Serum Albumin Research Articles

Impact of ligand structure and base bead pore size on host cell protein removal during monoclonal antibody purification using multimodal chromatography resin

Despite advancements in therapeutic monoclonal antibodies (mAbs) and cell line engineering, separating host cell proteins (HCPs) from mAbs during downstream purification remains challenging. Therefore, in this study, we developed a novel multimodal chromatography (MMC) resin to enhance HCP removal during mAb polishing processes. We evaluated the impact of both ligand structure and pore size of the MMC resin by purifying a post-protein A chromatography solution in flow-through mode. We observed that the efficiency of HCP clearance depended on the hydrophobic moiety structure of the ligand and predicted the mAb purification capability of MMC through linear salt-gradient elution experiments involving a mixture of transferrin, bovine serum albumin (BSA), and pepsin. Our findings revealed that the prototype immobilized 1,12-dodecanediamine via the formyl group exhibited the best performance attributed to its long alkyl chain. Furthermore, an investigation of effects of base bead pore size on HCP capacity using cellulose base beads of five different pore sizes showed that larger pore resin base beads had the highest HCP removal capacity. Specifically, MMC resins with a pore diameter exceeding 440 nm reduced the HCP level by three orders of magnitude under high mAb loading conditions (> 1000 mg/mL-resin). The MMC resin developed in this study, along with the insights gained into ligand structure and pore size, not only enhances mAb polishing efficiency but also contributes to improving downstream processes in mAb biopharmaceutical production.

Modification of polyvinylidene fluoride membrane with ciprofloxacin to improve the bacteriostatic performance

The common polyvinylidene fluoride (PVDF) membrane itself is susceptible to membrane fouling, especially biofouling, which is a serious threat. In this study, PVDF membrane was modified with ciprofloxacin (CIP) through co-blending to investigate the filtration properties, bacterial inhibition and fouling resistance. Modified membranes were prepared by adding 0.3 g (MC0.3), 0.6 g (MC0.6), 0.9 g (MC0.9) and 1.2 g (MC1.2) CIP per 100 g casting solution. Among these modified membranes, MC0.6 showed the best filtration performances, with the pure water flux stabilized at about 416.67 L/(m2·h) and bovine serum albumin (BSA) rejection of 92.0% at a trans-membrane pressure of 0.1 MPa. The pore size was reduced, the average roughness was reduced to 29.4 nm, the contact angle was lowered to 68.9°, and the hydrophilicity was greatly improved. The width of the inhibition circle produced by MC0.6 was 0.35-0.45 mm, and the modified membrane showed good inhibition of non-specific bacteria and algal removal during urban river water filtration. The rejection of BSA was increased by 16.32% compared to the base membrane and the adsorption rate for BSA was reduced by 68.45%. In addition, the removal of conventional pollutants in urban river water by the modified membranes for was also improved. Compared with that of the base membrane, the removal of TN, NH3-N, TP and COD by MC0.6 was increased by 10.58%, 12.45%, 15.44% and 13.53%. The results showed that CIP co-blending modified PVDF membrane could effectively improve membrane performances and has good value for water treatment.

Portal Venous and Hepatic Arterial Coefficients Predict Post-Hepatectomy Overall and Recurrence-Free Survival in Patients with Hepatocellular Carcinoma: A Retrospective Study.

