Serum Albumin Research Articles

3D printed subcutaneous implant for prolonged delivery of tenofovir with desired release capability, biocompatibility, and viability

This study introduced a progressive approach to address the challenges associated with drug delivery in hepatitis B by developing a 3D printed subcutaneous implant. The implant, composed of a polymer-reinforced bovine serum albumin hydrogel crosslinked with glutaraldehyde, exhibited shear thinning and thixotropic behavior. This allows it to undergo transformation into an implant using a semisolid extrusion-based 3D printing technique. Thorough analysis encompassing physical, thermal, and FTIR assessments ensured thermal and chemical stability of the implant. Scanning electron microscopy revealed the highly porous microstructures of the hydrogel implant, enhancing its suitability for drug encapsulation. Swelling study showed only 7.88 % swelling of the implant after 28 days, suggesting restricted water movement into the implant matrix, leading to a gradual release of the encapsulated drug. The biodegradable nature of the implant was confirmed using an enzyme degradation study. Notably, the hydrogel implant exhibited sustained release of tenofovir of approximately 63 % over a 28-day period, presenting a promising avenue for prolonged therapeutic interventions in both hepatitis B. Furthermore, the crosslinked hydrogel was found to be cytocompatible in human kidney and dermal fibroblast cells. Overall, this work signifies a notable advancement in implantable drug delivery systems utilizing both hydrogel and the emerging 3D printing technique, offering a potential solution for the sustained and localized treatment of chronic viral infections.

Validation and stability analysis of guanine deaminase assay kit

Guanine deaminase (GD)plays important roles in the diagnosis of liver function. However, there is no totally rapid and simple for the eatimation of GD activity in clinical application. Herein, we have constructed an enzymatic assay system with highly sensitive and strong stability for quantification of GD activity by highly double enzyme-coupling (xanthine oxidase and uric acid oxidase) and adding compound stabilizer in GD kit. In this study, we validated parameters, including reagent blank, sensitivity, accuracy, inter-batch difference, intra-batch difference, linear range. Furthermore, composite stabilizers, containing gentamicin sulfate, bovine serum albumin, and mannitol, were selected to improve stability of GD kit during long-term storage. The experimental results showed that the absorbance of the reagent blank was <0.2, the mean recovery rate was 103 %, the inter-batch and intra-batch diffeerence were <15 %, The linearity range was 0 U/L-50 U/L (R2 > 0.99). All indicators met the kit requirements for clinical applications. When gentamicin sulfate, bovine serum albumin, and mannitol were used as a stabilizer, the kit remained stable for 12 months without significant loss of enzymatic activity. These results indicated that GD kit possesses high sensitivity and strong stability, which can be used for routine biochemical applications and is of great significance for the diagnosis and differential diagnosis of liver diseases.

Tailoring the specificity of ficin versus large hemoglobin and small casein by co-immobilizing inert proteins on the immobilized enzyme layer and further modification with aldehyde dextran

Ficin has been immobilized at full loading on glyoxyl agarose beads. Then, ficin was blocked with 2,2′-dipyridyldisulfide. To be effective, the modification must be performed in the presence of 0.5 M urea, as the enzyme was not inhibited under standard conditions, very likely because the catalytic Cys was not fully exposed to the medium. Activity could be fully recovered by incubation with 1 M mercaptoethanol. This biocatalyst could hydrolyze hemoglobin and casein. The objective of this paper was to increase the enzyme specificity versus small proteins by generating steric hindrances to the access of large proteins. The step by step blocking via ionic exchange of the biocatalyst with aminated bovine serum albumin (BSA), aldehyde dextran and a second layer of aminated BSA produced a biocatalyst that maintained its activity versus small synthetic substrates, increased the biocatalyst stability, while reduced its activity to over 50 % versus casein. Interestingly, this treatment almost fully annulled the activity versus hemoglobin, more effectively at 37 °C than at 55 °C. The biocatalyst could be reused 5 times without changes in activity. The changes could be caused by steric hindrances, but it cannot be discarded some changes in enzyme sequence specificity caused by the modifications.

