Biochemical and Biomolecular Engineering Commons^™

Optimal Conditions Of Fermentation Of The Pichia Pastoris Yeast Expressing The Recombinant Pres2-S Protein (M-Hbsag) Of Hepatitis B Virus (Hbv), Sasmakоv Sobirdjan, Ashirоv Оybek, Abdurakhmanоv Jaloliddin, Xasanоv Shuhrat

CHEMISTRY AND CHEMICAL ENGINEERING

The aim of this study is determination of optimal fermentation conditions of the Pichia pastoris yeast expressing the recombinant Pres2-S protein of hepatitis B virus (HBV). For this purpose we investigated the main factors influencing for the cultivation of recombinant strain of the yeast Pichia pastoris pPIC3,5-S-HBsAg - the composition of the nutrient medium, pH and dissolved oxygen (DO). As a result, by enzyme-linked immunosorbent assay (ELISA) the optimal concentration of methanol ≈1.0 %, which is the initiator and the only source of carbon for protein expression in the Mut+ phenotype Pichia pastoris yeast cells was determined. In the selected …

Detection Of Nucleotides In Hydrated Ssdna Via 2-D H-Bn Nanopore With Ionic-Liquid/Salt-Water Interface, Jungsoo Lee

Multidisciplinary Studies Theses and Dissertations

Accomplishing slow translocation speed with high sensitivity has been the greatest mission for solid-state nanopore (SSN) to electrically detect nucleobases in single-stranded DNA (ssDNA). In this study, a method to detect nucleobases in ssDNA using a SSN is introduced by considerably slowing down the translocation speed and effectively increasing its sensitivity. The ultra-thin titanium dioxide (TiO₂) coated hexagonal boron nitride (h-BN) nanopore was fabricated, along with an ionic-liquid [bmim][PF₆]/2.0 M KCl aqueous (cis/trans) interfacial system, to increase both the spatial and the temporal resolutions. As the ssDNA molecules entered the nanopore, a …

Bridging Thermochemical And Biochemical Conversion: Impact Of Biochar Addition On The Anaerobic Digestion Of Aqueous Pyrolysis Condensate, Neha Batta

Electronic Thesis and Dissertation Repository

Profitable treatment of low value waste biomass is one of the biggest challenges of the industry. Where most of the current treatment strategies, such as pyrolysis, are efficient in complete breakdown of hard to synthesize, low value waste biomass such as lignocellulosic wastes, it leads to the generation of secondary waste thereby compromising its efficiency. The aim of this research is to utilize the aqueous pyrolysis condensate as a feedstock for anaerobic digestion, previously adapting the microbial consortia to the acidity and other potential organic inhibitors present in the APC, to produce energy in the form of biogas following the …

Micro-Physiological Models To Mimic Mucosal Barrier Complexity Of The Human Intestine In Vitro, Abhinav Sharma

Doctoral Dissertations

The mucosal barrier in the intestine is vital to maintain selective absorption of nutrients while protecting internal tissues and maintaining symbiotic relationship with luminal microbiota. This bio-barrier consists of a cellular epithelial barrier and an acellular mucus barrier. Secreted mucus regulates barrier function via in situ biochemical and biophysical interaction with luminal content that continually evolves during digestion and absorption. Increasing evidence suggests that a mucus barrier is indispensable to maintain homeostasis in the gastrointestinal tract. However, the importance of mucus barrier is largely underrated for in vitro mucosal tissue modeling. The major gap is the lack of experimental material …

Value-Added Lipids Extraction From Wet Microalgae Using Ionic Liquids, Yujie Zhang

Electronic Thesis and Dissertation Repository

Microalgae have gained interest as sources of renewable lipids in the biofuel sector due to their ability to sequester carbon dioxide into triacylglycerol (TAG), a biodiesel feedstock. However, industrial-scale production of microalgae exclusively for fuel production is limited by technical and economic challenges. Some marine microalgae can accumulate large amounts of polyunsaturated fatty acids (PUFAs) such as docosahexaenoic acid (DHA) and other unsaturated fatty acids, which are high-value compounds linked to the prevention of various cardiovascular diseases. This thesis therefore examines the extraction of lipids and DHA from two microorganisms (Chlorella vulgaris, a model organism for lipid production, …

