Bradford Assay Research Articles

Virulence and biofilm inhibition of 3-methoxycinnamic acid against Agrobacterium tumefaciens.

In the current study the anti-virulence and anti-biofilm activities of the cinnamic acid derivative, 3-methoxycinnamic acid, was investigated against Agrobacterium tumefaciens. Based on the disc diffusion test and β-galactosidase activity assay, 3-methoxycinnamic acid was shown to interfere with the quorum sensing (QS) system of A. tumefaciens. Crystal violet staining assay, phenol-sulfuric acid method, Bradford protein assay and confocal laser scanning microscopy (CLSM) revealed that the biofilm formation of A. tumefaciens was inhibited after the treatment of 3-methoxycinnamic acid. Employing high-performance liquid chromatography (HPLC) analysis of culture supernatant revealed that the production of 3-oxo-octanoylhomoserine lactone (3-oxo-C8-HSL) decreased concentration-dependently after treatment with 3-methoxycinnamic acid. Swimming and chemotaxis assays also indicated that 3-methoxycinnamic acid had a good effect on reducing the motility and chemotaxis of A. tumefaciens. In addition, the RT-qPCR, molecular docking and simulations further demonstrated that 3-methoxycinnamic acid could competitively inhibit the binding of 3-oxo-C8-HSL to TraR and down-regulate virulence-related genes. 3-Methoxycinnamic acid is proved to have good anti-virulence and anti-biofilm activities against A. tumefaciens. This is the first study that investigates the anti-virulence and anti-biofilm activities of 3-methoxycinnamic acid against A. tumefaciens. With its potential QS-related virulence and biofilm inhibitory activities, 3-methoxycinnamic acid is expected to be developed as a potent pesticide or adjuvant for the prevention and treatment of crown gall caused by A. tumefaciens.

Effects of Pre-Heating Between Controlled Temperature Microwave Heating and Hot Air Oven Heating in Protein Extraction Process of Soybeans

Abstract Heating is an important treatment before protein extraction process. Rapid heating of microwave heating (MW) was investigated to compare with hot air oven (HA) heating prior to protein extraction in soybeans. Heating temperature was controlled at 50°C, with the heating time of 15, 30, 45, 60 and 75 min. The control treatment was no heat treatment. The treated samples were analyzed i) the total protein using Kjeldahl method, ii) the protein soluble by Bradford method and iii) cells microscope investigated by scanning electron microscope (SEM). The results were found that the pretreatment of 75 min microwave (MW75) heating provided the highest protein with the amount of absorbance value and total protein of 3.0365 ± 0.004 mg mL-1 and 3.70 ± 0.01% respectively. The lowest protein extraction of 2.7707 ± 0.020 mg mL-1 and 3.38 ± 0.005% total protein value occurred in the condition of 15 mines hot air oven (HA15) heating. The investigation using scanning electron microscope (SEM) revealed that protein extraction correlated with the enlargement and breakage of cell wall and endosperm caused by microwave heating rather than hot air oven heating. The rapid heating of microwave and heating time influenced the amount of extracted protein. Keywords: Soybean, Protein, Microwave, Hot air, Heating

Portuino-A Novel Portable Low-Cost Arduino-Based Photo- and Fluorimeter.

A novel portable low-cost Arduino-controlled photo- and fluorimeter for on-site measurements has been developed. The device uses LEDs as a light source and a phototransistor as a light sensor. The circuit is based on the discharge of a capacitor with the photocurrent from the phototransistor. Validation experiments for absorbance measurements were performed by measuring protein concentration using the Bradford method and measuring phosphate ions in water using a commercial test kit. The emission light of the excited fluorescent dyes rhodamine 6G and calcofluor white was measured to validate the usability of the device as a fluorescence photometer. In all validation experiments, similar correlation coefficients and limit of detection could be achieved with the portable photo- and fluorimeter and a laboratory spectrometer and fluorimeter. Real sample analysis was performed, measuring phosphate concentration in freshwater and concentration of green fluorescent protein, extracted from Escherichia coli.

