Development of In-Vitro cell based assay for the determination of biological activity of FSH using a CHO based recombinant cell line

The In-Vitro bioassay was developed for the determination of biological activity of Follicle Stimulating Hormone (FSH) using recombinant Chinese hamster ovary (CHO) based cell line that expresses the human FSH receptor (CHOSI FSHR CRELuc B-5) as well as luciferase reporter construct. The signal measured here comes from the luciferase activity that is dependent on the FSH stimulation. The present work focuses on an In-Vitro receptor-binding assay in which binding of FSH to its receptors (expressed on CHO cells) gives a dose and/or activity-dependent production of cAMP quantified by luciferase assay system. This kind of In-Vitro bioassay is used to determine the biological potency of FSH molecule. The assay is simple, has high throughput (performed in 96 half well area plates), and shows reproducible dose-response curves in the concentration range of 10.5-0.02 mIU/mL. With overall repeatability 6.5% and spike recovery of 100.7% and the assay has high precision and accuracy. Specificity was demonstrated by the lake of activity of assaying a variety of proteins manufactured by Intas. The In-Vitro cell based reporter gene assay has the preferred characteristics which makes it suitable to determine the biological activity of Recombinant Human Follicle Stimulating Hormone rHuFSH

Mechanistic Investigation of a Silylation-Based Kinetic Resolution Using Linear Free Energy Relationships and Its Application to Substrate Expansion and Polymer Supported Kinetic Resolutions

This dissertation represents our preliminary mechanistic investigations on the silylation-based kinetic resolution of secondary alcohols, and how we use that knowledge for further expansion of this methodology. Chapter 2 describes how we came up with a preliminary mechanism of our silylation-based methodology using a linear free energy relationship and a rate study. In this chapter, several para substituted triphenylsilyl chlorides were prepared that varied electronically and sterically in order to understand the substituent effects on the rate and the selectivity of the reaction. Selectivity factors and initial rates were experimentally determined for the kinetic resolution reactions with the newly designed silyl chlorides. Linear free energy relationships were found to correlate both selectivity factors and initial rates. Chapter 3 covers our 29Si NMR studies in order to understand if a complex is forming between the catalyst (-)-tetramisole and Ph3SiCl. A variety of different techniques were used including 1H NMR titrations, 29Si NMR experiments, and 1H-29Si gHSQC 2D experiments. Finally, three different mechanisms were proposed for future study. Chapter 4 discusses our efforts to apply the silylation-based methodology developed by the Wiskur lab to allylic alcohols, homoallylic alcohols, and 2-arylcyclohexanols. No enantiodiscrimination was obtained with allylic alcohols and homoallylic alcohols while a moderate level of selectivity was achieved with 2-arylcyclohexanols. Employing Ph3SiCl substituted in the para position with an isopropyl group in the kinetic resolution reaction of 2-arylcyclohexanol resulted in a doubling of the selectivity factor. The synthesis of various substrates and employing them in our kinetic resolution is discussed. Chapter 5 describes the use of a polymer supported triphenylsilyl chloride in our kinetic resolution reaction. Different molecular weight polymers containing triphenylsilyl chloride were prepared and tested in the kinetic resolution of 4-chromanol. Similar selectivity factors were reported in all cases. Reaction optimization along with future work for this project is discussed. Finally, Chapter 6 explores our attempt toward the development of a kinetic resolution of amines by transforming them into imines and employing them in an asymmetric aza-Diels-Alder reaction. Various chiral Lewis acids were attempted to achieve selectivity in the aza-Diels-Alder reaction

Highly focused document retrieval in aerospace engineering : user interaction design and evaluation