CLN-617 Retains IL2 and IL12 in Injected Tumors to Drive Robust and Systemic Immune-Mediated Antitumor Activity.

Despite clinical evidence of antitumor activity, the development of cytokine therapies has been hampered by a narrow therapeutic window and limited response rates. Two cytokines of high interest for clinical development are interleukin 2 (IL2) and interleukin 12 (IL12), which potently synergize to promote the activation and proliferation of T cells and NK cells. However, the only approved human IL2 therapy, Proleukin, is rarely used in the clinic due to systemic toxicities, and no IL12 product has been approved to date due to severe dose-limiting toxicities. Here, we describe CLN-617, a first-in-class therapeutic for intratumoral (IT) injection that co-delivers IL2 and IL12 on a single molecule in a safe and effective manner. CLN-617 is a single-chain fusion protein comprised of IL2, leukocyte-associated immunoglobulin-like receptor 2 (LAIR2), human serum albumin (HSA), and IL12. LAIR2 and HSA function to retain CLN-617 in the treated tumor by binding collagen and increasing molecular weight, respectively. We found that IT administration of a murine surrogate of CLN-617, mCLN-617, eradicated established treated and untreated tumors in syngeneic models, significantly improved response to anti-PD1 checkpoint therapy, and generated a robust abscopal response dependent on cellular immunity and antigen cross-presentation. CLN-617 is being evaluated in a clinical trial in patients with advanced solid tumors (NCT06035744).

Encapsulation of 4-oxo-N-(4-hydroxyphenyl) retinamide in human serum albumin nanoparticles promotes EZH2 degradation in preclinical neuroblastoma models.

Neuroblastoma is the most prevalent and aggressive solid tumor that develops extracranially in children between the ages of 0-14 years, which accounts for 8-10% of all childhood malignancies and ∼15% of pediatric cancer-related mortality. The polycomb repressive complex 2 (PRC2) protein, EZH2, is overexpressed in neuroblastoma and mediates histone H3 methylation at lysine 27 (K27) positions through its methyl transferase activity and is a potential epigenetic silencer of many tumor suppressor genes in cancer. Phosphorylation of EZH2 decreases its stability and leads to proteasomal degradation. The 4-oxo-N-(4-hydroxyphenyl) retinamide (4O4HPR) promotes EZH2 degradation via activation of PKC-δ, but its limited solubility and physiological instability limit its application. In the current study, the encapsulation of 4O4HPR in Human Serum Albumin Nanoparticles (HSANPs) enhanced the solubility and physiological stability of the nanoformulation, leading to improved therapeutic efficacy through G2-M cell cycle arrest, depolarization of mitochondrial membrane potential, generation of reactive oxygen species and caspase 3 mediated apoptosis activation. The molecular mechanistic approach of 4O4HPR loaded HSANPs has activated caspase 3, which further cleaves PKC-δ into two fragments wherein the cleaved fragment of PKC-δ possesses the kinase activity that phosphorylates EZH2 and decreases the protein stability leading to its further ubiquitination in SH-SY5Y cells. Co-immunoprecipitation experiments revealed the direct interaction between PKC-δ and EZH2 phosphorylation, followed by ubiquitination. Moreover, 4O4HPR loaded HSANPs demonstrated improved in vivo biodistribution, greater dispersibility, and biocompatibility and exhibited enhanced protein instability and degradation of EZH2 in the neuroblastoma xenograft mouse model.

