Serum Albumin Protein Research Articles

Effects of dietary supplementation of postbiotic derived from Bacillus subtilis ACCC 11025 on growth performance, meat yield, meat quality, excreta bacteria, and excreta ammonia emission of broiler chicks

The primary aim of this study was to explore the impact of dietary supplementation with a postbiotic derived from Bacillus subtilis ACCC 11025 on growth performance, meat yield, meat quality, excreta bacterial populations, and excreta ammonia emissions of broiler chicks. A total of 480 day-old Arbor Acre broiler chicks, initially weighing 52.83 ± 1.38 g, were randomly allocated into 4 distinct groups. Each group was housed in 6 separate cages, each containing 20 birds. The experimental phase spanned 42 d, divided into 2 periods (d 1-21 and d 22-42). Dietary interventions were based on a basal diet, with postbiotic supplementation at levels of 0.000, 0.015, 0.030, or 0.045%. Our findings indicate that dietary supplementation with postbiotic had a positive influence on body weight gain (BWG) and feed efficiency. The most substantial improvements in BWG and feed efficiency were observed in the group of broiler chicks fed a diet containing 0.015% postbiotic. Furthermore, the inclusion of postbiotic in the diet led to an increase in the yield of breast and leg muscles, with a significant difference in meat yields observed between the control group and the group receiving 0.015% postbiotic supplementation. It's noteworthy that dietary manipulation did not exert any discernible impact on the quality of breast and leg muscle samples. Concurrently, we observed an elevation in serum albumin and total protein contents corresponding to the increasing postbiotic dosage in the diet. Additionally, dietary supplementation with postbiotic effectively controlled the emission of ammonia from excreta and reduced the abundance of Salmonella in excreta while enhancing the presence of Lactobacillus bacteria. The group receiving 0.015% postbiotic supplementation displayed the lowest levels of ammonia emission and the highest counts of Lactobacillus bacteria in excreta. In light of these results, we conclude that dietary supplementation with 0.015% postbiotic represents an efficacious strategy for increasing BWG and meat yield of broiler chicks by enhancing feed efficiency as well as mitigating ammonia emissions from excreta by modulating the composition of excreta bacterial communities.

Evaluating binding and interaction of selected pesticides with serum albumin proteins by Raman, 1H NMR, mass spectrometry and molecular dynamics simulation

Addressing the acute pesticide poisoning and toxicity to humans, is a global challenge of top priority. Serum albumin is the most abundant plasma protein, capable of binding with herbicide and pesticide residues. This study reports multifaceted approaches for in-depth and robust investigation of the molecular interactions of selected pesticides, including propanil (PPL), bromoxynil (BXL), metolachlor (MLR) and glyphosate (GPE) with bovine serum albumin (BSA) proteins using experimental (Raman and FTIR spectroscopy, native mass spectrometry and high field 1H NMR), molecular dynamics (MD) simulation and principal component analysis (PCA). The binding of pesticides with BSA resulted in BSA amide I and amide II Raman spectral shifts. PCA of Raman spectra of serum-pesticide complexes showed the grouping of pesticides on the score plot based on the similarities and differences in pesticides’ chemical structures. Native mass spectrometry results revealed strong adduct formation of the pesticides with the protein. The observed changes in chemical shifts, peak broadening or peak disappearance of characteristic proton signals of the pesticides, indicated altered chemical environments due to binding BSA-pesticides interactions. The results of MD simulation conducted for over 500 ns revealed strong pesticides interaction with LEU197, LEU218, LEU237, TRP213, SER286 and ILE289 residues to the site I of BSA. Free energy landscapes provided insights into the conformational changes in BSA on the binding of pesticides. Overall, the experimental and computational results are in consonant and indicate the binding of pesticides into the site I and site II (sub-domain IIA) of the BSA via hydrogen bonding, non-covalent and hydrophobic interactions. Communicated by Ramaswamy H. Sarma

Optimization of ultrasound-assisted extraction of galactomannan from carob seeds “ceratonia siliqua L” and evaluation of their functional properties and in vitro anti-inflammatory activity

