Total Protein Content Research Articles

Modification of the Protein Amino Acid Content in Hen Eggs as a Consequence of Different Concentrations of Lupine and Soy in Feed

The effect of the diet modification (soybean and lupine addition) on the content of protein and amino acids (AA) in eggs was studied. Both the sampling day and the diet influenced the total protein content. In albumen, the lowest protein content (10.6%) was noted after administering a diet containing 25% lupine; in the same egg the yolk contained the most proteins (16.7%). In the content of nonessential AA (NAA) in egg yolks, differences were noted only for cysteine, with its the highest content in the yolks of the control group. The stable content of essential yolk amino acids (EAA) was observed only for isoleucine, leucine, tryptophan and phenylalanine. The highest contents of EAA and NAA were recorded in the yolks of the control group (~47 and ~53 g/100 g of protein, respectively) and in the group with 25% additions of lupine (~42 and ~51 g/100 g of protein, respectively). AA with constant content in the tested albumens were methionine, tryptophan and alanine. The highest content of EAA (>~42 g/100 g of protein) and NAA (>~62 g/100 g of protein) were determined in albumen of eggs determined in the group with at least 20% additions of lupine. The highest content of EAA for humans delivered eggs from groups 4–6 (with the addition of soy into the diet ≤5%). The protein sources used in the hen diet significantly influenced the content of protein and individual AA in the produced eggs.

Продуктивные особенности коров голштинской породы разных генетических линий

Abstract. The purpose of the study is to investigate the productive features of Holstein cows of different genetic lines. Methods. The research was carried out in two breeding enterprises of the Sverdlovsk region on the livestock of Holstein cows. Groups of animals were formed by the method of pairs of analogues, taking into account the age of animals in lactation, physiological condition, live weight, lines of breeding bulls. The coefficient of milk content was estimated by the calculation method. The lactose, fat and protein content in milk was determined using a “Bentley 150” device. Somatic cells were determined on the “Somatos-M” device. Bacterial contamination was studied using a reductase assay. Prolactin was determined by enzyme immunoassay, cortisol by immunochemiluminescent method. Results. In terms of milk yield for 305 days of lactation and protein content in milk, Montwick Chieftain cows had an advantage with a significant difference between the groups (P < 0.05). The animals of the Reflection Sovering line were the best in terms of fat content, the difference between the groups was insignificant. The lowest content of somatic cells in milk and bacterial contamination was found in animals of the Reflection Sovering and Vis Back Idial lines. The best indicators of the leukocyte profile were determined in animals of the Vis Back Idial and Montwick Chieftain lines. In terms of glucose and total protein content in the blood, cows of the Montwick Chieftain line had an advantage, in terms of the amount of urea, animals of the Montwick Chieftain and Vis Back Idial line affiliation. The highest level of total lipids in the blood is typical for cows of the Reflection Sovering line. Scientific novelty. The research results indicate the relationship of biological characteristics and milk productivity with different lines of breeding bulls. The leukocyte profile of this herd of cows of different genetic lines in breeding enterprises of the Sverdlovsk region was studied for the first time. Practical significance. The results of the research can be used in breeding enterprises in order to increase the milk productivity of cows.

Proteomics analysis of carcinogenesis in a rat model of mammary cancer induced by DMBA (7,12-dimethylbenz[a]anthracene)

Background: Mammary cancer, called breast cancer in humans, results from the abnormal growth of cells in the mammary glands that attack the surrounding tissue. The process of carcinogenesis at the molecular level can be monitored through the production of proteins as biomarkers for carcinogenesis. 7,12-Dimethylbenz[a]anthracene (DMBA) is a known carcinogenic compound. This study aimed to analyze the proteomic profile as critical data regarding DMBA-induced carcinogenesis in Sprague‒Dawley rats. Methods: Experimental animals were divided into two groups: a treatment group given DMBA at a dose of 10 mg/kg (intramammary) at intervals of 48 hours for a total of 10 doses, and a negative control group that was not given any treatment. Measurement of the total protein concentration was carried out using a spectrophotometer, and the data were analyzed using a t-test, while the characterization of protein profiles was carried out based on molecular weight data using SDS‒PAGE. Mammary gland histopathology was evaluated by hematoxylin and eosin (H&E) staining. Results: The results showed a significant (p<0.05) increase of 27% in the total protein concentration in the rat mammary cancer model. The results of proteomic characterization showed a protein profile containing proteins of 187, 169, 68, 64, 53, 41, 24, 18, and 14 kDa, which were suspected to be HER-2, Nischarin, COX-2, Albumine, Vimentin, ACTB, TNF, p16, and fatty acid binding protein 3 (FABP3), respectively. Histopathology of the mammary glands showed an irregular and indistinct arrangement of the alveoli and extensive epithelial cell proliferation from the surface to the lumen of the mammary ducts, and the mammary stroma showed the formation of new epithelial cells, which were cancer cells that spread to surrounding tissue. Conclusions: The proteomic profile was strongly associated with morphological alterations in mammary carcinogenesis in a rat model of DMBA-induced mammary cancer.

