Determination Of Total Protein Research Articles

Ascorbic acid and glucose can cause significant interference on quantitative measurement of biochemistry analytes in urine.

Preanalytical errors due to interferences can lead to inaccurate results, necessitating an understanding of potential interferences for each test. This study explores the impact of elevated concentrations of ascorbic acid and glucose on urine analysis, a pivotal diagnostic tool. Conducted at the Clinical Institute of Chemistry, KBC Sestre milosrdnice, the research utilized a 24-hour urine sample. Parameters assessed included total proteins, albumin, amylase, sodium, potassium, chlorides, calcium, phosphates, magnesium, creatinine, urea, and uric acid. Various concentrations of added interferents were prepared for duplicate measurements using statistical analysis in Microsoft Excel. No statistically significant interferences were found in albumin, amylase, sodium, potassium, or phosphate concentrations. However, ascorbic acid interfered with chloride, calcium, and magnesium determinations. Conversely, elevated glucose affected total protein, calcium, magnesium, creatinine, urea, and uric acid determinations. Interference of ascorbic acid with chloride and interference of glucose with total proteins and uric acid displayed a linear relationship. Results suggest cautious analysis interpretation from certain parameters in patients with elevated glucose and/or ascorbic acid in urine. Whereas ascorbic acid interference may go unnoticed due to its infrequent measurement, routine determination of glucose in urine is crucial, especially for diabetes patients.

The interference and elimination of nitrite on determination of total urinary protein by Pyrogallol Red–Molybdate method

BackgroundSome urine samples showed negative values of total urinary protein by Pyrogallol red–molybdate (PRM) method. Interestingly, these samples showed notably high levels of nitrite in the urine dipstick test. MethodsA total of 120 urine samples were collected and categorized into four groups (0 - < 100, 100-<500, 500-<1000, ≥1000 mg/L) based on total urinary protein concentration. Various concentrations of nitrite (2, 10, 50, 100, and 200 mg/L) were added to urine samples to investigate potential interference of nitrite in total urinary protein measurement by the PRM method. Additionally, different concentrations of L-ascorbic acid (100, 500, 1000, and 2000 mg/L) were added to urine-nitrite mixtures to explore the possibility of reversing the interference effects. Results2 mg/L nitrite had no impact on the test results.10 mg/L nitrite only showed significant effects on the detection results among the groups with 0 - < 100 and 100-<500 mg/L (P < 0.05). 200 mg/L nitrite caused a noticeable decrease in the urinary protein detection results of the four groups (0 - < 100, 100-<500, 500-<1000, ≥1000 mg/L), and the concentrations were reduced to 2.7 %, 26.85 %, 75.22 %, and 89.33 % of their original levels, respectively. 500 mg/L L-ascorbic acid effectively eliminated the interference from 200 mg/L nitrite and almost had no effect on detection. ConclusionsNitrite had negative effect on the detection of total urinary protein by PRM method and L-ascorbic acid was effective in counteracting the interference caused by nitrite.

COMPARATIVE ANALYSIS OF METHODS FOR DETERMINING THE CONTENT OF THE TOTAL PROTEIN SARS-COV-2 OF THE COVID-19 VIRUS

One of the main requirements for the new vaccines being created is their safety in use, including the total protein content, which are one of the main indicators of the quality and safety of vaccines. This article presents the results of work on the comparative analysis of methods for determining the total protein in the active pharmaceutical substance (AFS) and also the bulk product of the vaccine "QazCovid-in ® .Determination of the total protein concentration was carried out by comparing the methods of Lowry, Bradford, commercial sets of Clini Test-BL and BSA Gold Protein. As a result of the determination of the total protein in the composition of the AFS by the classical Lowry method and the commercial CliniTest-BL kit, the highest values were obtained on average 616.67 and 589 μl/ml, and in the bulk product 51.11 and 47.38 μl/ml, respectively. The most minimal indicator of the total protein content of 38.39±0.000059 is shown by the method using the commercial BSA Gold Protein kit. The average values of total protein content showed the results obtained by the Bradford method and the determination of total protein by a commercial set of Clini Test-BL. The data obtained allow us to recommend the use of a set of Clini Test-BL for use in quality control, since this set is characterized by the ease and speed of the determination method, which is not inferior to the classical Lowry method. When determining the total protein content in the bulk product used to prepare the QazVac vaccine against COVID-19, the classical Lowry method showed the best result with a high degree of sensitivity.