Protein change in Chardonnay wine from Bulgaria after treatment with different types of bentonites

Protein profile change of Bulgarian Chardonnay wine under the influence of treatment with bentonite was studied. Protein concentration was determined using the Bradford assay, while fractionation was performed via sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The amount of protein found in the original wine was 147.1 mg/dm3, expressed as bovine serum albumin divided into 9 electrophoretic fractions. Low molecular weight fractions with molecular weights of 20, 22 and 24 kDa, were predominant whose share was equal to 65 % of the total electrophoretic fraction profile of wine. The examined wine was treated with four different types of bentonite-two sodium, one activated calcium and one sodium-calcium bentonites. They were added in increasing doses, covering the range of 0.2-3 g/dm3. At lower treatment doses (0.2-0.8 g/dm3), differences in protein concentration reduction were observed. Above the dose of 0.8 g/dm3, an equalization of their effect compared to the reduction of total protein content was reported. Affinity, however small, of the studied bentonites to the different molecular fractions of proteins was observed.

Correlations between the Circulating Level of Cell-Derived Microparticles and Surgical Variables in Heart Valve Surgery with Cardiopulmonary Bypass.

Cell-derived microparticles (MPs) as membrane vesicles are procoagulant. They play a role in surgical hemostasis. In this study, the correlations between the circulating level of cell-derived MPs and surgical variables in heart valve surgery were investigated. The present prospective case-series study was conducted in Rajaie Cardiovascular Medical and Research Center from January through March 2021. Forty patients undergoing heart valve surgery with cardiopulmonary bypass (CPB) were enrolled. Before the induction of anesthesia and 30 minutes after the administration of protamine sulfate, venous blood samples were collected. After MP isolation, the concentration of MPs was determined via the Bradford method. Flow cytometry analysis was performed to determine the MP count and phenotype. Intraoperative variables and postoperative routine coagulation tests were defined as surgical variables. Postoperative coagulopathy was defined as an activated partial thromboplastin time (aPTT) ≥48 seconds or an international normalized ratio (INR) >1.5. The total concentration of MPs and the MP count increased significantly after surgery compared with before surgery. The postoperative concentration of MPs was positively correlated with the CPB time (P=0.030, ρ=0.40). The preoperative concentration of MPs was significantly lower in patients with higher postoperative aPTT and INR (P=0.003, P= -0.50 and P=0.020, P= -0.40, respectively). In multivariate logistic regression analysis, the preoperative MP concentration (OR, 1.00; 95% CI, 1.00 to 1.01; P=0.017) was considered a risk factor for postoperative coagulopathy. The levels of MPs, especially platelet-derived MPs, rose after surgery, in correlation with the CPB time. Given the role of MPs in the induction of coagulation and inflammation, they can be considered therapeutic goals for preventing postoperative complications. In addition, the preoperative levels of MPs are a risk factor for predicting the occurrence of postoperative coagulopathy in heart valve surgery.

Analysis of Saliva Composition: Parathyroid Hormone-Related Protein, Total Protein, and Secretory Immunoglobulin A (sIgA) in Rattus norvegicus with Stunted Growth.

This study aimed to determine total protein, secretory immunoglobulin A (sIgA) and parathyroid hormone-related protein (PTHrP) levels in the saliva of rats with stunted growth. Experimental laboratory research with a pre-and posttest control group design was conducted. Seventeen albino rats (Rattus norvegicus) were divided into the control group (eight rats) and the treatment group (nine rats). Rats in the treatment group were exposed to aflatoxin B1 5µg/kg orally for 5 weeks. Anthropometry data (body length, body weight) and saliva of R. norvegicus were collected. The levels of PTHrP and sIgA in the saliva were measured using an enzyme-linked immunosorbent assay kit for rats and the Bradford test for total protein and analyzed using SPSS 25.0. Aflatoxin caused stunted growth in rats in the treatment group. There was a significant difference in body length, salivary flow, PTHrP, sIgA, and total protein in the treatment group compared with the control group. The average rat's body length change in the control group was 6.4 ± 1.1mm/5 weeks, while in the treatment group, the change was 3.7 ± 0.9 mm /5 weeks. There was no significant weight gain in the treatment group compared with the normal group. The average values of PTHrP, sIgA, and total protein in the control group were x̄0.9, x̄18, and x̄0.7 m./L, respectively, while in the treatment group, they measured x̄0.4, x̄10.7, and x̄0.5 mg/L, respectively. This study showed that salivary flow, PTHrP, sIgA, and total protein levels in the saliva were significantly lower in stunted rats compared with normal rats.

