PROCESS OPTIMIZATION AND VALIDATION OF AN ASSAY FOR HIGH-THROUGHPUT SCREENING

A biological assay is designed to set up a rapid and robust drug-screening system on a small scale. An assay is considered as a single unit of a platform to screen various compounds for aiding in drug discovery. Each assay is carried out in a 96-well plate, each of whose wells consists of the biological component called the Spheroids. The value of each assay lies in it facilitating for versatile screening applications. The spheroid is considered as a micro-structural product. And the addition of various compounds for testing is performed in each well (consisting of the spheroids). The focus has been to put forth the production principles and validation strategies to run the biological assay and test its efficacy to be used for screening in high volumes. The assay development illustrates processing and validation techniques. The goal is to develop optimized standards to process the assay, addressing various quality control issues, from the raw material to the end-product stage. Such an approach also brings interesting analogies of biological process in a manufacturing scenario. The developed system incorporates a value stream approach, by pulling the product from the customer end. The process involves simply encapsulating HUVECs (Human Umbelical Vein Endothelial cells) from the raw material stage, culturing to form the spheroid and transferring the component to assemblage in a 96-well format undergoing stages of heat treatments. The small scale screening system allows the use of small amounts of drug, which is especially essential for new drug synthesis or in rapid decision making to find out any unknown potent compounds. The design of optimal processes in product development of the spheroid assay is illustrated. Thus in light of the value of this assay, developing the production system has been pivotal so as to produce quality spheroids in the 96-well plate formats. The quantification of the stimulatory and inhibitory effects of the different agents is required to help understand the complex biological behavior involved. The goal is to validate the data using image analysis software. The image analysis helps determine the quantification to be accurate, objective, and consistent. The quality of the product is tested by the reproducibility and robustness of the assay

Microbial conversion of organic residues into acid rich process liquids and their use in bio-electrochemical systems