ABSTRACT Ultrasound-assisted extraction (UAE) of galactomannan from the endosperm of ripe carob seeds (LBG.S3) was investigated through response surface methodology (RSM). The optimized UAE conditions were an extraction time of 40 minutes, a temperature of 45°C, and a solid-liquid ratio of 1:50 g/mL. Under these settings, the experimental yield of galactomannan was 47.53 ± 2.55%, which is closely aligning with the expected value of 49.78 ± 3.15%. The yield of galactomannan extracted from unripe carob seeds (LBG.S2) under the same optimal conditions was significantly higher 56.73 ± 2.76% (p < 0.05) than the yield of LBG.S3. The infrared spectroscopy (FT-IR) characterization revealed the existence of various functional groups in the spectrum of galactomannan extracted from unripe seeds, not found in LBG.S3. The galactomannan from ripe carob seeds exhibited superior functional properties, including solubility, emulsion and stability capacity, water and oil holding capacity, and viscosity, compared to LBG.S2. In vitro, tests demonstrated notable anti-inflammatory activity for both LBG.S2 and LBG.S3, as observed through the inhibition of bovine serum albumin (BSA) protein denaturation by heat and the membrane stabilization test. Carob galactomannan is a potential source of bioactive polysaccharides with anti-inflammatory and functional properties that can be used in the food and pharmaceutical industries.

Citrus limon (L.) Osbeck Fruit Peel Extract Attenuates Carbon Tetrachloride-Induced Hepatocarcinogenesis in Sprague-Dawley Rats.

Traditional herbal medicine practitioners in the Ashanti region of Ghana use the fruit peels of Citrus limon (L.) Osbeck (C. limon) in preventive and curative treatment of many cancers including liver cancer. This ethnobotanical claim remains to be verified scientifically. Aim of the Study. This study investigated prophylactic hepatoprotective and anti-HCC effects of C. limon peel extract (LPE) in CCl4/olive oil-induced HCC-like rats. After preparation of LPE, it was subjected to phytochemical screening using standard phytochemical methods. A total of 30 healthy adult male Sprague-Dawley rats (weighing 150-200 g) were randomly assigned into six groups of 5 rats each. Rats in the control group received olive oil (5 mL/kg ip) twice weekly for 16 weeks. Rats in the model group received CCl4/olive oil (2 mL/kg, ip) twice weekly for 16 weeks. Rats in capecitabine (10 mg/kg po) and LPE (50, 100, and 200 mg/kg po) groups received CCl4/olive oil (2 mL/kg, i.p) in the morning and their respective treatments in the afternoon twice a week for 16 weeks. Rats in all groups had free access to food and water ad libitum. Body weight and survival rates were monitored. Rats were sacrificed under deep anesthesia, blood was collected, and liver and other organs were isolated. Aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), prothrombin time, bilirubin, C-reactive protein (CRP), alpha- (α-) fetoprotein (AFP), and liver histology were assessed. Alkaloids, tannins, flavonoids, terpenoids, and saponins were detected in LPE. Model rats demonstrated increased serum levels of AFP, CRP, ALP, GGT, ALT, and AST, prothrombin time, total bilirubin, direct bilirubin, blood lymphocyte, and monocyte counts, but decreased serum albumin and total protein compared to control rats. Unlike the control, model rats demonstrated fat accumulation in periportal and centrilobular hepatocytes and neoplastic transformation. Semiquantitation of periodic acid Schiff- (PAS-) stained liver sections showed decreased glycogen storage in hepatocytes of model rats compared to control rats. Compared to the model, LPE treatment protected against CCl4-induced hepatocarcinogenesis, which was evidenced by decreased AFP, CRP, liver enzymes, total and direct bilirubin, prothrombin time, and blood lymphocyte and monocyte counts; attenuation of fat accumulation; and increased glycogen storage, albumin, and total protein. LPE abates CCl4-induced hepatocarcinogenesis by attenuating liver inflammation and improving metabolic, biosynthetic, and detoxification functions of the liver. The prophylactic hepatoprotective and anti-hepatocarcinogenic effects of LPE are attributable to its phytochemical composition raising hopes of finding potential anticancer bioactive compounds from C. limon fruit peels.

Native mass spectrometry analysis of conjugated HSA and BSA complexes with various flavonoids.

Mass spectrometry was used to study the binding interaction between serum albumin proteins (BSA and HSA) and flavone dyes, which is known to induce large fluorescence signals for protein detection. By electrospray ionization mass spectrometry (ESI-MS), multiple charged species/states could be produced in ammonium acetate buffer, while preserving the native structures of the proteins. Subsequent introduction of a flavone dye into the buffered solution resulted in an immediate interaction, forming the respective protein-dye conjugates associated by non-covalent interactions. Formation of protein-dye conjugates induced a notable response in the ESI-MS spectra, including changes in both the charge states and molecular mass of the protein species. The resulting data pointed out that the protein-flavone dye maintained a 1 : 1 ratio in the conjugate, although multiple binding sites for drug molecules are present in albumin proteins.