Enzymatic hydrolysis of Moringa oleifera Lam flour using bromelain and fig by-products as sources of protease

Moringa oleifera Lamarck flour (MOF) has been recognized as a source of vegetable proteins; however, around 70.1±0.98% of protein can be called “insoluble protein”, for its resistance to gastrointestinal digestion. For protein to be absorbed in the human intestinal tract, it must be consumed in the form of a soluble protein. Proteases have shown to increase soluble proteins. The objective of this study was, hence, to investigate the increase in soluble proteins of MOF soluble and insoluble water fraction through the hydrolysis with proteases obtained from fig and pineapple by-products, ficin and bromelain, respectively. In conclusion, the protein of MOF had changes in their solubility by enzymatic effect. In particular, the hydrolysis of water insoluble Moringa oleifera Lam flour fraction (IMOF) with ficin contributed to a significant increase in soluble proteins, a result detectable through the different methods applied (colorimetric Bradford method, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and Reversed phase high-performance liquid chromatography (RP-HPLC)).

Exosomes from human adipose-derived stem cells promoted the expression of angiogenic factors in endothelial cells

Introduction: In regenerative medicine, human adipose tissue–derived mesenchymal stem cells (hadMSCs) have exhibited tremendous promise as an efficient approach to chronic cardiovascular diseases. Noble effects of mesenchymal stem cells (MSCs) have been acknowledged to contribute to not only cellular engraftment and response to the site of injury but also paracrine action via the release of extracellular vesicles, notably exosomes. This study aimed to investigate the effects of exosomes from hadMSCs (hadMSC-Exo) on the angiogenesis of endothelial cells. Methods: Firstly, hadMSCs were characterized and cultured at a density of 500,000 cells in exosome-free medium for 48 hours. The conditioned medium was ultracentrifuged for exosome isolation. Obtained exosomes were characterized with the expression of CD9, CD63, and CD81 markers by flow cytometry and imaged with a TEM microscope. Exosomes were added to a culture medium of human umbilical vein endothelial cells (HUVECs) to understand the effects of exosomes on HUVECs. The expression of some angiogenic genes in HUVECs was identified by RT-qPCR. Results: The concentration of total protein in isolated exosomes by Bradford assay was 0.51 ± 0.04 mg/mL. Exosomes were successfully isolated and characterized by their morphology and immunophenotype. The results showed that exosomes significantly increased the mRNA expression of TGF (11.56 ± 2.03, P < 0.05), Flk-1 (4.04 ± 0.01, P < 0.05), VE-cadherin (7.25 ± 2.30, P < 0.05), and CD31 (3.02 ± 1.13, P < 0.05). Conclusion: This study suggested that exosomes promoted the angiogenesis of endothelial cells in vitro.

Anticandidal Activity of Lactoferrin, Apolactoferrin, and Oligosaccharides on Mueller-Hinton and Sabouraud Dextrose Agar against Fluconazole Resistant-Candida Albicans

Background: The demand for novel, useful, potential, and safe antifungal drugs and rapid fungal susceptibility test methods due to antifungal resistance and high prevalence of Candida albicans infection are continuing. Therefore, this study aimed to assess and compare the antifungal activity of lactoferrin, apolactoferrin, and oligosaccharides isolated from human, bovine, goat, and formula milk against C. albicans on Mueller-Hinton agar supplemented with 2% glucose and 5 µg/mL methylene blue and sabouraud dextrose agar.Methods: Lactoferrin, apolactoferrin, and oligosaccharides were extracted from human, bovine, goat, and formula milk. Lactoferrin was identified using the Bradford test and Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis. Oligosaccharides were identified using Thin-Layer Chromatography. The antifungal activity of lactoferrin, apolactoferrin, and oligosaccharides against fluconazole-resistant Candida albicans ATCC 10231 was determined and compared using the disk diffusion method on Mueller-Hinton agar and sabouraud dextrose agar. The inhibition zone formed around the disk was observed after 24 hours of incubation.Results: Lactoferrin showed an inhibition zone on sabouraud dextrose agar against C. albicans, but not on Mueller-Hinton agar. Meanwhile, apolactoferrin and oligosaccharides showed no antifungal activity on both agar media.Conclusions: Different agar media in the diffusion disk test can give different results even though using the same test method and substance. These results could shed light and become the useful references on why some potential antifungals could yield a different results in in-vitro studies, in-vivo studies, or clinical trials.