The dominant artery blood supply is a characteristic of hepatocellular carcinoma (HCC). However, it is not known whether the blood supply can predict the post-hepatectomy prognosis of patients with HCC. This retrospective study investigated the prognostic value of the portal venous and arterial blood supply estimated on triphasic liver CT (as a portal venous coefficient, PVC, and hepatic arterial coefficient, HAC, respectively) in patients with HCC following hepatectomy. HCC patients who were tested by triphasic liver CT 2 weeks before hepatectomy and received R0 hepatectomy at the Second Affiliated Hospital, Kunming Medical University between January 1, 2016 and December 31, 2020, were retrospectively screened. Their PVC and HAC, and other variables were analyzed for the prediction of overall survival (OS) and recurrence-free survival (RFS) using the least absolute shrinkage and selection operator and Cox proportional hazard regression models. Four hundred and nineteen patients (53.2 ± 10.6 years of age and 370 men) were evaluated. A shorter OS was independently associated with higher blood albumin and total bilirubin grade [hazard ratio (HR) 2.020, 95% confidence interval (CI) 1.534-2.660], higher Barcelona Clinic Liver Cancer (BCLC) stage (HR 1.514, 95% CI 1.290-1.777), PVC ≤ 0.386 (HR 1.628, 95% CI 1.149-2.305), and HAC > 0.029 (HR 1.969, 95% CI 1.380-2.809). A shorter RFS was independently associated with male (HR 1.652, 95% CI 1.005-2.716), higher serum α-fetoprotein ≥ 400 ng/mL (HR 1.672, 95% CI 1.236-2.263), higher BCLC stage (HR 1.516, 95% CI 1.300-1.768), tumor PVC ≤ 0.386 (HR 1.641, 95% CI 1.198-2.249), and tumor HAC > 0.029 (HR 1.455, 95% CI 1.060-1.997). Tumor PVC or HAC before hepatectomy is valuable for independently predicting postoperative survival of HCC patients.

Intestinal permeability in migraine male patients upon omega-3 and tocopherol administration

Background. Gut microbiome may affect CNS, while an important neurotransmitter in migraine patients can be expressed in enteric neurons. Omega-3 PUFA (polyunsaturated fatty acid) and vitamin E are reported to decrease neuroinflammation, have beneficial effects on intestinal wall, and microbiota of the small intestine. This study aims to find the effect of omega 3 PUFA and vitamin E on intestinal permeability in migrainous, and their possible modulation effect on certain vitamins that have relations for both migraine and intestinal integrity. Methods. A total of 21 migrainous men are included in this study. Blood samples were drawn before and after intake of omega-3 PUFA and vitamin E supplementation softgel once daily for 30 days to compare with 13 apparently healthy men (control group). The blood samples are used to measure serum lipopolysaccharide, zonulin, amylase, vitamin B12, vitamin D, and albumin. serum lipopolysaccharide, and zonulin. Results. Body mass index, serum lipopolysaccharide and serum zonulin of migrainous men are higher, but vitamin B12 and vitamin D are lower than in the control group. Upon intake of supplement, serum zonulin and lipopolysaccharide are decreased in migrainous men indicating an improvement of intestinal integrity, but vitamin B12 and vitamin D showed no significant difference. Serum albumin and amylase showed no significant differences. Conclusion. Omega 3 PUFA and vitamin E supplements decreased intestinal permeability and can be a useful as an adjunct therapy for migrainous men because of its positive effect on intestinal integrity and gut brain axis, but it has no significant effect on vitamin B12 and vitamin D levels.

