A combined computational-experimental approach to define the structural origin of antibody recognition of sialyl-Tn, a tumor-associated carbohydrate antigen.

Anti-carbohydrate monoclonal antibodies (mAbs) hold great promise as cancer therapeutics and diagnostics. However, their specificity can be mixed, and detailed characterization is problematic, because antibody-glycan complexes are challenging to crystallize. Here, we developed a generalizable approach employing high-throughput techniques for characterizing the structure and specificity of such mAbs, and applied it to the mAb TKH2 developed against the tumor-associated carbohydrate antigen sialyl-Tn (STn). The mAb specificity was defined by apparent KD values determined by quantitative glycan microarray screening. Key residues in the antibody combining site were identified by site-directed mutagenesis, and the glycan-antigen contact surface was defined using saturation transfer difference NMR (STD-NMR). These features were then employed as metrics for selecting the optimal 3D-model of the antibody-glycan complex, out of thousands plausible options generated by automated docking and molecular dynamics simulation. STn-specificity was further validated by computationally screening of the selected antibody 3D-model against the human sialyl-Tn-glycome. This computational-experimental approach would allow rational design of potent antibodies targeting carbohydrates

Four-YearFollow-upof [F-18]Fluorodeoxyglucose Positron Emission Tomography-Based Parkinson's Disease-Related Pattern Expression in 20 Patients With Isolated Rapid Eye Movement Sleep Behavior Disorder Shows Prodromal Progression

Background: Isolated rapid eye movement sleep behavior disorder is known to be prodromal for alpha-synucleinopathies, such as Parkinson's disease (PD) and dementia with Lewy bodies. The [18F]fluorodeoxyglucose-positron emission tomography (PET)–based PD-related brain pattern can be used to monitor disease progression. Objective: We longitudinally investigated PD-related brain pattern expression changes in 20 subjects with isolated rapid eye movement sleep behavior disorder to investigate whether this may be a suitable technique to study prodromal PD progression in these patients and to identify potential phenoconverters. Methods: Subjects underwent two [18F]fluorodeoxyglucose-PET brain scans ~3.7 years apart, along with baseline and repeated motor, cognitive, and olfactory testing within roughly the same time frame. Results: At baseline, 8 of 20 (40%) subjects significantly expressed the PD-related brain pattern (with z scores above the receiver operating characteristic–determined threshold). At follow-up, six additional subjects exhibited significant PD-related brain pattern expression (70% in total). PD-related brain pattern expression increased in all subjects (P = 0.00008). Four subjects (20%), all with significant baseline PD-related brain pattern expression, phenoconverted to clinical PD. Conclusions: Suprathreshold PD-related brain pattern expression and greater score rate of change may signify greater shorter-term risk for phenoconversion. Our results support the use of serial PD-related brain pattern expression measurements as a prodromal PD progression biomarker in patients with isolated rapid eye movement sleep behavior disorder

A Novel, Multifunctional c-Cbl Binding Protein in Insulin Receptor Signaling in 3T3-L1 Adipocytes

The protein product of the c-Cbl proto-oncogene is prominently tyrosine phosphorylated in response to insulin in 3T3-L1 adipocytes and not in 3T3-L1 fibroblasts. After insulin-dependent tyrosine phosphorylation, c-Cbl specifically associates with endogenous c-Crk and Fyn. These results suggest a role for tyrosine-phosphorylated c-Cbl in 3T3-L1 adipocyte activation by insulin. A yeast two-hybrid cDNA library prepared from fully differentiated 3T3-L1 adipocytes was screened with full-length c-Cbl as the target protein in an attempt to identify adipose-specific signaling proteins that interact with c-Cbl and potentially are involved in its tyrosine phosphorylation in 3T3-L1 adipocytes. Here we describe the isolation and the characterization of a novel protein that we termed CAP for c-Cbl-associated protein. CAP contains a unique structure with three adjacent Src homology 3 (SH3) domains in the C terminus and a region showing significant sequence similarity with the peptide hormone sorbin. Both CAP mRNA and proteins are expressed predominately in 3T3-L1 adipocytes and not in 3T3-L1 fibroblasts. CAP associates with c-Cbl in 3T3-L1 adipocytes independently of insulin stimulation in vivo and in vitro in an SH3-domain-mediated manner. Furthermore, we detected the association of CAP with the insulin receptor. Insulin stimulation resulted in the dissociation of CAP from the insulin receptor. Taken together, these data suggest that CAP represents a novel c-Cbl binding protein in 3T3-L1 adipocytes likely to participate in insulin signaling