In 2016, 2.01 billion tonnes of solid waste were generated worldwide. The volume of waste is expected to grow to 3.40 billion tonnes by 2050. Worldwide, most solid waste is disposed of in landfills or dumps. Due to improper treatment and disposal of solid waste, nearly 1.6 billion tonnes of CO2 equivalents of greenhouse gas emissions were generated worldwide in 2016. This amount is expected to rise to 2.6 billion tonnes of CO2 equivalents per year by 2050. It will therefore become increasingly important in the future not only to treat waste sustainably, but also to use it as an alternative to fossil fuels. Different waste-to-energy concepts are used, particularly for the treatment of OFMSW. As an alternative to the previously dominant biogas production, intensive research is currently being carried out into technologies for the recycling of organic residual materials, including so-called bio-electric systems (BES). In contrast to biogas production, this technology enables the treatment of a wide range of wastes to produce different end products, e.g. electrical energy, hydrogen or methane, can be preferred in BES depending on the selected process parameters. Despite numerous advances in research, considerable additional optimization is still required in order to be able to use the systems in large-scale power generation. In order to use solid organic waste in BES systems, fermentative digestion is required to convert the organic components into dissolved short-chain organic acids (Volatile Fatty Acids (VFA)) and alcohols. In the course of the investigations, the solid waste residues were first digested to acid-rich hydrolysate in a hydrolysis reactor at pH-values of 5.5 and 6.0. However, this hydrolysate also contains particles that are inert to a subsequent degradation step leading to technical process disturbances. These inert particles can be removed by means of a membrane filtration step; a particle-free permeate is produced, which can be fed to the BES reactors. Within the scope of the present work, the basics of the utilization of OFMSW via microbial digestion, membrane filtration and utilization in BES should be investigated. Lab-scale BES reactors were developed and batch tests were carried out. The vegetable waste residues from hydrolysis could be efficiently converted into hydrolysate. At a pH value of 6.0, higher organic acid concentrations were achieved than at pH 5.5. At pH 6.0, based on the added organic dry matter, these were approx. 350 g kg-1 (oDMadded) and at pH 5.5 approx. 215 g kg-1 oDMadded. Likewise, the concentration of chemical oxygen demand (COD) of the hydrolysate at pH 6.0 was 21.85 % higher than at pH 5.5. However, the COD degradation rates in the AF used were insufficient because the inert particles present in the hydrolysate could not be completely microbially degraded. The subsequent integration of ceramic cross-flow membrane filtration into the two-stage system produced a particle-free permeate and significantly the increased microbial degradability. Clear differences could be shown depending on the substrate used (plant waste and grass/maize silage). The filtration step resulted in a significant improvement of the specific methane yield of permeate by 40% (vegetable waste) and 24.5% (grass/maize silage) compared to hydrolysate; proof that inert particles were separated efficiently. Finally, the process liquids hydrolysate and permeate produced by the hydrolysis of maize silage and the subsequent membrane filtration were fed to the anode chamber of two mixed-culture BES reactors. The investigations showed that all organic acids in both process liquids could be completely degraded in the BES. The highest COD (87%) and TOC degradation rates (88%) were achieved with permeate. However, the hydrolysate with added acetic acid yielded the highest current density of 470 µA/cm². Increasing the pH-value of the process liquids from 5.75 to 6.8 also significantly improved the current production and degradation rates. In this batch studies, relatively low Coulomb efficiencies of less than 10% were achieved due to the use of a mixed cultures. The promising results show that at high pH-values (pH 6.0) in hydrolysis organic residues can be efficiently converted into a hydrolysate with high concentrations of organic acids and that the system can be further optimized by coupling membrane filtration. The utilization of the permeate in BES enables, a sustainable production of bioenergy and platform chemicals with permeate enables, depending on the BES reactor configuration. In summary, it was described for the first time that the combination of the fermentative biomass degradation process with filtration via ceramic membranes and the use of permeate in BES systems is possible.Im Jahr 2016 fielen weltweit 2,01 Milliarden Tonnen an festen Abfällen an. Es wird erwartet, dass das Abfallaufkommen bis 2050 auf 3,40 Milliarden Tonnen anwächst. Weltweit werden die festen Abfälle überwiegend über Deponien oder Müllhalden entsorgt. Aufgrund unsachgemäßer Behandlung und Entsorgung wurden weltweit in 2016 nahezu 1,6 Milliarden Tonnen CO2-Äquivalente an Treibhausgasemissionen verursacht, mit weiter steigender Tendenz. Die nachhaltige Verwertung der Abfälle und deren Nutzung zur Energiegewinnung wird in Zukunft weiter an Bedeutung gewinnen. Insbesondere zur Behandlung der organischen Bestandteile des Hausmülls (Organic Fraction of Municipal Solid Waste (OFMSW)) kommen unterschiedliche Waste-to-Energy-Konzepte zum Einsatz. Alternativ zur bisher dominierenden Biogasgewinnung wird derzeit intensiv an neuen Technologien geforscht, u.a. an sogenannten bio-elektrischen Systemen (BES). In BES interagieren exo-elektrogene Mikroorganismen, die als Biofilm auf Elektroden aufwachsen, mit diesen und ermöglichen die Konversion gelöster organischer Verbindungen zu unterschiedlichen Endprodukten, z.B. elektrische Energie, Wasserstoff oder Methan. Trotz zahlreicher Fortschritte in der Forschung besteht noch erheblicher zusätzlicher Optimierungsbedarf, um die Systeme im technischen Maßstab einsetzen zu können. Um feste organische Abfälle in BES-Systemen einsetzten zu können, bedarf es zunächst eines fermentativen Aufschlusses zur Überführung der organischen Bestandteile in gelöste kurzkettige organische Säuren (VFA) und Alkohole. Dieses Hydrolysat enthält jedoch auch Partikel, die gegen einen nachfolgenden Abbauschritt inert sind und zu technischen Prozessstörungen führen können. Diese inerten Partikel können mithilfe eines Membranfiltrationsschritts entfernt werden; es entsteht ein partikelfreies Permeat, welches den BES-Reaktoren zugeführt werden kann. Im Rahmen der vorliegenden Arbeit sollten die Grundlagen der Verwertung von OFMSW über mikrobiellen Aufschluss, Membranfiltration und Verwertung in BES untersucht werden. Dazu wurden BES-Reaktoren im Labormaßstab entwickelt und Batch-Tests durchgeführt. Die pflanzlichen Reststoffe konnten fermentativ effizient in Hydrolysat überführt werden. Bei einem pH-Wert von 6,0 wurde höhere organischen Säurekonzentrationen und -erträge erreicht als bei pH 5,5. Die gebildete Gesamtmasse der kurzkettigen organischen Säuren betrugen bei pH 6,0, bezogen auf die zugeführte organische Trockenmasse, ca. 350 g kg-1 (oDMadded) und bei pH 5,5 ca. 215 g kg-1 oDMadded. Ebenso war die Konzentration an Chemischem Sauerstoffbedarfs (CSB) des Hydrolysats bei pH 6,0 um 21,85 % höher als bei pH 5,5. Die CSB-Abbauraten im genutzten Festbettreaktor waren jedoch unzureichend, da die im Hydolysat enthaltenen inerten Partikel nicht vollständig mikrobiell abgebaut werden konnten. Durch die anschließende Integration einer keramischen Cross-Flow- Membranfiltration in das zweistufige System konnte ein partikelfreies Permeat erzeugt werden und die mikrobielle Abbaubarkeit erheblich gesteigert werden. Dabei konnten deutliche Unterschiede in Abhängigkeit des eingesetzten Substrats (Pflanzenabfälle und Gras-/Maissilage) aufgezeigt werden. Durch den Filtrationsschritt konnte eine signifikante Verbesserung der spezifischen Methanausbeute von Permeat um 40% (pflanzliche Abfälle) und 24,5% (Gras-/Maissilage) im Vergleich zu Hydrolysat erreicht werden; ein Beleg dafür, dass inerte Partikel effizient abgetrennt wurden. Schließlich wurden die aus Maissilage erzeugte Prozessflüssigkeit Hydrolysat und dass durch die Membranfiltration gewonnene Permeat der Anodenkammer zweier BES-Reaktoren, die mit einer Mischkultur angeimpft waren, zugeführt. Die Untersuchungen zeigten, dass alle organischen Säuren in beiden Prozessflüssigkeiten im BES vollständig abgebaut werden können. Die höchsten CSB- (87%) und TOC-Abbauraten (88%) wurden mit Permeat erreicht. Das Hydrolysat mit zugesetzter Essigsäure ergab hingegen die höchste Stromdichte von 470 µA/cm². Die Erhöhung des pH-Wertes der Prozessflüssigkeiten von 5,75 auf 6,8 verbesserte auch die Produktions- und Abbauraten deutlich. In diesen Batchstudien wurden aufgrund der Verwendung von Mischkulturen relativ niedrige Coulomb-Wirkungsgrade von weniger als 10% erreicht. Die vielversprechenden Ergebnisse zeigen, dass bei hohen pH-Werten von 6,0 in der Hydrolyse organische Reststoffe effizient in ein Hydrolysat mit hohen Konzentrationen an organischen Säuren überführt werden können und mit der Kopplung einer Membranfiltration das System weiter optimiert werden kann. Die Verwertung des Permeats in BES ermöglicht, je nach Konfiguration der Reaktoren, eine nachhaltige Erzeugung von Bioenergie- und Plattformchemikalien. Zusammenfassend wurde erstmalig beschrieben, dass die Kombination des fermentativen Biomasse-Abbauprozesses mit der Filtration über keramische Membranen und Nutzung des Permeats in BES-Systemen möglich ist