Analysis of Oral Albumin Use and Factors Affecting the Success of its Therapy

Hypoalbuminemia is a risk factor for medical treatment. Albumin has been shown to be a predictor in a number of studies in various settings. Several studies examined albumin as a marker of nutritional status and its association with outcomes (eg, mortality and length of hospital stay). In critical illness, the Acute Physiology and Chronic Health Evaluation (APACHE III) risk score includes serum albumin levels and shows a strong inverse correlation between serum albumin and mortality in critically ill patients. This research was conducted observationally to analyze the use of oral albumin and the factors that influence the success of therapy in the Lung Inpatient Room at Dr Soetomo Regional Hospital for the period 1 January – 31 October 2022. Patients diagnosed with Community Pneumonia, Lung Cancer and Lung Tuberculosis. This research was carried out by collecting secondary data via e-medical records. Data taken were patient demographic data (name, gender, weight, age, length of stay) and albumin data before and after receiving oral albumin. From the research results, the number of samples was 87. The number of samples was 87 patients. Of the 87 patients, 66 male patients (75.86%) were 21 female patients (24.14%), the largest age range was 51 - 60 years, 21 patients (24.14%). The highest body weight ranged from 61-70 kg for 50 patients (57.47%), the longest length of stay ( Length of Stay ) was 10-20 days for 45 patients (51.72%). The most common primary diagnosis was new tuberculosis in 42 patients (48.28%). The highest use of oral albumin was more than 100 capsules, as many as 20 patients (22.99%) with the highest duration of use being 10-19 days, namely 38 patients (43.68%). The statistical results show that there is no significant difference between before receiving oral albumin and after receiving oral albumin with a value of P = 0.541. Factors that influence the success of oral albumin therapy are length of stay with a value of P = 0.001 * Keywords: Oral albumin, pulmonary tuberculosis, lung cancer, pneumonia, length of stay

An Optofluidic Young Interferometer for Electrokinetic Transport-Coupled Biosensing

Label-free optical biosensors, such as interferometers, can provide a comparable limit of detection to widely used enzyme-linked immunosorbent assays while minimizing the number of steps and reducing false positives/negatives. In 2020, the authors reported on a novel optofluidic Young interferometer (YI) that could provide real-time spatial information on refractive index changes occurring along the length of the sensor and reference channels. Herein, we exploit these features of the YI to study interactions of biomolecules with recognition elements immobilized in selected regions of agarose gel in the sensor channel. We show that the YI is well suited for the biosensing of an exemplar biomolecule, streptavidin, in the absence and presence of the bovine serum albumin interferent. Equally, we couple the YI with electrokinetic transport to reduce the time needed for biosensing.

Freezing Diluted Bovine Serum Albumin Standards Does Not Significantly Affect Standard Curves.

Total protein isolation followed by quantitation is a common protocol in many laboratories. Quantitation is often done using a colorimetric assay such as the bicinchoninic acid (BCA) assay in which a change in the color of the BCA reagent is related to protein concentration. Extracted protein samples are compared to a standard curve made with dilutions of a protein standard such as bovine serum albumin (BSA) to determine their concentrations. A series of experiments was designed to determine the most reproducible and accurate method for quantifying protein concentrations of samples in an experimental series over time. The effect of freezing on diluted standards was investigated. Standards were frozen at -20°C or -80°C and serially thawed and refrozen up to three times prior to their use in a BCA assay. Thawing and refreezing the standards had no significant effect on protein concentration and the resulting standard curves. Inter-person and intra-person variability in the preparation of standards was also investigated. Protein concentration differences due to inter-person and intra-person variability were greater than protein concentration variability resulting from freezing and thawing, regardless of the freezing temperature. The most reproducible and accurate method for determining the protein concentration of extracted samples in an experimental series over time is diluting a large batch of BSA standards and freezing them at either -20°C or -80°C. Reproducibility was maintained with up to three freeze-thaws.

The Modified Albumin–Bilirubin (ALBI) Grade Reflect the Fate of Limb Prognosis in Patients with Chronic Limb-Threatening Ischemia