Integration of highly sensitive large-area graphene-based biosensors in an automated sensing platform

Graphene-based biosensors, featuring exceptional electronic, mechanical, and surface properties, have emerged as frontrunners in advanced sensing technologies. However, to achieve widespread industrial adoption, advancements in the fabrication and integration of large-area graphene devices are essential. Critical parameters such as enhanced sensitivity, scalable production methods, economic viability, integration capabilities, and consistent uniformity must be meticulously addressed. In this work, we demonstrate that our ultra-clean, chemical wet transfer protocol of large-area graphene enables a scalable, smooth integration of graphene into an established assay platform for transporter protein drug discovery. Furthermore, we demonstrate sensitive detection of electrolytic buffers, varying pH, bovine serum albumin (BSA) and single-stranded DNA (ssDNA) adsorption, using our large-area graphene solution-gated field-effect transistor (SGFET) sensors, thereby proving their robust and reliable performance. The sensors’ biocompatibility and ion sensitivity, down to the picomolar range, substantiate their suitability for the investigation of electroactive transport in ion channels and membrane transporters.

Reconstruction for extensive sacrococcygeal defects in complex tumor patients with personalized customized gluteus maximus myocutaneous flaps

AimTo evaluate the clinical effects of personalized customized gluteus maximus myocutaneous flaps (GMMF) for reconstruction of extensive sacrococcygeal soft tissue defects in complex tumor patients. MethodsA retrospective chart review was conducted on 8 patients who underwent personalized customized GMMF reconstruction for large sacrococcygeal defect from December 2021 to August 2023. The personalized customized GMMF were designed based on the variations of tissue defect in location, shape and volume of different dead spaces. The principle of the personalized GMMF is to ensure that the rotation point of the flap can reach the farthest end of the defect. Patient demographics, operative characteristics, and perioperative risk factors were analyzed. Clinical outcomes were assessed, focusing on complications such as flap necrosis, wound dehiscence, infection, seroma, and hematoma. ResultsSix patients with rectal cancer and two with sacral tumors underwent personalized customized GMMF reconstruction for extensive sacrococcygeal defects. The average volume of the wound cavity was 104 mL, with a mean vertical depth was 10.8 cm. Six patients had low serum albumin (<35 g/L). After a mean follow-up of 15.5 months, no major complications occurred, except for one seroma that resolved within 2 weeks. ConclusionThe personalized customized GMMF described in this study is an effective method for reconstructing large sacrococcygeal wounds with significant depth in complex tumor patients. It allows for greater rotation of the muscle flap into the sacrococcygeal wound defect and provides adequate blood supply by utilizing the bulk of muscle tissue to obliterate dead space.

High prevalence of short telomeres in idiopathic porto-sinusoidal vascular disorder.

Telomeres prevent damage to coding DNA as end-nucleotides are lost during mitosis. Mutations in telomere maintenance genes cause excessive telomere shortening, a condition known as short telomere syndrome (STS). One hepatic manifestation documented in STS is porto-sinusoidal vascular disorder (PSVD). As the etiology of many cases of PSVD remains unknown, this study explored the extent to which short telomeres are present in patients with idiopathic PSVD. This monocentric cross-sectional study included patients with histologically defined idiopathic PSVD. Telomere length in 6 peripheral blood leukocyte subpopulations was assessed using fluorescent in situ hybridization and flow cytometry. Variants of telomere-related genes were identified using high-throughput exome sequencing. In total, 22 patients were included, of whom 16 (73%) had short (9/22) or very short (7/22) telomeres according to age-adjusted reference ranges. Fourteen patients (64%) had clinically significant portal hypertension. Shorter telomeres were more frequent in males (p = 0.005) and patients with concomitant interstitial lung disease (p < 0.001), chronic kidney disease (p < 0.001), and erythrocyte macrocytosis (p = 0.007). Portal hypertension (p = 0.021), low serum albumin level (p < 0.001), low platelet count (p = 0.007), and hyperbilirubinemia (p = 0.053) were also associated with shorter telomeres. Variants in known STS-related genes were identified in 4 patients with VSTel and 1 with STel. Short and very short telomeres were highly prevalent in patients with idiopathic PSVD, with 31% presenting with variants in telomere-related genes. Telomere biology may play an important role in vascular liver disease development. Clinicians should consider measuring telomeres in any patient presenting with PSVD.