Using pre-fermented sugar beet pulp as a growth medium to produce Pleurotus ostreatus mycelium for meat alternatives

This study aimed to determine the compatibility of pre-fermented sugar beet pulp to support the growth of Pleurotus ostreatus mycelium in submerged fermentation. The goal was to create a meat alternative based on mycelial-fermented pulp. It was further explored whether pre-fermentation with lactic acid bacteria (LAB) on the pulp increased meat-like properties, such as aroma, springiness, and hardness, in the final product. Three strains were selected from a high throughput screening of 105 plant-derived LAB based on their acidification and metabolite production in the pulp. Two homofermentative strains (Lactococcus lactis) and one heterofermentative strain (Levilactobacillus brevis) were selected based on their low ethanol production, high lactic acid production, and overall acidification of the pulp. Mycelium of P. ostreatus was grown in submerged fermentations on the pre-fermented pulp, and the biomass was removed by centrifugation. The fungal strain consumed all available sugars and acids and released arabinose to the media. Volatiles were detected using GC–MS, and a large increase in concentrations of hexanal, 1-octen-3-ol, and 2-octenal was measured. Concentration of 1-octen-3-ol was lower in the pre-fermented samples vs. the non-pre-fermented. LC-MS amino acid analysis showed the presence of all essential amino acids on day 0 and 7 of fermentation. The highest concentration of amino acids was for glutamic acid/glutamine and aspartic acid/asparagine. A decrease in all amino acids after 7 days of fungal fermentation was measured for all fermentations. The decrease was more significant for pre-fermented samples. This was also confirmed through a total protein determination, except for samples pre-fermented with Lactococcus lactis strain NFICC142 which increased in total protein content after fungal fermentation. The protein digestibility increased after fungal fermentation, and the highest increase was seen for non-pre-fermented samples. The springiness of the fermented product indicated similarities to meat alternatives, while the hardness was much lower than other meat alternatives. The results indicate that dried sugar beet pulp can be used for submerged cultivation of P. ostreatus, but that pre-fermentation does not improve the physical or nutritional properties of the end product significantly, except for an increased protein content for NFICC142 pre-fermented media. This is the first known attempt to use LAB and P. ostreatus in mixed fermentation to produce fungal mycelium, as well as the first attempt at using SBP in a liquid fermentation for mycelial production of P. ostreatus.

Determination of total protein and sugar content in soy-based beverages using variable selection methods applied to ATR-FTIR spectroscopy

This study explores the potential of using ATR-FTIR coupled with partial least squares (PLS) and random forest (RF) for predicting total protein (TP), total sugar (TS), reducing sugars (RS), and non-reducing sugars (NRS) in soy-based beverages (SBBs). Employing variable selection techniques such as interval PLS (iPLS), synergy interval PLS (siPLS), uninformative variable elimination (UVE), ordered predictors selection (OPS), bat algorithm (BA), genetic algorithm (GA), and particle swarm optimization (PSO). The OPS-PLS emerges as the optimal model for TS prediction, yielding an RMSEP of 0.114 wt% and R2p of 0.934. GA-PLS excels in TP, RS, and NRS prediction, achieving RMSEP values of 0.024 wt%, 0.273 wt%, and 0.339 wt%, with corresponding R2p values of 0.880, 0.847, and 0.450. In RF modeling, GA-RF is identified as the superior model for all properties. These findings underscore the potential of regression models and ATR-FTIR as effective and environmentally friendly tools for analyzing SBB composition.

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.

