as well as with HLA-DQ 8/8 and 8/X subjects (P = 0.013); 3) RBA detected LV antibodies correlated with young age at diagnosis (P < 0.001) and with insulin autoantibodies (P < 0.001) especially in young HLA-DQ8 subjects (P = 0.004); 4) LV-peptide-VP1_31-60-IgG correlated to RBA LV antibodies (P = 0.009); 5) HPeV3-peptide-IgM and -IgG showed inter-peptide correlations (P < 0.001) but only HPeV3-VP1_1-30-IgG (P < 0.001) and VP1_95-124-IgG (P = 0.009) were related to RBA LV antibodies without relation to insulin autoantibody positivity (P = 0.072 and P = 0.486, respectively). Both exploratory suspension multiplex IgG to LV-peptide VP1_31-60 and RBA detected LV antibodies correlated with insulin autoantibodies and HLA-DQ8 suggesting possible role in type 1 diabetes. It remains to be determined if cross-reactivity or concomitant exposure to LV and HPeV3 contributes to the seroprevalence.

Doubly Reactive INS-IGF2 Autoantibodies in Children with Newly Diagnosed Autoimmune (type 1) Diabetes.

The splice variant INS-IGF2 entails the preproinsulin signal peptide, the insulin B-chain, eight amino acids of the C-peptide and 138 unique amino acids from an ORF in the IGF2 gene. The aim of this study was to determine whether levels of specific INS-IGF2 autoantibodies (INS-IGF2A) were related to age at diagnosis, islet autoantibodies, HLA-DQ or both, in patients and controls with newly diagnosed type 1 diabetes. Patients (n = 676), 0-18 years of age, diagnosed with type 1 diabetes in 1996-2005 and controls (n = 363) were analysed for specific INS-IGF2A after displacement with both cold insulin and INS-IGF2 to correct for non-specific binding and identify double reactive sera. GADA, IA-2A, IAA, ICA, ZnT8RA, ZnT8WA, ZnT8QA and HLA-DQ genotypes were also determined. The median level of specific INS-IGF2A was higher in patients than in controls (P < 0.001). Irrespective of age at diagnosis, 19% (126/676) of the patients had INS-IGF2A when the cut-off was the 95th percentile of the controls (P < 0.001). The risk of INS-IGF2A was increased among HLA-DQ2/8 (OR = 1.509; 95th CI 1.011, 2.252; P = 0.045) but not in 2/2, 2/X, 8/8, 8/X or X/X (X is neither 2 nor 8) patients. The association with HLA-DQ2/8 suggests that this autoantigen may be presented on HLA-DQ trans-heterodimers, rather than cis-heterodimers. Autoantibodies reactive with both insulin and INS-IGF2A at diagnosis support the notion that INS-IGF2 autoimmunity contributes to type 1 diabetes.

Antibodies to islet cell autoantigens, rotaviruses and/or enteroviruses in cord blood and healthy mothers in relation to the 2010-2011 winter viral seasons in Israel: a pilot study.

AIMS: To determine whether antivirus and/or islet cell antibodies can be detected in healthy pregnant mothers without diabetes and/or their offspring at birth in two winter viral seasons. METHODS: Maternal and cord blood sera from 107 healthy pregnant women were tested for islet cell autoantibodies using radioligand binding assays and for anti-rotavirus and anti-CoxB3 antibody using an enzyme-linked immunosorbent assay. RESULTS: Glutamic acid decarboxylase (GAD)65 autoantibodies and rotavirus antibodies, present in both maternal and cord blood sera, correlated with an odds ratio of 6.89 (95% CI: 1.01-46.78). For five, 22 and 17 pregnancies, antibodies to GAD65, rotavirus and CoxB3, respectively, were detected in cord blood only and not in the corresponding maternal serum. In 10 pregnancies, rotavirus antibody titres in the cord blood exceeded those in the corresponding maternal serum by 2.5-5-fold. Increased antibody titres after the 20(th) week of gestation suggested CoxB3 infection in one of the 20 pregnancies and rotavirus in another. CONCLUSION: The concurrent presence of GAD65 antibodies in cord blood and their mothers may indicate autoimmune damage to islet cells during gestation, possibly caused by cross-placental transmission of viral infections and/or antivirus antibodies. Cord blood antibody titres that exceed those of the corresponding maternal sample by >2.5-fold, or antibody-positive cord blood samples with antibody-negative maternal samples, may imply an active in utero immune response by the fetus.