Impaired Cholesterol Efflux Capacity rather than Low HDL-C Reflects Oxidative Stress under Acute Myocardial Infarction.

Acute myocardial infarction (AMI) causes irreversible damage to cardiomyocytes due to the discontinuation of oxygen supply and leads to systemic oxidative stress. It has been reported that high-density lipoprotein (HDL) particles have antioxidant capacity, and reduced antioxidant capacity is associated with decreased cholesterol efflux capacity (CEC). The purpose of this study was to clarify the usefulness of CEC measurement in patients with AMI. We investigated the association between CEC and oxidative stress status in a case-control study. This study included 193 AMI cases and 445 age- and sex-matched controls. We examined the associations of CEC with HDL-cholesterol (HDL-C) and oxidized human serum albumin (HSA), an index of systemic oxidative stress status, and the effect of aldehyde dehydrogenase 2 (ALDH2) rs671 polymorphism, which has been reported to affect HDL-C level and risk for MI, on these associations. Both bivariable and multivariable analyses showed that CEC was positively correlated with HDL-C levels in both AMI cases and controls, with a weaker correlation in AMI cases than in controls. In AMI cases, oxidized HSA levels were associated with CEC in both bivariable and multivariable analyses, but not with HDL-C. These associations did not differ among the ALDH2 genotypes. CEC, but not HDL-C level, reflects systemic oxidative stress status in patients with AMI. CEC measurement for patients with AMI may be useful in that it provides information on systemic oxidative stress status as well as atherosclerosis risk.

Structure-affinity-pharmacokinetics relationships of 111In-labeled PSMA-targeted ligands with different albumin binders

IntroductionProstate-specific membrane antigen (PSMA) is a promising target for treating metastatic castration-resistant prostate cancer. Our previous report presented 111In- or 225Ac-labeled PSMA-NAT-DA1 (PNT-DA1) as a PSMA-targeted ligand. To improve its therapeutic efficiency, PNT-DA1 contains 4-(p-iodophenyl)butyric acid (IPBA), which is known as an albumin binder (ALB) moiety. However, few reports have examined the relationship between the chemical modification of the ALB moiety and pharmacokinetics of PSMA-targeted radioligands. To assess this relationship, we designed, synthesized, and evaluated four [111In]In-PNT-DA1 analogues with ALB moieties different from IPBA. MethodsThe [111In]In-PNT-DA1 analogues were synthesized from their corresponding precursors through ligand substitution reaction. The stability of [111In]In-PNT-DA1 analogues in mouse plasma, their affinity for human serum albumin (HSA), their binding to mouse plasma proteins, and their affinity for PSMA were evaluated in vitro. The tissue distribution profile of the radioligands was assessed in biodistribution studies using LNCaP tumor-bearing nude mice. ResultsAll [111In]In-PNT-DA1 analogues were obtained at a high radiochemical yield and purity. These analogues were highly stable in mouse plasma after 24 h. The binding affinity to HSA significantly varied among the different ALB moieties. Moreover, high affinity for mouse plasma proteins was observed for all [111In]In-PNT-DA1 analogues compared with their counterparts without an ALB moiety. The affinity for PSMA was comparable for all radioligands. In the biodistribution assay, the pharmacokinetics of [111In]In-PNT-DA1 analogues varied markedly depending on the type of ALB moiety. In particular, tumor area under the curve (AUC) values were increased for radioligands with higher blood retention, while some previous studies reported that compounds with moderate blood retention exhibited the highest tumor AUC values. ConclusionThe introduction of an appropriate ALB moiety into the ligand may lead to the development of more useful PSMA-targeted radioligands with higher tumor accumulation.

Association between serum vitamin D levels and skeletal muscle indices in an older Japanese population: The SONIC study.