Evaluation of portable refractometr as an alternative method to biuret assay for measurement of serum total protein in horse

The measurement of serum total protein (TP) is commonly performed by the biuret method. Refractometery which is less expensive and faster method also determines serum protein; however its accuracy is controversial. Therefore the primary objective of this study was to investigate portable refractometer as an alternative method to the biuret assay for measuring total serum proteins in horse serum. Serum from 50 male horses admitted to the society of the protection of animals’ abroad (SPANA) clinic was collected and measurement of serum total protein determined using portable refractometer and biuret method. The statistical differences between both methods were analyzed using a paired t-test and the correlation was determined by spearman correlation test. The results of this study demonstrate that there were no statistically significant variations in the mean total protein measured with the refractometer and biuret method (P > 0.05). The correlation between two methods using spearman correlations revealed high correlation between refractometry and biuret assay measurements of TP concentration r = 0.855. Bland–Altman plots indicated that biases indicating the analytic error was (4/50) 8% which is less than allowable total error for total protein determination (< 10%). Therefore portable refractometery can be used as an alternative to biuret method for serum total protein in horse as it is easy to use, quick turnaround time, and is inexpensive.

Effects of Ethyl Acetate Fraction of Mangifera haden Seeds on Carbon Tetrachloride Induced Hepatotoxicity in Albino Rats

The present study investigated the effects of ethyl acetate fraction of Mangifera haden seeds on carbon tetrachloride-induced liver damage in wistar albino rats. The phytochemical compositions and acute toxicity studies of crude extract and Mangifera haden seed fraction were determined using standard methods. Albino rats (60) randomly grouped into twelve of five rats each were used for this study. Determination of total bilirubin and protein, lipid peroxidation, liver enzymes marker, antioxidant enzymes, glutathione, lipid profile analysis and histological examination were carried out using standard methods. Phytochemical constituents of crude extract and ethyl acetate fraction of Mangifera haden seed showed that they contain relative amount of flavonoids, phenolics, saponins, tannins, terpenoids, triterpenes, alkaloids and coumarins. Liver injury was induced with carbon tetrachloride (CCl4) at a dose of 1.0 ml/kg body weight of the animals. All the CCl4 intoxicated rats displayed hyperlipidemia as shown by their elevated levels of serum total cholesterol (TC), triacylglycerol (TAG), low density lipoprotein (LDL), and reduction in high density lipoprotein (HDL) levels. The administration of ethyl acetate fraction of Mangifera haden seeds significantly (P = 0.05) lowered the levels of serum TC, TAG, LDL, and increased HDL levels. The level of serum malondialdehyde (MDA) significantly (P = 0.05) increased in the CCl4 group as compared to the normal and treated groups. The elevated level of serum MDA decreased (P = 0.05) significantly at the treatment of rats with 200, 400 and 600 mg/kg of ethyl acetate fraction. The rats treated with CCl4 produced significant (P = 0.05) increase in serum ALT, AST and ALP when compared to the normal group. Animals treated with varying doses of ethyl acetate fraction produced significant (P = 0.05) reduction of serum ALT, AST and ALP activities when compared to the CCl4-untreated group. The activities of hepatic antioxidant enzymes (SOD, CAT and GPx) and glutathione concentration significantly (P = 0.05) decreased in CCl4­-untreated group compared to the normal group. The decreased activities of the antioxidant enzymes and glutathione concentration in CCl4 intoxicated rats were ameliorated/modulated more effectively in rats treated with 200, 400 and 600 mg/kg ethyl acetate fraction of Mangifera haden seed. Total bilirubin (TB) concentrations significantly (P = 0.05) increased in CCl4-untreated group when compared to the normal group. Oral treatment of rats with varying doses (200, 400, 600 mg/kg) of ethyl acetate fraction caused a significant (P = 0.05) decrease in the serum TB levels. Administration of CCl4 showed a reduction in serum total proteins. Treatment of the animals with 200, 400 and 600 mg/kg of ethyl acetate fraction caused significant (P = 0.05) increase in serum total potein. Histological examination showed hepatoprotective properties of plant’s fractions. The compounds found in the different fractions (ethyl acetate and n-hexane) of Mangifera haden seeds are 9-Octadecenoic acid (Z)-, methyl ester, (Z) –Oleic acid and so on. Therefore the medicinal importance of the plant is been established.