Synthesis of an Aryl-Semicarbazone-Based Cu(II) Complex for DNA and BSA Interaction and Anti-Cancer Activity against Human Cervix Uteri Carcinoma

The current study provides an in-depth analysis of the biological properties of a Cu(II) complex (C22H24Cu2N6O10) obtained from an aryl-semicarbazone ligand derived (L) from the condensation of 2,4-dihydroxy acetophenone and semicarbazide. The binding behavior of this complex with calf thymus DNA (CT-DNA) and bovine serum albumin (BSA) protein was explored using a combination of experimental and theoretical approaches. The results suggest that the complex binds with CT-DNA via a partial intercalation, and hydrophobic interaction. However, the complex binds to BSA protein predominantly through hydrogen bonding or van der Waals interactions rather than hydrophobic interactions. The molecular docking methodology was carried out to substantiate the experimental finding. Furthermore, the in vitro cytotoxicity study was conducted on human cervix uteri carcinoma (SiHa cancerous cell) lines upon exposure to the complex, and the findings reveal a considerable decrease in cell viability, when compared to the control. Overall, this study provides a comprehensive understanding of the biological potential of the Cu(II) complex and its potential as an anti-cancer agent.

Copper(II) coordination compounds based on bis-hydrazones of 2,6-diacetylpyridine: synthesis, structure, and cytotoxic activity.

By reacting a series of 2,6-diacetylpyridine bis-hydrazones containing pyrimidine (H2L1), benzimidazole (H2L2) and phthalazine (H2L3) heterocyclic fragments with copper(II) chloride and bromide, a variety of pentacoordinated complexes of the composition [Cu(H2L1)X]X, [Cu(HL2)X] and [Cu(HL3)X], where X = Cl-, Br-, are formed. The properties and structure of the compounds were studied by means of NMR, IR, UV-vis, ESR, and X-ray absorption spectroscopy, cyclic voltammetry and X-Ray single crystal diffraction methods. It was shown that complexes of the cationic type [Cu(H2L1)X]X have an asymmetric structure with a distorted square-pyramidal geometry of the coordination unit. The coordination polyhedron of metal chelates [Cu(HL2)X] and [Cu(HL3)X] is an almost ideal square pyramid. Investigations of the cytotoxic activity of the obtained compounds in vitro on human hepatocellular carcinoma (HepG2) and non-tumor human lung fibroblast (MRC-5) cell lines demonstrated that complexes show higher activity compared with the well-known anticancer agent cisplatin. In addition, metal chelates [Cu(H2L1)Cl]Cl, [Cu(HL2)Cl], [Cu(HL2)Br] and [Cu(HL3)Cl] were less toxic to non-tumor cells MRC-5. A study of the binding of complexes to bovine serum albumin (BSA) protein using fluorescence spectroscopy showed that copper complexes are strongly bound to BSA. To study the mechanism of interaction of the complexes with the DNA of cancer cells, molecular dynamics simulation of the compound [Cu(HL3)Cl] was carried out. It was shown that the complex enters into π-stacking interactions predominantly with adenine and thymine bases.

Angiotensin II Type 2 Receptor Antibodies in Glomerular Diseases.

We evaluated the concentration of AT2R antibodies in 136 patients with primary and secondary glomerular diseases: membranous nephropathy (n = 18), focal and segmental glomerulosclerosis (n = 25), systemic lupus erythematosus (n = 17), immunoglobulin A (IgA) nephropathy (n = 14), mesangial (non-IgA) proliferative nephropathy (n = 6), c-ANCA vasculitis (n = 40), perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA) vasculitis (n = 16), and compared it with a healthy control group (22 patients). Serum creatinine levels, proteinuria, serum albumin, and total protein concentrations were prospectively recorded for 2 years. The mean levels of AT2R antibodies in the lupus nephropathy group were significantly higher compared to the control group, 64.12 ± 26.95 units/mL and 9.72 ± 11.88 units/mL, respectively. There was no association between this level and the clinical course of the disease. The AT2R levels in other kinds of glomerular disease were no different from the control group. We found significant correlations between AT1R and AT2R in patients with membranous nephropathy (r = 0.66), IgA nephropathy (r = 0.61), and c-ANCA vasculitis (r = 0.63). Levels of AT2R antibodies in systemic lupus erythematosus are higher compared to other types of glomerulonephritis, vasculitis, and a healthy control group. Levels of AT2R antibodies correlate with AT1R antibodies in the groups of patients with membranous nephropathy, IgA nephropathy, and c-ANCA vasculitis. These kinds of AT2R antibodies have a stimulative effect on AT2R, but we have not found the influence of these antibodies on the clinical course of glomerular diseases.