Vitamin D (VD) affects skeletal muscles. The high prevalence of VD deficiency in Japan may lead to decreased skeletal muscle mass and strength, increasing the prevalence of sarcopenia. Therefore, we aimed to investigate the association between serum VD levels and skeletal muscle indices in a Japanese community-dwelling older population. We extracted data from the Septuagenarians, Octogenarians, Nonagenarians Investigation with Centenarians (SONIC) study. We analyzed the data for participants in the 70s and 90s age groups. Skeletal mass index (SMI) using bioimpedance analysis, grip strength, walking speed, and serum VD levels using 25-hydroxyvitamin D [25(OH)D] were measured. We analyzed the data of 310 participants in their 70s and 48 in their 90s. Mean serum 25(OH)D levels were 21.6 ± 5.0 ng/mL in the 70s group and 23.4 ± 9.1 ng/mL in the 90s group. In the 70s group, serum 25(OH)D levels correlated with SMI (r = 0.21, P < 0.0001) and grip strength (r = 0.30, P < 0.0001). Serum 25(OH)D levels were independently associated with SMI after adjusting for sex, body mass index, and serum albumin levels. In the 90s group, serum 25(OH)D levels were correlated with SMI (r = 0.29, P = 0.049) and grip strength (r = 0.34, P = 0.018). However, the multivariate analysis showed no independent association between SMI, grip strength, and serum 25(OH)D levels. In a cross-sectional analysis of an older population, serum VD levels were associated with SMI and grip strength, and this association was more pronounced in the 70s group than in the 90s group. Our results suggest that serum VD levels maintain skeletal muscle mass and grip strength. Geriatr Gerontol Int 2024; ••: ••-••.

Dysregulated serum concentrations of fat-soluble vitamins in dogs with chronic enteropathy.

In inflammatory bowel disease (IBD) of humans, nutrient malabsorption can result in fat-soluble vitamin deficiency, especially of vitamin D. In veterinary species, decreased concentrations of vitamin D are relatively common in dogs with chronic enteropathy (CE), but data on the status of other fat-soluble vitamins (FSVs) is lacking. Determine the serum concentrations of retinol, vitamin D, and α-tocopherol in dogs with CE compared with healthy dogs and compare clinical, clinicopathologic variables between CE and healthy dogs to detect associations with decreased FSVs concentrations. Eighteen client-owned dogs with CE and 33 healthy dogs. Serum 25-hydroxyvitamin D (25[OH]D), serum retinol and α-tocopherol concentrations were compared between groups. Correlations and multiple regression modeling were used to examine the relationship between serum 25(OH)D, retinol, and α-tocopherol concentrations and clinical and clinicopathological variables. Dogs with low serum albumin concentrations were more likely to have lower 25(OH)D concentrations than dogs with normal serum albumin concentration. Dogs with CE had higher serum concentrations of retinol, and variable α-tocopherol concentrations. The cause of these dysregulated vitamin concentrations is unclear and requires further study. Dogs with severe forms of CE should be monitored for decreased concentrations of 25(OH)D. Additional studies are needed to evaluate the clinical relevance and the possible benefit of vitamin D supplementation in these patients.

Dual synergies of functionalized hydrophilic MOFs based poly (aryl ether ketone sulfone) ultrafiltration membranes: Electrostatic action and pore size screening

