One for All and All in One: Modified Silica Kit-based Protocol for simultaneous sample-specific Extraction of DNA from a Variety of Source Materials

Protocols utilized for the extraction of DNA vary significantly with regards to steps involved and duration of the overall procedure due to material-specific requirements for ensuring the highest possible yield in recovery of DNA. This variation mostly affects aspects of sample preparation and digestion steps required to release the DNA from the sample material.In contexts such as the development of new PCR-based assays - which always includes a test of species-specificity - reference samples from a number of species are utilized, requiring extraction of DNA from a variety of source materials, each with their specific conditions for effective isolation of DNA.The method presented here follows the strategy of synchronizing sample material-specific aspects such as sample preparation and digestion in such a way that one common protocol can be utilized for the actual extraction and purification of the DNA, allowing for an overall more efficient extraction process, while maintaining optimized conditions for DNA recovery.

Comparative Plasmid-Mediated Molecular Resistance Status of Diarrheic Escherichia coli Isolates from Human and Goat Kids

Globally antibiotic resistance has become a major concern, which warrants the real time monitoring for resistance in very common pathogenic organisms. E. coli is normal micro flora in humans, but sometimes it can be pathogenic. For the observation and increment of antimicrobial resistance among pathogen, E. coli has been one of the important pathogens. It is present everywhere in fecal, water, food etc., if resistant E. coli will present in the environment that it can be transferrable anywhere through water, fecal food, animals and humans. This is very dangerous to living beings. This study was designed on status of antibiotic resistance in E. coli isolates in human kids and animal kids, both. Newborns are affected more because of poor or lack of immune system. In this study, fecal materials were used as sample material collected from goat kids (0-3 months) and human children (up to 3 years) residing in same local area. Fifteen fecal samples were collected from human children (up to 3 years) and goat kids (0-3 months) in each case to study the risk of transmission of resistance in E. coli isolates. PCR was conducted on genomic DNA isolates for the presence of usp A gene of E. coli. Multiplex PCR were conducted on plasmid DNA isolates for the resistance specific genes. Molecular resistance results in goat kids isolates showed resistance to antibiotics with tetracycline, sulphonamide, gentamycin, streptomycin and cephalothin to the level of 93.33, 53.33, 46.66, 13.33 & 6.66% respectively, whereas, human E. coli isolates were showed the highest resistance to sulphonamide, Tetracycline and β-lactams were as 53.33, 46.66 and 13.33% respectively but no resistance with gentamycin and streptomycin. Here, we concluded that humans and animals both were refractory to the various groups of antibiotics. This study will help in making the strategy for prevention or reduction of resistance in public

Determination of the optimum ball radius for researching materials using ball indenting

The article discusses the study of the effect of a change in the radius of the ball in the injecting of the sample on the curve in the coordinates «load – indentation depth», the deviation of the indentation depth during elastoplastic indentation from the indentation depth with the elastic indentation and the amount of the axial deformation of the ball. The study was conducted using the Ansys Mechanical APDL program implementing the fenite element method. In the process of the study, it was found that with a change in the radius of the ball, there is no obvious change in the behavior of the sample material, and the deviation of the indentation depth during the elastoplastic indulgence from the indentation depth during the elastic indentation is not dependent on the size of the ball radius. There was also an effect of changing the radius of the ball on the size of the axial deformation of the ball and proposed a formula for determining the size of the axial deformation of the ball for the ball of any diameter, which will determine the actual depth of the ball into the ball when using the balls of different radius.

Different Phenomena Encountered during Dilatometry of Low-density Steels

In recent decades, highly alloyed low-density steels are being developed to reduce the weight of different automotive parts. Dilatometry can be a very useful experimental technique to understand phase transformations during heating or cooling of new low-density steel alloys. When performing dilatometry measurements some assumptions are made such as the homogeneity of the sample material tested during the experiment. In this study, dilatometry tests were performed for two different low-density steels, and the variations of the composition between the surface and the inner part of the sample were analyzed. The migration of manganese by diffusion from the interior of the samples and finally its evaporation on the surface under vacuum were observed. This compositional gradient generated in the samples may influence the veracity and interpretation of the results obtained in dilatometry when working with high manganese steel alloys. The detachment of surface grains created by this compositional change near the surface of the samples is also investigated.

The COMPOND COMPOSITION OF AIRLESS TIRE TREAD (AIRLES TYRE)

The tire without air is a tire that has a very extraordinary influence on the units of vehicle users, both public vehicles and tactical vehicles in the military. In addition to being used in the field of duty, this airless tire can be used for other supporting vehicles of army so there are not many obstacles during operations. Therefore, it takes good materials and proper calculations in making this airless tire tread so that in the future there are no obstacles when using it. The research methods used are experimental methods and field studies. The study was conducted repeatedly and the data was taken directly in the field. After mechanical testing of the rubber material is done to find out the physical properties of the strength of the material. Furthermore, the entire printing and assembly process is carried out and tested on the vehicle, for the conclusion of static analysis of rubber sample material that has been done has determined the proper hardness for the material connected with polyurethane, the appropriate hardness for this rubber is 80 KN/mm. The process has been tested from each sample of rubber hardness variations and the results are determined, in addition to rheometer testing or maturation process takes 5 minutes and is given a temperature of 100 °C and is assisted by hydraulic so that air from volcanic rubber is lost with the help of hydraulic pressure.