Symmetry breaking perturbations and strange attractors

The asymmetrically forced, damped Duffing oscillator is introduced as a prototype model for analyzing the homoclinic tangle of symmetric dissipative systems with \textit{symmetry breaking} disturbances. Even a slight fixed asymmetry in the perturbation may cause a substantial change in the asymptotic behavior of the system, e.g. transitions from two sided to one sided strange attractors as the other parameters are varied. Moreover, slight asymmetries may cause substantial asymmetries in the relative size of the basins of attraction of the unforced nearly symmetric attracting regions. These changes seems to be associated with homoclinic bifurcations. Numerical evidence indicates that \textit{strange attractors} appear near curves corresponding to specific secondary homoclinic bifurcations. These curves are found using analytical perturbational tools

Brucellosis remains a neglected disease inthe developing world: a call forinterdisciplinary action

Brucellosis places significant burdens on the human healthcare system and limits the economic growth of individuals, communities, and nations where such development is especially important to diminish the prevalence of poverty. The implementation of public policy focused on mitigating the socioeconomic effects of brucellosis in human and animal populations is desperately needed. When developing a plan to mitigate the associated consequences, it is vital to consider both the abstract and quantifiable effects. This requires an interdisciplinary and collaborative, or One Health, approach that consists of public education, the development of an infrastructure for disease surveillance and reporting in both veterinary and medical fields, and campaigns for control in livestock and wildlife species

Histomorphology of Salivary Glands after Ligature and Reconstruction of Common Carotid Artery in Rabbits

Introduction. In occlusive diseases of carotid arteries may be involved salivary glands which play important role in oral and general homeostasis.Aim of the study. The purpose of this study was histomorphological evaluation of rabbit’s salivary glands after regional reduction af arterial circulation and reconstruction of ligated common carotid arteries comparing with normal histological sight.Materials and methods. The experiments were authorized by the Animal Ethics Committee of the Latvian Food and Veterinary Service. 20 male Californian rabbits weighing approximately 3 kg were used. Under intravenous general anesthesia and local anesthesia the ligature of a. carotis communis dextra was done. After 28 days 10 rabbits were sacrificed and samples were taken from all major salivary glands and histological measured acinar dimensions and acinar epithelial cell.For rest 10 rabbits under the general anesthesia were performed revascularization of a. carotis communis dextra. After 28 days the rabbits were sacrificed with intrapulmonal injection of T61. The acini dimensions and acinar epithelial cell diameter in microns in all glands were measured except buccal glands were measured. Results. In ischemic conditions all glands react with decrease of acinar epithelial cell diameter and also diametr of functional unit of salivary gland.After revascularisation reestablishment of acini and secretory epithelial cells was observed. Conclusions. Rabbit’s salivary glands have potential to recovery in diminished after ligation of ipsilateral common carotid artery size of acini and secretory epithelium after 4 week ischemia if the blood supply is reestabilished.publishersversionPeer reviewe

Applicability of a short/rapid 13C-urea breath test for Helicobacter pylori: retrospective multicenter chart review study

