Extraction and separation of astaxanthin with the help of pre-treatment of Haematococcus pluvialis microalgae biomass using aqueous two-phase systems based on deep eutectic solvents.

The microalgae Haematococcus pluvialis are the main source of the natural antioxidant astaxanthin. However, the effective extraction of astaxanthin from these microalgae remains a significant challenge due to the rigid, non-hydrolyzable cell walls. Energy savings and high-efficiency cell disruption are essential steps in the recovery of the antioxidant astaxanthin from the cysts of H. pluvialis. In the present study, H. pluvialis microalgae were first cultured in Bold's Basal medium under certain conditions to reach the maximum biomass concentration, and then light shock was applied for astaxanthin accumulation. The cells were initially green and oval, with two flagella. As the induction time increases, the motile cells lose their flagellum and become red cysts with thick cell walls. Pre-treatment of aqueous two-phase systems based on deep eutectic solvents was used to decompose the cell wall. These systems included dipotassium hydrogen phosphate salt, water, and two types of deep eutectic solvents (choline chloride-urea and choline chloride-glucose). The results of pre-treatment of Haematococcus cells by the studied systems showed that intact, healthy cysts were significantly ruptured, disrupted, and facilitated the release of cytoplasmic components, thus facilitating the subsequent separation of astaxanthin by liquid-liquid extraction. The system containing the deep eutectic solvent of choline chloride-urea was the most effective system for cell wall degradation, which resulted in the highest ability to extract astaxanthin. More than 99% of astaxanthin was extracted from Haematococcus under mild conditions (35% deep eutectic solvent, 30% dipotassium hydrogen phosphate at 50 °C, pH = 7.5, followed by liquid-liquid extraction at 25 °C). The present study shows that the pre-treatment of two-phase systems based on deep eutectic solvent and, thus, liquid-liquid extraction is an efficient and environmentally friendly process to improve astaxanthin from the microalgae H. pluvialis.

Surface modification of hollow gold nanoparticles conducted by incorporating cancer cell membrane and AS1411 aptamer, aiming to achieve a dual-targeted therapy for colorectal cancer.

Due to its inherent membrane structure, a nanostructure enveloped by an active cell membrane possesses distinctive characteristics such as prolonged presence in the bloodstream, precise identification capabilities, and evasion of immune responses. This research involved the production of biomimetic nanoparticles, specifically hollow gold nanoparticles (HGNPs) loaded with methotrexate (MTX), which were further coated with cancer cell membrane. These nanoparticles were then adorned with AS1411 aptamer to serve as a targeting agent (Apt-CCM-HG@MTX). The nanoplatform demonstrated precise targeting towards cancer cells due to its dual-targeting characteristic (AS1411 aptamer and C26 cancer cell membrane), exhibiting uniformity in distribution. It also displayed a desirable response to photothermal stimulation, controlled release of drugs, and exceptional properties for fluorescence imaging. The system was composed of spherical HGNPs measuring 51.33 ± 5.70 nm in diameter, which were effectively loaded with MTX using a physical absorption method. The encapsulation efficiency achieved was recorded at 79.54 %, while the loading efficiency reached 38.21 %. The targeted formulation demonstrated a noteworthy mortality of approximately 45 % in the nucleolin positive cell line, C26, as determined by in vitro cytotoxicity assays. As a result of the functionalization process applied to the homologous binding adhesion molecules found in cancer cell membranes and targeting ability of AS1411 aptamer, Apt-CCM-HG@MTX demonstrated a substantial enhancement in targeting tumors and facilitating cellular uptake during in vivo experiments. Furthermore, under NIR radiation the photothermal effect exhibited by Apt-CCM-HG@MTX in the tumor area was notably robust due to the distinctive attributes of HGNPs. The conclusions obtained from this study have the potential to assist in adopting a bioinspired strategy that will significantly improve the effective management of MTX and therapy for individuals with colorectal cancer.

Fabrication of Bacterial Cellulose/Chitosan-MIL-100(Fe) Composite for Adsorptive Removal of Dacarbazine.