Purpose – This paper seeks to describe the preliminary studies (on both users and data), the design and evaluation of the K-Search system for searching legacy documents in aerospace engineering. Real-world reports of jet engine maintenance challenge the current indexing practice, while real users’ tasks require retrieving the information in the proper context. K-Search is currently in use in Rolls-Royce plc and has evolved to include other tools for knowledge capture and management. Design/methodology/approach – Semantic Web techniques have been used to automatically extract information from the reports while maintaining the original context, allowing a more focused retrieval than with more traditional techniques. The paper combines semantic search with classical information retrieval to increase search effectiveness. An innovative user interface has been designed to take advantage of this hybrid search technique. The interface is designed to allow a flexible and personal approach to searching legacy data. Findings – The user evaluation showed that the system is effective and well received by users. It also shows that different people look at the same data in different ways and make different use of the same system depending on their individual needs, influenced by their job profile and personal attitude. Research limitations/implications – This study focuses on a specific case of an enterprise working in aerospace engineering. Although the findings are likely to be shared with other engineering domains (e.g. mechanical, electronic), the study does not expand the evaluation to different settings. Originality/value – The study shows how real context of use can provide new and unexpected challenges to researchers and how effective solutions can then be adopted and used in organizations.</p

Design of 5.1 GHz ultra-low power and wide tuning range hybrid oscillator

The objective of the proposed work is to demonstrate the use of a hybrid approach for the design of a voltage-controlled oscillator (VCO) which can lead to higher performance. The performance is improved in terms of the tuning range, frequency of oscillation, voltage swing, and power consumption. The proposed hybrid VCO is designed using an active load common source amplifier and current starved inverter that are cascaded alternatively to achieve low power consumption. The proposed VCO achieves a measured phase noise of -74 dBc/Hz and a figure of merit (FOM) of -152.6 dBc/Hz at a 1 MHz offset when running at 5.1 GHz frequency. The hybrid current starved-current starved VCO (CS-CS VCO) consumes a power of 289 µW using a 1.8 V supply and attains a wide tuning range of 96.98%. Hybrid VCO is designed using 0.09 µm complementary metal–oxide–semiconductor (CMOS) technology. To justify the robustness, reliability, and scalability of the circuit different corner analysis is performed through 500 runs of Monte-Carlo simulation

AMMI and GGE biplots for G×E analysis of wheat genotypes under rain fed conditions in central zone of India

The highly significant environments, genotypes and G×E interaction observed by AMMI analysis of 17 wheat genotypes evaluated at 8 locations in the central zone of the country. Environments(E), genotypes -environment interaction(GE) and genotypes explained 68.8%, 17.6% and 3.2% of the total sum of squares respectively. First four interaction principal components accounted 33.7%, 30.2%, 14.6% and 12.6% of the G×E interaction variation, respectively. The highest positive IPCA1 score of genotype G8 followed by G11 and G10 supported by yield higher than the grand mean 21.8q/ha. Environments E4 (Jabalpur) and E8 (Partapgarh) recorded maximum yield 32.6q/ha and 28.4q/ha while lowest yield was realized in E1 (Arnej). GGE biplot analysis under polygon view indicated that G13 was better in E6 (Sagar), whereas G1 was better in E7 (Bilaspur) and E8 (Partapgarh). The genotype G1, at the centre of concentric circles, was the ideal genotype in terms of yield performance as compared to the other genotypes. In addition, G15 and G12, located on the next consecutive concentric circle, may be regarded as desirable genotypes

Preparation and characterization of rufinamide HP-β-cyclodextrin complexes prepared by the kneading method for solubility enhancement