ObjectiveTo examine the influence of liver function on patients with chronic limb-threatening ischemia (CLTI), we classified patients with CLTI after revascularization according to their modified ALBI grades. MethodsWe retrospectively analyzed single-center data of patients who underwent revascularization for CLTI between 2015 and 2020. Patients were classified with modified albumin–bilirubin (ALBI) grades 1, 2a, and 2b & 3 according to the ALBI score, which was calculated based on serum albumin and total bilirubin levels. The endpoints were the two-year amputation-free survival (AFS) and one-year wound healing rates. ResultsWe included 190 limbs in 148 patients, and 50, 54, and 86 cases were assigned as grade 1, 2a, and 2b & 3, respectively. The two-year AFS rates for the grade 1, 2a, and 2b & 3 groups were 79 ± 6%, 66% ± 7%, and 45 ± 6%, respectively (P < 0.01). One-year cumulative wound healing rates for grade 1, 2a, and 2b & 3 groups were 68 ± 7%, 69% ± 6%, and 48% ± 5%, respectively (P = 0.01). Multivariate Cox proportional hazard analyses identified age (≥75 years), dependent ambulatory status, and modified ALBI grades 2b & 3 compared with grades 1 and 2a as significant independent predictors of AFS. The dependent ambulatory status and WIfI stage 4 were significant negative predictors of wound healing. ConclusionsMany patients with CLTI had high modified ALBI grades and impaired liver function classified as modified ALBI grade 2b or 3 is a robust negative predictor of amputation-free survival.

Saps1–3 Antigens in Candida albicans: Differential Modulation Following Exposure to Soluble Proteins, Mammalian Cells, and Infection in Mice

The secreted aspartic peptidases (Saps) of Candida albicans play crucial roles in various steps of fungal–host interactions. Using a flow cytometry approach, this study investigated the expression of Saps1–3 antigens after (i) incubation with soluble proteins, (ii) interaction with mammalian cells, and (iii) infection in immunosuppressed BALB/c mice. Supplementation strategies involving increasing concentrations of bovine serum albumin (BSA) added to yeast carbon base (YCB) medium as the sole nitrogenous source revealed a positive and significant correlation between BSA concentration and both the growth rate and the percentage of fluorescent cells (%FC) labeled with anti-Saps1–3 antibodies. Supplementing the YCB medium with various soluble proteins significantly modulated the expression of Saps1–3 antigens in C. albicans. Specifically, immunoglobulin G, gelatin, and total bovine/human sera significantly reduced the %FC, while laminin, human serum albumin, fibrinogen, hemoglobin, and mucin considerably increased the %FC compared to BSA. Furthermore, co-cultivating C. albicans yeasts with either live epithelial or macrophage cells induced the expression of Saps1–3 antigens in 78% (mean fluorescence intensity [MFI] = 152.1) and 82.7% (MFI = 178.2) of the yeast cells, respectively, compared to BSA, which resulted in 29.3% fluorescent cells (MFI = 50.9). Lastly, the yeasts recovered from the kidneys of infected immunosuppressed mice demonstrated a 4.8-fold increase in the production of Saps1–3 antigens (MFI = 246.6) compared to BSA, with 95.5% of yeasts labeled with anti-Saps1–3 antibodies. Altogether, these results demonstrated the positive modulation of Saps’ expression in C. albicans by various key host proteinaceous components, as well as by in vitro and in vivo host challenges.

Association of Bovine Respiratory Disease during the Pre-Weaning Period with Blood Cell Counts and Circulating Concentration of Metabolites, Minerals, and Acute Phase Proteins in Dairy Calves Transported to a Calf Raising Facility.

Our objective was to investigate the association of bovine respiratory disease (BRD) occurring within the first 56 days of life with blood cell counts and the circulating concentration of metabolites, minerals, and acute phase proteins throughout the pre-weaning period in dairy calves transported to a heifer raising facility within their first week of life. Data from 305 calves transported from dairies in Minnesota to a calf raising facility in New Mexico within their first four days of life were used in this retrospective cohort study. Blood samples were collected at 7, 17, 34, and 56 days of life for the analysis of blood cell counts, biochemistry, and the concentration of acute phase proteins. Blood urea nitrogen, albumin, GLDH, CK, P, Na, K, Cl, Zn, Hp, SAA, and monocyte counts were associated with BRD status throughout or at least at one of the time points evaluated in this study. In conclusion, several hematological variables were associated with BRD status in dairy calves that underwent transportation stress in early life.