Approaches To Studying Bacterial Biofilms In The Bioeconomy With Nanofabrication Techniques And Engineered Platforms., Michelle Caroline Halsted

Doctoral Dissertations

Studies that estimate more than 90% of bacteria subsist in a biofilm state to survive environmental stressors. These biofilms persist on man-made and natural surfaces, and examples of the rich biofilm diversity extends from the roots of bioenergy crops to electroactive biofilms in bioelectrochemical reactors. Efforts to optimize microbial systems in the bioeconomy will benefit from an improved fundamental understanding of bacterial biofilms. An understanding of these microbial systems shows promise to increase crop yields with precision agriculture (e.g. biosynthetic fertilizer, microbial pesticides, and soil remediation) and increase commodity production yields in bioreactors. Yet conventional laboratory methods investigate these micron-scale …

Rapid Design, Construction, And Validation Of Synthetic Metabolic Pathways In A Modular Escherichia Coli (Chassis) Cell, Jong-Won Lee

Doctoral Dissertations

Current strain development has been hindered by the vast biochemical space in nature. The concept of modular cell design has been invented to enable rapid and predictable construction of multiple optimal production strains for efficient production of a large variety of biochemicals with minimal experimental effort. While modular cell design principles have been successfully validated in some cases, its development is still limited by the small library of the production modules demonstrated.

The goals of this thesis are i) to establish a framework for rapid design, construction, and validation of production modules to explore a large space of molecules (e.g., …

Atherosclerosis: Pathology, Role Of Sd-Ldl, Particle Size Characterization Of Sd-Ldl With Multiangle Light Scattering, Akshay Sudhakar Ukey

Masters Theses

Atherosclerosis is an inflammatory disease due to the accumulation of lipids in the inner wall of arteries. As per the report of Center of Diseases Control and Prevention (CDC), 1 in every 4 deaths is due to heart diseases each year. Atherosclerosis occurs when artery walls harden by the buildup of cholesterol, forming multiple atherosclerotic plaques within the aorta (4). Out of the modern medicines available to control cholesterol, statin is amongst the foremost widely used because it is both safe and effective in lowering high risk patients (9). However, usage of high statins dosage typically leads to some mild …

The Use Of Nanoparticles And Electrospun Fibers For Intravaginal Delivery To Treat Viral And Bacterial Infections And Electrophysiological Measurements Of Synthetic Chloride Channels., Farnaz Minooei

Electronic Theses and Dissertations

Female reproductive viral and bacterial infections affect millions of women worldwide. Given the diversity and magnitude of these unmet reproductive health challenges, topical administration of antiretrovirals (ARVs) and antibiotics have emerged as promising approaches to maintain and restore reproductive health. However, currently available intravaginal dosage forms often suffer from low user adherence and the need for frequent, daily administration to achieve therapeutic effect. To address these challenges, the broad goal of this research was to focus on the development of new localized nanoparticle (NP) and electrospun fiber dosage forms to prolong the delivery and enhance the efficacy of active agents …

Incorporation Of Lignin In Natural And Synthetic Biomaterials To Alter Mechanical And Biochemical Properties For Enhanced Wound Healing, Jorge Alfonso Belgodere

LSU Doctoral Dissertations

It is estimated that chronic, non-healing wounds affect more than 6.5 million Americans annually, with an estimated healthcare cost beyond $14 billion. Here, we attempted to create composites of natural (collagen type I or gelatin-methacrylate) or synthetic (poly(ethylene glycol) polymers incorporating a natural plant component, lignin, to combat the costs and limitations current wound healing methods face. Three-dimensional matrices of collagen type I (Col I) are widely used in tissue engineering applications for its abundance in many tissues, bioactivity with many cell types, and excellent biocompatibility. Inspired by the structural role of lignin in plant tissue, we found that sodium …

Calcium Sulfate-Containing Glass Polyalkenoate Cement For Revision Total Knee Arthroplasty Fixation, Leyla Hasandoost, Adel Alhalawani, Omar Rodriguez, Alireza Rahimnejad Yazdi, Paul Zalzal, Emil H. Schemitsch, Stephen D. Waldman, Marcello Papini, Mark R. Towler