The Expression of Plasma Protein in Bali-polled Bulls Using 1D-SDS-PAGE

The fertility rate of bulls in a breeding program is not only described by the quantity and quality of semen. Factors, such as the interstice factor of the sperm and the plasma component of semen, affect the fertility rate of bulls. The fertility rate can also be determined by identifying the protein content of semen plasma. Therefore, the current study aimed to identify the relationship between seminal plasma protein molecular weight and semen quality of Bali-polled bulls. The study was conducted at the Laboratory of Semen Processing, Faculty of Animal Science, Hasanuddin University, Makassar, Indonesia, the Research Center for Applied Zoology, National Research and Innovation Agency, Cibinong, Indonesia and the Laboratory of Animal Biotechnology Center, IPB University, Bogor, Indonesia from November 2021 to January 2022. The samples came from 5 Bali-polled and 5 Bali-horned bulls. Semen collection was conducted twice a week using an artificial vagina. The concentration of seminal plasma protein was determined by the Bradford method of 1D-SDS-PAGE. The study results showed that fresh semen of Bali-polled and Bali-horned bulls was considered a normal category. Seminal plasma proteins of Bali-polled and Bali-horned bulls were classified using 8 bands to categorize molecular weight; 150 kD (IGF-1), 110 kD (A-kinase anchoring protein 3), 93 kD (A-kinase anchoring protein 4), 54-87 kD (Arylsulfatase-a), 44-62 kD (N-Acetyl-ß-Guicosaminidase), 44kD (Phosphoglycerate kinase), 15-30 kD (BSP A1/A2, BSP-A3 and BSP-30 [BSP1, BSP3, and BSP5]) and 12-14 kD (Acidic seminal fluid proteins). The findings indicated that both Bali-polled and Bali-horned bulls could have a high reproductive rate. In conclusion, protein analysis based on molecular weight using 1D-SDS-PAGE can be used as a biomarker for semen quality in Bali-polled bulls. Therefore, evaluating the semen quality with a molecular basis as an additional indicator of superior bull in the selection process is an alternative method.

Comparative study of the most commonly used methods for total protein determination in milk of different species and their ultrafiltration products.

Milk ultrafiltration is a widely used membrane filtration process that allows the recuperation of whey proteins in a concentrate high in total solids, which can later be transformed in multiple healthy dairy products with great prospects for the food industry. Protein content is a decisive factor for the technological performance of milk concentrates and currently, the ISO standard method for its determination is Kjeldahl, which is time-consuming and requires specific instrumentation. For this reason, the use of rapid methods to quantify protein would greatly facilitate the monitoring of the milk ultrafiltration process. In this study, the bicinchoninic acid assay (BCA), the detergent compatible Bradford assay and the Dumas method were compared to Kjeldahl protein determination to select a quick and accurate methodology suitable for milk of different species and its ultrafiltration products (retentates and permeates). The protein content obtained from Bradford assay and Dumas method in origin milk and retentate samples was consistent with Kjeldahl values. In contrast, BCA protein levels were significantly different when compared to Kjeldahl and no method was proved to be suitable for protein determination in permeate samples. The use of sodium dodecyl sulfate was also examined to improve protein measurements without success. In comparison with the official method, Bradford assay quantitatively provided the best results, and it would be recommended for a quick, economic and easy determination of total protein content in milk and retentate samples.