<p>Abstract</p> <p>Background</p> <p>Carbon labeled urea breath tests usually entail a two point sampling with a 20 to 30-minute gap. Our aim was to evaluate the duration of time needed for diagnosing <it>Helicobacter pylori </it>by the BreathID^®System.</p> <p>Methods</p> <p>This is a retrospective multicenter chart review study. Test location, date, delta over baseline, and duration of the entire test were recorded. Consecutively ¹³C urea breath tests results were extracted from the files over a nine year period.</p> <p>Results</p> <p>Of the 12,791 tests results, 35.1% were positively diagnosed and only 0.1% were inconclusive. A statistically significant difference in prevalence among the countries was found: Germany showing the lowest, 13.3%, and Israel the highest, 44.1%. Significant differences were found in time to diagnosis: a positive diagnosis had the shortest and an inconclusive result had the longest. Overall test duration averaged 15.1 minutes in Germany versus approximately 13 minutes in other countries. Diagnosis was achieved after approximately 9 minutes in Israel, Italy and Switzerland, but after 10 on average in the others. The mean delta over baseline value for a negative diagnosis was 1.03 ± 0.86, (range, 0.9 - 5), versus 20.2 ± 18.9, (range, 5.1 - 159.4) for a positive one.</p> <p>Conclusions</p> <p>The BreathID^®System used in diagnosing <it>Helicobacter pylori </it>can safely shorten test duration on average of 10-13 minutes without any loss of sensitivity or specificity and with no test lasting more than 21 minutes.</p

Mitochondrial Mutations in Adenoid Cystic Carcinoma of the Salivary Glands

Background: The MitoChip v2.0 resequencing array is an array-based technique allowing for accurate and complete sequencing of the mitochondrial genome. No studies have investigated mitochondrial mutation in salivary gland adenoid cystic carcinomas. Methodology: The entire mitochondrial genome of 22 salivary gland adenoid cystic carcinomas (ACC) of salivary glands and matched leukocyte DNA was sequenced to determine the frequency and distribution of mitochondrial mutations in ACC tumors. Principal Findings: Seventeen of 22 ACCs (77%) carried mitochondrial mutations, ranging in number from 1 to 37 mutations. A disproportionate number of mutations occurred in the D-loop. Twelve of 17 tumors (70.6%) carried mutations resulting in amino acid changes of translated proteins. Nine of 17 tumors (52.9%) with a mutation carried an amino acid changing mutation in the nicotinamide adenine dinucleotide dehydrogenase (NADH) complex. Conclusions/Significance: Mitochondrial mutation is frequent in salivary ACCs. The high incidence of amino acid changing mutations implicates alterations in aerobic respiration in ACC carcinogenesis. D-loop mutations are of unclear significance

Increase in immune cell infiltration with progression of oral epithelium from hyperkeratosis to dysplasia and carcinoma

In the present study, epithelium derived lesions of various pathological manifestations were examined histologically and immunohistochemically for mononuclear cell infiltration. The infiltrate under the transformed epithelium of oral lesions, was examined for differences in the composition of immune mononuclear cells as the epithelium moves from hyperkeratosis through various degrees of dysplasia to squamous cell carcinoma. The study was performed on 53 human tongue tissues diagnosed as hyperkeratosis (11 cases), mild dysplasia (nine cases), moderate and severe dysplasia (14 cases) and squamous cell carcinoma (19 cases). A similar analysis was performed on 30 parotid gland tissues diagnosed as pleomorphic adenoma (14 cases) and carcinoma ex-pleomorphic adenoma (16 cases). Immunohistochemical analysis of various surface markers of the tumour infiltrating immune cells was performed and correlated with the transformation level as defined by morphology and the expression of p53 in the epithelium. The results revealed that, in the tongue lesions, the changes in the epithelium from normal appearance to transformed were accompanied by a corresponding increase in the infiltration of CD4, CD8, CD14, CD19+20, and HLA/DR positive cells. The most significant change was an increase in B lymphocytes in tongue lesions, that was in accordance with the transformation level (P<0.001). In the salivary gland, a significant number of cases did not show an infiltrate. In cases where an infiltrate was present, a similar pattern was observed and the more malignant tissues exhibited a higher degree of immune cell infiltration

Comedian Hosts and the Demotic Turn

Podcasting is a showcase for what cultural studies scholar Graeme Turner coined “the demotic turn” or the increasing visibility of the ordinary person in the today’s media landscape. Collins argues that the emergence of a particular breed of podcasts – comedian-hosted interviews with celebrities – function in an “off-label” manner as a form of self-help or vicarious therapy. The emergence and rapid growth of this genre can attributed to three main factors: a confessional culture, the triumph of experience over expertise, and the democratization allowed by the form’s technology. She explores the link between emotional intimacy and comedy, and analyzes podcasts like Marc Maron’s WTF that are, in expression, a rejection of the pedestal version of stardom