In this paper, amino monomer (4Am-PH) and the sulfonated poly (arylene ether ketone sulfone) involving amino groups (Am-SPAEKS) were successfully prepared by diazotization reaction and direct polycondensation, respectively. The polyethyleneimine-modified UiO-66-NH2 (P-MOFs) were prepared as fillers. Composite membranes were fabricated by incorporating different contents of P-MOFs into Am-SPAEKS polymer matrix using non-solvent-induced phase separation. The morphology, chemical structure and properties of the polymers, fillers and composite membranes were proved by FT-IR, 1H NMR, FE-SEM, XPS and etc. The final experimental results showed that the introduction of hydrophilic fillers, P-MOFs, substantially improved the hydrophilicity, water flux and antifouling capabilities of the membranes. The water contact angle decreased from 78.5° to 56.4°. The flux of water from a pure membrane was 333.68 L/m2 h. When the content of P-MOFs was 0.1 %, the water flux could attain to 830.23 L/m2 h, the retention rate of bovine serum albumin (BSA) solution could attain to 98.6 % and the flux recovery rate (FRR) values of the composite membrane remained high (86.1 %) for BSA solution. In addition, after three cycles, the pure water flux of the composite membrane could still attain 595.49 L/m2 h. The results demonstrated that P-MOFs have significant potential for utilization in the preparation of composite ultrafiltration membranes and provided an effective idea for the modification and design of other hydrophilic MOFs applied in the study area of ultrafiltration membranes.

Fe3O4@silica-thermolysin: A robust, advantageous, and reusable microbial nanobiocatalyst for proteolysis and milk-clotting

Thermolysin (TLN) is a microbial highly-priced thermostable metallo-endoprotease with complementary substrate specificity to those of proteases widely used in science and industry for protein digestion and milk-clotting. This study is the first to immobilize TLN on aminated superparamagnetic nanoparticles (Fe3O4@silica-NH2) aiming for higher stability, recoverability, reusability, and applicability in proteolysis and as a microbial rennet-like milk-clotting enzyme. The nanobiocatalyst developed (Fe3O4@silica-TLN) displays hydrolytic activity on a synthetic TLN substrate and, apparently, was fully recovered from reaction media by magnetic decantation. More importantly, Fe3O4@silica-TLN retains TLN catalytic properties in the presence of calcium ions even after exposure to 60 °C for 48 h, storage at 4 °C for 80 days and room temperature for 42 days, use in proteolyses, and in milk-clotting for up to 11 cycles. Its proteolytic activity on bovine milk casein in 24 h furnished 84 peptides, of which 29 are potentially bioactive. Also, Fe3O4@silica-TLN catalyzed the digestion of bovine serum albumin. In conclusion, Fe3O4@silica-TLN showed to be a new, less autolytic, thermostable, non-toxic, magnetically-separable, and reusable nanobiocatalyst with highly attractive properties for both science (peptide/protein chemistry and structure, proteomic studies, and the search for new bioactive peptides) and food industry (cheese manufacture).

Highly miscible bio-based poly(lactic acid)/acetylated cellulose ether ultrafiltration membranes with improved water permeances

Biopolymer-based membranes are eco-friendly alternatives to petroleum-based membranes. However, biopolymers often require harsh membrane fabrication conditions due to their low solubilities, which are incompatible with general non-solvent-induced phase separation (NIPS). Thus, establishing mild blending conditions for NIPS membrane production is needed. This study developed ultrafiltration membranes by blending poly (lactic acid) (PLA) and acetylated cellulose ether (ACE), both of which are derived from renewable resources, using NIPS under mild conditions. We investigated the effects of PLA proportion on the structural, chemical, and thermal properties of the blend membranes and assessed polymer miscibility using theoretical models. Results indicated excellent miscibility with a single glass transition temperature. The 1:1 PL A/ACE membrane demonstrated the lowest water contact angle of 45°, 35 % lower than that of the pure PLA membrane. Additionally, its pure water permeance was 179.4 LMH/bar, significantly higher than 103.4 LMH/bar of the pure PLA membrane. Flux recovery ratio at pH = 4 was 69.5 for the blend membrane as compared to 70.6 for the PLA membrane. At pH = 7, all membranes could be fully recovered via physical washing, even those fouled with bovine serum albumin. This study successfully fabricates PLA/ACE-based membranes with outstanding properties under mild conditions.

Prediction model for treatment response of primary membranous nephropathy with nephrotic syndrome.

