The Development and Characterisation of Peptides to Image αvβ6 in Cancer.

PhDIntroduction The epithelial-specific integrin αvβ6 usually is undetectable on normal, adult, human tissue but is upregulated during tissue re-modelling and carcinogenesis. Objective To develop and characterise agents for targeting αvβ6 in cancer. Methods A series of peptides was generated from known high-affinity αvβ6 ligands. These peptides were then assessed for their ability to inhibit αvβ6-dependent cell adhesion. The lead 20 mer peptide, A20FMDV2, was identified. Flow cytometry, with two genetically identical cell lines that differed only in their expression of αvβ6 – A375P puro(negative) and A375Pβ6(positive)-, confirmed specificity for binding of A20FMDV2 to v6. The A20FMDV2 sequence was synthesised with both a biotin and DTPA-chelator group added. This was radiolabelled with Indium-111 and purity and structural integrity analysed by HPLC. Prior to in vivo experiments the endogenous expression of αvβ6 in nu/nu athymic mice was determined by examining 20 different organs (in triplicate) by immunohistochemistry. For in vivo experiments, athymic mice were inoculated subcutaneously with A375Pβ6 (right shoulder) and A375P puro (left shoulder). A radiolabelled variant of A20FMDV2 (111In-DTPA-A20FMDV2) was injected into xenografted mice. Some mice were imaged on a NanoSPECT/CT imager and biodistribution confirmed by determining the radioactivity associated with excised organs. 12 Results Immunohistochemistry of mouse tissues revealed, as expected, strong expression of v6 in the A375Pβ6 tumour but none in the A375Ppuro tumour. In addition, expression of vβ6 was found in normal murine hair follicles, gallbladder, urinary bladder, secretory endometrium, the stomach and along the GI tract. Flow cytometry with A375Ppuro and A375Pβ6 cells confirmed that DTPA-chelate addition to A20FMDV2 did not affect its specificity or affinity for v6. HPLC confirmed that DTPA-biotinylated-A20FMDV2 was labelled successfully with Indium-111 resulting in a single radiolabelled peak. Ex-vivo gamma counts revealed seven-fold higher uptake of the radiolabelled-peptide in the A375Pβ6 tumour compared with the A375Ppuro tumour. Biodistribution data were consistent with the endogenous αvβ6 expression as determined by immunohistochemistry. NanoSPECT/CT imaging clearly identified that the A375P6 tumour retained more radioactive peptide than the A375Ppuro tumour at 30, 60 and 120 minutes after injection. Images also showed uptake in the GI tract. Conclusion We have determined that the peptide DTPA-A20FMDV2 is specific for v6, can be labelled with Indium-111 and selectively localises in vivo to v6-positive tissues, including human cancer. This is the first peptide described for the successful imaging, by SPECT, of v6-positive cancers.Cancer Research U

Origin of anomalous breakdown of Bloch's rule in the Mott-Hubbard insulator MnTe2_2

We reinvestigate the pressure dependence of the crystal structure and antiferromagnetic phase transition in MnTe$_2$ by the rigorous and reliable tool of high pressure neutron powder diffraction. First-principles density functional theory calculations are carried out in order to gain microscopic insight. The measured N\'eel temperature of MnTe$_2$ is found to show unusually large pressure dependence of $12$ K GPa$^{-1}$. This gives rise to large violation of Bloch's rule given by $\alpha=\frac{d\log T_N}{d\log V}=-\frac{10}{3} \approx -3.3$, to a $\alpha$ value of -6.0 $\pm$ 0.1 for MnTe$_2$. The ab-initio calculation of the electronic structure and the magnetic exchange interactions in MnTe$_2$, for the measured crystal structures at different pressures, gives the pressure dependence of the Ne\'el temperature, $\alpha$ to be -5.61, in close agreement with experimental finding. The microscopic origin of this behavior turns to be dictated by the distance dependence of the cation-anion hopping interaction strength

Optical conductivity study of screening of many-body effects in graphene interfaces

Theoretical studies have shown that electron-electron (e-e) and electron-hole (e-h) interactions play important roles in many observed quantum properties of graphene making this an ideal system to study many body effects. In this report we show that spectroscopic ellipsometry can enable us to measure this interactions quantitatively. We present spectroscopic data in two extreme systems of graphene on quartz (GOQ), an insulator, and graphene on copper (GOC), a metal which show that for GOQ, both e-e and e-h interactions dominate while for GOC e-h interactions are screened. The data further enables the estimation of the strength of the many body interaction through the effective fine structure constant, $\alpha_{g}^{*}$. The $\alpha_{g}^{*}$ for GOQ indicates a strong correlation with an almost energy independent value of about 1.37. In contrast, $\alpha_{g}^{*}$ value of GOC is photon energy dependent, is almost two orders of magnitude lower at low energies indicating very weak correlation.Comment: Main Article (4 pages, 4 figures); Supporting Online Material (12 pages, 9 figures