Evolution of the structure and interaction in the surfactant-dependent heat-induced gelation of protein.

The addition of a surfactant and/or an increase in temperature disrupt the native structure of proteins, where high temperature further results in protein gelation. However, in a mixed protein-surfactant system, surfactant concentration and temperature have been observed to exhibit both mutually associative and counter-balancing effects towards heat-induced gelation of protein-surfactant dispersion. This study is conducted on globular bovine serum albumin (BSA) protein and cationic surfactant dodecyl trimethyl ammonium bromide (DTAB), which interact strongly owing to their oppositely charged nature. The findings reveal that the BSA-DTAB suspension undergoes gelation with increasing temperature but only at lower concentrations of DTAB, where the presence of the surfactant facilitates gelation (associative effect). Conversely, as the surfactant concentration increases beyond a critical value, temperature-driven gelation of the BSA-DTAB system is completely inhibited, despite surfactant-induced protein denaturation (counter-balancing effect). To conceptualize these results, we compared them with observations made in a system comprising protein and a similarly charged surfactant, sodium dodecyl sulfate (SDS). It has been further demonstrated that the anionic surfactant (SDS) can restrict protein gelation at much lower concentration compared to the cationic surfactant (DTAB). The evolution of the structure and interaction during gel formation/inhibition has been examined to understand the underlying mechanism guiding these sol-gel transitions. We present a comprehensive phase diagram, encompassing the solution/gel states of the protein-surfactant dispersion, with respect to the dispersion temperature, surfactant concentration, and ionic behavior (anionic or cationic) of the surfactants.

Integrated magneto-plasmonic nanostructures-based immunoassay for galectin-3 detection.

Cardiovascular diseases remain a leading cause of global mortality, highlighting the need for accurate diagnostic tools and the detection of specific cardiac biomarkers. Surface-enhanced Raman scattering (SERS) spectroscopy has proved to be a promising alternative diagnostic tool to detect relevant biomarkers compared to traditional methods. To our knowledge, SERS methodology has never been used to detect galectin-3 (Gal-3), a crucial biomarker for cardiovascular conditions. Our study aimed to develop plasmonic and magneto-plasmonic nanoplatforms for the sensitive immunodetection of Gal-3 using SERS. Spherical gold nanoparticles (AuNPs) were synthesized and functionalized with 11-mercaptoundecanoic acid (MUDA) to enable antibody binding and 4-mercaptobenzoic acid (4MBA) that served as a Raman reporter due to its intense Raman signal. Following bioconjugation with Gal-3 antibody, the AuNPs were employed in the immunodetection of Gal-3 in phosphate-buffer saline (PBS) solution, offering a limit of detection (LOD) of 12.2 ng mL-1 and a working range up to 120 ng mL-1. Furthermore, our SERS-based immunosystem demonstrated selectivity for Gal-3 (40 ng mL-1) in the presence of other biomolecules such as α-amylase, bovine serum albumin and human C-reactive protein. As a proof of concept, we developed magneto-plasmonic nanoparticles composed of silica-coated magnetite decorated with the bioconjugated AuNPs aimed at enhancing the uptake and detection of Gal-3 via SERS coupled with Raman imaging. Our findings underscore the potential of SERS-based techniques for the sensitive and specific detection of biomarkers, holding significant implications for improved diagnosis and surveillance of cardiovascular diseases. Future research will focus on further optimizing these nanoplatforms and their translation into clinical settings.

Enhancing the sequence coverage of nanodiamond-extracted early secretory proteins from the Mycobacterium tuberculosis complex.