CRUDE PROTEIN CONTENT IN PIGEON PEA GRAIN LINES

Pigeon pea (Cajanus cajan (L.) Millspaugh) is a multipurpose legume, which can be used as a grain producer, fodder and soil improver. Its nutritional composition has high protein, on average 22%, characterizing a relevant species in several emerging countries. Although it is an important crop in semi-arid regions of other countries, in Brazil has been little used and studied, so that it is still considered a subsistence culture. The goal of the present study was to evaluate the protein content in 21 lines of pigeon pea, from the breeding program of Embrapa Semiárido, including the control variety ‘guandu Petrolina’. The experiments were conducted in Barbalha/CE, Petrolina/PE and Juazeiro/BA, years of 2019, 2020 and 2021, distributed in four environments, three under irrigated cultivation and one under rainfed cultivation. The experiments were arranged in plots with two planting lines measuring 2.4m x 2.5m, spaced between rows by 1.2m and between plants by 0.5m, with two plants per hole, using a randomized block design with three replications. The cultural treatments consisted of manual weeding, without any type of fertilization in the experimental area. The determination of total nitrogen and crude protein was performed using the Kjeldahl method. The genotype × environment interaction was highly significant (p&lt;0.01) for protein content. There was no significant difference between the levels of protein under irrigated and rainfed cultivation. After joint analysis of variance among the four environments, the control cultivar 'Petrolina presented the highest average, with 24.60%. The lowest protein average was presented by treatment 17 (ICPL 900053 x Anagé), 20.12%. Lines 12 (ICPL 90053 x D2 Type), 9 (ICPL 90045 x ICPL 89027) and 19 (ICPL 89027 x D3 Type) expressed acceptable values for protein content, 22.35, 21.99 and 22.72%, respectively, associated with high productivity, broad adaptability and good predictability, showing great potential to be new cultivars to the Brazilian semiarid region.

Hemolysis in blood samples

Hemolysis is one of the interfering factors that can limit the results of various biochemical tests, and this study aimed to assess the interference of hemolysis in the reliability of biochemical tests conducted in clinical-surgical procedures in the veterinary hospital routine. The methodology was retrospective, based on the analysis of records of animals treated at the Veterinary Hospital Unit (SUHVU) of the Federal University of Fronteira, Campus Realeza (Paraná) from January of 2020 to December of 2022. The tests conducted in the Veterinary Clinical Analysis laboratory included serum determination of total proteins, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), creatinine and urea. Based on the biochemical results, animals were divided into three groups according to the degree of hemolysis observed in the serum: mild hemolysis (HD); moderate hemolysis (HM) and intense hemolysis (HI). The control group (GC) was determined based on reference values specific to the species. Data were tabulated in spreadsheets and analyzed using Pearson's correlation test (p). Mild hemolysis was predominant and, in dogs, did not affect the analyzed biochemical values in comparison to the reference averages for the species, regardless of the degree. Isolated, most measurements increased. In cats, changes were more significant concerning the group average, with ALP showing the greatest variation, not consistent with the elevation of hemolysis. None of the parameters had a strong correlation of interference between hemolyses, only moderate negative and positive correlations in ALP and urea.

Determination of moisture and total protein and phosphorus contents in powdered chicken egg samples using digital images, NIR spectra, data fusion, and multivariate calibration