Atorvastatin attenuates NS1 (Non-structural protein-1) of dengue type-2 serotype-induced expressions of matrix metalloproteinases in HL-60 cells, differentiated to neutrophils: Implications for the immunopathogenesis of dengue viral disease

BackgroundThe dengue is a vector borne viral infection in humans. Bite of mosquito infected with a dengue virus transmits the disease. The neutrophils support more to the innate immune response by switching to infected tissues and triggering immunomodulatory mechanisms including the release of proteases and host defence peptides. MethodsCell viability by MTT and trypan blue dye exclusion assay, bright field microscopy for assessment of cell morphology, cytokines measurements by ELISA, estimation of protein by Bradford assay were done. Assessments of matrix metalloproteinase genes mRNA expressions were done using real-time PCR. ResultsIn the present study, we have for the first time unveiled that, NS1 antigen of dengue type-2 serotype, induce and stimulate the neutrophils cells to express high levels of matrix metalloproteases. NS1 exposure of HL-60 cells differentiated to neutrophils affected cell morphology and in 24 h of exposure. We have demonstrated that, the NS1 antigen has induced MMP-2, MMP-14 and MMP-9 expressions in neutrophils in a 24hrs exposure time. NS1 exposure has also further upregulated MMP-1, MMP-13, and MMP-8 expressions in neutrophils in a 24hrs exposure time. Notably, treatment with atorvastatin concentrations downregulated the expression profile of the all matrix metalloprotease significantly. Importantly, NS1 antigen has significantly increased the IL-6, IL-13 release by the HL,60 cells which was reversed by atorvastatin. On the other hand, NS1 exposure enhanced the mRNA expressions of VEGF-A and VEGF-D which was reversed by atorvastatin. However, we found that, NS1 exposure reduced the mRNA expressions profile of VEGF-C, which was reversed by atorvastatin. ConclusionIn conclusion, we report that, neutrophils associated matrix metalloprotease are involved in the pathogenesis of dengue viral disease. VEGF growth factors may also be released by the neutrophils which may subsequently participate in the endothelial dysfunctions leading to dengue shock syndrome.

Abciximab coated albumin nanoparticles of rutin for improved and targeted antithrombotic effect

In this work, we fabricated rutin (RTN)-loaded albumin nanoparticles (RTN-ALB-NPs) and abciximab (ABX) coated RTN-loaded albumin nanoparticles (ABX-RTN-ALB-NPs) for the targeted prevention and treatment of thrombosis. The prepared albumin nanoparticles were of 130–200 nm in particle size range with smooth and spherical surface morphology. The entrapment efficiency was achieved up to 73.08% with an in-vitro release rate of 34.49% in the first 4 h and 85.99% by the end of 72 h. The degree of ABX conjugation on the nanoparticle surface about 62% which was further confirmed by X-ray photoelectron spectroscopy analysis, energy-dispersive X-ray spectroscopy analysis and Bradford assay. The serum stability and biocompatibility study demonstrated that the antithrombotic property of RTN was maintained, indicative of good serum stability and hemocompatibility. Moreover, the in-vitro targeting efficiency study showed approximately 10 to 20-fold much higher intracellular fluorescence intensities of targeted formulation than non-targeted formulation towards the activated platelets. The in-vitro platelet aggregation assay and blood clot assay also revealed that the inhibition of thrombus formation was significantly higher when treated with ABX-RTN-ALB-NPs than RTN-ALB-NPs. The findings showed that the developed nanoformulation is a potential site-specific delivery system of RTN for targeted prevention and treatment of thrombosis.

RETRACTED: Chitosan-alginate nanoparticles of cabazitaxel: Design, dual-receptor targeting and efficacy in lung cancer model

Cabazitaxel (CZT) loaded chitosan-alginate based (CSA) nanoparticles were developed with dual targeting functions of both folate receptor and epidermal growth factor receptor (EGFR) using ionic gelation technique. The chitosan-folate conjugate was synthesized, and characterized by using FTIR, NMR and Mass spectroscopy. The physicochemical parameters and morphology of all CSA nanoparticles were examined. The degree of conjugation of folic acid and cetuximab (CTXmab) was determined by UV–Visible spectroscopy and Bradford assay, respectively. Moreover, XPS analysis also supported the presence of the ligands on nanoparticles. The cellular-uptake study performed on A-549 cells demonstrated a significant enhancement in the uptake of dual-receptor targeted CSA nanoparticles than non-targeted and single-receptor targeted CSA nanoparticles. Further, CZT-loaded dual receptors targeted CSA nanoparticles also showed significantly lower IC50 values (~38 folds) than the CZT control against A-549 cells. Further, in-vivo histopathological evaluations of dual receptor-targeted CSA nanoparticles have demonstrated better safety in Wistar rats. Moreover, its treatment on the Benzo(a)pyrene (B(a)P) induced lung cancer mice model has showed the enhanced anticancer efficacy of CZT with a prolonged survival rate.