The unambiguous identification of protein species requires high sequence coverage. In this study, we successfully improved the sequence coverage of early secretory 10 kDa cell filtrate protein (CFP-10) and 6 kDa early secretory antigenic target (ESAT-6) proteins from the Mycobacterium tuberculosis complex (MTC) in broth culture media with the use of the 4-chloro-α-cyanocinnamic acid (Cl-CCA) matrix. Conventional matrices, α-cyano-hydroxy-cinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB), were also used for comparison. After nanodiamond (ND) extraction, the sequence coverage of the CFP-10 protein was 87% when CHCA and DHB matrices were used, and the ESAT-6 protein was not detected. On the other hand, the sequence coverage for ND-extracted CFP-10 and ESAT-6 could reach 94% and 100%, respectively, when the Cl-CCA matrix was used and with the removal of interference from bovine serum albumin (BSA) protein and α-crystallin (ACR) protein. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) was also adopted to analyze the protein mass spectra. A total of 6 prominent ion signals were observed, including ESAT-6 protein peaks at mass-to-charge ratios (m/z) of ∼7931, ∼7974, ∼9768, and ∼9813 and CFP-10 protein peaks at m/z of ∼10 100 and ∼10 660. The ESAT-6 ion signals were always detected concurrently with CFP-10 ion signals, but CFP-10 ion signals could be detected alone without the ESAT-6 ion signals. Furthermore, the newly found ESAT-6 peaks were also confirmed using a Mag-Beads-Protein G kit with an ESAT-6 antibody to capture the ESAT-6 protein, which was also consistent with the sequence coverage analysis.

The hepatoprotective effects of sitagliptin against cyclophosphamide-induced hepatotoxicity in rat.

Hepatotoxicity is a serious side effects of cyclophosphamide. Thus, the present research investigates the protective properties of sitagliptin against cyclophosphamide-induced hepatotoxicity. Fifty male rats were randomly divided into five groups. They were pre-treated with either sitagliptin or normal saline once a day for the first ten days of the study. To induce acute hepatotoxicity, cyclophosphamide (200 mg/kg, i.p) was injected only one time and 45 min after the last dose of sitagliptin. The rats were sacrificed on the 11th day, and their blood and liver were collected for biochemical, gene expression, and histopathological assessments. Our results showed that cyclophosphamide induced obvious liver toxicity as marked by an increase in serum levels of alanine transaminase and aspartate transaminase, reduced serum albumin and total protein levels, in addition to histopathological changes. The malondialdehyde, tumor necrosis factor-α, and interleukin-6 levels were also elevated and total antioxidant capacity declined in serum and hepatic homogenates. Sitagliptin magnificently attenuated the cylophosphamide-induced histological alterations, improved liver function tests, enhanced total antioxidant capacity, and decreased malondialdehyde, tumor necrosis factor-α, and interleukin-6 in the blood and hepatic tissues. These findings suggest that sitagliptin has hepatoprotective activity against cyclophosphamide toxicity, which may be due to its antioxidant and anti-inflammatory effects.

Unraveling mechanisms of protein encapsulation and release in coacervates via molecular dynamics and machine learning.

Coacervates play a pivotal role in protein-based drug delivery research, yet their drug encapsulation and release mechanisms remain poorly understood. Here, we utilized the Martini model to investigate bovine serum albumin (BSA) protein encapsulation and release within polylysine/polyglutamate (PLys/PGlu) coacervates. Our findings emphasize the importance of ingredient addition sequence in coacervate formation and encapsulation rates, attributed to preference contact between oppositely charged proteins and poly(amino acid)s. Notably, coacervates composed of β-sheet poly(amino acid)s demonstrate greater BSA encapsulation efficiency due to their reduced entropy and flexibility. Furthermore, we examined the pH responsiveness of coacervates, shedding light on the dissolution process driven by Coulomb forces. By leveraging machine learning algorithms to analyze simulation results, our research advances the understanding of coacervate-based drug delivery systems, with the ultimate goal of optimizing therapeutic outcomes.

In situ Fourier transform infrared-attenuated total reflection spectroscopy and modeling investigation of protein adsorption: Case of expanded bovine serum albumin on titanium dioxide anatase.