In this research, a polymeric composite based on a chitosan/bacterial cellulose (CS/BC) matrix filled with MIL-100(Fe) particles was prepared to solve the recyclability of issue MIL-100(Fe) particles and utilized as an efficient adsorbent for removing dacarbazine (DTIC) from wastewater. The adsorption capacity of the composite (CS/BC-MIL) was higher than both MIL-100(Fe) and the CS/BC polymeric matrix. The adsorption performance of the fabricated composite was evaluated through kinetics and isotherm studies. While isotherm studies revealed that the adsorption of DTIC onto the adsorbent can be well described by the Freundlich model, kinetics studies indicated that a combination of factors, rather than a single rate-limiting factor, are responsible for the adsorption rate. Thermodynamics investigation showed that the adsorption of DTIC to CS/BC-MIL composite is exothermic and occurs spontaneously. Additionally, due to the negative entropy change, it was established that the adsorption is governed by the enthalpy change. Exploring the solution chemistry revealed that the optimum pH for the adsorption process was about 4. Moreover, the CS/BC-MIL can selectively adsorb DTIC in the presence of other pharmaceuticals like doxorubicin (DOX). Furthermore, regeneration investigations disclosed that the composite holds its structural features and has an acceptable adsorption capacity after several cycles of adsorption/desorption.

Cu-vitamin B3 donut-like MOFs incorporated into TEMPO-oxidized bacterial cellulose nanofibers for wound healing.

In this study, a novel multifunctional nanocomposite wound dressing was developed, consisting of TEMPO-oxidized bacterial cellulose (TOBC) nanofibers functionalized with donut-like copper-based metal-organic frameworks (CuVB3 MOFs). These CuVB3 MOFs were constructed using copper nodes linked by vitamin B3 molecules, resulting in a copper nicotinate crystal structure as confirmed by X-ray diffraction. Electron microscopy confirmed the presence of donut-like microstructures with uniform element distribution in the synthesized MOFs. Through the incorporation of CuVB3 MOFs into the TOBC nanofibers, innovative TOBC-CuVB3 nanocomposites were created. Biocompatibility testing using the MTT assay demonstrated enhanced cell viability of over 115% for the TOBC-CuVB3 nanocomposite. Acridine Orange staining revealed a ratio of 88-92% live cells on the wound dressings. Furthermore, fibroblast cells cultured on TOBC-CuVB3 exhibited expanded morphologies with long filopodia. The agar diffusion method exhibited improved antibacterial activity against both Gram-positive and Gram-negative bacterial strains, correlating with increased CuVB3 concentration in the samples. In vitro cellular scratch assays demonstrated excellent wound healing potential, with a closure rate of over 98% for wounds treated with the TOBC-CuVB3 nanocomposite. These findings underscore the synergistic effects of copper, vitamin B3, and TOBC nanofibers in the wound healing process.

Assessment of the properties of natural-based chiral deep eutectic solvents for chiral drug separation: insights from molecular dynamics simulation.

The structural and physicochemical properties of chiral deep eutectic solvents (DESs) consisting of racemic mixtures of menthol and acetic acid (DES1), racemic mixtures of menthol and lauric acid (DES2), and racemic mixtures of menthol and pyruvic acid (DES3) for enantioselective extraction processes are investigated. Structural results, such as the radial distribution function (RDF) and the combined distribution function (CDF), indicate that the hydroxyl hydrogen of menthol has a dominant interaction with the carbonyl oxygen of the acids in the considered DESs. The number of hydrogen bonds and non-bonded interaction energies formed between S-menthol and HBDs are larger than those with R-menthol, resulting in the self-diffusion coefficient of S-menthol being larger than that of R-menthol. Therefore, it can be said that the proposed DESs are good candidates for the separation of drugs with S chirality. The effects of acid type on the density and isothermal compressibility of DESs show the behaviour of DES2 > DES3 > DES1 and DES1 > DES3 > DES2, respectively. Our results provide a better perspective on new chiral DESs at the molecular level for enantioselective processes.

A Novel Silver-Based Metal-Organic Framework Incorporated into Nanofibrous Chitosan Coatings for Bone Tissue Implants.

In this work, new multi-layer nanocomposite coatings comprised of chitosan (CS) nanofibers functionalized using an innovative silver-based metal-organic framework (SOF) were developed. The SOFs were produced via a facile process using green and environmental-friendly materials. The CS-SOF nanocomposites were coated on hierarchical oxide (HO) layers fabricated on titanium substrates by an innovative two-step etching process. X-ray diffraction revealed fruitful production of the SOF NPs and their stable crystalline structure within the nanocomposite coatings. Energy-dispersive x-ray spectroscopy approved uniform SOFs distribution in the CS-SOF nanocomposites. Atomic force microscopy indicated more than 700% increased nanoscale roughness for the treated surfaces compared to the bare sample. In vitro MTT assay revealed proper cell viabilities on the samples, however, high SOFs concentration led to less biocompatibility. All coatings demonstrated positive cell proliferation rates up to 45% after 72 h. Antibacterial studies showed significant inhibition zones against Escherichia coli and Staphylococcus aureus bacteria with 100-200% effective antibacterial activities. Electron microscopy exhibited excellent cell-implant integration for the CS-SOF nanocomposite surfaces due to the attached cells with expanded morphologies and long filopodia. The prepared coatings showed high apatite formation capability and bone bioactivity.