Graphite foils as potential skin and epithelium dosimeters at therapeutic photon energies.

This work builds upon a prior study, examining the dosimetric utility of pencil lead and thin graphitic sheets, focusing upon the measurement of skin doses within the mammographic regime. In recognizing the near soft-tissue equivalence of graphite and the earlier-observed favourable thermoluminescence yield of thin sheets of graphite, this has led to present study of 50 µm thick graphite for parameters typical of external beam fractionated radiotherapy and skin dose evaluations. The graphite layers were annealed and then stacked to form an assembly of 0.5 mm nominal thickness. Using a 6 MV photon beam and delivering doses from 2- to 60 Gy, irradiations were conducted, the assembly first forming a superficial layer to a solid water phantom and subsequently underlying a 1.5 cm bolus, seeking to circumvent the build-up to electronic equilibrium for skin treatments. Investigations were made of several dosimetric properties arising from the thermoluminescence yield of the 50 µm thick graphite slabs, in particular proportionality and sensitivity to dose. The results show excellent sensitivity within the dose range of interest, the thermoluminescence response varying with increasing depth through the stacked graphite layers, obtaining a coefficient of determination of 90%. Acknowledging there to be considerable challenge in accurately matching skin thickness with dose, the graphite sheets have nevertheless shown considerable promise as dosimeters of skin, sensitive in determination of dose from the surface of the graphite through to sub-dermal depth thicknesses.

Antigenicity and epitope specificity of ZnT8 autoantibodies in type 1 diabetes.

The single nucleotide polymorphism (SNP) rs13266634 encodes either an Arginine (R) or a Tryptophan (W) (R325W) at the amino acid position 325 in the Zinc Transporter 8 (ZnT8) protein. Autoantibodies (Ab) that recognize ZnT8R, ZnT8W or both at the polymorphic site are common in newly diagnosed type 1 diabetes (T1D) patients. The epitope specificity and affinity of ZnT8Ab are poorly understood, but may be of importance for the prediction and clinical classification of T1D. Therefore, the aims were to 1) determine the immunogenicity of short (318-331) ZnT8 peptides in mice and 2) test the affinity of short and long (268-369) ZnT8 proteins in T1D patients positive for either ZnT8RAb or ZnT8wAb. Sera from BALB/cByJ mice immunized with short R, W or Q (Glutamine) ZnT8 peptides were tested for ZnT8-peptide antibodies in ELISA and radiobinding assay (RBA). Using reciprocal permutation experiment, short synthetic ZnT8R and ZnT8W (318-331) and long in vitro transcription translation ZnT8R and ZnT8W (268-369) proteins were tested in competitive RBA with R- and W-monospecific T1D sera samples. All mouse sera developed non-epitope-specific peptide antibodies in ELISA and only 6/12 mice had ZnT8-RWQ antibodies in RBA. Both long ZnT8R and ZnT8W (268-369), but not any short, proteins displaced labelled ZnT8 (268-369) proteins in binding to T1D ZnT8Ab-specific sera. The reciprocal cross-over tests showed that half-maximal displacement varied 2- to 11-fold indicating variable affinity of patient ZnT8Ab, signifying crucial autoantibody epitope spreading. The present approach should make it possible to dissect the importance of the R325W ZnT8 autoantigen epitope in the T1D pathogenesis.

Triple specificity of ZnT8 autoantibodies in relation to HLA and other islet autoantibodies in childhood and adolescent type 1 diabetes.

OBJECTIVE: To establish the diagnostic sensitivity of and the relationships between autoantibodies to all three Zinc transporter 8 (Zinc transporter 8 autoantibody to either one, two, or all three amino acid variants at position 325, ZnT8A) variants to human leukocyte antigen (HLA)-DQ and to autoantibodies to glutamic acid decarboxylase (GADA), insulinoma-associated protein 2 (IA-2A), and insulin (IAA). METHODS: We analyzed 3165 patients with type 1 diabetes (T1D) in the Better Diabetes Diagnosis study for HLA-DQ genotypes and all six autoantibodies (ZnT8RA, arginine 325 Zinc transporter 8 autoantibody; ZnT8WA, tryptophan 325 Zinc transporter 8 autoantibody; ZnT8QA, glutamine 325 Zinc transporter 8 autoantibody; GADA, IA-2A, and IAA). RESULTS: ZnT8A was found in 65% of the patients and as many as 108 of 3165 (3.4%) had 1-3 ZnT8A alone. None had ZnT8QA alone. Together with GADA (56%), IA-2A (73%), and IAA (33%), 93% of the T1D patients were autoantibody positive. All three ZnT8A were less frequent in children below 2 yr of age (p < 0.0001). All three ZnT8A were associated with DQA1-B1*X-0604 (DQ6.4) and DQA1-B1*03-0302 (DQ8). ZnT8WA and ZnT8QA were negatively associated with DQA1-B1*05-02 (DQ2). CONCLUSIONS: Analysis of ZnT8A increased the diagnostic sensitivity of islet autoantibodies for T1D as only 7% remained islet autoantibody negative. The association between DQ6.4 and all three ZnT8A may be related to ZnT8 antigen presentation by the DQ6.4 heterodimer.