The purpose of this experimental and modeling research is to study the pH effect and to determine the surface coverage plus the adsorption constant (Ka) of bovine serum albumin (BSA) protein adsorbed on TiO2 anatase surface, respectively. In situ Fourier transform infrared-attenuated total reflection spectroscopy in a flow-through cell was used to study the BSA adsorption on porous TiO2 anatase films. The experiments were performed in water solution, under different pH values, at a concentration of 10-6 mol/l. Theoretically, we extended the two-state model, based on a system of coupled differential equations, by adding a desorption parameter Kd2, for unfolded state. The model was solved taking into account the adsorption (Ka), desorption (Kd1,2), transformation (Kf) coefficients, and the initial solution protein concentration (C0). The findings clearly illustrated that the solution pH drastically changed the behavior of BSA adsorption, whereas the mathematical analytical solutions allowed us to determine the native state (θ1), the unfolded state (θ2), and the full one (θ) surface coverages. Finally, a good application of the approximated model on the experimental work, expanded BSA adsorbed on TiO2 anatase at pH = 1.7, indicated a value of Ka = (408.36 ± 0.996) × 102 mol-1 l min-1.

Towards a better understanding of the effect of protein conditioning layers on microbial adhesion: a focused investigation of fibronectin and bovine serum albumin layers on SiO2 surfaces.

The interaction of foreign implants with their surrounding environment is significantly influenced by the adsorption of proteins on the biomaterial surfaces, playing a role in microbial adhesion. Therefore, understanding protein adsorption on solid surfaces and its effect on microbial adhesion is essential to assess the associated risk of infection. The aim of this study is to evaluate the effect of conditioning by fibronectin (Fn) or bovine serum albumin (BSA) protein layers of silica (SiO2) surfaces on the adhesion and detachment of two pathogenic microorganisms: Pseudomonas aeruginosa PAO1-Tn7-gfp and Candida albicans CIP 48.72. Experiments are conducted under both static and hydrodynamic conditions using a shear stress flow chamber. Through the use of very low wall shear stresses, the study brings the link between the static and dynamic conditions of microbial adhesion. The results reveal that the microbial adhesion critically depends on: (i) the presence of a protein layer conditioning the SiO2 surface, (ii) the type of protein and (iii) the protein conformation and organization in the conditioning layer. In addition, a very distinct adhesion behaviour of P. aeruginosa is observed towards the two tested proteins, Fn and BSA. This effect is reinforced by the amount of proteins adsorbed on the surface and their organization in the layer. The results are discussed in the light of atomic force microscopy analysis of the organization and conformation of proteins in the layers after adsorption on the SiO2 surface, as well as the specificity in bacterial behaviour when interacting with these protein layers. The study also demonstrates the very distinctive behaviours of the prokaryote P. aeruginosa PAO1-Tn7-gfp compared to the eukaryote C. albicans CIP 48.72. This underscores the importance of considering species-specific interactions between the protein conditioning layer and different pathogenic microorganisms, which appear crucial in designing tailored anti-adhesive surfaces.

Two populations of protein molecules detected by small-angle neutron and X-ray scattering (SANS and SAXS) in lyophilized protein:lyoprotector (disaccharide) systems.

Two protein interaction peaks are observed in pharmaceutically-relevant protein (serum albumin) : disaccharide 1 : 1 and 1 : 3 (w/w) freeze-dried systems for the first time. In samples with a higher disaccharide content, the protein-protein distances are longer for both populations, while the fraction of the protein population with a shorter protein-protein distance is lower. Both factors would favor better stability against aggregation for disaccharide-rich protein formulations. This study provides direct experimental support for a "dilution" hypothesis as a potential stabilization mechanism for freeze-dried protein formulations.

Hepatoprotective efficacy of Lagenaria siceraria seeds oil against experimentally carbon tetrachloride-induced toxicity.