This work uses digital images, near-infrared (NIR) spectra, data fusion, and multivariate calibration to develop fast, non-laborious, and cheap methods for determining moisture and total protein and phosphorus contents in powdered chicken egg samples. These analyzed samples were obtained after breaking, homogenizing, lyophilizing, and pulverizing whole chicken eggs. Digital images were captured employing a scanner, while NIR spectra were recorded using benchtop and portable spectrometers. The proposed multivariate calibration methods were better in terms of correlation coefficient (r) and ratio performance deviation of prediction (RPD) for predicting total protein content using digital images (r = 0.995 and RPD = 9.55) than those obtained by benchtop and portable NIR spectra. On the other hand, in predicting moisture (r = 0.974 and RPD = 4.35) and total phosphorus content (r = 0.98 and RPD = 4.3), the benchtop NIR produced better results. The analytical performance of data fusion methods was slightly poorer than the best methods using preprocessed NIR spectra or digital images separately. Therefore, these proposed methods proved to be a good alternative tool for destructive chemical analysis, given that powdered chicken egg samples could be analyzed without using chemicals and without generating waste harmful to health and the environment, following the basic principles of Green Chemistry.

Determination of human serum total protein via electrophoresis titration and capacitively coupled contactless conductivity detection

Serum total protein refers to the sum of all proteins in the serum, and its content determination is relevant to human health monitoring and disease diagnosis. However, existing detection techniques present a number of limitations; for example, the Kjeldahl method suffers from the negative effects of interfering substances such as non-protein nitrogen (NPN). Although the electrophoresis titration (ET) method has solved interference problems to some extent, the current ET technique relies on optical detection methods, which increases the tediousness of the operation. This study addresses the challenge of accurate serum total protein detection by combining the traditional ET technique with capacitively coupled contactless conductivity detection (C4D). The research contributions of this work are multifold. First, it presents the first development of an ET-C4D detection system, which consists of six components: an ET power module, an ET chip, a C4D sensing module, a detection module, a data acquisition card, and software. The developed system can capture the conductivity of substances in the channel using the software developed by our laboratory during ET. The detection system can be used to quantify the total protein content in human serum without the addition of specific labeling reagents or using optical detection equipment, and its running time is approximately 300 s. Second, this research proposes the corresponding principle of the system. Under an electric field, ion migration results in different pH levels before and after the boundary, leading to a protein surface charge difference. The maintenance of the electrical neutrality of the substances in the detection channel is related to the protein surface charge; therefore, the ion concentration distribution of the substances in the detection channel changes as the protein surface charge varies. A plot of conductivity as a function of running time showed an "inverted clock shape", first falling and then rising. Owing to the addition of different types and concentrations of proteins, the microenvironment of the entire system changes, resulting in different changes in conductivity. Third, the performance of the detection system was tested using human serum albumin (HSA) standard protein, which was mixed with polyacrylamide gel (PAG) mother liquor, riboflavin, etc., and irradiated under ultraviolet light for 10 min to form a gel. The ET experiments were then carried out. The shape of the conductivity curve was consistent with the proposed principle, and the higher the HSA concentration, the lower the conductivity curve trough, followed by a lagged time of the trough. Quantitative analysis of the conductivity signals showed that the linear range was 0.25-3.00 g/L, with a linearity of up to 0.98. The limit of detection (LOD) was 0.01 g/L, the relative standard deviation (RSD) was 1.90%, and the relative error of the test values was <7.20%, indicating the good detection stability and sensitivity of the system. Clinical samples collected from healthy volunteers were used as target blood samples for serum total protein content measurement using our detection system. Blood samples from a volunteer were used to obtain a standard curve, and the serum samples of other four volunteers were selected for ET-C4D and biuret detection. The results showed that the relative errors between the two methods were within 4.43%, indicating the accuracy and reliability of the detection system. The advantages of the ET-C4D detection system proposed in this paper are as follows: (i) ET-C4D realizes the rapid detection of total serum protein content based on the ET technique; (ii) compared with the traditional protein ET technique, the ET-C4D method does not rely on specific labeling components or optical detection equipment, thereby reducing the complexity of the operation; and (iii) the output signal of ET-C4D can be used for quantitative analysis with excellent analytical performance and high accuracy. These merits highlight the potential of the developed system for clinical application and biochemical analysis.