Isothiocyanate-Corticosteroid Conjugates against asthma: Unity makes strength

Asthma is a major noncommunicable disease, affecting both children and adults, and represents one of the major causes leading to high health care costs due to the need for chronic pharmacological treatments. The standard gold therapy of inflammation in asthmatic patients involves the use of glucocorticoids even if their chronic use is often related to serious adverse effects. Growing evidence suggests the biological relevance of hydrogen sulfide (H2S) in the pathogenesis of airway diseases. Hence, aiming to associate the beneficial effects of steroidal anti-inflammatory drugs (SAIDs) to H2S biological activity, we designed and synthesized novel multi-target molecules by chemically combining a group of glucocorticoids, usually employed in asthma treatment, with an isothiocyanate moiety, well-known for its H2S releasing properties. Firstly, the synthesized compounds have been screened for their H2S-releasing profile using an amperometric approach and for their in vitro effects on the degranulation process, using RBL-2H3 cell line. The physicochemical profile, in terms of solubility, chemical and enzymatic stability of the newly hybrid molecules, has been assessed at different physiological pH values and in esterase-rich medium (bovine serum albumin, BSA). The selected compound 5c, through both its corticosteroid and H2S releasing component, has been evaluated in vivo in experimental model of asthma. The compound 5c inhibited in vivo all asthma features with a significative effect on the restoration of pulmonary structure and reduction of lung inflammation.

Crystal Structure, Theoretical Analysis, and Protein/DNA Binding Activity of Iron(III) Complex Containing Differently Protonated Pyridoxal-S-Methyl-Isothiosemicarbazone Ligands.

Pyridoxal-S-methyl-isothiosemicarbazone (PLITSC) is a member of an important group of ligands characterized by different complexation modes to various transition metals. In this contribution, a new complex containing two differently protonated PLITSC ligands ([Fe(PLITSC-H)(PLITSC)]SO4)∙2.5H2O was obtained. The crystal structure was solved by the X-ray analysis and used further for the optimization at B3LYP/6-311++G(d,p)(H,C,N,O,S)/def2-TZVP(Fe) level of theory. Changes in the interaction strength and bond distance due to protonation were observed upon examination by the Quantum Theory of Atoms in Molecules. The protein binding affinity of [Fe(PLITSC-H)(PLITSC)]SO4 towards transport proteins (Bovine Serum Albumin (BSA) and Human Serum Albumin (HSA)) was investigated by the spectrofluorimetric titration and molecular docking. The interactions with the active pocket containing fluorescent amino acids were examined in detail, which explained the fluorescence quenching. The interactions between complex and DNA were followed by the ethidium-bromide displacement titration and molecular docking. The binding along the minor groove was the dominant process involving complex in the proximity of DNA.

Molecular dynamics simulations of betaine-based deep eutectic solvents with varying iso-alcohols chain lengths

Betaine-based (BET) deep eutectic solvents (DESs) made with isoalcohols of varying hydrocarbon chain lengths, namely triethylene glycol (TEG), 1, 2-ethanediol (ETD), 1, 3-propanediol (PPD) and 1, 4-butanediol (BTD) were modelled. Their effects on the formation of aqueous biphasic system (ABS) with 2 M dipotassium phosphate salt solution (K2HPO4) as well as the extraction of common protein molecules, bovine serum albumin (BSA), were studied using molecular dynamics simulation with Gromacs. Qualitative and quantitative data were used to analyse the simulation results, which includes visualised systems in equilibrium, root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration, radial distribution function (RDF) and hydrogen bond analysis. The results and literature research validated the role of inorganic salt solute concentration on the phase forming ability as well as the possibility of protein denaturation. More investigations and experimental works are to be carried out regarding the type of inorganic salt and its concentration for aqueous biphasic systems (ABSs) with alcohol based DESs and salt solution for protein extraction purposes. This is essential to understand the salting-out effect and to isolate the effects of hydrocarbon chain length from other influencing factors. Additionally, inorganic salt concentrations, presence of ether linkage as well as hydrocarbon chain length is shown to play a role in the dynamics between molecules in the system as well as protein conformational changes. Overall, ABS incorporating PPD and BTD with hydrocarbon chain lengths not exceeding C4 demonstrated superior performance in terms of protein conformational stability and interaction, showcasing their efficacy in protein extraction process.

