Electrophoresis-Aided Biomimetic Mineralization System Using Graphene Oxide for Regeneration of Hydroxyapatite on Dentin

Graphene oxide (GO) is an emerging luminescent carbon nanomaterial with the ability to foster hydroxyapatite (HA). A specially designed electrophoresis system can be used to accelerate the mineralization process. The aim of this study was to promote HA crystal growth on demineralized dentin using a GO incorporated electrophoresis system. GO was successfully synthesized by carbonization of citric acid and its presence was confirmed by Fourier transform infrared and UV-visible spectrophotometry evaluation. Dentin slices were placed in demineralized solution and divided into control (without the electrophoresis system) and experimental group. Demineralized dentin slices in the experimental group were remineralized using the electrophoresis system for 8 h/1.0 mA, with one subgroup treated without GO and the other with GO. Energy dispersive spectroscopy evaluation showed that the calcium/phosphate ratio of the crystal formed in control and experimental group with addition of GO was close to natural hydroxyapatite. However, scanning electron microscopy evaluation showed that the exposed dentinal tubules were occluded with rod-like crystals, which is similar to native enamel morphology, in the experimental group with addition of GO compared to the flake-like crystal in the control group. Mechanical evaluation revealed that the nanohardness and modulus of remineralized dentin were significantly higher in the experimental group. In conclusion, GO is a promising material to remineralize dentin and the introduction of an electrophoresis system can accelerate its process.

RNA Quality Control Non-Denaturing Agarose "Bleach" Gel v1

To evaluate the quality of RNA without the use of toxic chemicals, samples are run in a non-denaturing agarose "bleach" gel and product segregation is used to estimate RNA integrity. Be aware that in a non-denaturing gel, the RNA will not segregate strictly on particle size due to secondary structures of the molecules. This method is for an estimation of quality only and the location of the banding in relation to a base pair ladder does not allow confident determination of the size of the RNA fragment. If true fragment size determinations are required, RNA quality should be evaluated on a denaturing gel electrophoresis system.

An Automated Hydrodynamically Mediated Technique for Preparation of Calibration Solutions via Capillary Electrophoresis System as a Promising Alternative to Manual Pipetting

In this paper, a novel procedure for preparing calibration solutions for capillary electrophoresis (CE)-based quantitative analysis is proposed. Our approach, named the automated hydrodynamically mediated technique (AHMT), uses a capillary and a pressure system to deliver the expected amount of working solution and diluent directly to a sample vial. As a result, calibration solutions are prepared automatically inside the CE instrument, without any or with minimal manual operation. Two different modes were tested: forward and reverse, differing in the direction of hydrodynamic flow. The calibration curves obtained for a model mixture of analytes using AHMT were thorough compared to the standard procedure based on manual pipetting. The results were consistent, though the volume of obtained calibration solutions and the potential risk of random errors were significantly minimized by AHMT. Its effectiveness was further enhanced by the application of SCIEX® nanoVials, reducing the actual volume of calibration solutions down to 10 μL.

Development of an In-Line Enzyme Reactor Integrated into a Capillary Electrophoresis System

The goal of this paper was to develop an in-line immobilized enzyme reactor (IMER) integrated into a capillary electrophoresis platform. In our research, we created the IMER by adsorbing trypsin onto the inner surface of a capillary in a short section. Enzyme immobilization was possible due to the electrostatic attraction between the oppositely charged fused silica capillary surface and trypsin. The reactor was formed by simply injecting and removing trypsin solution from the capillary inlet (~1–2 cms). We investigated the factors affecting the efficiency of the reactor. The main advantages of the proposed method are the fast, cheap, and easy formation of an IMER with in-line protein digestion capability. Human tear samples were used to test the efficiency of the digestion in the microreactor.

Evaluation of Aqueous Biphasic Electrophoresis System Based on Halide-Free Ionic Liquids for Direct Recovery of Keratinase

Aqueous biphasic electrophoresis system (ABES) incorporates electric fields into the biphasic system to separate the target biomolecules from crude feedstock. Ionic liquid (IL) is regarded as an excellent candidate as the phase-forming components for ABES because of the great electrical conductivity, which can promote the electromigration of biomolecules in ABES, and thereby enhances the separation efficiency of the target biomolecules from crude feedstock. The application of electric fields to the conventional biphasic system expedites the phase settling time of the biphasic system, which eases the subsequent scaling-up steps and reduces the overall processing time of the recovery process. Alkyl sulphate-based IL is a green and economical halide-free surfactant when compared to the other halide-containing IL. The feasibility of halide-free IL-based ABES to recover Kytococcus sedentarius TWHK01 keratinase was studied. Optimum partition coefficient (Ke = 7.53 ± 0.35) and yield (YT = 80.36% ± 0.71) were recorded with IL-ABES comprised of 15.0% (w/w) [EMIM][ESO4], 20.0% (w/w) sodium carbonate and 15% (w/w) crude feedstock. Selectivity (S) of 5.75 ± 0.27 was obtained with the IL-ABES operated at operation time of 5 min with 10 V voltage supplied. Halide-free IL is proven to be a potential phase-forming component of IL-ABES for large-scale recovery of keratinase.

β-globin variants present in Western Sudan

Objective: A variety of observations of the hemoglobin D (Hgb D) phenotype has occurred in association with family studies of patients with sickle cell disease. Very little is known about the occurrence and prevalence of the Hgb D variant and its impact on blood profiles among Sudanese. This study was aimed at determining the percentage of Hb D in North Darfur State, West Sudan, whose population has been shown to have hemoglobin-S (Hb S) disease. Methods: From December 2017 to August 2018, this descriptive community-based investigation was conducted. Six hundred and sixty-six (666) people were randomly selected to participate in this study. With each participant's there was verbal consent. A questionnaire was designed to collect personal details. 5 mL of venous blood was gathered in EDTA containers. The Hb D variant was checked using Sebia Minicap Automated Capillary Electrophoresis System- USA and frequency was calculated using version 21.0 of the software package for social science (SPSS). Result: The prevalence of Hb variants was as follows: AD=0.6%, SS=2%, AS=10.5 %, and AA=86.9%. Conclusion: The prevalence of Hb D variant was 4 (0.6 %) in 666 participants from four western Sudanese tribes, beside reported of Hb SS and Hb AS.

Prevalence of Sickle Cell Anemia in Association of Plasmodium falciparum from Rajnandgaon, India

Sickle cell anemia is a blood disorder resulting from the inheritance of abnormal genes from parents. It is caused due to mutation in the β-globulin gene. Sickle cell anemia is widespread across the world and in Central India. The present study was undertaken to study the prevalence of the disorder in the Rajnandgaon district of Central India. A random sampling of 6088 people was done to test the sickle cell anemia problem by slide test method and a total of 249 (4.09%) people were found sickled positive. Further electrophoresis test was performed for all 249 of which 67 were found homozygous (HbSS) and 182 were found heterozygous (HbAS) positive. Besides the above analysis, chloroquine prophylaxis associated with a high prevalence of Plasmodium falciparum Pfcrt K76T mutation in people (n=26) with sickle cell anemia was also analyzed. The genotype of the subject was screened using the hemoglobin electrophoresis system and the P. falciparum Pfcrt genotyping was carried out using PCR-Restriction Fragment Length Polymorphism (RFLP). The prevalence rate of Pfcrt K76T mutant gene was proportionately found higher in the hemoglobin SS (n=40, m=32, r=0.67) genotype individuals than the hemoglobin AS (n=52, m= 27, r=0.519) and AA (n=182, m= 68, r= 0.37).