Evaluation of the effects of radiation doses from computed tomography on the biochemical parameters of the hepatorenal organs of albino rats

Background Aims: Computed tomography (CT) uses X-rays which are ionizing radiation. Interaction of ionizing radiation with the body may result in harmful effects on the organism depending on the radiation dose. This study aimed to evaluate the impact of different doses of CT radiation on the hepatorenal organs of male albino rats using biochemical parameters as the pathological response of the organs to radiation. Materials and Methods: Thirty healthy male Wistar albino rats weighing 180–200 g were assigned into five groups of six rats each. Rats in groups B, C, D, and E underwent noncontrast helical total body CT irradiation and received varying doses of CT radiation while group A received sham irradiation and served as control. At 72 h postirradiation, blood was collected using conventional methods, and serum was harvested for the determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TB), total protein (TP), albumin (Alb), urea, uric acid, and creatinine. Results: The mean serum levels of TP in the control (group A) and in the irradiated groups B, C, D, and E were 8.21 ± 0.34, 6.08 ± 0.30, 5.86 ± 0.42, 5.22 ± 0.30, and 4.72 ± 0.16, respectively. The mean values of TP in the irradiated groups decreased significantly in a dose-dependent manner compared to the control. The mean serum levels of ALP in the group A and in the irradiated groups B, C, D, and E were 75.20 ± 3.14, 111.60 ± 1.60, 117.60 ± 1.12, 124.40 ± 2.48, and 133.60 ± 2.46, respectively. The mean serum levels of AST in the group A and in the irradiated groups B, C, D, and E were 33.80 ± 0.58, 54.40 ± 2.29, 52.60 ± 1.78, 58.60 ± 2.64, and 63.60 ± 0.81, respectively. The mean serum levels of ALT in the group A and in the irradiated groups B, C, D, and E were 21.20 ± 0.86, 36.00 ± 0.84, 40.40 ± 1.17, 37.40 ± 0.93, and 41.60 ± 1.03, respectively. The mean serum levels of TB in the group A and in the irradiated groups B, C, D, and E were 2.78 ± 0.28, 3.70 ± 0.18, 4.77 ± 0.40, 5.88 ± 0.11, and 6.34 ± 0.04, respectively. The mean serum levels of Alb in the group A and in the irradiated groups B, C, D, and E were 3.68 ± 0.16, 5.96 ± 0.21, 6.65 ± 0.26, 8.15 ± 0.23, and 8.91 ± 0.17, respectively. The mean serum levels of urea in the group A and in the irradiated groups B, C, D, and E were 42.40 ± 0.17, 54.40 ± 1.33, 56.00 ± 1.76, 54.60 ± 1.72, and 57.80 ± 1.39, respectively. The mean serum levels of uric acid in the group A and in the irradiated groups B, C, D, and E were 5.65 ± 0.36, 8.32 ± 0.46, 8.87 ± 1.11, 9.13 ± 0.29, and 9.91 ± 0.29, respectively. The mean serum levels of creatinine in the group A and in the irradiated groups B, C, D, and E were 2.05 ± 0.13, 4.44 ± 0.32, 5.81 ± 1.17, 6.31 ± 0.03, and 6.63 ± 0.40, respectively. The mean values of ALP, AST, ALT, TB, Alb, urea, uric acid, and creatinine increased significantly in the irradiated groups compared to the control group. Conclusion: From the lowest to the highest doses of CT irradiation in the study, alterations in the biochemical parameters were evident and significant. Moreover, this suggests a possible deleterious biological effect of CT radiation on the hepatorenal organs probably due to the effects of radiation-induced free radicals which adversely affected the hepatorenal organs.