3-T MRI with phased-array surface coil in the local staging of rectal cancer.

PURPOSE: This study sought to evaluate the diagnostic accuracy of surface-coil 3T magnetic resonance (MR) imaging in the preoperative study of patients with rectal cancer. MATERIALS AND METHODS: Thirty patients with histologically proven rectal cancer underwent surface-coil 3T MR imaging with sagittal, paracoronal and para-axial T2-weighted turbo spin echo (TSE) sequences. Slice thickness was 3 mm without gap, field of view 24 cm, matrix 400 × 512. Images were assessed for infiltration of the rectal wall, perirectal fat and pelvic structures. Tumours were staged according to the TNM system, and the MR imaging results were correlated with histopathology. RESULTS: In the patients who underwent MR imaging before and after radiotherapy (group 1), the diagnostic accuracy of 3T MR imaging was 88% for T2, 94% for T3 and 88% for T4 cancers. In those who underwent surgical treatment without preoperative radiotherapy (group 2), the diagnostic accuracy was 90% for T2, 87% for T3 and 87% for T4 cancers. CONCLUSIONS: The high signal-to-noise ratio coupled with a large field of view enables surface-coil 3T MR imaging to achieve high levels of diagnostic accuracy in the local staging of rectal cancer, and in particular in assessing infiltration of mesorectum and mesorectal fascia.

Myasthenia gravis associated with autoimmune thyroid disease: a report of two patients.

Myasthenia gravis (MG) is an acquired autoimmune disorder causing skeletal muscle fatigue and weakness. This is a report of one woman and her daughter presenting with myasthenia and gravis and Grave's disease. It highlights possible hereditary component of this condition which has not been commonly reported in our setting.

Magnetic actuation and feedback cooling of a cavity optomechanical torque sensor.

Cavity optomechanics has demonstrated remarkable capabilities, such as measurement and control of mechanical motion at the quantum level. Yet many compelling applications of optomechanics-such as microwave-to-telecom wavelength conversion, quantum memories, materials studies, and sensing applications-require hybrid devices, where the optomechanical system is coupled to a separate, typically condensed matter, system. Here, we demonstrate such a hybrid optomechanical system, in which a mesoscopic ferromagnetic needle is integrated with an optomechanical torsional resonator. Using this system we quantitatively extract the magnetization of the needle, not known a priori, demonstrating the potential of this system for studies of nanomagnetism. Furthermore, we show that we can magnetically dampen its torsional mode from room-temperature to 11.6 K-improving its mechanical response time without sacrificing torque sensitivity. Future extensions will enable studies of high-frequency spin dynamics and broadband wavelength conversion via torque mixing.

Factor H binds to washed human platelets.

BACKGROUND: Factor H regulates the alternative pathway of complement. The protein has three heparin-binding sites, is synthesized primarily in the liver and copurifies from platelets with thrombospondin-1. Factor H mutations at the C-terminus are associated with atypical hemolytic uremic syndrome, a condition in which platelets are consumed. Objectives The aim of this study was to investigate if factor H interacts with platelets. METHODS: Binding of factor H, recombinant C- or N-terminus constructs and a C-terminus mutant to washed (plasma and complement-free) platelets was analyzed by flow cytometry. Binding of factor H and constructs to thrombospondin-1 was measured by surface plasmon resonance. RESULTS: Factor H bound to platelets in a dose-dependent manner. The major binding site was localized to the C-terminus. The interaction was partially blocked by heparin. Inhibition with anti-GPIIb/IIIa, or with fibrinogen, suggested that the platelet GPIIb/IIIa receptor is involved in factor H binding. Factor H binds to thrombospondin-1. Addition of thrombospondin-1 increased factor H binding to platelets. Factor H mutated at the C-terminus also bound to platelets, albeit to a significantly lesser degree. CONCLUSIONS: This study reports a novel property of factor H, i.e. binding to platelets, either directly via the GPIIb/IIIa receptor or indirectly via thrombospondin-1, in the absence of complement. Binding to platelets was mostly mediated by the C-terminal region of factor H and factor H mutated at the C-terminus exhibited reduced binding.