Chemical Profile and In Vitro Gut Microbiota Modulation of Wild Edible Mushroom Phallus atrovolvatus Fruiting Body at Different Maturity Stages

Phallus atrovolvatus, a wild edible mushroom, has attracted increasing interest for consumption due to its unique taste and beneficial health benefits. This study determined the chemical components in the so-called fruiting body during the egg and mature stages and investigated its gut microbiota-modulating activities. The egg stage contained higher total carbohydrates, dietary fiber, glucans, ash, and fat, while the total protein content was lower than in the mature stage. Two consumption forms, including cooked mushrooms and a mushroom aqueous extract from both stages, were used in this study. An in vitro gut fermentation was performed for 24 h to assess gut microbiota regulation. All mushroom-supplemented fermentations increased short-chain fatty acid (SCFA) production compared to the blank control. Furthermore, all mushroom supplementations promoted the growth of Bifidobacterium and Streptococcus. Samples from the mature stage increased the relative abundance of Clostridium sensu stricto 1, while those from the egg stage increased the Bacteroides group. The inhibition of harmful bacteria, including Escherichia-Shigella, Klebsiella, and Veillonella, was only observed for the mature body. Our findings demonstrate that P. atrovolvatus exhibits potential benefits on gut health by promoting SCFA production and the growth of beneficial bacteria, with the mature stage demonstrating superior effects compared to the egg stage.

Economic and financial viability of the production of Brazilian somalis sheep fed with different nutritional plans.

The objective of this study was to evaluate the economic and financial viability of finishing of two sexes of Brazilian Somalis sheep in feedlot receiving diets with 0 and 15% reduction in the total digestible nutrients and crude protein contents. A completely randomized design was adopted, in a 2 × 2 factorial arrangement, with treatments consisting of two sexes (non-castrated males (21.13 ± 5.00kg) and females (17.99 ± 3.79kg) and two diets (0 and 15% reduction in the levels of total digestible nutrients and crude protein). Four simulations of productive scenarios were carried out: non-castrated males receiving the diet with 0% reduction; non-castrated males receiving the diet with 15% reduction; females receiving the diet with 0% reduction and females receiving the diet with 0% reduction, for a period of 70 days in feedlot. The highest costs of the system were with feeding and other costs, mainly the acquisition of animals. For both sexes, the 0% reduction diet showed the highest total cost, total revenue, break-even point. Non-castrated males fed a non-reduced diet and females fed a 15% reduction diet had higher net income, rate of return, total productivity, net present value, internal rate of return, profitability index and profitability rate. Less favorable situations were not profitable for any scenario. However, the increase in sales price favorability makes the activity viable. The reduction of total digestible nutrients and crude protein affects the economic and financial viability of Brazilian Somalis sheep. For non-castrated males it is recommended the diet without reduction and for females the diet with 15% reduction of nutrients.

Evaluation of four novel non-grain protein sources completely replacing soybean meal on growth performance, serum biochemistry, amino acid transport and intestinal health of grass carp (Ctenopharyngodon idella) at different water temperatures

