Prenatal Exclusion of Lamellar Ichthyosis Based on Identification of Two New Mutations in the Transglutaminase 1 Gene

Lamellar ichthyosis is a severe, generalized, autosomal recessive genodermatosis characterized clinically by large, parchment-like scales and histologically by acanthosis and marked hyperkeratosis. Genetic heterogeneity in lamellar ichthyosis has been recognized with reports of two linked loci (on chromosomes 14q11 and 2q33–35). In a cohort of four small families with lamellar ichthyosis we found confirmatory evidence for linkage (p ≤ 0.01) to D14S275, a microsatellite marker close to transglutaminase 1 on chromosome 14q11. We also identified two novel transglutaminase 1 mutations in an affected sibling pair from one of these families. The paternal mutation in exon 3, 1387insCAGC, causes a frameshift predicted to result in premature termination of translation within the same exon. The maternal mutation in exon 8, 4561delAC, also causes a frameshift and a premature stop codon in this exon. The mother of these siblings recently became pregnant with twins. Genotyping and direct sequencing of DNA isolated from fetal amniotic fluid cultures revealed the presence of the paternal but the absence of the maternal mutation, thus predicting a normal skin phenotype. Both twins were born with normal-appearing skin. Our findings demonstrate that mutations of both alleles of the transglutaminase 1 gene are the cause of lamellar ichthyosis in this family, and illustrate an emerging clinical application of molecular genetics in dermatology

CMR2009: 5.04: Post-mortem analysis of gadolinium distribution in NSF subjects

No Abstract.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/64915/1/323_ftp.pd

Luminescent osmium(II) bi-1,2,3-triazol-4-yl complexes: photophysical characterisation and application in light-emitting electrochemical cells

The series of osmium(II) complexes [Os(bpy)3-n(btz)n][PF6]2 (bpy = 2,2’-bipyridyl, btz = 1,1’-dibenzyl-4,4’-bi-1,2,3-triazolyl, 1 n = 0, 2 n = 1, 3 n = 2, 4 n = 3), have been prepared and characterised. The progressive replacement of bpy by btz leads to blue-shifted UV-visible electronic absorption spectra, indicative of btz perturbation of the successively destabilised bpy-centred LUMO. For 4, a dramatic blue-shift relative to the absorption profile for 3 is observed, indicative of the much higher energy LUMO of the btz ligand over that of bpy, mirroring previously reported data on analogous ruthenium(II) complexes. Unlike the previously reported ruthenium systems, heteroleptic complexes 2 and 3 display intense emission in the far-red/near-infrared (λmax = 724 and 713 nm respectively in aerated acetonitrile at RT) as a consequence of higher lying, and hence less thermally accessible, 3MC states. This assertion is supported by ground state DFT calculations which show that the dσ* orbitals of 1 to 4 are destabilised by between 0.60 and 0.79 eV relative to their Ru(II) analogues. The homoleptic complex 4 appears to display extremely week room temperature emission, but on cooling to 77 K the complex exhibits highly intense blue emission with λmax 444 nm. As complexes 1 to 3 display room temperature luminescent emission and readily reversible Os(II)/(III) redox couples, light-emitting electrochemical cell (LEC) devices were fabricated. All LECs display electroluminescent emission in the deep-red/near-IR (λmax = 695 to 730 nm). Whilst devices based on 2 and 3 show inferior current density and luminance than LECs based on 1, the device utilising 3 shows the highest external quantum efficiency at 0.3 %

Current Murine Models and New Developments in H3K27M Diffuse Midline Gliomas

Diffuse Midline Gliomas with Histone 3-Lysine-27-Methionine (H3K27M) mutation constitute the majority of Diffuse Intrinsic Pontine Glioma (DIPG), which is the most aggressive form of pediatric glioma with a dire prognosis. DIPG are lethal tumors found in younger children with a median survival <1 year from diagnosis. Discovery of the characteristic H3K27M mutations offers opportunity and hope for development of targeted therapies for this deadly disease. The H3K27M mutation, likely through epigenetic alterations in specific H3 lysine trimethylation levels and subsequent gene expression, plays a significant role in pathogenesis of DIPG. Animal models accurately depicting molecular characteristics of H3K27M DIPG are important to elucidate underlying pathologic events and for preclinical drug evaluation. Here we review the past and present DIPG models and describe our efforts developing patient derived cell lines and xenografts from pretreated surgical specimens. Pre-treated surgical samples retain the characteristic genomic and phenotypic hallmarks of DIPG and establish orthotopic tumors in the mouse brainstem that recapitulate radiographic and morphological features of the original human DIPG tumor. These models that contain the H3K27M mutation constitute a valuable tool to further study this devastating disease and ultimately may uncover novel therapeutic vulnerabilities

Combination CTLA-4 Blockade and 4-1BB Activation Enhances Tumor Rejection by Increasing T-Cell Infiltration, Proliferation, and Cytokine Production