Unsupervised methodology to unveil content delivery network structures

A method for analyzing a content delivery network. The method includes obtaining network traffic flows corresponding to user nodes accessing contents from a set of servers of the content delivery network, extracting a timing attribute from each network traffic flow associated with a server, where the timing attribute is aggregated into a timing attribute dataset of the server based on all network traffic flows associated with the server, generating a statistical measure of the timing attribute dataset as a portion of a feature vector representing the server, where the feature vector is aggregated into a set of feature vectors representing the set of servers, analyzing the set of feature vectors based on a clustering algorithm to generate a set of clusters, and generating, based on the set of clusters, a representation of server groups in the content delivery network

Using pneumococcal and rotavirus surveillance in vaccine decision-making: A series of case studies in Bangladesh, Armenia and the Gambia.

Pneumonia and diarrhea are the leading causes of child morbidity and mortality globally and are vaccine preventable. The WHO-coordinated Global Rotavirus and Invasive Bacterial Vaccine-Preventable Disease Surveillance Networks support surveillance systems across WHO regions to provide burden of disease data for countries to make evidence-based decisions about introducing vaccines and to demonstrate the impact of vaccines on disease burden. These surveillance networks help fill the gaps in data in low and middle-income countries where disease burden and risk are high but support to sustain surveillance activities and generate data is low. Through a series of country case studies, this paper reviews the successful use of surveillance data for disease caused by pneumococcus and rotavirus in informing national vaccine policy in Bangladesh, Armenia and The Gambia. The case studies delve into ways in which countries are leveraging and building capacity in existing surveillance infrastructure to monitor other diseases of concern in the country. Local institutions have been identified to play a critical role in making surveillance data available to policymakers. We recommend that countries review local or regional surveillance data in making vaccine policy decisions. Documenting use of surveillance activities can be used as advocacy tools to convince governments and external funders to invest in surveillance and make it a priority immunization activity

HST observations of the Local Group dwarf galaxy Leo I

We present deep HST F555W (V) and F814W (I) observations of a central field in the Local Group dwarf spheroidal (dSph) galaxy Leo I. The resulting color-magnitude diagram (CMD) reaches I \simeq 26 and reveals the oldest ~10-15 Gyr old turnoffs. Nevertheless, a horizontal branch is not obvious in the CMD. Given the low metallicity of the galaxy, this likely indicates that the first substantial star formation in the galaxy may have been somehow delayed in Leo I in comparison with the other dSph satellites of the Milky Way. The subgiant region is well and uniformly populated from the oldest turnoffs up to the 1 Gyr old turnoff, indicating that star formation has proceeded in a continuous way, with possible variations in intensity but no big gaps between successive bursts, over the galaxy's lifetime. The structure of the red-clump of core He-burning stars is consistent with the large amount of intermediate-age population inferred from the main sequence and the subgiant region. In spite of the lack of gas in Leo I, the CMD clearly shows star formation continuing until 1 Gyr ago and possibly until a few hundred Myrs ago in the central part of the galaxy.Comment: 26 pages with 8 figures (fig 2 not available electronically). To be published in ApJ, April 1 1999 (vol.514, #2

Catecholase-Like Activity and Theoretical Study in Solid State of a New Ru(III)-Schiff Base Complex.

A new ruthenium(III) complex of molecular formula [Ru(PPh3)Cl2(L)] (1) has been synthesized using the Schiff base ligand obtained from 5-chlorosalicylaldehyde and N,N-dimethylethylenediamine and characterized by FT-IR, UV-Vis, cyclic voltammetry and single crystal X-ray structural analysis. The metal ion exhibits a slightly distorted octahedral environment where the chelating Schiff base ligand contributes with its NNO donor set. The coordination geometry around the Ru(III) ion is completed by a PPh3 ligand and two chloride anions, and the charge balance is assured by the phenoxo oxygen of the Schiff base. With the aim to analyse the energy related to the halogen bonding interactions in solid state, a theoretical study has been performed on complex 1, by using the MEP and NCl plot computational tools. Furthermore, complex 1 shows catecholase-like activity in conversion of the model substrate 3,5-di-tert-butylcatechol (3,5-DTBC) to the corresponding 3,5-di-tert-butylquinone (3,5-DTBQ) under aerobic condition. The parameters regarding the enzymatic kinetics have been evaluated from the Lineweaver-Burk plot using the Michaelis-Menten approach of enzyme catalysis. A significant high T.O.N value (2.346 × 103 h–1) indicates that complex 1 has a very good catalytic efficiency towards 3,5-DTBC