To investigate the effects of non-grain protein source and water temperature on growth and feed utilization differences of grass carp, the effects of different protein sources on the growth performance, serum biochemistry, digestive enzymes, amino acid transport and intestinal health of grass carp were studied at 24°C, 28°C and 32°C. In this study, a total of 1350 grass carp (5.00 ± 0.02 g) were selected, and Clostridium autoethanogenum protein (CAP), Tenebrio molitor meal (TMM), cottonseed protein concentrate (CPC) and Chlorella powder (CHP) were used as a single protein source to completely replace soybean meal for 56 days. The results showed that the final body weight (FBW), weight gain rate (WGR), specific growth rate (SGR) and protein efficiency ratio (PER) of grass carp increased significantly with the increasing temperature (P < 0.001). The CHP and SBM groups showed no significant differences in FBW, WGR, SGR and PER (P > 0.05), which were higher than the CAP, TMM and CPC groups (P < 0.001). The alanine transaminase (ALT), aspartate aminotransferase (AST), total protein (TP) and triglyceride (TG) concentrations of grass carp at 32°C were significantly lower than those at 24°C and 28°C (P < 0.001). The acid phosphatase (ACP) activity decreased significantly with the increase of temperature (P = 0.001). The amylase (AMS) activity of the TMM, CPC and CHP groups was significantly lower than that of the SBM and CAP groups (P < 0.001), and the ACP and lipase (LPS) activities in the TMM group were significantly lower than those in the SBM group (P < 0.001). In addition, the interaction between temperatures and protein sources significantly affected the gene expression levels of amino acid transport including solute carrier family 1 member 3 (SLC1A3), solute carrier family 7 member 1 (SLC7A1), solute carrier family 7 member 5 (SLC7A5), solute carrier family 15 member 1b (SLC15A1b), solute carrier family 7 member 7 (SLC7A7), target of rapamycin (TOR), 4E binding protein 1 (4E-BP1) and ribosomal protein S6 kinase 1 (S6K1), intestinal inflammatory including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-8 (IL-8), interleukin-10 (IL-10) and tight junction proteins (occludin, claudin1, claudin3, claudin7 and claudin11) (P ≤ 0.001). Collectively, our results indicated that CHP could be a potential protein source in the case of complete replacement of soybean meal in grass carp.

INDUCED PLURIPOTENT STEM CELL-DERIVED MESENCHYMAL STEM CELLS-DERIVED EXTRACELLULAR VESICLES ATTENUATE LPS-INDUCED LUNG INJURY AND ENDOTOXEMIA IN MICE.

Introduction: We hypothesized extracellular vesicles (EVs) from preconditioned human-induced pluripotent stem cell-derived mesenchymal stem cells (iMSCs) attenuate LPS-induced acute lung injury (ALI) and endotoxemia. Methods: iMSCs were incubated with cell stimulation cocktail (CSC) and EVs were isolated. iMSC-EVs were characterized by size and EV markers. Biodistribution of intratracheal (IT), intravenous, and intraperitoneal injection of iMSC-EVs in mice was examined using IVIS. Uptake of iMSC-EVs in lung tissue, alveolar macrophages, and RAW264.7 cells was also assessed. C57BL/6 mice were treated with IT/IP iMSC-EVs or vehicle ± IT/IP LPS to induce ALI/acute respiratory distress syndrome and endotoxemia. Lung tissues, plasma, and bronchoalveolar lavage fluid (BALF) were harvested at 24 h. Lung histology, BALF neutrophil/macrophage, cytokine levels, and total protein concentration were measured to assess ALI and inflammation. Survival studies were performed using IP LPS in mice for 3 days. Results: iMSC-EV route of administration resulted in differential tissue distribution. iMSC-EVs were taken up by alveolar macrophages in mouse lung and cultured RAW264.7 cells. IT LPS-treated mice demonstrated marked histologic ALI, increased BALF neutrophils/macrophages and protein, and increased BALF and plasma TNF-α/IL-6 levels. These parameters were attenuated by 2 h before or 2 h after treatment with IT iMSC-EVs in ALI mice. Interestingly, the IT LPS-induced increase in IL-10 was augmented by iMSC-EVs. Mice treated with IP LPS showed increases in TNF-α and IL-6 that were downregulated by iMSC-EVs and LPS-induced mortality was ameliorated by iMSC-EVs. Administration of IT iMSC-EVs 2 h after LPS downregulated the increase in proinflammatory cytokines (TNF-α/IL-6) by LPS and further increased IL-10 levels. Conclusions: iMSC-EVs attenuate the inflammatory effects of LPS on cytokine levels in ALI and IP LPS in mice. LPS-induced mortality was improved with administration of iMSC-EVs.

