Automated Protein Quantification with the Biomek-FX Liquid Handler System v2

This protocol details steps to perform the protein quantification (Lowry-based) assay by using a Biomek FX liquid handler system. It is optimized to assay a full 96-well plate of protein samples in duplicate with a separate (control) plate for BSA standards. You will need a plate reader to measure the samples and standards. This protocol works best as part of a full proteomic sample preparation workflow with: Automated Chloroform-Methanol Protein Extraction on the Biomek-FX Liquid Handler System and Automated Protein Normalization and Tryptic Digestion on a Biomek-FX Liquid Handler System

T-Shaped Control Plate Effect on Flow past a Square Cylinder at Low Reynolds Numbers

In this study, the influence of the T-shaped control plate on the fluid flow characteristics around a square cylinder for a low Reynolds numbers flow is systematically presented. The introduction of upstream attached T-shaped control plate is novel of its kind as T-shaped control plate used for the first time rather than the other passive control methods available in the literature. The Reynolds numbers (Re) are chosen to be Re = 100, 150, 200, and 250, and the T-shaped control plate of the same width with varying length is considered. A numerical investigation is performed using the single-relaxation-time lattice Boltzmann method. The numerical results reveal that there exists an optimum length of T-shaped control plate for reducing fluid forces. This optimum length was found to be 0.5 for Re = 100, 150, and 200 and 2 for Re = 250. At this optimum length, the fluctuating drag forces acting on the cylinder are reduced by 134%, 1375, 133%, and 136% for Re = 100, 150, 200, and 250, respectively. Instantaneous and time-averaged flow fields were also presented for some selected cases in order to identify the three different flow regimes around T-shaped control plate and square cylinder system.

Numerical Simulation Studies on Stall Suppression of a NACA0015 Airfoil

This study aims to achieve an improved airfoil performance at low Reynolds number, and to determine the optimum position and size of rectangular cross-section burst control plate (BCP) to suppress stall in airfoil. The type of airfoil used in the present study is NACA0015 (National Advisory Committee for Aeronautics) airfoil with 200 mm of chord (c) length. Here, rectangular cross-section burst control plates with different sizes and at different locations are investigated numerically at the low Reynolds number of 1.6×105. Total of three positions (0.05c, 0.1c and 0.2c from the leading edge of airfoil), and four sizes (with heights 0.3 mm, 0.7mm, 1mm and 1.5 mm, and constant width 4 mm) of rectangular BCPs are simulated in ANSYS Fluent software using Transition SST model. The results indicate that the rectangular cross-section burst control plate is an effective device in the suppression of airfoil stall. For 0.7 mm and 1 mm height BCPs, the stall angle is postponed by 2° for all positions, while for 0.3 mm and 1.5 mm height BCPs, the reduction in sudden fall of lift can be observed but at the cost of reduction in maximum lift coefficient. Among various configurations, the 1mm height BCP located at 0.2c position is found to be most effective in the suppression of stall.

Automated Protein Quantification with the Biomek-FX Liquid Handler System v1

This protocol details steps to perform the protein quantification (Lowry-based) assay by using a Biomek FX liquid handler system. It is optimized to assay a full 96-well plate of protein samples in duplicate with a separate (control) plate for BSA standards. You will need a plate reader to measure the samples and standards. This protocol works best as part of a full proteomic sample preparation workflow with: Automated Chloroform-Methanol Protein Extraction on the Biomek-FX Liquid Handler System and Automated Protein Normalization and Tryptic Digestion on a Biomek-FX Liquid Handler System

In vitro control of Botrytis cinerea and Penicillium italicum by antagonistic yeasts

Gray mold in grapes (caused by Botrytis cinerea) and blue mold in citrus (caused by Penicillium italicum) are post-harvest diseases that cause major losses in these crops. The control of these diseases is based on the use of synthetic chemical fungicides. The increase in regulatory policies and demand to reduce the application of pesticides, due to harmful effects on the environment and humans, have led to the search for more ecofriendly alternatives, such as biological control agents. Thus, the present work aims to verify the antagonistic potential of four yeast strains, Pichia caribbica (CCMA 0759), Hanseniaspora opuntiae (CCMA 0760), Pichia manshurica (CCMA 0762), and Lachancea thermotolerans (CCMA 0763), against of B cinerea and P. italicum. To assess the antagonism of volatile compounds, Petri plates with two divisions containing potato-dextrose-agar (PDA) were used by placing a fungal mycelial disc and yeast suspension (3.0 × 106 cells mL-1) on opposite sides of the plate. The colony diameter and mycelial growth rate index of the fungi were evaluated via comparisons with the control plate without yeast. For the evaluation of the antagonism of diffusible substances in the medium, yeasts were striated 3 cm from the center of the plates containing PDA. After 48 h, a mycelial disc of each phytopathogen was placed in the center of the plates. The colony growth, inhibition halo, and mycelial growth rate index were evaluated via comparisons with the control plate. All yeast strains showed an antagonistic effect on the mycelial growth of B. cinerea in both tests. In the volatile compounds test, H. opuntiae, L. thermotolerans, P. caribbica, and P. manshurica inhibited mycelial growth by approximately 82%, 75%, 72%, and 50%, respectively. In the antagonism test of the diffusible substances in the medium, P. caribbica and P. manshurica inhibited mycelial growth by 58% and 33%, respectively. However, these yeast strains did not show an antagonistic effect against P. italicum. Thus, all isolates demonstrated potential to be tested as biocontrol agents of gray mold in post-harvest grape fruits.

Carbamate, Organophosphate and Organochlorine Pesticidal Effect on the Qualitative Occurrence and Survivability of Blue-green Algae

Blue-green algae make a valuable contribution to the nitrogenous soil fertility by fixing atmospheric nitrogen and are of great agricultural importance as biofertilizer. Four commercial grade pesticides, Carbamate, Furadan and Sevin and Organophosphate, Rogor and Organochlorine, Endotaf were used to study their effect on the survivability and qualitative occurrence of blue-green algae by taking survival on the control plate as 100%. Stock solutions of these pesticides were prepared freshly for experiments in the sterilized BG-11 media and added to the culture media to obtain the desired concentrations of 100, 250, 500 and 1000 ppm. After 30 days of incubation, the blue-green algal forms appeared in the culture flask were identified using standard monographs. The pragmatic results indicated that, soil blue-green algae show variable resistance to pesticide treatments. The survival percentage of the tested blue-green algae was reduced upto 50 percent at 500 ppm of carbamate pesticides, Furadan and Sevin. While with Rogor only 10.52% and Endotaf 5.26% survivability was noticed at 500 ppm dose level. Furthermore, the organochlorine pesticide Endotaf was more toxic than the organophosphate, Rogor and the carbamates, Sevin and Furadan in sequence to the blue-green algae. It was cocluded that higher doses of pesticide application in the crop fields i.e. more than 100 ppm of Furadan, Sevin and Rogor and even at 100 ppm of Endotaf, qualitative and quantitative occurrence of heterocystous and non- heterocystous blue-green algae was decreased considerably. This suggests that, indiscriminate use of these pesticides may cause adverse effects on the nitrogen fixing blue-green algae of various crop fields, which has a direct influence on total productivity. In general, the sensitiveness of different blue-green algae to pesticide application was found to be more in sheathless heterocystous and unicellular forms than the heterocystous ensheathed and non- heterocystous ensheathed forms.