Biomimetic gold nanocages incorporating copper-human serum albumin for tumor immunotherapy via cuproptosis-lactate regulation

Cancer immunotherapy remains a significant challenge due to insufficient proliferation of immune cells and the sturdy immunosuppressive tumor microenvironment. Herein, we proposed the hypothesis of cuproptosis-lactate regulation to provoke cuproptosis and enhance anti-tumor immunity. For this purpose, copper-human serum albumin nanocomplex loaded gold nanocages with bacterial membrane coating (BAu-CuNCs) were developed. The targeted delivery and disassembly of BAu-CuNCs in tumor cells initiated a cascade of reactions. Under near infrared (NIR) laser irradiation, the release of copper-human serum albumin (Cu-HSA) was enhanced that reacted with intratumoral glutathione (GSH) via a disulfide exchange reaction to liberate Cu2+ ions and exert cuproptosis. Subsequently, the cuproptosis effect triggered immunogenic cell death (ICD) in tumor by the release of damage associated molecular patterns (DAMPs) to realize anti-tumor immunity via robust production of cytotoxic T cells (CD8+) and helper T cells (CD4+). Meanwhile, under NIR irradiation, gold nanocages (AuNCs) promoted excessive reactive oxygen species (ROS) generation that played a primary role in inhibiting glycolysis, reducing the lactate and ATP level. The combine action of lower lactate level, ATP reduction and GSH depletion further sensitized the tumor cells to cuproptosis. Also, the lower lactate production led to the significant blockage of immunosuppressive T regulatory cells (Tregs) and boosted the anti-tumor immunity. Additionally, the effective inhibition of breast cancer metastasis to the lungs enhanced the anti-tumor therapeutic impact of BAu-CuNCs + NIR treatment. Hence, BAu-CuNCs + NIR concurrently induced cuproptosis, ICD and hindered lactate production, leading to the inhibition of tumor growth, remodeling of the immunosuppressive tumor microenvironment and suppression of lung metastasis. Therefore, leveraging cuproptosis-lactate regulation, this approach presents a novel strategy for enhanced tumor immunotherapy.

Transport of Zinc-Phthalocyanine to Cancer Cells using Myoglobin-Albumin Fusion Protein for Photodynamic Therapy.

Photodynamic therapy (PDT) is a noninvasive approach to cancer treatment, wherein cell death is initiated by singlet oxygen (1O2) production via energy transfer from excited photosensitizers to ground-state O2. Effective clinical photosensitizers necessitate water solubility for in vivo administration. Hydrophobic dyes, such as phthalocyanines, cannot be used directly as photosensitizers. Herein, we synthesized a myoglobin-(human serum albumin) fusion protein reconstituted with zinc-phthalocyanine (ZnPc), termed ZnPcMb-HSA. The photophysical properties of ZnPcMb-HSA closely resemble those of ZnPc-substituted Mb. Notably, ZnPc dissociates from ZnPcMb-HSA and selectively accumulates within cancer cells, while the protein components remain extracellular. Treatment of four distinct cell lines with ZnPcMb-HSA, followed by red-light irradiation, effectively induced apoptosis. The half-maximal inhibitory concentrations (IC50) against these cancer cell lines ranged between 0.1-0.5 μM. Reconstituted Mb-HSA emerges as a promising carrier for transporting various water-insoluble porphyrinoid photosensitizer to target cancer cells in PDT applications.

Posteruptive Loss of Enamel Proteins Concurs with Gain in Enamel Hardness.