The liver is crucial for maintaining normal metabolism in the body. Various substances, such as toxic chemicals, drugs, and alcohol, can damage hepatocyte cells, leading to metabolic imbalances. The experiment aimed to determine the efficacy of Lagenaria siceraria seed oil (LSS) as a hepatoprotective agent against acute hepatotoxicity triggered by carbon tetrachloride (CCl4). A total of 20 rats were randomly separated into four groups. The control group: rats received 2 ml of distilled water orally, followed by 1.25 ml of olive oil intraperitoneally (i.p.) after 30 minutes. CCL4 group: rats were given a single intraperitoneal dose of 1.25 ml/kg b.w. of CCl4 in a 1:1 mixture with olive oil. Silymarin group: received 100 mg of silymarin per kg of b.w. diluted in 2 ml of distilled water orally, followed by CCl4 treatment after 30 minutes. LSS oil group: received LSS oil at 3g/kg b.w. orally, followed by CCl4 treatment after 30 minutes. Blood samples were collected to assess liver enzymes (AST, ALT, and ALP), proteins and bilirubin fractions, and redox status (catalase, reduced glutathione (GSH), and malondialdehyde (MDA)) were assessed in hepatic tissues. Changes in liver histopathological examination were also evaluated. In CCl4-treated rats, there was a significant increase in serum liver marker enzyme activity (ALP, AST, and ALT) along with a significant elevation (p < 0.05) in total bilirubin, indirect bilirubin, and direct bilirubin compared to the control rats. However, all these parameters decreased in the CCl4+ Silymarin and CCl4+LSS groups compared to CCl4-treated rats. There was a significant decline in total protein level and serum albumin in all experimental groups compared to the control, while globulin levels significantly increased in all experimental groups. There was a significant (p < 0.05) reduction in the level of GSH and catalase, with an increase in MDA level in CCl4 rats compared to other rats. Histopathological investigation of the LSS-treated group showed a hepatoprotective effect against CCl4. The study revealed that LSS oil has antioxidant activity against CCl4-induced toxicity.

Proteins denaturation upon interaction of heavy metals and cosolvent glycerol assisted refolding: in-vitro study with human serum albumin.

Human blood serum protein albumin (HSA) is well studied protein owing its importance being a carrier molecule for endogenous molecules, toxins, xenobiotics and other small ligands like metal ions.

A highly sensitive triazole-based perfectly water soluble novel bis-Schiff base reversible fluorescent-colorimetric chemosensor for fast detection of Pb2+ ions.

A reversible fluorescent-colorimetric azino bis-Schiff base receptor for the detection of Pb2+ in aqueous medium has been developed for the first time. Receptor L exhibits an excellent selective and rapid fluorescent-colorimetric response towards Pb2+. The sensitivity of the fluorescent-based assay (0.53 nM) and colorimetric assay (1.0 nM) for Pb2+ is sufficiently good in comparison to previously reported literature. From 1H NMR data, Job plot measurement and the ESI-MS spectrum, a 1 : 2 stoichiometric complexation between L and Pb2+ has been established. Receptor L shows a remarkable detection ability in a wide pH range of 4-8 and it has been successfully utilised in the determination of Pb2+ in aqueous solution of bovine serum albumin protein and in real samples. The geometry of L has been optimized by both DFT studies and NMR, FTIR and mass spectra. Moreover, we have studied molecular docking of the probe L.

A prototype of ultrasensitive time-resolved fluoroimmunoassay for the quantitation of lead in plasma using a fluorescence-enhanced europium chelate label for the detection system.

This study describes the prototype of a novel ultra-sensitive time-resolved fluoroimmunoassay (TRFIA) for the quantification of lead (Pb) in plasma. The assay procedures were conducted in 96-microwell plates and involved the competitive binding format. The assay used a mouse monoclonal antibody, designated as 2C33, that specifically recognized the diethylenetriamine pentaacetic acid chelate of Pb (Pb-DTPA) but did not recognize Pb-free DTPA chelator. The antigen used for coating onto the inner surfaces of assay plate microwells was Pb-DTPA conjugated with bovine serum albumin protein (Pb-DTPA-BSA). The competitive binding reaction occurred between Pb-DTPA chelates, formed in the sample solutions by treating the samples with an excess DTPA, and the coated Pb-DTPA-BSA for a limited quantity of 2C33 antibody binding sites. The antigen-antibody complex formed in the plate wells was quantified by a europium-DTPA-labeled secondary antibody and a fluorescence enhancement solution. The conditions of the assay were refined, and its optimum procedures were established. The TRFIA was validated following the immunoassay validation guidelines, and all of the validation criteria were acceptable. The working range of the assay was 20-300 pg mL-1 and its limit of quantitation was 20 pg mL-1. Metals that are commonly encountered in blood plasma did not interfere with Pb in the analysis by the proposed TRFIA. The assay was applied to the quantitation of Pb in plasma samples with satisfactory accuracy and precision. The results were compared favorably with those obtained by atomic emission spectroscopy. In conclusion, the present study represents the first TRFIA for the quantitation of Pb in plasma. The assay is superior to the existing atomic spectrometric methods and other immunoassays for Pb in terms of sensitivity, convenience, and analysis throughputs. The proposed TRFIA is anticipated to effectively contribute to assessing Pb concentrations and controlling the exposure of humans to its potential toxicity.