Chemical and Biochemical Engineering Faculty Research & Creative Works

Poly (methyl methacrylate) (PMMA) bone cement is used as a minor void filler in revision total knee arthroplasty (rTKA). The application of PMMA is indicated only for peripheral bone defects with less than 5 mm depth and that cover less than 50% of the bone surface. Treating bone defects with PMMA results in complications as a result of volumetric shrinkage, bone necrosis, and aseptic loosening. These concerns have driven the development of alternative bone cements. We report here on novel modified glass polyalkenoate cements (mGPCs) containing 1, 5 and 15 wt% calcium sulfate (CaSO₄) and how the modified …

Physical And Chemical Characteristics Of Supramolecular Complexes Of Glycirrizinic Acid With Phytohormones, Djuraev Tulqin, Esanov Rakhmatillo, Gafurov Makhmudjon, Kushiev Khabibjon

CHEMISTRY AND CHEMICAL ENGINEERING

The process of formation of supramolecular complexes of glycyrrhizinic acid with some phytohormones (indolylacetic acid, indolylbutyric acid, α-naphthyl acetic acid and kinetin) were studied with the aim of creation the plant growth stimulator. The complexes structure was studied using IR and UV spectroscopy methods. The prepared compounds described by some physicochemical parameters. The stability constants and stoichiometric composition of complexes were determined in an aqueous solution at pH of 7,2 using UV spectrometry method. In result, all complexes have composition as 1:1 ratio, except the kinetin complex which has 2:1 ratio. The stability constants were equal: KGA:IAA=1,86±1×104; KGA:IBA=2,51±1×103; KGA:NAA =2,70±1×103 …

Study Of The Process For The Removal Of Dyeing Matters Of The Plant Occimum Basilicum L. Lamiaceae, Usmonjonova Khulkar, Atkhamova Saida, Dodaev Kuchkor

CHEMISTRY AND CHEMICAL ENGINEERING

The aim of the work is dedicated isolation of anthocyanins by ethanol from herbal plant of Occimum basilicum L. Lamiaceae basis. The were studied influencing of temperature, concentration of ethanol to isolation of anthocyanins. There were passed investigations on studying influence level of heating and hydromodule system of rav material-solvent to maximal isolating of anthocyanins. The colors define by fotokolorometrical method at wave length 400-600 nm. There are shown at 540 nm maximal peak of anthocyanins and it is geven as a grapfical diagram. From the obtained depends on content of alcohol in aquatic solution from outlet of colors is …

Study Of Recombinant Protein Fractions From The Bombyx Mori And Pichia Pastoris Using "Native Page" Method, Abdurhakhmanov Jaloliddin, Sasmakov Sobirdjan, Khasanov Shuhrat, Ashirov Оybek

CHEMISTRY AND CHEMICAL ENGINEERING

We have studied the recombinant protein fractions of ̴ 24-34 kDa obtained from Bombyx mori - silkworm larvae and yeast Pichia pastoris. Fractions, containing the recombinant proteins by gel-electrophoresis using the “Native PAGE” method, which allows for differentiation of proteins according to their natural (surface) charge and conformational properties were studied. The results showed that the recombinant protein fraction from Bombyx mori (̴ 34 kDa) consisted of four components and fraction from Pichia pastoris (̴ 24 kDa) consisted one component. The most optimal conditions for the “Native PAGE”- the main gel: Tris HCl (pH-8.8) 244 mM, Bis-Acrylamide 10%, TEMED 0.08%, …

Novel Application Of Graphene Oxide Based-Biosensor Utilizing Crispr/Dcas9 For Rapid Detection Of Antibiotic Resistance Genes, Joel Omage

Masters Theses & Specialist Projects

Antibiotic resistance is one of the biggest public health challenges globally. To combat this threat, we have designed a method for the facile and rapid detection of antibiotic resistance gene (ARG) in bacteria. We utilized an emerging tool termed the clusters of regularly interspaced short palindromic repeats (CRISPR) in complex with deactivated CRISPR associated proteins (dCas9) on conjugated 2-dimensional graphene oxide (GO) sheets. Nuclease-deactivated Cas9 in complex with customized single guided RNA (sgRNA) was used for the target DNA recognition. The sgRNA was easy to design and we demonstrated that it is complementary to its target sequence by performing a …

Bio-Oils From Vacuum Ablative Pyrolysis Of Torrefied Tobacco Residues, Nattawut Khuenkaeo, Blake Macqueen, Thossaporn Onsree, Sangu Daiya, Nakorn Tippayawong, Jochen A. Lauterbach