To investigate the predictors and establish a nomogram model for the prediction of the response to treatment in primary membranous nephropathy (PMN) with nephrotic syndrome (NS). The clinical, laboratory, pathological and follow-up data ofpatients withbiopsy-proven membranous nephropathy at the Affiliated Hospital of QingdaoUniversity were collected. A total of 373 patients were randomly assigned into development group (n = 262) and validation group (n = 111). Logistic regression analysis was performed in the development group to determine the predictors of treatment response. A nomogram model was established based on the multivariate logistic regression analysis and validated in the validation group. The C-index and calibration plots were used for the evaluation of the discrimination and calibration performance, respectively. Serum albumin levels (OR = 1.151, 95% CI 1.078-1.229, P < 0.001) and glomerular C3 deposition (OR = 0.407, 95% CI 0.213-0.775, P = 0.004) were identified as independent predictive factors for treatment response in PMN with NS, then a nomogram was established combining the above indicators and treatment regimen. The C-indices of this model were 0.718 (95% CI 0.654-0.782) and 0.789 (95% CI 0.705-0.873) in the development and validation groups, respectively. The calibration plots showed that the predicted probabilities of the model were consistent with the actual probabilities (P > 0.05), which indicated favorable performance of this model in predicting the treatment response probability. Serum albumin levels and glomerular C3 deposition were predictors for treatment response of PMN with NS. A novel nomogram model with good discrimination and calibration was constructed to predict treatment response probability at an early stage.

Preparation of an adsorptive membrane of polyvinylidene fluoride incorporated functionalized boron nitride nanosheets for arsenic removal

Incorporating nanomaterials into membranes will enhance wastewater treatment efficiency with their unique characteristics, such as higher permeability, thermal stability, surface roughness, hydrophilicity, and fouling control. In this study, the surface modified boron nitride with phosphoric acid 2-hydroxyethyl methacrylate ester (PA/BN) was grafted with polyethylene glycol (PEG) via conventional grafting. The PEG grafted PA/BN (PEG-g-PA/BN) melt blended with polyvinylidene fluoride (PVDF) resin by using an internal mixer at different mass percentages (100% PVDF (PVDF), 3% PEG-g-PA/BN + 97% PVDF (97:3 BN), 5% PEG-g-PA/BN + 95% PVDF (95:5 BN), 7% PEG-g-PA/BN + 93% PVDF (93:7 BN), and 7% PEG-g-PA/BNNS + 93% PVDF (93:7 BNNS). Phase inversion technique was used to cast the blended mixture into a thin membrane. The prepared membranes were analyzed with different characterization techniques to determine chemical composition, crystallinity, morphology, and thermal properties. The prepared composite membrane was evaluated in terms of water permeability, anti-fouling resistance, and solute rejection efficiency with deionized water, bovine serum albumin, and arsenic solution as well. PVDF membranes show high water flux and porosity. The water flux and porosity of the blends decrease as the percentage of PEG-g-PA/BN increases. However, the highest removal capacity for arsenic was observed at 93:7 BN. The adsorption of arsenic ions takes place via complexation with PA/BN in the PVDF matrix. This was confirmed with field emission scanning electron microscopy–energy-dispersive X-ray analysis and X-ray photoelectron spectroscopy analyses.

Strategies for improving expression of Recombinant Human Chorionic Gonadotropin in Chinese Hamster Ovary (CHO) cells.