Hypophyseal acetylcholinesterase, and growth hormones of broiler chicken fed Ficus carica and vitamin C supplemented diets

The research was conducted to investigate hypophyseal acetylcholinesterase and growth hormones of broiler chickens fed Ficus carica and Vitamin C supplemented diets. 160-day-old chicks were randomly assigned to 4 treatments supplemented with Ficus carica and vitamin C in 4 replicates and 10 birds per replicate. At the end of the 8th week, the brains and hypophyses of 3 randomly selected birds per treatment were removed and tested for Acetylcholinesterase activities and total protein determination while blood samples were collected for hormonal assay. Results showed that total protein of the whole brain (0.50 mg/dl) and cerebellum (0.12 mg/dl) were significantly (p&lt;0.05) lower in birds fed control diet while that of the olfactory lobe was significantly higher than that of the control (0.25 mg/dl). However, the acetylcholinesterase activities (µmol/g tissue) of the optic lobe (0.36), Cerebellum (0.22), and medulla (0.25) of birds fed the control diet were significantly (p&lt;0.05) higher than those fed supplemented diets. Also, all hormonal assays tested were significantly influenced by the treatments with the exception of thyroxine. Birds fed diet 4 had the highest record for most growth hormonal assay. From the results obtained, Diet IV could be recommended in broiler chickens’ diets for superior growth promotion.

Plant-Based Total Protein Determination of Soybeans Using Fluorescent Technique

Plant-Based Total Protein Determination of Soybeans Using Fluorescent Technique

Comparison of calf morbidity, mortality, and future performance across categories of passive immunity: A retrospective cohort study in a dairy herd

Four categories of transfer of passive immunity (TPI) were recently proposed in response to the widespread high preweaning morbidity and mortality risks in calves with adequate TPI when a dichotomous classification was used. Hitherto, however, the risks of preweaning morbidity and mortality and future performance among these TPI categories have not been compared. Thus, the objective of this retrospective cohort study was to compare dairy calf morbidity, mortality, growth until weaning, and reproductive efficiency until first calving among the categories of poor (<5.1 g/dL total protein), fair (5.1-5.7 g/dL), good (5.8-6.1 g/dL), and excellent (>6.2 g/dL) TPI. For this, the records from 4,336 dairy calves (2,272 female, 2,064 male) born January 2014 to April 2017 on a commercial dairy farm in Michigan were analyzed. These calves had been randomly selected for weekly serum total protein determination on calves 2 to 7 d old. Data from both sexes were used to evaluate preweaning health and mortality, whereas only the female's data were used to investigate average daily gain (ADG), reproductive performance, and first-lactation milk yield. For each calf, data regarding disease status, growth, and reproductive parameters were obtained from the farm's software database. Associations of TPI categories with disease events (diarrhea or pneumonia), reproduction indices (age at first insemination, successful insemination, and calving, and number of inseminations), first-lactation milk yield and ADG at weaning were evaluated by survival analysis and mixed models. Compared with calves with excellent TPI, calves in the inferior TPI categories showed increased risk of diarrhea: poor [hazard ratio (HR) = 1.49; 95% CI: 1.22-1.82], fair (HR = 1.32; 95% CI: 1.16-1.51), good (HR = 1.14; 95% CI: 1.02-1.29). However, the risk of pneumonia differed only between the calves in the poor and excellent TPI groups (HR = 1.39; 95% CI: 1.05-1.84). The preweaning mortality risk was also higher in calves with poor TPI (HR = 4.29; 95% CI: 1.98-9.27) compared with excellent TPI. However, mortality risks were not statistically different between calves with fair or good TPI and those with excellent TPI. Similarly, calves with poor TPI had a 64, 55, and 24% lower risk of reaching first insemination, successful insemination, or first calving, respectively. However, there were no differences in ADG, number of inseminations, or first-lactation 305-d mature equivalent milk production across TPI groups. Our results confirm the positive effects of optimal TPI in calf preweaning health and postweaning reproductive efficiency. The 4 proposed categories of TPI can assist in decreasing the incidence of diseases that occur in the first weeks of life (i.e., diarrhea), but their effect on other diseases or future performance might be more limited. Although conducted in one herd, this study can be used to illustrate the effect of TPI on future calf performance.