Faculty Publications

Fast pyrolysis, in combination with torrefaction pretreatment, was used to convert tobacco residues to value-added bio-fuels and chemicals. Tobacco plant residues were torrefied at 220, 260, and 300 °C, before being pyrolyzed at 450, 500, 550, and 600 °C in a rotating blade ablative reactor under vacuum conditions to test the effects on product yields. With torrefaction, tobacco residues thermally decomposed 20-25% w/w at low temperatures. Torrefaction and pyrolysis temperatures were found to markedly affect pyrolytic product yields of bio-chars and bio-oils, while having no effect on gas-phase products. Bio-oil yields exhibited a direct relation with pyrolysis temperature and an …

Bio-Oils From Vacuum Ablative Pyrolysis Of Torrefied Tobacco Residues, Nattawut Khuenkaeo, Blake Macqueen, Thossaporn Onsree, Sangu Daiya, Nakorn Tippayawong, Jochen A. Lauterbach

Faculty Publications

Fast pyrolysis, in combination with torrefaction pretreatment, was used to convert tobacco residues to value-added bio-fuels and chemicals. Tobacco plant residues were torrefied at 220, 260, and 300 °C, before being pyrolyzed at 450, 500, 550, and 600 °C in a rotating blade ablative reactor under vacuum conditions to test the effects on product yields. With torrefaction, tobacco residues thermally decomposed 20-25% w/w at low temperatures. Torrefaction and pyrolysis temperatures were found to markedly affect pyrolytic product yields of bio-chars and bio-oils, while having no effect on gas-phase products. Bio-oil yields exhibited a direct relation with pyrolysis temperature and an …

Challenges To Levulinic Acid And Humins Valuation In The Sugarcane Bagasse Biorefinery Concept, Emília Savioli Lopes, Jean Felipe Leal Silva, Elmer Ccopa Rivera, Alana Petrina Gomes, Melina Savioli Lopes, Rubens Maciel Filho, Laura Plazas Tovar

Faculty Publications

© 2020, Springer Science+Business Media, LLC, part of Springer Nature. Levulinic acid (LA) is currently one of the most promising chemicals derived from biomass. However, its large-scale production is hampered by the challenges in biomass hydrolysis and the poor selectivity due to the formation of humins (HUs). This study addresses these challenges using the biorefinery concept of biomass fractionation. A three-step process (pretreatment, delignification, and acid-catalyzed conversion) was optimized to produce LA from SCB considering the yield (YLA), efficiency (ELA), and concentration of LA (CLA) as functions of temperature, reaction time, acid concentration, and solids loading. By means of a …

Sequence Control Of Complex Coacervation, Li-Wei Chang

Doctoral Dissertations

Complex coacervation is a liquid-liquid phase separation driven by the complexation of oppositely charged polyelectrolytes. The resulting coacervate phase has been used for many applications, such as underwater adhesives, drug delivery, food and personal care products. There also has been increasing interest in coacervate-like droplets occurring in biological systems. The majority of these “membraneless organelles” involve a combination of intrinsically-disordered proteins and RNA, and phase separate due to long-range charge effects and short-range hydrophobic effects. While evolution has optimized the self-assembly of these types of biological polymers, our ability to design such materials remains limited, in part because the relevant …

Improved Fermentation Design And Screening Devices For Biobutanol Production, Garret Christopher Munch

Electronic Thesis and Dissertation Repository

A worldwide increase in demand for renewable fuels has revived interest in fermentatively produced butanol. However, butanol fermentation suffers from low product yields and productivity. The work presented in this thesis addresses part of these research and development needs at three levels: innovative fermentation process design; genetic manipulation for strain enhancement; and the development of a new tool for anaerobic process characterization and optimization.