Optimizations of the gene expression cassette combined with the selection of an appropriate signal peptide are important factors that must be considered to enhance heterologous protein expression in Chinese Hamster Ovary (CHO) cells. In this study, we investigated the effectiveness of different signal peptides on the production of recombinant human chorionic gonadotropin (r-hCG) in CHO-K1 cells. Four optimized expression constructs containing four promising signal peptides were stably transfected into CHO-K1 cells. The generated CHO-K1 stable pool was then evaluated for r-hCG protein production. Interestingly, human serum albumin and human interleukin-2 signal peptides exhibited relatively greater extracellular secretion of the r-hCG with an average yield of (16.59 ± 0.02 μg/ml) and (14.80 ± 0.13 μg/ml) respectively compared to the native and murine IgGκ light chain signal peptides. The stably transfected CHO pool was further used as the cell substrate to develop an optimized upstream process followed by a downstream phase of the r-hCG. Finally, the biological activity of the purified r-hCG was assessed using in vitro bioassays. The combined data highlight that the choice of signal peptide can be imperative to ensure an optimal secretion of a recombinant protein in CHO cells. In addition, the stable pool technology was a viable approach for the production of biologically active r-hCG at a research scale with acceptable bioprocess performances and consistent product quality.

Hazelnut proteins detection in cookies using an immersible label-free photonic chip sensor

Rapid and sensitive methods to detect allergenic proteins in foods at the point-of-need could help to protect allergic consumers from life-threatening accidental exposure. In this context, the development of a label-free optical immunosensor for detecting hazelnut proteins in cookies is presented. The sensor is based on silicon photonic chips containing two integrated Mach-Zehnder interferometers (MZI) with light input and output on one side of the chip, and sensing window openings on the other side. Coupling with a white-light LED and a spectrometer for signal recording is achieved via a bifurcated fiber. The sensing window of one of the MZIs is modified with a mouse monoclonal antibody against hazelnut proteins, while the other sensing window is modified with bovine serum albumin to serve as blank. The assay follows a two-step sandwich immunoassay format, involving a 10-min reaction with the hazelnut proteins calibrator or cookie sample, followed by a 2-min reaction with a mixture of monoclonal antibodies. All reactions were performed by immersing the chip side where the MZIs windows are located into the solutions. The recorded spectrum is processed in real-time to transform the spectrum shifts due to immunoreactions taking place on the sensing windows of the two MZIs into phase shifts. The developed assay detects hazelnut proteins at concentrations as low as 25 ng/mL in calibrators prepared in buffer or in aqueous extract of hazelnut-free cookies. Additionally, cookies containing hazelnuts were analyzed demonstrating the sensor ability for fast and sensitive detection of the hazelnut proteins in commercial products.

Evaluation of the association between predictive factors and the development of immune-related adverse events and prognostic factors for chemoimmunotherapy in patients with non-small cell lung cancer: A multicenter retrospective study.

Chemoimmunotherapy is widely used as the first-line management of advanced non-small cell lung cancer (NSCLC) in clinical settings. However, predictive factors associated with the development of immune-related adverse events (irAEs) and prognostic factors for NSCLC patients undergoing chemoimmunotherapy remains largely unexplored. Therefore, in this study, we aimed to evaluate predictive factors for irAE development and prognostic factors associated with chemoimmunotherapy in NSCLC patients. This study enrolled 199 patients with advanced and recurrent NSCLC who underwent chemoimmunotherapy across eight institutions in Nagano prefecture from December 2018 to January 2023. We examined predictive factors associated with irAE development and prognostic factors associated with overall survival (OS). Among the patients, 106 experienced irAEs, while 93 patients did not. A total of 44 (22.1%) patients developed multiple irAEs. High serum albumin levels (Alb >3.5 g/dL) emerged as an independent predictive factor associated with irAE development in logistic regression analysis (odds ratio; 2.35, 95% confidence interval 1.27-4.34, p = 0.007). Furthermore, the development of multiple irAEs (p = 0.016), lower lactate dehydrogenase level (<223 U/L, p = 0.002), and decreased neutrophil-to-lymphocyte ratio (<3, p = 0.049) were identified as independent favorable prognostic factors associated with OS in multivariate Cox hazard analyses. The study results suggest that high serum Alb is a predictive factor for irAE development and that the presence of multiple irAEs is a favorable prognostic indicator for NSCLC patients undergoing chemoimmunotherapy.