ACKNOWLEDGEMENT: The author(s) are thankful to Ramanbhai Patel College of Pharmacy, Gujarat for providing facility to carry out research work.Aims: The present investigation concerns the preparation and characterization of Rufinamide HP-β-cyclodextrin complexes prepared by the kneading method. Material & methods: Rufinamide was procured as a gift sample from Torrent Pharmaceuticals limited. HP-β-cyclodextrin (HP-β-CD) was purchased from Himedia, India. Methanol and Hydrochloric Acid were purchased from S. D. Fine Chem. Pvt. Ltd., India. kneading method was selected to prepare inclusion complexation of Rufinamide. Phase solubility study was performed to check formation of inclusion complex. Prepared complex were characterize by different methods like DSC study, FTIR study, X-RPD study & in-vitro dissolution study Results: It was found that there is a formation of 1:1 inclusion complex between HP-β-CD as stability constant was found to be 221.27 M-1. DSC study, FTIR study had given supporting data for formation of inclusion complex. Amorphous nature of the complex was confirmed from the X-RPD study. Conclusions: From in-vitro dissolution study it was found that 1:1.5 complex showed around 50% drug released in 30 min & more than 70% of Drug release in 60 mins.Objetivos: La presente investigación se refiere a la preparación y caracterización de complejos de rufinamida HP-β-ciclodextrina preparados por el método de amasado. Material y métodos: La rufinamida fue donada por la empresa Torrent Pharmaceuticals limitado. HP-β-ciclodextrina (HP-β-CD) se adquirió de Himedia, India. Metanol y ácido clorhídrico se obtuvieron de SD Fine Chem. SA. Ltd., India. Se utilizó el método de amasado para preparar complejos de inclusión de rufinamida. El estudio de la fase de solubilidad se realizó para comprobar la formación de complejos de inclusión. Los complejos preparados se caracterizaron por diferentes métodos como DSC, FTIR, X-RPD y ensayo de disolución in vitro. Resultados: Se encontró que se producían formación de complejos en la relación 1:1. La constante de estabilidad encontrada fue de 221,27 M-1. Los estudios de DSC, FTIR confirmaron la formación del complejo de inclusión. Mediante X-RPD se confirmó la naturaleza amorfa del complejo. Conclusiones: El estudio de disolución in vitro mostró que la proporción 1:1.5 liberaba alrededor de 50% de fármaco en 30 min y a los 60 minutos se consiguió una liberación del 70%

Evidence of variable bacterial colonization on coloured elastomeric ligatures during orthodontic treatment : an intermodular comparative study

Besides, other factors, the choice of materials used as orthodontic ligatures could be one of the many tools to counter the effects of microbial adhesion, that culminates into dental ailments. Therefore, we assessed bacterial adhesion on elastomeric ligatures with special reference to coloured elastomeric rings during orthodontic treatment. A split mouth study, involving 240 samples of different elastomeric ligatures from forty orthodontic patients possessing good oral hygiene was carried out. The archwire was ligated to the brackets on both arches with elastomeric rings (superslick, clear transparent , blue and pink) at predetermined quadrants. After six weeks, ligatures from second premolars were removed and processed for bacterial enumeration using standard techniques. Bacterial counts were also determined for stimulated saliva samples taken at 0 and 6 weeks. A statistically significant difference in bacterial counts was obtained amongst different elastomeric modules used. Maximum bacterial counts were found on conventional pigmented elastomeric modules, followed by Superslick module and clear module. More number of bacteria associated with the conventional pink as compared to the conventional blue pigmented modules, however it was not statistically significant. The three bacterial genera Streptococcus Staphylococcus and Aerobic Lactobacilli adhered to elastomeric modules in following predominant pattern i.e. Conventional pink>Conventional Blue>Superslick>Clear. The studies evidenced colour and material dependent bacterial colonization on orthodontic modules and could be an indicator of bacterial biofilm forming potential based on surface chemistries and a clinically efficacious tool to redesign conventional and modified elastomeric rings as orthodontic ligation accessories

Advances in Treatment and Outcomes of Patients with <em>Legionella</em> Infection

Manifestations of Legionella infections range from benign, mild disease to a more severe form with increased morbidity and mortality, especially in untreated patients. Despite diagnostic advances, clinical diagnosis remains elusive. Macrolides and respiratory fluoroquinolones remain the antibiotics of choice for treatment of Legionella; however, several new antibiotics are currently under development or in clinical trials. The recommended duration of antibiotics is 5–7 days; although, some critically ill or immunosuppressed patients may require longer treatment. In vivo resistance to these antibiotics is rare, and there is no evidence that combination therapy is more beneficial than monotherapy. Early suspicion, diagnosis, and treatment are paramount for improving outcomes