BACKGROUND: The co-inhibitory receptor Cytotoxic T-Lymphocyte Antigen 4 (CTLA-4) attenuates immune responses and prevent autoimmunity, however, tumors exploit this pathway to evade the host T-cell response. The T-cell co-stimulatory receptor 4-1BB is transiently upregulated on T-cells following activation and increases their proliferation and inflammatory cytokine production when engaged. Antibodies which block CTLA-4 or which activate 4-1BB can promote the rejection of some murine tumors, but fail to cure poorly immunogenic tumors like B16 melanoma as single agents.METHODOLOGY/PRINCIPAL FINDINGS: We find that combining ?CTLA-4 and ?4-1BB antibodies in the context of a Flt3-ligand, but not a GM-CSF, based B16 melanoma vaccine promoted synergistic levels of tumor rejection. 4-1BB activation elicited strong infiltration of CD8+ T-cells into the tumor and drove the proliferation of these cells, while CTLA-4 blockade did the same for CD4+ effector T-cells. Anti-4-1BB also depressed regulatory T-cell infiltration of tumors. 4-1BB activation strongly stimulated inflammatory cytokine production in the vaccine and tumor draining lymph nodes and in the tumor itself. The addition of CTLA-4 blockade further increased IFN-? production from CD4+ effector T-cells in the vaccine draining node and the tumor. Anti 4-1BB treatment, with or without CTLA-4 blockade, induced approximately 75% of CD8+ and 45% of CD4+ effector T-cells in the tumor to express the killer cell lectin-like receptor G1 (KLRG1). Tumors treated with combination antibody therapy showed 1.7-fold greater infiltration by these KLRG1+CD4+ effector T-cells than did those treated with ?4-1BB alone.CONCLUSIONS/SIGNIFICANCE: This study shows that combining T-cell co-inhibitory blockade with ?CTLA-4 and active co-stimulation with ?4-1BB promotes rejection of B16 melanoma in the context of a suitable vaccine. In addition, we identify KLRG1 as a useful marker for monitoring the anti-tumor immune response elicited by this therapy. These findings should aid in the design of future trials for the immunotherapy of melanoma

Evaluating Surveillance Strategies for the Early Detection of Low Pathogenicity Avian Influenza Infections

In recent years, the early detection of low pathogenicity avian influenza (LPAI) viruses in poultry has become increasingly important, given their potential to mutate into highly pathogenic viruses. However, evaluations of LPAI surveillance have mainly focused on prevalence and not on the ability to act as an early warning system. We used a simulation model based on data from Italian LPAI epidemics in turkeys to evaluate different surveillance strategies in terms of their performance as early warning systems. The strategies differed in terms of sample size, sampling frequency, diagnostic tests, and whether or not active surveillance (i.e., routine laboratory testing of farms) was performed, and were also tested under different epidemiological scenarios. We compared surveillance strategies by simulating within-farm outbreaks. The output measures were the proportion of infected farms that are detected and the farm reproduction number (Rh). The first one provides an indication of the sensitivity of the surveillance system to detect within-farm infections, whereas Rh reflects the effectiveness of outbreak detection (i.e., if detection occurs soon enough to bring an epidemic under control). Increasing the sampling frequency was the most effective means of improving the timeliness of detection (i.e., it occurs earlier), whereas increasing the sample size increased the likelihood of detection. Surveillance was only effective in preventing an epidemic if actions were taken within two days of sampling. The strategies were not affected by the quality of the diagnostic test, although performing both serological and virological assays increased the sensitivity of active surveillance. Early detection of LPAI outbreaks in turkeys can be achieved by increasing the sampling frequency for active surveillance, though very frequent sampling may not be sustainable in the long term. We suggest that, when no LPAI virus is circulating yet and there is a low risk of virus introduction, a less frequent sampling approach might be admitted, provided that the surveillance is intensified as soon as the first outbreak is detected

Acoustic Correlates of Information Structure.

This paper reports three studies aimed at addressing three questions about the acoustic correlates of information structure in English: (1) do speakers mark information structure prosodically, and, to the extent they do; (2) what are the acoustic features associated with different aspects of information structure; and (3) how well can listeners retrieve this information from the signal? The information structure of subject-verb-object sentences was manipulated via the questions preceding those sentences: elements in the target sentences were either focused (i.e., the answer to a wh-question) or given (i.e., mentioned in prior discourse); furthermore, focused elements had either an implicit or an explicit contrast set in the discourse; finally, either only the object was focused (narrow object focus) or the entire event was focused (wide focus). The results across all three experiments demonstrated that people reliably mark (1) focus location (subject, verb, or object) using greater intensity, longer duration, and higher mean and maximum F0, and (2) focus breadth, such that narrow object focus is marked with greater intensity, longer duration, and higher mean and maximum F0 on the object than wide focus. Furthermore, when participants are made aware of prosodic ambiguity present across different information structures, they reliably mark focus type, so that contrastively focused elements are produced with greater intensity, longer duration, and lower mean and maximum F0 than noncontrastively focused elements. In addition to having important theoretical consequences for accounts of semantics and prosody, these experiments demonstrate that linear residualisation successfully removes individual differences in people's productions thereby revealing cross-speaker generalisations. Furthermore, discriminant modelling allows us to objectively determine the acoustic features that underlie meaning differences