An investigation of extracellular vesicles in bovine colostrum, first milk and milk over the lactation curve

Extracellular vesicles (EVs) in milk have claimed benefits ranging from conveying immunological privilege to infants to being suitable as natural delivery vehicles for therapeutic drugs. However, a longitudinal study of bovine EVs quantities and characteristics in colostrum (COL), first milk (FM) and throughout the lactation curve of mature milk (MM) had never been performed and so was our aim. COL, FM and 9 months of MM samples were collected. Caseins -overlapping size with EVs- were removed. EVs were collected by density gradient ultracentrifugation and characterised by SDS-PAGE, Bradford assay, nanoparticle tracking analysis, immunoblotting, imaging flow cytometry analysis, and transmission electron microscopy. COL and FM had substantially more EVs than MM, with COL enriched in small EVs. No significant differences were observed between months 1–9 of MM. Altogether, although COL and FM are particularly rich sources of EVs, mature milk throughout the lactation curve is also an abundant source of intact EVs.

Biochemical characterization of alkaline phosphatase in midgut of Cry2Ab dosed, pink bollworm, Pectinophora gossypiella (Saunders)

Aim: The holistic management of pink bollworm is possible only after uncovering the underlying mechanism/s of pink bollworm resistance. The present study was carried out for investigating the effect of Cry2Ab toxin on alkaline phosphatase activity and expression of its isoforms in midgut of Pectinophora gossypiella. Methodology: Resistant and susceptible pink bollworm larval populations were subjected to Cry2Ab bioassays using F1 larvae in a 21 days diet-incorporation method. The total midgut protein was estimated by Bradford method. Alkaline phosphatase activity was estimated in midgut homogenates, Non-denaturing SDS-PAGE and Native polyacrylamide gel electrophoresis was performed by the following standard procedures. Results: The study showed increased alkaline phosphatase activity in pink bollworm midgut is correlated with insect resistance in response to Cry2Ab. The non-denaturing SDS-PAGE analysis of pink bollworm midgut alkaline phosphatase isozymes had differential banding patterns in resistant and susceptible populations. Yavatmal (R1), Hingoli (R4) and Parbhani (R5) showed similar banding patterns ranging from 65-95 kDa; Guntur (R2) and Hingoli (R3) showed near similar banding patterns ranging from 70-95 kDa; whereas susceptible population showed banding pattern in the range of 50-70 kDa. Interpretation: The expression profiles of alkaline phosphatase isomers in resistant and susceptible populations can be utilized as a biomarker to aid for the screening of PBW infected Bt -cotton fields and development of pink bollworm management strategies. Key words: Alkaline Phosphatase, Cry2Ab, Insect resistance, Pectinophora gossypiella, SDS-PAGE

Hypothyroidism correlates with dyslipidemia and protein contents in patients with various metabolic disorders.

ObjectivesThe present study aimed to investigate the status of abnormalities in thyroid-related hormones, lipid profile parameters and total proteins in the sera of patients suffering from various metabolic disorders.MethodsTo analyze the study parameters, enzyme-linked immunosorbent assays, Bradford assays and standard clinical kits and methods were applied. Data were analyzed through the appropriate statistical tests.ResultsIn all subjects except those with hypotension, thyroid-stimulating hormone, total cholesterol and low-density lipoprotein were elevated, whereas triiodothyronine levels were downregulated. Thyroxin was downregulated in subjects with diabetes and symptomatic thyroiditis but upregulated in patients with hypertension. High-density lipoprotein was upregulated in men who were diabetic only, and total protein was downregulated in those with hypotension only. Hypothyroidism in patients with diabetes, symptomatic thyroiditis and hypertension was correlated with dyslipidemia. In subjects with hypertension, it was correlated with total protein.ConclusionThis study revealed a link between hypothyroidism, dyslipidemia and total protein in patients with various metabolic disorders.