Natural variation in the nutritional composition of African baobab (Adansonia digitata L.) from two ecological sites in Northern Malabar, Kerala, India

Adansonia digitata L., commonly known as the African baobab, is an important plant within the Malvaceae family. It is a highly nutritious forest tree and holds a special place in several African countries. Despite its prevalence in Africa, its presence is very limited in India, and only a few individual plants of this species are reported from different parts of India, including Kerala. This study focuses on the plants growing at Ajanur and Thalassery in the Kasaragod and Kannur districts of Kerala, respectively. This study reports natural variation in phytochemicals, minerals, and the proximate components of its flowers and leaves at these locations. The results of this study show that total carbohydrate (28.79%), crude fibre (10.76%), and protein (53.09%) content was higher in leaves at Ajanur. In contrast, chlorophyll a, b, and carotenoid content were highest in leaves at Thalassery and showed a value of 39.03 mg/g, 16.65 mg/g, and 13.21 mg/g, respectively. The highest quantity of elements viz. Ca, Zn, K, Fe, Cu, Ni, and Cr were reported in leaves from Thalassery, whereas Mg and Mn were abundant in the leaves from Ajanur. Flowers from Ajanur demonstrated a superior nutritional index, while Thalassery leaves exhibited a higher mineral index. Assessment of antioxidant activity indicated the highest DPPH scavenging activity in the leaves from Ajanur (IC50 = 102.57 μg/mL) and the highest ABTS scavenging activity in its flowers from Thalassery (IC50 = 62.74 μg/mL). The differences in nutritional components and antioxidant properties of the baobab from these two ecological sites underscore the role of ecological factors in its natural variation. Although it is an important forest food tree in the African continent, its potential contribution towards food security from the Indian subcontinent remains untapped.

Supported ionic liquids to purify microbial L-Asparaginase

L-Asparaginase (ASNase) is a versatile enzyme that converts L-asparagine into ammonia and aspartic acid. This enzyme has applications in the food industry and health sector. However, high purity ASNase is required, resulting in high production costs. Therefore, in this work, two supported ionic liquids (SILs), specifically silica modified with dimethylbutylpropylammonium chloride ([Si][N3114]Cl) or triethylpropylammonium chloride ([Si][N3222]Cl), were investigated as alternative adsorption materials to purify ASNase. Different conditions were evaluated to improve enzyme purity, including total protein content in the cell extract, contact time, and SIL/cell extract ratio (w/v). Under optimal conditions using [Si][N3114]Cl, a maximum ASNase purification of 6.1-fold is achieved in a single step, resulting from the preferential attachment of other proteins on [Si][N3114]Cl SIL. According to the results, hydrophobic interactions rule the selective adsorption of protein impurities from the cell extract by the SIL, thereby increasing the ASNase purification levels. This approach offers a significant advantage, not requiring the desorption and elution of the target enzyme, while envisioning the application of SILs in a flow-through elution approach. The protonation state of protein surface was calculated by computational analysis, revealing that positively charged amino acids such as arginine and lysine block the effective binding of the enzyme to the SILs. Overall, if properly designed, SILs are promising alternative supports for the downstream processing of ASNase from cell extracts.

Effect of Seed Hardening and Foliar Application of Growth Substances on Morpho-biochemical Parameters of Groundnut (Arachis hypogaea L.)

Design with three replications was used for an experiment that included seed hardening as one factor with nine levels while foliar spray of Chlorocholine Chloride @500 mg/L as another factor with two levels. Leaf dry weight and stem dry were significantly maximum with GA3-150 mg/L seed hardening treatment. Meanwhile, total dry weight, protein content, oil content and anti-oxidant activity were found significantly higher in seed hardening with GA3-100 mg/L while these morphological parameters were found significantly positive result after application of foliar spray of CCC @500 mg/L as compared to control which helps to increases the quality of groundnut.

Effects of fermented jujube powder on growth performance, rumen fermentation, and antioxidant properties of simmental bulls

IntroductionFermented jujube powder (FJP) promotes a balance between the intestinal microflora and immune factors in animals. In this study, we aimed to investigate the effects of FJP on the production performance, nutrient digestion, rumen fermentation, and antioxidant properties of bulls.MethodsForty Simmental bulls were randomly divided into four groups based on body weight and fed a basal diet with [5, 7.5, or 10% dry matter (DM)] or without FJP. The experimental period was 20 d for adaptation and 60 d for the feeding trial.ResultsDietary FJP supplementation did not affect DM intake (P &amp;gt; 0.05) but increased the average daily gain quadratically (P = 0.049) and decreased the feed conversion ratio linearly (P = 0.042). FJP quadratically enhanced DM and crude protein digestibility (P = 0.026 and P = 0.041, respectively) and linearly enhanced acid detergent fiber digestibility (P = 0.048). It also increased the total volatile fatty acid concentration quadratically (P = 0.037), acetate molar percentage, and acetate-to-propionate ratio linearly (P = 0.002 and 0.001), and reduced the ammonia nitrogen concentration linearly (P = 0.003). Additionally, xylanase and protease activities and Ruminococcus flavefaciens abundance increased linearly (P = 0.006, 0.018, and 0.009, respectively), and total bacteria, Ruminococcus albus, and Ruminobacter amylophilus abundance increased quadratically (P = 0.047, 0.011, and 0.021, respectively). FJP linearly increased serum total protein concentration and antioxidant capacity (P = 0.003 and 0.018, respectively) and decreased malonaldehyde content (P = 0.006).DiscussionFJP supplementation (7.5%) enhanced production performance, nutrient digestion, rumen fermentation, and serum antioxidant capacity in bulls. The improved nutrient digestion may be due to an increase in ruminal microorganisms and total volatile fatty acids from the FJP. High blood antioxidant levels indicate that FJP may preserve proteins, thereby boosting the production performance of bulls.