Short HPLC gradient method for 20-Hydroxyecdysone (20E) quantification in malaria vectors v1

Ecdysteroids are arthropod steroid hormones that are primarily involved in insect moulting. In arthropod vectors, especially in mosquitoes, ecdysteroids of interest include mainly ecdysone (E) and 20-hydroxyecdysone (20E). These two compounds are involved in several important biological processes. Targeting these compounds and their regulatory pathways could lead to the characterisation of novel genetic tools towards implementing new malaria control strategies. To date, there are two main methods for quantifying E and 20E. These methods include an enzymatic method (Enzyme-Linked Immunosorbent Assay (ELISA)) and a chromatographic method (High-Performance Liquid Chromatography (HPLC)). However, for ecdysteroids quantification, the HPLC methods available in literature go from 30 minutes to one hour. Here, we developed a short HPLC gradient method for 20-hydroxyecdysone quantification in the malaria vector Anopheles gambiae. This method was developed specifically when sample material is limited as well as to save time and cost.

Optimized Protocol for Chelex-based Extraction of DNA from Historical Skeletal Remains and Forensic Trace Samples

PCR-based analysis of skeletonized human remains is a common aspect in both forensic human identification as well as Ancient DNA research. In this, both areas not merely utilize very similar methodology, but also share the same problems regarding quantity and quality of recovered DNA and presence of inhibitory substances in samples from excavated remains. To enable amplification based analysis of the remains, development of optimized DNA extraction procedures is thus a critical factor in both areas.The method paper here presents an optimized protocol for DNA extraction from ancient skeletonized remains using Chelex-100, with improved effectively in yielding amplifiable extracts from sample material excavated after centuries in a soil environment, which consequently have high inhibitor content and overall limited DNA preservation. Further studies showed that the optimized protocol can likewise be utilized for extraction of DNA from common and trace Forensic sample material.

The relationship of the parameters of the optical density of the sample and the absorbance of the sample material

The relationship of the parameters of the optical density of the sample and the absorbance of the sample material is studied, which is relevant for the analysis and differentiation of measurement results by devices with light flux filtration and spectral devices (densitometers and spectrophotometers). To identify the analytical dependence of the obtained values of the optical density of the sample and the absorbance of the sample material, a formula is derived for calculating the diffuse optical transmittance density if the detected radiation contains directional and diffuse components. The first term in the obtained formula characterizes the absorbance of the sample material, the second is a correction due to the diffuse component of the radiation, and is a function of the ratio of the diffuse and regular transmittances. Expressions are obtained that allow functional and quantitative determination of the relationship of the absorbance of the sample material and the parameters of the optical transmittance density of the sample. The results of calculating the absorbance of the sample material, the component of diffuse radiation, diffuse optical transmittance density and corrections of these values are presented.

Dimethylsulfoniopropionate and dimethylsulfoxide in Posidonia oceanica

The present work aims at determining the natural variability of dimethylsulfoniopropionate (DMSP) and dimethylsulfoxide (DMSO) contents in the seagrass Posidonia oceanica, which is the largest producer of these molecules reported to data among coastal autotrophs. Samples were collected during a period of 3.5 years in the pristine Revellata Bay (Calvi, northwestern Corsica, France). The DMSP content ranged from 25 to 265 µmol.gfw−1; DMSO from 1.0 to 13.9 µmol.gfw−1. The dynamics of the two molecules were closely linked, the DMSO content being equivalent to 3.5 % of the DMSP content, all leaf samples considered (n = 423 samples and 414 DMSP(O) data pairs). The annual growth cycle of the seagrass diluted the initial stocks of the two molecules. Temperature indirectly affected molecule content dynamics through their direct effect on the seagrass productivity and biomass. Inter-annual variations in DMSP(O) content in relation to shallow water temperature might further indicate that DMSP(O) could have been involved in the physiological response of P. oceanica to heat-stress. Finally, middle-aged leaf tissues with an organosulfur molecule content similar to the average value calculated for the seagrass leaf bundle appeared to be the best choice of sample material to study DMSP and DMSO in that species. More research is needed to elucidate the biosynthetic pathways of these molecules in seagrasses, the evolutionary reasons for such a high production in P. oceanica and the physiological functions they play.

Description of the concrete carbonation process with adjusted depth-resolved thermogravimetric analysis

The thermal gravimetric analysis (TG) is a common method for the examination of the carbonation progress of cement-based materials. Unfortunately, the thermal properties of some components complicate the evaluation of TG results. Various hydrate phases, such as ettringite (AFt), C–S–H and AFm, decompose almost simultaneously in the temperature range up to 200 °C. Additionally, physically bound water is released in the same temperature range. In the temperature range between 450 °C and 600 °C, the decomposition of calcium hydroxide and amorphous or weakly bound carbonates takes place simultaneously. Carbonates, like calcite, from limestone powder or other additives may be already contained in the noncarbonated sample material. For this research, an attempt was made to minimise the influence of these effects. Therefore, differential curves from DTG results of noncarbonated areas and areas with various states of carbonation of the same sample material were calculated and evaluated. Concretes based on three different types of cement were produced and stored under accelerated carbonation conditions (1% CO2 in air). The required sample material was obtained by cutting slices from various depths of previously CO2-treated specimen and subsequent grinding. During the sample preparation, a special attention was paid that no additional carbonation processes took place. As reference method for the determination of the carbonation depth, the sprayed application of phenolphthalein solution was carried out. Microscopic analysis was examined to confirm the assumptions made previously. Furthermore, the observed effect of encapsulation of calcium hydroxide by carbonates caused by the accelerated carbonation conditions was examined more closely.