Robot Path Planning with IGA-MMAS and MMAS-IGA

Path Planning of mobile robots is one of the essential tasks in robotic research and studies with intelligent technologies. It helps in determining the path from a source to the destination. It has extended its roots from classic approaches to further improvements over time, such as evolutionary approaches. Ant Colony Optimization (ACO) and Genetic algorithm are well known evolutionary approaches in effective path planning. This research work focuses on the Max-Min Ant System (MMAS) derived from the ACO evolutionary approach of Ant System (AS) and Improved Genetic Algorithm (IGA) which is efficient over the classical Genetic Algorithm. In-order to study robot path planning two methods are combined in this research work combining MMAS and IGA as two-hybrid methods MMAS-IGA and IGA-MMAS . The results of the two-hybrid methods will be deriving the near optimal solution, demonstrated in the experimental study of this work. Grid maps are used for simulating the robot path planning environment which is modeled using the grid method. Genetic operators of IGA are combined with MMAS for the enhancement of the overall result of the methods IGA-MMAS and MMAS-IGA. The effectiveness of these two methods will be determined in the simulation modeled using MATLAB environment. The experimental results of these methods are done in a static environment and the results of MMAS-IGA and IGA-MMAS are compared to the path planning method GA-ACO

Characterization of Parallel Application Runtime Sensitivity on Multi-core High Performance Computing Systems