Torque-mixing magnetic resonance spectroscopy.

A universal, torque-mixing method for magnetic resonance spectroscopy is presented. In analogy to resonance detection by magnetic induction, the transverse component of a precessing dipole moment can be measured in sensitive broadband spectroscopy, here using a resonant mechanical torque sensor. Unlike induction, the torque amplitude allows equilibrium magnetic properties to be monitored simultaneously with the spin dynamics. Comprehensive electron spin resonance spectra of a single-crystal, mesoscopic yttrium iron garnet disk at room temperature reveal assisted switching between magnetization states and mode-dependent spin resonance interactions with nanoscale surface imperfections. The rich detail allows analysis of even complex three-dimensional spin textures. The flexibility of microelectromechanical and optomechanical devices combined with broad generality and capabilities of torque-mixing magnetic resonance spectroscopy offers great opportunities for development of integrated devices.

Prospective evaluation of glutamine and phospholipids levels in first degree relatives of patients with Type 1 Diabetes from a multiethnic population.

BACKGROUND: A dysregulation in the metabolism of lipids may be an early marker of autoimmunity in Type 1 Diabetes (T1D). It would be of general importance to identify metabolic patterns that would predict the risk for T1D later in life. The aim of this study was to perform a prospective evaluation of glutamine and phospholipids levels in Brazilian first degree relatives (FDR) of patients with T1D in a mean interval of 5 years. FINDINGS: Brazilian FDR (n = 30) of patients with T1D were evaluated and blood was sampled to measure the levels of glutamine and phospholipids in the fasting serum by quantitative colorimetric method. The tests were repeated after a mean interval of 5 years and compared to a control group (n = 20). The FDR presented lower levels of phospholipids than controls (p = 0.028), but not of glutamine (p = 0.075). Phospholipids levels decreased over time (p = 0.028) in FDR and were associated with Glutamic acid decarboxylase autoantibody (GADA) titers (p = 0.045), autoantibody positivity (p = 0.008) and PTPN22 polymorphisms (p = 0.014). CONCLUSIONS: In this Brazilian multiethnic population, there was a significant decrease in phospholipids levels in FDR in patients with T1D during a 5-year prospective follow-up, as well as a significant association between these metabolite, GADA and PTPN22 polymorphisms. For Glutamine no difference was found. These findings suggest that a dysregulation in the metabolism of lipids may precede the onset of the autoimmunity in T1D.

Correlation among [h-3] thymidine, flow-cytometry and proliferating cell nuclear antigen indexes in colorectal tumors.

In colorectal cancer and other tumors, proliferating cell nuclear antigen (PCNA) has been found to be a useful marker in immunohistochemical studies of cell proliferation, The presence of a correlation among PCNA labeling index (PCNA LI) and values of cell proliferation expressed by [H-3]thymidine labeling index (TLI) and flow cytometry (FC), the most common techniques used in the evaluation of proliferative activity in colorectal cancer were studied. In 50 operable colorectal cancer patients, TLI, PCNA LI and determination of S-phase fraction by FC were carried out in order to evaluate the correlation among these three indices. A good correlation was obtained between PCNA LI and both TLI (r=0.667; P=0.0001) and percent of cells in S-phase as determined by FC (r=0.819; P=0.0001). It is concluded that PCNA immunohistochemistry can be a reliable marker of the proliferative activity in colorectal rumours. Furthermore, because of its technical simplicity and lower cost it can be more advantageous than TLI or FC.

Children's subjective identification with the group and in-group favoritism.

Recent developments in social psychology have explained children's preference for members of the in-group in terms of processes of self-categorization and identification with the in-group. In contrast, this study, addressing nationality self-conceptions, examines the possibility that even before subjective identification with the group has occurred, as de facto group members, children will have been exposed to a great deal of positive information about their own national group, which is likely to encourage group-serving judgments. Children who had failed to identify themselves as members of their national group were required in this study to make evaluative judgments about 5 national groups, including their own. Significant preference for the in-group emerged on 2 of 3 measures. It is concluded that subjective identification with the in-group is not a necessary precondition for in-group favoritism.

Contested identities and schisms in groups: opposing the ordination of women as priests in the Church of England.