Therapeutic targeting of integrin αvβ6 in breast cancer

BACKGROUND: Integrin ?v?6 promotes migration, invasion, and survival of cancer cells; however, the relevance and role of ?v?6 has yet to be elucidated in breast cancer.METHODS: Protein expression of integrin subunit beta6 (?6) was measured in breast cancers by immunohistochemistry (n &gt; 2000) and ITGB6 mRNA expression measured in the Molecular Taxonomy of Breast Cancer International Consortium dataset. Overall survival was assessed using Kaplan Meier curves, and bioinformatics statistical analyses were performed (Cox proportional hazards model, Wald test, and Chi-square test of association). Using antibody (264RAD) blockade and siRNA knockdown of ?6 in breast cell lines, the role of ?v?6 in Human Epidermal Growth Factor Receptor 2 (HER2) biology (expression, proliferation, invasion, growth in vivo) was assessed by flow cytometry, MTT, Transwell invasion, proximity ligation assay, and xenografts (n ? 3), respectively. A student's t-test was used for two variables; three-plus variables used one-way analysis of variance with Bonferroni's Multiple Comparison Test. Xenograft growth was analyzed using linear mixed model analysis, followed by Wald testing and survival, analyzed using the Log-Rank test. All statistical tests were two sided.RESULTS: High expression of either the mRNA or protein for the integrin subunit ?6 was associated with very poor survival (HR = 1.60, 95% CI = 1.19 to 2.15, P = .002) and increased metastases to distant sites. Co-expression of ?6 and HER2 was associated with worse prognosis (HR = 1.97, 95% CI = 1.16 to 3.35, P = .01). Monotherapy with 264RAD or trastuzumab slowed growth of MCF-7/HER2-18 and BT-474 xenografts similarly (P &lt; .001), but combining 264RAD with trastuzumab effectively stopped tumor growth, even in trastuzumab-resistant MCF-7/HER2-18 xenografts.CONCLUSIONS: Targeting ?v?6 with 264RAD alone or in combination with trastuzumab may provide a novel therapy for treating high-risk and trastuzumab-resistant breast cancer patients.<br/

Streptococcus pneumoniae Serotype-2 Childhood Meningitis in Bangladesh: A Newly Recognized Pneumococcal Infection Threat

BACKGROUND: Streptococcus pneumoniae is a leading cause of meningitis in countries where pneumococcal conjugate vaccines (PCV) targeting commonly occurring serotypes are not routinely used. However, effectiveness of PCV would be jeopardized by emergence of invasive pneumococcal diseases (IPD) caused by serotypes which are not included in PCV. Systematic hospital based surveillance in Bangladesh was established and progressively improved to determine the pathogens causing childhood sepsis and meningitis. This also provided the foundation for determining the spectrum of serotypes causing IPD. This article reports an unprecedented upsurge of serotype 2, an uncommon pneumococcal serotype, without any known intervention. METHODS AND FINDINGS: Cases with suspected IPD had blood or cerebrospinal fluid (CSF) collected from the beginning of 2001 till 2009. Pneumococcal serotypes were determined by capsular swelling of isolates or PCR of culture-negative CSF specimens. Multicenter national surveillance, expanded from 2004, identified 45,437 patients with suspected bacteremia who were blood cultured and 10,618 suspected meningitis cases who had a lumber puncture. Pneumococcus accounted for 230 culture positive cases of meningitis in children <5 years. Serotype-2 was the leading cause of pneumococcal meningitis, accounting for 20.4% (45/221; 95% CI 15%-26%) of cases. Ninety eight percent (45/46) of these serotype-2 strains were isolated from meningitis cases, yielding the highest serotype-specific odds ratio for meningitis (29.6; 95% CI 3.4-256.3). The serotype-2 strains had three closely related pulsed field gel electrophoresis types. CONCLUSIONS: S. pneumoniae serotype-2 was found to possess an unusually high potential for causing meningitis and was the leading serotype-specific cause of childhood meningitis in Bangladesh over the past decade. Persisting disease occurrence or progressive spread would represent a major potential infection threat since serotype-2 is not included in PCVs currently licensed or under development