A commonly seen behavior of parallel applications is that their runtime is influenced by network communication load. The way a parallel application is run in a network and the presence of other applications and processes in the network can contribute to a wide range of variations in the runtime. Therefore, in order to achieve consistent and optimal runtimes, it is important to understand and characterize the runtime sensitivity of parallel applications with respect to execution under the presence of network communication load. In this research, runtime sensitivities for various parallel applications were studied by applying additional network communication load. In particular, the focus was on the runtime sensitivity of parallel applications on a multi-core multi-processor (MCMP) system where less network switching and routing are involved compared to single-core single-processor machines. The objective of this work was to determine if a previously developed sensitivity model for single-core single-processor machines still holds good for multi-core machines. For this purpose, previously developed tools (PACE and PARSE) were used to perturb the communication sub-system while executing several parallel application benchmarks such as the NAS benchmarks and PSTSWM. Runtime variations of these parallel applications were studied, under a specific network communication load, for different test cases by changing computing core allocation. A 10-node 80-core cluster was used as the test bed for this research purpose. Several test cases were explored using a variety of core allocations (process locations) for the application under test (AUT) to simulate job scheduler fragmentation. To ensure statistical significance, several iterations (trial runs) were executed in each test case. Results indicate that the idea of application sensitivity to communication sub-system performance degradation holds for MCMP architectures

Eelam Poetry and the beginning of Penmozhi

Writing is the key to changing society. All those who have been excluded as unclean are exposed today. Women's liberation is not about leaving the door, it is only possible in poetry to feel the expansion of the boundary to break the lock of the language. On the one hand, the achievement of the female language, the thrill of womanhood. No, this is an 'alternative politics', on the one hand. However, the purpose of feminist poetry is to face the future through language. The restrictions imposed by society on the female body, such as the summary laws of the outer space, etc. The illusions that invented later traditions based on traditional examples of female image (for example, the female image of Sita, Kannagi, Nalaini, etc., recorded the female tradition. Mother Karaikal was conducted with a bone towards Kayil, and the mother's consciousness was dried with devotional dye.) The female language is the expression of the imposition through the way. The study indicates that the impact of the female language being constructed by tamil women poets Malathi Maitri, Lena Manimegalai, Kuttilathi and Sugirtha Rani followed the beginning of Eelam

Genomic distribution of histone H1 in budding yeast (Saccharomyces cerevisiae) : yeast chromosome III

Includes bibliographical references.The linker histone HI binds to the nucleosome and is essential for the organization of nucleosomes into the 30-nm filament of the chromatin. This compaction of DNA has a well-characterized effect on DNA function. In Saccharomyces cerevisiae, HHO 1 encodes a putative linker histone with very significant homology to histone HI. In vitro chromatin assembly experiments with recombinant Hho 1 p have shown that it is able to complex with the dinucleosomes in a similar manner to histone HI. It has also been reported that disruption of HHOl has little affect on RNA levels. A longstanding issue concerns the location of Hho 1 p in the chromatin and studies have shown using immunoprecipitation technique with anti-HA antibody, that Hho 1 p shows a preferential binding to rDNA sequences. In this project we have tried to confirm the above results in wild type cells, using immunopurifi ed anti rHho 1 p antibody

Karisalkattu Ecology in the Narrative of Ki.Ra.

Ki.Rajanarayanan is a sculptor of karisal forest literature who is revered as the father of regional literature. Ki.Ra. completely dissolves his writing brush into the karisal soil and converses within the reader's mind as it is contextualized. Karisalkattu ecology is naturally threaded in the Koballa village narrative. "Is it possible to destroy the forest if you want this after the lace?". The voice of the narrator, who sounds in the storytelling, is an initiative of natural curiosity.  The story points to the hard work of the Kammawar people who migrated to the karisal land and the nuances of preparing the karisal earth for yield. Ki.Ra. Karisal also regretted the environmental pollution that occurs in the region while recording its normal life

Pulmonary Function Test in Rheumatoid Arthritis Patients without Extra Articular Manifestations