Product yield could be increased through traditional fermentation engineering. Co-fermentation of butyric acid with glycerol increased the butanol yield from 0.45 mol/mol (mols C in butanol / mol C in substrates) to 0.51 mol/mol. In building …

Physical And Chemical Characteristics Of Supramolecular Complexes Of Glycirrizinic Acid With Phytohormones, Djuraev Tulqin, Esanov Rakhmatillo, Gafurov Makhmudjon, Kushiev Khabibjon

CHEMISTRY AND CHEMICAL ENGINEERING

The process of formation of supramolecular complexes of glycyrrhizinic acid with some phytohormones (indolylacetic acid, indolylbutyric acid, α-naphthyl acetic acid and kinetin) were studied with the aim of creation the plant growth stimulator. The complexes structure was studied using IR and UV spectroscopy methods. The prepared compounds described by some physicochemical parameters. The stability constants and stoichiometric composition of complexes were determined in an aqueous solution at pH of 7,2 using UV spectrometry method. In result, all complexes have composition as 1:1 ratio, except the kinetin complex which has 2:1 ratio. The stability constants were equal: KGA:IAA=1,86±1×104; KGA:IBA=2,51±1×103; KGA:NAA =2,70±1×103 …

Study Of The Process For The Removal Of Dyeing Matters Of The Plant Occimum Basilicum L. Lamiaceae, Usmonjonova Khulkar, Atkhamova Saida, Dodaev Kuchkor

CHEMISTRY AND CHEMICAL ENGINEERING

The aim of the work is dedicated isolation of anthocyanins by ethanol from herbal plant of Occimum basilicum L. Lamiaceae basis. The were studied influencing of temperature, concentration of ethanol to isolation of anthocyanins. There were passed investigations on studying influence level of heating and hydromodule system of rav material-solvent to maximal isolating of anthocyanins. The colors define by fotokolorometrical method at wave length 400-600 nm. There are shown at 540 nm maximal peak of anthocyanins and it is geven as a grapfical diagram. From the obtained depends on content of alcohol in aquatic solution from outlet of colors is …

Study Of Recombinant Protein Fractions From The Bombyx Mori And Pichia Pastoris Using "Native Page" Method, Abdurhakhmanov Aloliddin, Sasmakov Sobirdjan, Khasanov Shuhrat, Ashirov Оybek

CHEMISTRY AND CHEMICAL ENGINEERING

We have studied the recombinant protein fractions of ̴ 24-34 kDa obtained from Bombyx mori - silkworm larvae and yeast Pichia pastoris. Fractions, containing the recombinant proteins by gel-electrophoresis using the “Native PAGE” method, which allows for differentiation of proteins according to their natural (surface) charge and conformational properties were studied. The results showed that the recombinant protein fraction from Bombyx mori (̴ 34 kDa) consisted of four components and fraction from Pichia pastoris (̴ 24 kDa) consisted one component. The most optimal conditions for the “Native PAGE”- the main gel: Tris HCl (pH-8.8) 244 mM, Bis-Acrylamide 10%, TEMED 0.08%, …

Metabolic Network Analysis Of Filamentous Cyanobacteria, Daniel Alexis Norena-Caro

LSU Doctoral Dissertations

Cyanobacteria were the first organisms to use oxygenic photosynthesis, converting CO₂ into useful organic chemicals. However, the chemical industry has historically relied on fossil raw materials to produce organic precursors, which has contributed to global warming. Thus, cyanobacteria have emerged as sustainable stakeholders for biotechnological production. The filamentous cyanobacterium Anabaena sp. UTEX 2576 can metabolize multiple sources of Nitrogen and was studied as a platform for biotechnological production of high-value chemicals (i.e., pigments, antioxidants, vitamins and secondary metabolites). From a Chemical engineering perspective, the biomass generation in this organism was thoroughly studied by interpreting the cell as a microbial …

The Role Of Poly(Methyl Methacrylate) In Management Of Bone Loss And Infection In Revision Total Knee Arthroplasty: A Review, Leyla Hasandoost, Omar Rodriguez, Adel Alhalawani, Paul Zalzal, Emil H. Schemitsch, Stephen D. Waldman, Marcello Papini, Mark R. Towler

Chemical and Biochemical Engineering Faculty Research & Creative Works

Poly (methyl methacrylate) (PMMA) is widely used in joint arthroplasty to secure an implant to the host bone. Complications including fracture, bone loss and infection might cause failure of total knee arthroplasty (TKA), resulting in the need for revision total knee arthroplasty (rTKA). The goals of this paper are: (1) to identify the most common complications, outside of sepsis, arising from the application of PMMA following rTKA, (2) to discuss the current applications and drawbacks of employing PMMA in managing bone loss, (3) to review the role of PMMA in addressing bone infection following complications in rTKA. Papers published between …