Schisms constitute a common characteristic of human groups. Nevertheless, they have been neglected by social psychology, mainly because social psychological theories either dismiss group consensus or else depict groups as monolithic. This study proposes a social psychological approach to schism which integrates recent developments of self-categorization theory (SCT) with work on category argumentation. According to SCT, shared group identification leads to a process by which members should reach agreement. However, it is suggested that where members construe the positions of others as fundamentally altering group identity, then consensus is impossible. The corollary of assuming that groups will be consensual is that lack of consensus indicates the existence of different groups. This idea is examined through an analysis of a video and booklet produced for a rally organized by Forward in Faith, an organization opposed to the ordination of women as priests within the Church of England. It is shown that the existence of women priests is construed as changing the essence of the Church both on a structural level (by dividing it from the rest of the Christian community and turning it into a sect) and doctrinally. Such changes are seen as threatening the very existence of the Church of England and therefore demanding all out opposition. However, it is also shown that the decision of whether opponents fight the changes from inside the Church or by splitting from it depends upon the perception of whether they will be accorded the opportunity to advance their position from within. Thus schism is associated with both a perceived 'change of essence' and also with 'lack of voice'.

Skin cancer attitudes: a cross-national comparison.

A questionnaire concerning attitudes towards skin cancer, sun exposure and general environmental issues was administered to 132 holiday-makers on a beach in south-west England and (in translation) to 142 visitors to another beach in north-west Italy. Following the Janis & Mann (1977) classification of strategies for coping with decision conflicts, subscales were derived measuring tendencies to 'avoid' thinking about environmental issues, to 'bolster' prior attitudes (by playing down the seriousness of the risk of skin cancer while attending to the pleasures of sunbathing), and to be 'vigilant' concerning risk information and the need for specific protective behaviour (e.g. sunscreen use). The British scored higher than the Italians, and women higher than men, on vigilance, but there were no gender or nationality differences on the other subscales considered as a whole. Responses were also related to the covariates of age and self-reported vulnerability to sunburn. Those who showed less concern with environmental issues also tended to play down the risks of skin cancer and be less vigilant with regard to self-protection. It is suggested that health promotion should address both cultural norms concerning exposure to the sun and people's intuitive notions about their relative personal vulnerability.

Effect of carbon to nitrogen ratio on the composition of microbial extracellular polymers in activated sludge.

Effect of carbon to nitrogen ratio (C/N) on the sludge extracellular polymer composition is studied in synthetically fed semi-continuous reactors with 8 days of sludge age. Results show that C/N ratio influences the relative distribution of polymer carbohydrate and protein. At low C/N ratio of 5, polymer extracts have high protein and low carbohydrate content. As the C/N ratio is increased to 17.5 and then to 40, carbohydrate concentration increases sharply and protein concentration decreases.

Children's conceptions of characteristic features of category members.

The authors investigated children's conceptions of the characteristic features of category members and found that their conceptions underwent qualitative developmental changes. They hypothesized that (a) children initially tend to perceive category members in terms of individual characteristics (i.e., internal dispositions and behavioral patterns) and (b) only later do they conceive of category members in terms of their shared beliefs and values. They explored these hypotheses in 2 studies: In Study 1, they investigated how Protestant and Catholic children in Northern Ireland understood the religious intergroup context in their own country; in Study 2, children in Italy were presented with a fictional scenario of intergroup conflict and asked to explain the causes of the conflict. The results of both studies confirmed the 2 hypotheses.

[Efficacy of erythropoietin in anemia of patients with immuno-lymphoproliferative disease]. / Efficacia dell'eritropoietina nell'anemia dei pazienti con malattie immuno-linfoproliferative.

Anemia is a common complication of lymphoproliferative syndromes. The exact pathogenic mechanism of this anemia is unclear. Many patients require progressive and persistent blood transfusions. We treated 10 patients (8 with multiple myeloma, 1 with non Hodgkin Lymphoma, 1 with chronic lymphocytic leukemia) by administering low doses of recombinant human erythropoietin (60 U/kg 3 times a week s.c.). All patients presented anemia with hemoglobin levels less than 10 gr/dl; renal function was not impaired (serum creatinine levels less than 1.2 mg/dl or creatinine clearance greater than 60 ml/min). A response was defined as an increase of hemoglobin level of at least 2 gr/dl or stop of red-cell transfusion within the first 3 months of treatment. Nine patients (90%) responded to treatment with a significant increase in the hemoglobin concentration. Two patients presented a cerebral stroke not correlated with erythropoietin administration.