House dust mite-induced asthma exacerbates Alzheimer's disease changes in the brain of the AppNL-G-F mouse model of disease.

Alzheimer's disease (AD) is an age-related neurodegenerative disorder characterized by the accumulation of amyloid-β (Aβ) plaques, neuroinflammation, and neuronal death. Besides aging, various comorbidities increase the risk of AD, including obesity, diabetes, and allergic asthma. Epidemiological studies have reported a 2.17-fold higher risk of dementia in asthmatic patients. However, the molecular mechanism(s) underlying this asthma-associated AD exacerbation is unknown. This study was designed to explore house dust mite (HDM)-induced asthma effects on AD-related brain changes using the AppNL-G-F transgenic mouse model of disease. Male and female 8-9 months old C57BL/6J wild type and AppNL-G-F mice were exposed to no treatment, saline sham, or HDM extract every alternate day for 16 weeks for comparison across genotypes and treatment. Mice were euthanized at the end of the experiment, and broncho-alveolar lavage fluid (BALF), blood, lungs, and brains were collected. BALF was used to quantify immune cell phenotype, cytokine levels, total protein content, lactate dehydrogenase (LDH) activity, and total IgE. Lungs were sectioned and stained with hematoxylin and eosin, Alcian blue, and Masson's trichrome. Serum levels of cytokines and soluble Aβ1-40/42 were quantified. Brains were sectioned and immunostained for Aβ, GFAP, CD68, and collagen IV. Finally, frozen hippocampi and temporal cortices were used to perform Aβ ELISAs and cytokine arrays, respectively. HDM exposure led to increased levels of inflammatory cells, cytokines, total protein content, LDH activity, and total IgE in the BALF, as well as increased pulmonary mucus and collagen staining in both sexes and genotypes. Levels of serum cytokines increased in all HDM-exposed groups. Serum from the AppNL-G-F HDM-induced asthma group also had significantly increased soluble Aβ1-42 levels in both sexes. In agreement with this peripheral change, hippocampi from asthma-induced male and female AppNL-G-F mice demonstrated elevated Aβ plaque load and increased soluble Aβ 1-40/42 and insoluble Aβ 1-40 levels. HDM exposure also increased astrogliosis and microgliosis in both sexes of AppNL-G-F mice, as indicated by GFAP and CD68 immunoreactivity, respectively. Additionally, HDM exposure elevated cortical levels of several cytokines in both sexes and genotypes. Finally, HDM-exposed groups also showed a disturbed blood-brain-barrier (BBB) integrity in the hippocampus of AppNL-G-F mice, as indicated by decreased collagen IV immunoreactivity. HDM exposure was responsible for an asthma-like condition in the lungs that exacerbated Aβ pathology, astrogliosis, microgliosis, and cytokine changes in the brains of male and female AppNL-G-F mice that correlated with reduced BBB integrity. Defining mechanisms of asthma effects on the brain may identify novel therapeutic targets for asthma and AD.

Biochar with KMnO4-hematite modification promoted foxtail millet growth by alleviating soil Cd and Zn biotoxicity

