Development Of Tier Research Articles

Development of Tier 2 LC-MRM-MS protein quantification methods for liquid biopsies

In the pursuit of personalized diagnostics and tailored treatments, quantitative protein tests contribute to a more precise definition of health and disease. The development of new quantitative protein tests should be driven by an unmet clinical need and performed in a collaborative effort that involves all stakeholders. With regard to the analytical part, mass spectrometry (MS)-based platforms are an excellent tool for quantification of specific proteins in body fluids, for example focused on cancer. The obtained readouts have great potential in determining tumor aggressiveness to facilitate treatment decisions, and can furthermore be used to monitor patient response. Internationally standardized TNM classifications of malignant tumors are beneficial for diagnosis, however treatment outcome and survival of cancer patients is poorly predicted. To this end, the importance of the tumor microenvironment has endorsed the introduction of the tumor-stroma ratio as a prognostic parameter in solid primary tumor types. Currently, the stromal content of tumor tissues is determined via routine diagnostic pathology slides. With the development of liquid chromatography (LC)-MS methods we aim at quantification of tumor-stroma specific proteins in body fluids. In this mini-review the analytical aspect of this developmental trajectory is further detailed.

Vaccine-Elicited Tier 2 HIV-1 Neutralizing Antibodies Bind to Quaternary Epitopes Involving Glycan-Deficient Patches Proximal to the CD4 Binding Site.

Eliciting broad tier 2 neutralizing antibodies (nAbs) is a major goal of HIV-1 vaccine research. Here we investigated the ability of native, membrane-expressed JR-FL Env trimers to elicit nAbs. Unusually potent nAb titers developed in 2 of 8 rabbits immunized with virus-like particles (VLPs) expressing trimers (trimer VLP sera) and in 1 of 20 rabbits immunized with DNA expressing native Env trimer, followed by a protein boost (DNA trimer sera). All 3 sera neutralized via quaternary epitopes and exploited natural gaps in the glycan defenses of the second conserved region of JR-FL gp120. Specifically, trimer VLP sera took advantage of the unusual absence of a glycan at residue 197 (present in 98.7% of Envs). Intriguingly, removing the N197 glycan (with no loss of tier 2 phenotype) rendered 50% or 16.7% (n = 18) of clade B tier 2 isolates sensitive to the two trimer VLP sera, showing broad neutralization via the surface masked by the N197 glycan. Neutralizing sera targeted epitopes that overlap with the CD4 binding site, consistent with the role of the N197 glycan in a putative “glycan fence” that limits access to this region. A bioinformatics analysis suggested shared features of one of the trimer VLP sera and monoclonal antibody PG9, consistent with its trimer-dependency. The neutralizing DNA trimer serum took advantage of the absence of a glycan at residue 230, also proximal to the CD4 binding site and suggesting an epitope similar to that of monoclonal antibody 8ANC195, albeit lacking tier 2 breadth. Taken together, our data show for the first time that strain-specific holes in the glycan fence can allow the development of tier 2 neutralizing antibodies to native spikes. Moreover, cross-neutralization can occur in the absence of protecting glycan. Overall, our observations provide new insights that may inform the future development of a neutralizing antibody vaccine.

A Study on the Prospects and Potential Threats to the Hospitality Industry in India

The hospitality sector in India plays a vital role in terms of contributions to the economy of the country as an employment generator. The much wanted growth in the tourism sector of the country is in fact dependent on the growth and development of the hospitality sector. However, the ability of the hospitality sector to remain viable, profitable and competitive in the years to come will depend on its ability to foresee emerging trends and then capitalize on them. This is most essential, given that India, despite having high tourist potential, attracts far lesser foreign tourists as compared to smaller countries like Singapore, Thailand and Malaysia. This paper examines the key trends that are set to emerge in the hospitality sector in the country over the next decade. An exploratory methodology was used in this research including both theoretical and empirical methods. From the theoretical studies, it was found that these key trends include the growth of domestic tourism, the emergence of medical tourism as the key attraction in India, the development of tier 2 and tier 3 cities as important tourist hubs and the shift in tourist traffic away from luxury formats which have hitherto dominated the hotel industry and towards economically priced mid – level and budget hotels.

Development of Tiers 2 and 3 methane emission factors for enteric fermentation and manure management of cattle and sheep using Hillsborough herd data and calorimetric methane measurements

Development of Tiers 2 and 3 methane emission factors for enteric fermentation and manure management of cattle and sheep using Hillsborough herd data and calorimetric methane measurements

Exposure scenarios for workers

The new European chemicals legislation REACH (Registration, Evaluation, Authorisation and restriction of Chemicals) requires the development of Exposure Scenarios describing the conditions and risk management measures needed for the safe use of chemicals. Such Exposure Scenarios should integrate considerations of both human health and the environment. Specific aspects are relevant for worker exposure. Gathering information on the uses of the chemical is an important step in developing an Exposure Scenario. In-house information at manufacturers is an important source. Downstream users can contribute information through direct contact or through their associations. Relatively simple approaches (Tier 1 tools, such as the ECETOC Targeted Risk Assessment and the model EASE) can be used to develop broad Exposure Scenarios that cover many use situations. These approaches rely on the categorisation of just a few determinants, including only a small number of risk management measures. Such approaches have a limited discriminatory power and are rather conservative. When the hazard of the substance or the complexity of the exposure situation require a more in-depth approach, further development of the Exposure Scenarios with Tier 2 approaches is needed. Measured data sets of worker exposure are very valuable in a Tier 2 approach. Some downstream user associations have attempted to build Exposure Scenarios based on measured data sets. Generic Tier 2 tools for developing Exposure Scenarios do not exist yet. To enable efficient development of the worker exposure part of Exposure Scenarios a further development of Tier 1 and Tier 2 tools is needed. Special attention should be given to user friendliness and to the validity (boundaries) of the approaches. The development of standard worker exposure descriptions or full Exposure Scenarios by downstream user branches in cooperation with manufacturers and importers is recommended.

Development of Tier I Neurobehavioral Testing Capabilities for Incorporation into Pivotal Rodent Safety Assessment Studies

A functional observational battery and an automated test of motor activity have been developed in this laboratory in response to present and future needs for routine assessment of behavioral function in acute and repeated dose rodent safety assessment studies. Data have been collected for untreated 42–49-day-old animals of both sexes in order to gain information on normal ranges of responses in animals of this age as well as to determine the relative variability of the various test measures. Data will continue to be added to this database, both for animals of this age range and for other age ranges. A validation strategy has been outlined that emphasizes the characterization of behavioral profiles of both pharmacologic agents and chemically related neurotoxins, as well as the importance of determining how well these results translate to those observed in humans exposed to the same test agents. The logistical and economic factors associated with neurobehavioral testing and the importance of intralaboratory standardization are discussed in detail. Both controllable and difficult to control environmental and experimental variables that can affect the outcome of a behavioral study are identified. The problems associated with neurobehavioral data interpretation and the importance of a multisystem approach to interpreting functional changes in the context of toxicology studies are discussed.