Effect of sample fractionation and normalization when immunoblotting for human muscle Na+/K+-ATPase subunits and glycogen synthase

Immunoblotting is widely used in muscle physiology to determine protein regulation and abundance. However, research groups use different protocols, which may result in differential outcomes. Herein, we investigated the effect of various homogenization procedures on determination of protein abundance in human m. vastus lateralis biopsies. Furthermore, we investigated differences in abundance between young healthy males (n = 12) and type-2 diabetics (n = 4), and the effect of data normalization. Fractionated lysates had the lowest variation in total protein determination as compared to non-fractionated homogenates. Abundance of NKAα2, NKAβ1, FXYD1, and glycogen synthase was higher (P < 0.05) in young healthy than in type-2 diabetics determined in both fractionated and non-fractionated samples for which normalization to the stain-free signal and/or standard curve did not affect outcomes. Precision and reliability of protein abundance determination between sample types showed a moderate to good reliability for these proteins, whereas the commonly used house-keeping protein, actin, showed poor reliability. In conclusion, fractionated and non-fractionated immunoblotting samples yield similar data for several sarcolemmal and cytosolic proteins, except for actin, which, therefore appears inappropriate for data normalization in immunoblotting of human skeletal muscle. Thus, fractionation does not seem to be a major source of bias when immunoblotting for NKA subunits and GS.

Determination of total protein and lipid contents in some soybean varieties

Soybean seeds consist of a lot of necessary nutrients for health, they are considered a complete source of protein andvegetable lipids for the human body. In this study, six soybean varieties (DT22, Coc Chum B, black soybean, Kwangan, Cuc Ha Bac and DT84) were evaluated for total protein, lipids contents and acid and iodine indexes. Total protein content was determined by the Kjeldahl method. Oil was extracted from mature seeds using the Soxhlet system with diethyl ether solvent. The results showed that levels of total protein and lipids are relatively high in tested soybean varieties. The total protein content ranged from 29.36 to 37.32%, and lipids ranged from 19.8 to 21.2%. The free acid index was from 0.12 to 1.69 mg KOH/g, the iodine value was about 52.43 to 68.64 g I2/100 g. Varieties with high protein content were black soybean and Cuc Ha Bac. Nutritionally, these varieties are all sources of high nutritional value in providing protein and essential fatty acids for humans.

STUDIES ON BOVINE TRAUMATIC RETICULO- PERITONITIS IN BAGHDAD

A total of fourty-five cases of traumatic- reticulo- peritonitis in cows (16 cows ill for 24-48 hrs and 29 cows ill for 72-120 hrs). in Baghdad were diagnosed clinically, laboratory and autopical findings. The most valuable diagnostic signs were increased rectal temperature (39.9 C), respiratory rates (36/min.) and heart beats (95/min.), decreased in milk production (100.00 %), ruminal atony (95.56%), abdominal pain (93.33%), disturbed appetite (91.11%), decrease manure production (84.44%) and recurrent tympany (84. 44%). The total and differential leucocyte counts provided good diagnostic and prognostic data. The determination of total plasma protein and fibrinogen were used as aids to the diagnosis of traumatic reticulo-peritonitis in this study.

Effect of Monosodium Glutamate on the Digestibility of Different Nutrients Using Standardized Static In vitro Digestion Model

Monosodium glutamate (MSG) is a flavor enhancer and food additive with a unique umami taste. Due to its widespread use in humans, this study focused on the cytotoxicity, anti-diabetic effect, and interaction with protein digestion by performing a standardized static in vitro digestion model and lipid digestion by estimating free fatty acids released from 0.5 g of olive oil during intestinal lipolysis. The study showed that monosodium glutamate has an apparent cytotoxic effect on the Caco-2 cell line in a dose-dependent manner. MSG glutamate also showed low inhibitory activity on alpha-glucosidase enzyme even at high concentrations (16.3 % at 1800 ppm). By performing simulated in vitro digestion to study the interaction between MSG and protein digestion, followed by MTT study, total protein determination, and pH drop method, all results concluded that MSG affected proteolysis. Finally, the impact of MSG on lipolysis was studied through a free fatty acid release test. The results of the study demonstrated that MSG harmed fat digestibility in a concentration-dependent manner. As a result, it is essential to conduct further studies, especially in vivo studies, to determine the potential negative effects of MSG on human health.