Tooth enamel maturation requires the removal of proteins from the mineralizing enamel matrix to allow for crystallite growth until full hardness is reached to meet the mechanical needs of mastication. While this process takes up to several years in humans before the tooth erupts, it is greatly accelerated in in the faster developing pig. As a result, pig teeth erupt with softer, protein-rich enamel that is similar to hypomineralized human enamel but continues to harden quickly after eruption.Proteins, such as albumin, that bind to enamel crystals and prevent crystal growth and enamel hardening have been suggested as cause for hypomineralized human enamel that does not naturally harden after eruption. However, albumin is abundant in pig enamel. It is unclear whether fast posteruptive enamel hardening in pigs occurs despite the high protein content or requires a facilitated protein loss to allow for crystal growth. This study asked how the protein content in porcine enamel changes after eruption in relation to saliva. Based on previous data demonstrating the high albumin content in erupted porcine enamel, we hypothesize that following pre-eruptive maturation, enamel and saliva derived enzymes facilitate protein removal from porcine enamel after eruption. We analyzed enamel and the saliva proteome at three critical timepoints: at the time of tooth eruption, 2 weeks after eruption, and enamel 6 weeks after eruption. We used only fourth deciduous premolars and saliva samples from animals sacrificed at the respective time points to determine the organic content in tooth enamel, saliva, and saliva proteins within enamel. We found a decrease in the number of proteins and their abundancy in enamel with posteruptive time, including a decrease in serum albumin within enamel. The rapid decrease in the first two weeks is in line with previously reported rapid increase in mineral density of porcine enamel after eruption. In addition to the enamel proteases KLK-4 and MMP-20, we identified serine-, cysteine-, aspartic-, and metalloproteases. Some of these were only identified in enamel, while almost half of the enzymes are in common with saliva at all timepoints. Our findings suggest that the fast posteruptive enamel maturation in the porcine model coincides with saliva exchange and influx of saliva enzymes into porous enamel.

The potential ameliorative effects of Moringa oleifera in emamectin benzoate induced toxicity in freshwater Labeo rohita fish

Emamectin benzoate pesticide (EMB) is life-threatening chemical causing harmful effects on aquatic organisms specially fish species. Current study was aimed to understand the ameliorative role of Moringa oleifera against the toxic effects of emamectin benzoate on hematological, biochemical and changes in the soft tissues of Labeo rohita. Four groups were made. First group without chemicals is the control group, while 2nd, 3rd and 4th groups were exposed to 3.64µg/L, 7.28µg/L and 14.56µg/L EMB respectively for 30 days. Results revealed that the number of RBCs, HGB, HCT, MCV, MCH, MCHC, and WBCs were decreased while lymphocytes, monocytes, eosinophils and platelets were increased significantly. Biochemical parameters of fish including serum cholesterol, triglycerides, LDL, VLDL, albumin, globulin, blood glucose, ALT, AST, TSH, urea, creatinine, and blood urea nitrogen were significantly increased while HDL, total protein, T3 and T4 were decreased. Histological changes revealed that EMB causes degenerative effects in soft tissues of exposed fish. After using Moringa oleifera, all the effects were reversed and the fish was moved towards recovery. The overall results of the study showed that emamectin benzoate can alter the histological, hematological, and biochemical processes of Labeo rohita, thus toxic to aquatic organisms.

Effects of Dried khat (Catha edulis) Leaves as a Natural Feed Additive on the White Leghorn Layers Performance