The excessive accumulation of Cd and Zn in soil poisons crops and threatens food safety. In this study, KMnO4-hematite modified biochar (MnFeB) was developed and applied to remediate weakly alkaline Cd-Zn contaminated soil, and the heavy metal immobilization effect, plant growth, and metal ion uptake of foxtail millet were studied. MnFeB application reduced the phytotoxicity of soil heavy metals; bioavailable acid-soluble Cd and Zn were reduced by 57.79% and 35.64%, respectively, whereas stable, non-bioavailable, residual Cd and Zn increased by 96.44% and 32.08%, respectively. The chlorophyll and total protein contents and the superoxide dismutase (SOD)activity were enhanced, whereas proline, malondialdehyde, the H2O2 content, glutathione reductase (GR), ascorbate peroxidase (APX) and catalase (CAT) activities were reduced. Accordingly, the expressions of GR, APX, and CAT were downregulated, whereas the expression of MnSOD was upregulated. In addition, MnFeB promoted the net photosynthetic rate and growth of foxtail millet plants. Furthermore, MnFeB reduced the levels of Cd and Zn in the stems, leaves, and grains, decreased the bioconcentration factor of Cd and Zn in shoots, and weakened the translocation of Cd and Zn from roots to shoots. Precipitation, complexation, oxidation-reduction, ion exchange, and π-π stacking interaction were the main Cd and Zn immobilization mechanisms, and MnFeB reduced the soil bacterial community diversity and the relative abundance of Proteobacteria and Planctomycetota. This study provides a feasible and effective remediation material for Cd- and Zn-contaminated soils.

Subcritical Water Extraction of Undaria pinnatifida: Comparative Study of the Chemical Properties and Biological Activities across Different Parts

The subcritical water extraction of Undaria pinnatifida (blade, sporophyll, and root) was evaluated to determine its chemical properties and biological activities. The extraction was conducted at 180 °C and 3 MPa. Root extracts exhibited the highest phenolic content (43.32 ± 0.19 mg phloroglucinol/g) and flavonoid content (31.54 ± 1.63 mg quercetin/g). Sporophyll extracts had the highest total sugar, reducing sugar, and protein content, with 97.35 ± 4.23 mg glucose/g, 56.44 ± 3.10 mg glucose/g, and 84.93 ± 2.82 mg bovine serum albumin (BSA)/g, respectively. The sporophyll contained the highest fucose (41.99%) and mannose (10.37%), whereas the blade had the highest galactose (48.57%) and glucose (17.27%) content. Sporophyll had the highest sulfate content (7.76%). Key compounds included sorbitol, glycerol, L-fucose, and palmitic acid. Root extracts contained the highest antioxidant activity, with IC50 values of 1.51 mg/mL (DPPH), 3.31 mg/mL (ABTS+), and 2.23 mg/mL (FRAP). The root extract exhibited significant α-glucosidase inhibitory activity with an IC50 of 5.07 mg/mL, indicating strong antidiabetic potential. The blade extract showed notable antihypertensive activity with an IC50 of 0.62 mg/mL. Hence, subcritical water extraction to obtain bioactive compounds from U. pinnatifida, supporting their use in functional foods, cosmetics, and pharmaceuticals is highlighted. This study uniquely demonstrates the variation in bioactive compound composition and bioactivities across different parts of U. pinnatifida, providing deeper insights. Significant correlations between chemical properties and biological activities emphasize the use of U. pinnatifida extracts for chronic conditions.

The Influence of Nitrogen and Sulfur Fertilization on Oil Quality and Seed Meal in Different Genotypes of Winter Oilseed Rape (Brassica napus L.)

Adequate nitrogen (N) and sulfur (S) fertilization of oilseed rape crops is necessary to obtain good-quality oil and post-extraction rapeseed meal. The aim of this study was to determine the effect of different doses of N fertilization (100, 160 and 220 kg ha−1) and S (0, 30, 60 and 90 kg ha−1) on the value of seeds of three winter oilseed rape genotypes. Two winter oilseed rape genotypes obtained by mutagenesis (cultivar Polka and breeding genotype PN440) were characterized by changed fatty acid profile. The cultivar Polka, type HO (high oleic), had a high content of oleic acid (C18:1, 78.0%) and the breeding genotype PN440, type HOLL (high-oleic and low-linolenic), had a high content of oleic acid (C18:1, 75.0%) and a low content of linolenic acid (C18:3, 3.0%). We also used the canola type of winter oilseed rape cultivar, Monolit. The analysed winter oilseed rape genotypes responded similarly to the N and S fertilization factors with regard to the content of crude fat and total protein in the seeds and the composition of fatty acids in the oil. N fertilization increased the content of glucosinolates (GLS-alkenyl, indole and total) in seeds, whereas S application decreased the content of saturated fatty acids (stearic acid-C18:0) in oil and increased the content of alkenyl and total glucosinolates (GLSs) in seeds. A significant interaction between N and S was observed for crude-fat and total-protein content. This study suggests that ensuring an adequate supply of both nitrogen and sulfur in the soil is essential for optimizing meal and oil quality in different types of winter oilseed rape cultivars. Proper management of these nutrients can lead to improved oil content and overall crop performance.