INTRODUCTION: Rheumatoid Arthritis is one of the commonest Auto immune disorder affecting females commonly and smoking males. Asians are more proned. HLA DR4 is the MHC involved. The triggering factors are smoking, porphyromonas gingivalis (bacteria) and post mycoplasma pneumonia. In RA, there occurs Acute synovitis in the early followed by chronic synovitis and finally to erosive arthritis which is irreversible. The mechanism is mediated by release of all proteolytic enzymes causing fibrous deposition and finally leading to ankylosis which is a fixed deformity. One of the important extraarticular manifestation is involvement of pulmonary system. The manifestations observed so far are pleuritis followed by transudate pleural effusion and interstitial lung disease (particularly smokers). Hence a PULMONARY FUNCTION TEST is carried among the affected individuals of Rheumatoid Arthritis to get a inference. The detailed analysis of this test help us in understanding the incidence of pulmonary manifestation of rheumatoid arthristis, its effect on respiratory volumes and aid us in further management. AIMS AND OBJECTIVES: 1. To know the incidence of pulmonary manifestation of Rheumatoid Arthritis. 2. To know its effects over pulmonary system. 3. To assess further improvement in treating our patients. METHODOLOGY: This prospective cross sectional study is carried out in RHEUMATOID ARTHRIATIS patients (clinically and serologically confirmed) with respect to inclusion and exclusion criteria, who attend the Rheumatology outpatient clinic or in General Medicine ward of Tirunelveli Medical College and Hospital between April 2010 and April 2019Written informed consent was obtained from the patients selected for the study. They have been elaborated history and subjected to clininal examination. Routine investigations and RA factor were done. Then the patient subjected to perform Pulmonary function testing in the Thoracic Department Tirunelveli Medical College. RESULTS: Out of 100 patients studied 83% were females and 17% were males.32% had pulmonary abnormalities on screening with pulmonary function tests. Mild restriction is the most common pulmonary abnormality seen followed by mild obstruction. Mean age for development of pulmonary abnormality is 45.3 years and is significant by unpaired test. Pulmonary function test in patients with additional risk factors like diabetes, smoking is found to be abnormal. CONCLUSION: Our study highlighted the pulmonary function abnormalities in RA population without any pulmonary disease. The commonest defect pattern seen is mild restrictive pattern followed by obstructive pattern. In most patients the ventilatory defects were mild. There was increased prevalence of abnormal pulmonary function test in patients with additional risk factor like diabetes and hypertension. Rheumatoid disease activity by CDAI score, Age, Duration of disease, Duration of treatment, Diabetes, Hypertension, Smoking were found as predictors of pulmonary impairment as determined by Spirometry

A multivariate analysis of factors affecting the management outcome in rhegmatogenous retinal detachment.

Retinal detachment is a condition in which fluid from the vitreous cavity passes through a full thickness retinal defect into the sub retinal space to cause separation of the neural retina from the underlying Retinal pigment epithelium. The term detachment of the retina is a misnomer. In the first place the retina as a whole is not involved, but a cleavage occurs between the two primitive retinal layers, the pigmentary epithelium remaining in position attached to the membrane of Bruch, while the inner neural layer becomes separated from it due to the opening of the potential space of the primary optic vesicle. The condition of the RD is not a pathological entity but an anatomical accident. In the spontaneous variety, the detachment itself is essential catastrophe and the casual lesion may be so slight as to escape notice even after the most careful search. In the secondary variety, the RD is an incident epiphenomenon in a clinical picture dominated by a neoplasm, inflammation or other pathological features of obvious importance - Sir Stewart Duke Elder. Over centuries the concepts and our approach towards retinal detachment has constantly changed a lot. An effort to trace it takes us through a journey into the evolution of modern ophthalmology itself. This study recommends the recognition of risk factors, proper patient education, periodic detailed examination and early detection as valuable tools to tackle rhegmatogenous retinal detachment and the importance of screening patients with known risk factors. So much is stressed upon the early diagnosis of glaucoma and diabetic retinopathy. But this study clearly emphasizes the importance of duration of retinal detachment in successful treatment outcome. So increased awareness about the symptoms of this condition like flashes and floaters should be provided to the general public. All patients who come for refraction should undergo a proper indirect ophthalmoscope examination under full dilatation with special reference to myopes. In modern cataract surgery refined techniques and instrumentation if meticulously used could greatly prevent the occurrence of posterior capsule rent. This can go a long way in preventing retinal detachment as a delayed complication in such cases. All patients who undergo cataract surgery especially those who undergo surgery in mobile eye camps in rural areas who will not be able to come to regular follow up should be made aware of the symptoms of retinal detachment at the time of final glass prescription. The benefit of YAG capsulotomy especially in the early post operative period in any case of posterior capsular opacification should be weighed against the risk it induces in the occurrence of retinal breaks. And if it has to be done it should be preferably done after six months post operative. Any patient with ocular trauma should undergo detailed indirect ophthalmoscope examination and should be educated about the symptoms of retinal detachment and report immediately if such symptoms occur

A New Sensor Based Communication for Environmental Monitoring

This paper presents the development of a new sensor based communication for environmental monitoring at far off locations. In this proposed system the parameters of sensors are sent to any device having internet by using wireless LAN based on IEEE 802.11b/g standards. The parameters are also sent to mobile phones using GSM. This system includes embedded systems, sensor networks, coordination and management processes and services to capture physical data and to act on the physical environment, all integrated under a intelligent decision system. The system eliminates large amount of solutions, provides the data where network coverage exists