Khat contains many bioactive compounds that are beneficial for chickens’ health. Most of the chemical constituents found in Khat are biologically active and are used worldwide for the treatment of many diseases. However, there are few studies on the use of Khat in poultry as a nutrition, antioxidant, and antimicrobial activity. Therefore, the present study evaluated the effects of dried Khat leave (DKL) as a natural feed additive on the White Leghorn layer's performance, serum chemistry, and hematology. A total of 180 White Leghorn chickens, aged 168 days were randomly divided into four groups consisting of 45 chickens in three replications. Thirteen layers and two cocks were assigned to each replication and reared on a deep litter system. The diets of layers in T1, T2, T3, and T4 were supplemented by DKL at proportion of 0%, 0.2%, 0.4%, and 0.6%, respectively. The layers were evaluated for feed intake, body weight gain, egg production, feed conversion ratio, and some blood parameters, such as total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, albumin, globulin, red blood cells, hemoglobin, packed cells volume, white blood cells, and total protein. Feed intake in T1 (103.5 g) was significantly higher than T4 (98.5 g) and T3 (98.9 g). Layers in T1 significantly gained higher body weight than layers in other treatments. The use of DKL in layer chickens’ diet did not affect egg production, egg mass, and feed conversion ratio. An increase in the level of DKL in the layers diet significantly reduced the serum high-density lipoprotein, cholesterol, and albumin. The present study showed that chickens that received a higher proportion of dried khat leaf (0.6%) significantly recorded lower levels of low-density lipoprotein and total cholesterol in their blood.

Bilateral Unsymmetrical Disulfurating Reagent Design for Polysulfide Construction.

Polysulfides are significant compounds in life science, pharmaceutical science, and material science. Therefore, polysulfide construction is in great demand. The controllable sequential installation on both ends of S-S motif faces an enormous challenge, due to the reversible covalent S-S bond. A library was established with two divergent mask groups for bilateral unsymmetrical disulfurating reagents (R1O-SS-SO2R2). Sequential coupling was successfully achieved with preferential activation of S-SO2 bond (37.6 kcal/mol) and controllable activation of S-O bond (54.8 kcal/mol) superior to S-S bond (62.0 kcal/mol), enabled untrammeled installation on the bilateral sides of S-S motif to afford unsymmetrical disulfides and trisulfides, even for natural products, pharmaceuticals, peptides, and protein (bovine serum albumin) cross-linkage successively.

A Comparative Study on the Interaction Between Protein and PET Micro/Nanoplastics: Structural and Surface Characteristics of Particles and Impacts on Lung Carcinoma Cells (A549) and Staphylococcus aureus.

The interaction between particles and proteins is a key factor determining the toxicity responses of particles. Therefore, this study aimed to examine the interaction between the emerging pollutant polyethylene terephthalate micro/nanoplastics from water bottles with bovine serum albumin. The physicochemical characteristics of micro/nanoplastics were investigated using nuclear magnetic resonance, x-ray diffraction, Fourier transform infrared, dynamic light scattering, and x-ray energy dispersive spectroscopy after exposure to various concentrations and durations of protein. Furthermore, the impact of protein-treated micro/nanoplastics on biological activities was examined using the mitochondrial activity and membrane integrity of A549 cells and the activity and biofilm production of Staphylococcus aureus. The structural characteristics of micro/nanoplastics revealed an interaction with protein. For instance, the assignment of protein-related new proton signals (e.g., CH2, methylene protons of CH2O), changes in available protons s (e.g., CH and CH3), crystallinity, functional groups, elemental ratios, zeta potentials (-11.3 ± 1.3 to -12.4 ± 1.7 to 25.5 ± 2.3 mV), and particle size (395 ± 76 to 496 ± 60 to 866 ± 82 nm) of micro/nanoplastics were significantly observed after protein treatment. In addition, the loading (0.012-0.027 mM) and releasing (0.008-0.013 mM) of protein also showed similar responses with structural characteristics. Moreover, the cell-based responses were changed regarding the structural and surface characteristics of micro/nanoplastics and the loading efficiencies of protein. For example, insignificant mitochondrial activity (2%-10%) and significant membrane integrity (12%-28%) of A549 cells increased compared with control, and reductions in bacterial activity (5%-40%) in many cases and biofilm production specifically at low dose of all treatment stages (13%-46% reduction) were observed.