Basal levels of biochemical biomarkers in the freshwater prawn Palaemon argentinus and their alterations due to the exposure of both insecticides cypermethrin and spirotetramat

The aim of this study was to evaluate the sensitivity of the prawn Palaemon argentinus to the pyrethroid cypermethrin (CYP) and the tetramic acid spirotetramat (STM). These treatments were compared with prawns collected at a reference site to define their basal physiological state. Initially, physicochemical parameters and several pollutants at the selected site were analyzed. The LC50-96 h was determined in adult prawns. Then, prawns were exposed for 96 h to sublethal concentrations of CYP (0.0005 μg/l) and STM (0.44 mg/l) to evaluate the effects on some biochemical endpoints. A treatment combining both pesticides was also added at 5 % of these values. Controls with and without solvent (acetone) were included. The LC50-96 h values were 0.005 μg/l and 4.43 mg/l for CYP and STM, respectively. Moreover, some biomarkers linked to oxidative and energy metabolism were analyzed in the hepatopancreas and muscle of both essayed prawns and those at the basal state. The STM caused a significant decrease in total protein content (32 %) in contrast to the increase of protein carbonyl content (71 %) (p < 0.05). Also, glutathione S-transferase (52 %) and catalase (61 %) activities in the hepatopancreas of exposed prawns were higher compared to both the control and state basal groups (p < 0.05). In muscle, only a significant decrease in the lactate content (69 %) was caused by STM (p < 0.05). In addition, CYP caused a significant increase in the lactate dehydrogenase activity (110 %) in muscle and triacylglycerol content (73 %) in the hepatopancreas (p < 0.05). The integrated biomarker index (IBRv2) analysis showed that STM caused greater damage than CYP. Besides, the combined treatment showed an antagonistic interaction between both insecticides. The differential response of biomarkers to both CYP and STM exposure with respect to their basal levels shows a high sensitivity of P. argentinus demonstrating its potential role as a bioindicator organism.

Intramammary administration of lipopolysaccharides at parturition does not affect the transfer of passive immunity in goat kids.

This study evaluated the effect of feeding colostrum obtained from an intramammary administration (IA) of LPS from Escherichia coli (O55:B5) to dairy goats at parturition, on goat kids performance, biochemical parameters (i.e., calcium, LDH, glucose, total proteins, albumin, and urea) and immune status (i.e., IgG and IgM) during the first month of life. At birth, goat kids were weighted (d 0) and immediately allocated into either the LPS group (n = 15) or the CON group (n = 21) based on the experimental group of the dam. At parturition, 20 multiparous dairy goats were allocated in one of the 2 experimental groups (LPS vs. CON). The LPS group received an IA of saline solution (2 mL) containing 50 µg of LPS in each half udder whereas goats in the CON group received an IA of saline solution (2 mL) without LPS. Goat kids were bottle-fed dam colostrum equivalent to 10% of the birth BW divided in 2 meals (i.e., at 3 and 12 h relative to birth), and then fed twice daily with milk replacer ad libitum. Individual milk intake (MI) and BW were recorded on d 7, 15, 21 and 30 of life. Blood samples were collected on d 0, 1, 2, 4, 7, 15, 21 and 30 after birth. Data was analyzed using the MIXED procedure of SAS (9.4). The model included IA, time (T) and the interaction (IA x T) as fixed effects and sex and litter size as random effects. Both groups showed similar MI, except on d 7 relative to birth as the LPS group showed higher MI than the CON group (910.5 ± 69.77 and 683.9 ± 59.64 mL, respectively). No differences in BW or rectal temperature were observed between groups, neither in plasma IgG nor IgM concentrations. Despite the IA did not affect calcium, glucose, LDH, total protein, and albumin concentrations an interaction between the IA and T was observed for urea concentration, showing the LPS group higher urea concentrations than the CON group on d 0 (20.1 ± 1.34 and 20.0 ± 1.25 mg/dL, respectively). In conclusion, feeding colostrum from goats that received an IA of LPS at parturition does not affect goat kid performance, plasma immunoglobulin concentrations and serum metabolites during the first month of life.