Boosting bone cell growth using nanofibrous carboxymethylated cellulose and chitosan on titanium dioxide nanotube array with dual surface charges as a novel multifunctional bioimplant surface.

One of the most vital aspects of the orthopedic implant field has been the development of multifunctional coatings that improve bone-implant contact while simultaneously preventing bacterial infection. The present study investigates the fabrication and characterization of multifunctional polysaccharides, including carboxymethyl cellulose (CMCn) and carboxymethyl chitosan nanofibers (CMCHn), as a novel implant coating on titania nanotube arrays (T). Field emission scanning electron microscopy (FESEM) images revealed a nanofibrous morphology with a narrow diameter for CMCn and CMCHn, similar to extracellular matrix nanostructures. Compared to the T surface, the roughness of CMCn and CMCHn samples increased by over 250 %. An improved cell proliferation rate was observed on CMCHn nanofibers with a positively charged surface caused by the amino groups. Furthermore, in an antibacterial experiment, CMCn and CMCHn inhibited bacterial colony formation by 80 % and 73 %, respectively. According to the results, constructed modified CMCn and CMCHn increased osteoblast cell survival while inhibiting bacterial biofilm formation owing to their surface charge and bioinspired physicochemical properties.

Expression Optimizing of Recombinant Oxalyl-CoA Decarboxylase in Escherichia coli.

Background: One of the most common diseases of the urinary tract is stones of this system, including kidney stones. About 70%-80% of kidney stones are calcium oxalate. Oxalyl-CoA decarboxylase is a single polypeptide included of 568 amino acids which play a key role in oxalate degradation. Materials and Methods: The aim of current study is high-level expression of oxalyl-CoA decarboxylase in Escherichia coli BL21 (DE3). To achieve this aim, oxalyl-CoA decarboxylase gene was cloned upon pET-30a (+) with T7 promoter. The vector containing the oxalyl-CoA decarboxylase gene was transformed into E. coli and the expression of the gene was examined on a laboratory scale and fermentor. Atfirst, the effect of temperature, culture medium, and induction time on oxalyl-CoA decarboxylase expression at three levels was examined. Results: The obtained data showed that the highest expression was related to the terrific broth culture medium and temperature of 32°C with an inducer concentration of 1 mM. Under this situation the ultimate cells dry weight and the final oxalyl-CoA decarboxylase expression were 2.46 g/l and 36% of total protein, respectively. Then induction time was optimized in a bench bioreactor and productivity of oxalyl-CoA decarboxylase was calculated. Under optimized condition the cell density, biomass productivity and oxalyl-CoA decarboxylase concentration reached 4.02 g/l, 0.22 g/l/h, and 0.7 g/l which are one of the highest reported rates. Conclusion: This study demonstrated that high levels of oxalyl-CoA decarboxylase can be achieved by optimizing the expression conditions.

Recent Advances on Immune Targeted Therapy of Colorectal Cancer Using bi-Specific Antibodies and Therapeutic Vaccines.

Colorectal cancer (CRC) is a universal heterogeneous disease that is characterized by genetic and epigenetic alterations. Immunotherapy using monoclonal antibodies (mAb) and cancer vaccines are substitute strategies for CRC treatment. When cancer immunotherapy is combined with chemotherapy, surgery, and radiotherapy, the CRC treatment would become excessively efficient. One of the compelling immunotherapy approaches to increase the efficiency of CRC therapy is the deployment of therapeutic mAbs, nanobodies, bi-specific antibodies and cancer vaccines, which improve clinical outcomes in patients. Also, among the possible therapeutic approaches for CRC patients, gene vaccines in combination with antibodies are recently introduced as a new perspective. Here, we aimed to present the current progress in CRC immunotherapy, especially using Bi-specific antibodies and dendritic cells mRNA vaccines. For this aim, all data were extracted from Google Scholar, PubMed, Scopus, and Elsevier, using keywords cancer vaccines; CRC immunotherapy and CRC mRNA vaccines. About 97 articles were selected and investigated completely based on the latest developments and novelties on bi-specific antibodies, mRNA vaccines, nanobodies, and MGD007.