A Review Of The Latest Insights Into The Mechanism Of Action Of Strontium In Bone, Daniella Marx, Alireza Rahimnejad Yazdi, Marcello Papini, Mark R. Towler

Chemical and Biochemical Engineering Faculty Research & Creative Works

Interest in strontium (Sr) has persisted over the last three decades due to its unique mechanism of action: it simultaneously promotes osteoblast function and inhibits osteoclast function. While this mechanism of action is strongly supported by in vitro studies and small animal trials, recent large-scale clinical trials have demonstrated that orally administered strontium ranelate (SrRan) may have no anabolic effect on bone formation in humans. Yet, there is a strong correlation between Sr accumulation in bone and reduced fracture risk in post-menopausal women, suggesting Sr acts via a purely physiochemical mechanism to enhance bone strength. Conversely, the local administration of …

Design, Fabrication And Applications Of Efficient Conductive Polymers For Photocatalytic Antimicrobials, Chunbo Liu

Electronic Thesis and Dissertation Repository

Designing new antimicrobial surfaces which are effective under visible light irradiation without leaching toxic ions is a current challenge for effective disinfection. A new polymeric system poly[2,11’-thiophene-ethylene-thiophene-alt-2,5-(3-carboxyl) -thiophene] (PTET-T- COOH) with broad light absorption was synthesized. Its photocatalytic disinfection performance against staphylococcus aureus (S. aureus) and streptococcus suis (S. suis) was evaluated, showing over 99.999% inactivation (higher than 5-log inactivation) in 2 h for both bacteria, under visible light irradiation at a low concentration of PTET-T-COOH (0.1 mg/mL). In addition, a PTET-T-COOH/polyurethane (PU) polymeric coating was designed and fabricated. Chemical attachment was confirmed between PTET-T-COOH and PU using various thermophysical …

Kinetic Insights Into The Lignocellulosic Biomass-Based Levulinic Acid Production By A Mechanistic Model, Emília Savioli Lopes, Elmer Ccopa Rivera, Julio César De Jesus Gariboti, Luis Henrique Zimmermann Feistel, João Vitor Dutra, Rubens Maciel Filho, Laura Plazas Tovar

Faculty Publications

© 2020, Springer Nature B.V. Abstract: In this work, a mechanistic model was developed to simulate the kinetics of the production of levulinic acid (LA) from sugarcane bagasse (SCB), rice husk (RH) and soybean straw (SS). The production of LA from those agro-industrial wastes followed the methodology of biorefining in three stages. Experimental data from the third stage (catalytic depolymerization of cellulose) obtained under a wide range of operating conditions were used to estimate the parameters of the model. An optimization procedure based on a genetic algorithm was used to determine the optimal parameter values. The prediction of the concentrations …

Peptoid And Antibody-Based Gfp Sensors, Solomon Isu

Graduate Theses and Dissertations

In this work, we have made and characterized a pair of immunobiosensors for detecting the green fluorescent protein (GFP) in an aqueous matrix. An anti-GFP antibody-based biosensor was assembled to detect GFP, while a novel peptoid (N-substituted oligomers of glycine designated as IOS-1) biosensor was also assembled for GFP detection. A quartz crystal microbalance (QCM) gold sensor was used as the supporting substrate for self-assembly of the immunobiosensors. Gravimetric measurements of the QCM gold sensor during immunobiosensor construction and operation were available in real-time using a QCM instrument. X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, and Fluorescence microscopy were used …

Solution To Exide Technologies Inbatec Inefficiency, Shixuan Hou, Reagan Gilker, Mary Fairley, Spencer Christian, Covenson Latouche

Chemical Engineering Undergraduate Honors Theses

The inefficiency of the battery charging time in Inbatec Units 1 and 2 in the Exide Technologies Fort Smith was initially investigated by a student team in Fall 2019, and the investigation was continued into Spring 2020. The Exide Technologies facility in Fort Smith, Arkansas utilizes 13 Inbatec units to charge the lead-acid batteries. The Inbatec systems circulate the sulfuric acid solution through a cooling tower to maintain to optimal charging temperature. Previous analysis of the charging process for Inbatec Units 1 and 2 show the cooling tower have the capacity to quickly remove the excess heat in the sulfuric …