Green silver nanoparticles against Helicoverpa armigera and its effects on biochemical, morphological and histological aspects

The aim of this study was to evaluate the effect of green synthesized silver nanoparticles using aqueous leaf extract of Ricinus communis (castor plant) on the life cycle Helicoverpa armigera. The synthesized AgNPs were identified using different microscopic techniques: UV-Visible spectroscopy, Scanning electron microscopy (SEM), Energy-disruptive X-ray spectroscopy (EDX) and Transmission electron microscopy (TEM). Insecticidal activity of AgNPs was studied using leaf dipping method. The biochemical studies were analyzed at LC50 of AgNPs using different methods (total carbohydrate using phenol-sulphuric acid reaction of Dubois method, total protein using Bradford method, total lipid using sulfo-phospho vanillin of Knight method and non-specific esterases using Van-Asperan method, respectively). UV-vis spectrophotometer confirmed the spectra at 445 nm of AgNPs. Spherical shaped AgNPs of 8.96 nm size were confirmed by TEM and SEM. EDX spectra confirm the chemical composition AgNPs. Higher mortality was observed in 6th instar larvae (70-80% at LC50 175 ppm) than the other instars. The pupa and adult of H. armigera also exhibited toxicity against the AgNPs. Effect of AgNPs treatment was investigated on the midgut of 5th instar larva of H. armigera using the transmission electron microscope. Accumulation of AgNPs in cell organelles was confirmed by ultra structural studies of insect midgut cell using TEM. The biochemical components of 5th instar larvae showed that, the amount of total carbohydrate, total protein, total lipid and non-specific esterases were significantly decreased After 24 h treatment at LC50 of AgNPs. Now, we can summarise that, synthesis of AgNPs using R. communis is a green route to control pest population. The synthesized nanoparticles could be used for the improvement of new botanical nano-insecticide or nano-formulation after successful field trials.

Antioxidant and Antimicrobial Activity of Microalgae of the Filinskaya Bay (Baltic Sea).

Microalgae are rich in proteins, carbohydrates, lipids, polyunsaturated fatty acids, vitamins, pigments, enzymes, and other biologically active substances. This research aimed to study the composition and antioxidant and antimicrobial activity of proteins, lipids, and carbohydrates of microalgae found in the Filinskaya Bay of the Kaliningrad region. The chemical composition of Scenedesmus intermedius and Scenedesmus obliquus microalgae biomass was studied. Ultrasound was used to isolate valuable components of microalgae. It was found that microalgae are rich in protein and contain lipids and reducing sugars. To confirm the accuracy of the determination, the protein content was measured using two methods (Kjeldahl and Bradford). Protein content in S. intermedius and S. obliquus microalgae samples did not differ significantly when measured using different methods. Protein extraction by the Kjeldahl method was found to be 63.27% for S. intermedius microalgae samples and 60.11% for S. obliquus microalgae samples. Protein content in S. intermedius samples was 63.46%, compared to 60.07% in S. obliquus samples, as determined by the Bradford method. Lipids were 8.0–8.2 times less abundant than protein in both types of microalgae samples. It was determined that the content of reducing sugars in the samples of the studied microalgae was 5.9 times less than the protein content. The presence of biological activity (antioxidant) in proteins and lipids obtained from biomass samples of the studied microscopic algae was established, which opens up some possibilities for their application in the food, chemical, and pharmaceutical industries (as enzymes, hormones, vitamins, growth substances, antibiotics, and other biologically active compounds).

RGBradford: Accurate measurement of protein concentration using a smartphone camera and the blue to green intensity ratio

The color shift caused by the interaction between proteins and Coomassie Brilliant Blue in the Bradford assay can be recorded with a smartphone camera. Color data can then be extracted from pictures and processed as analytical signals. Here, I present a simple procedure to accurately measure protein levels using color data from pictures of microplates. Plotting the ratio of blue to green intensity (RGB scale) as a function of protein concentration results in a linear (R2 ≥ 0.99) relationship, from which protein levels in biological samples can be calculated with no significant difference from values obtained using absorbance data (RGB versus absorbance curves).