Resolving the Current Controversy of Use and Reuse of Housekeeping Proteins in Ageing Research: Focus on Saving People’s Tax Dollars

The use of housekeeping genes and proteins to normalize mRNA and protein levels in biomedical research has faced growing scrutiny. Researchers encounter challenges in determining the optimal frequency for running housekeeping proteins such as β-actin, Tubulin, and GAPDH for nuclear-encoded proteins, and Porin, HSP60, and TOM20 for mitochondrial proteins alongside experimental proteins. The regulation of these proteins varies with age, gender, disease progression, epitope nature, gel running conditions, and their reported sizes can differ among antibody suppliers. Additionally, anonymous readers have raised concerns about peer-reviewed and published articles, creating confusion and concern within the research and academic institutions. To clarify these matters, this minireview discusses the role of reference housekeeping proteins in Western blot analysis and outlines key considerations for their use as normalization controls. Instead of Western blotting of housekeeping proteins, staining of total proteins, using Amido Black and Coomassie Blue can be visualized the total protein content on a membrane. The reducing repeated Western blotting analysis of housekeeping proteins, will save resources, time and efforts and in turn increase the number of competitive grants from NIH and funding agencies. We also discussed the use of dot blots over traditional Western blots, when protein levels are low in rare tissues/specimens and cell lines. We sincerely hope that the facts, figures, and discussions presented in this article will clarify the current controversy regarding housekeeping protein(s) use, reuse, and functional aspects of housekeeping proteins. The contents presented in our article will be useful to students, scholars and researchers of all levels in cell biology, protein chemistry and mitochondrial research.

Total Protein Determination by Recovery Fluorescence of Trinitrotoluene-Quenched Protein-Functionalized Gold Nanoclusters.

Determination of total protein content is very important for clinical, pharmaceutical and food chemistry, and the simplicity and rapidity of the existing total protein content assays should be improved. Here, a novel fluorescent sensor for total protein content measurement was described using near-infrared emitting fluorescent gold nanoclusters, which were prepared with egg whites. Trinitrotoluene served as highly efficient quencher to quench the fluorescence of gold nanoclusters. Interestingly, the trinitrotoluene-protein complex that formed by premixing of trinitrotoluene and protein-containing real samples maintained the fluorescence of gold nanoclusters. These new findings provided a novel mechanism to design a fluorescence sensor for determination of total protein levels in various real samples. With available and low-cost bovine serum albumin as reference, the obtained standard curve for total protein determination indicated a linear range of 0.1-3.5g L- 1 with a detection limit of 0.06g L- 1 (3σ rule) and a correlation efficiency (R2) of 0.9850. The applicability of the proposed sensor was validated by determination of total protein levels in real samples. Comparison with the existing spectrophotometric methods, the proposed sensor is advantageous of simplicity, rapidity, and cost-effectiveness.

Role of Nannochloropsis Oculata supplement in improving performance, antioxidant status, blood metabolites, and egg quality of laying hens under hot environmental conditions

The increase in environmental temperature led to economic losses in the poultry industry, urging the use of feed supplements to mitigate the negative effects on chick's welfare and performance. Therefore, this study aimed to examine the effects of marine microalgae (Nannochloropsis Oculata, N. Oculata) additive on commercial Brown Lohmann laying hen's performance, blood metabolites, and antioxidant status under hot environmental conditions. One hundred and eighty birds (34 weeks old with an initial body weight of 1885 ± 23.5 g) were used till 47 weeks. The birds were divided into three equal groups (birds in each group were distributed into four equal replicates 15 hens/ replicate). The 1st group was the control (CON) and was fed the basal diet, while the 2nd (TR1) and 3rd (TR2) groups were fed the basal diet supplemented with 0.5 and 1.0% of N. Oculata, respectively. The results showed that total protein and globulin concentrations increased (P < 0.05) in treated groups compared with the control group, whereas, albumin concentration increased (P < 0.05) in TR2 compared to the control group. The concentration of ALT and AST decreased in hens fed N. Oculata. Supplementing with N. Oculata reduced serum cholesterol and creatinine concentrations, while glucose concentration increased (P < 0.05) in the treated groups compared to the control group. Serum calcium, total antioxidant capacity (TAC), Triiodothyronine (T3), and progesterone increased (P < 0.05) in hens fed N. Oculata. N. Oculata supplement improved production performance through a positive effect on egg number, egg weight, egg mass, feed conversion ratio, and mortality rate. In addition, the overall mean of shell thickness increased (P < 0.05) in hens fed N. Oculata. It can be concluded that the supplementation of 1.0% N. Oculata to the laying hens' diet enhanced productive performance, serum constituents, and antioxidant status under hot environmental conditions.