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1.
Chem Commun (Camb) ; 60(72): 9704-9732, 2024 Sep 05.
Article in English | MEDLINE | ID: mdl-39132917

ABSTRACT

Cellulosic biomass is the most abundantly available natural carbon-based renewable resource on Earth. Its widespread availability, combined with rising awareness, evolving policies, and changing regulations supporting sustainable practices, has propelled its role as a crucial renewable feedstock to meet the escalating demand for eco-friendly and renewable materials, chemicals, and fuels. Initially, biorefinery models using cellulosic biomass had focused on single-product platform, primarily monomeric sugars for biofuel. However, since the launch of the first pioneering cellulosic plants in 2014, these models have undergone significant revisions to adapt their biomass upgrading strategy. These changes aim to diversify the bioproduct portfolio and improve the revenue streams of cellulosic biomass biorefineries. Within this area of research and development, enzyme-based technologies can play a significant role by contributing to eco-design in producing and creating innovative bioproducts. This Feature Article highlights our strategies and recent progress in utilizing the biological diversity and inherent selectivity of enzymes to develop and continuously optimize sustainable enzyme-based technologies with distinct application approaches. We have advanced technologies for standalone platforms, which produce various forms of cellulose nanomaterials engineered with customized and enhanced properties and high yields. Additionally, we have tailored technologies for integration within a biorefinery concept. This biorefinery approach prioritizes designing tailored processes to establish bionanomaterials, such as cellulose and lignin nanoparticles, and bioactive molecules as part of a new multi-bioproduct platform for cellulosic biomass biorefineries. These innovations expand the range of bioproducts that can be produced from cellulosic biomass, transcending the conventional focus on monomeric sugars for biofuel production to include biomaterials biorefinery. This shift thereby contributes to strengthening the Bioeconomy strategy and supporting the achievement of several Sustainable Development Goals (SDGs) of the 2030 Agenda for Sustainable Development.


Subject(s)
Biomass , Cellulose , Cellulose/chemistry , Cellulose/metabolism , Biofuels
2.
Int J Biol Macromol ; 278(Pt 1): 134602, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39127282

ABSTRACT

This study evaluates the feasibility of using enzymatic technology to produce novel nanostructures of cellulose nanomaterials, specifically cellulose nanospheres (CNS), through enzymatic hydrolysis with endoglucanase and xylanase of pre-treated cellulose fibers. A statistical experimental design facilitated a comprehensive understanding of the process parameters, which enabled high yields of up to 82.7 %, while maintaining a uniform diameter of 54 nm and slightly improved crystallinity and thermal stability. Atomic force microscopy analyses revealed a distinct CNS formation mechanism, where initial fragmentation of rod-like nanoparticles and subsequent self-assembly of shorter rod-shaped nanoparticles led to CNS formation. Additionally, adjustments in process parameters allowed precise control over the CNS diameter, ranging from 20 to 100 nm, highlighting the potential for customization in high-performance applications. Furthermore, this study demonstrates how the process framework, originally developed for cellulose nanocrystals (CNC) production, was successfully adapted and optimized for CNS production, ensuring scalability and efficiency. In conclusion, this study emphasizes the versatility and efficiency of the enzyme-based platform for producing high-quality CNS, providing valuable insights into energy consumption for large-scale economic and environmental assessments.


Subject(s)
Cellulase , Cellulose , Nanospheres , Cellulose/chemistry , Hydrolysis , Nanospheres/chemistry , Cellulase/chemistry , Cellulase/metabolism , Endo-1,4-beta Xylanases/chemistry , Endo-1,4-beta Xylanases/metabolism
4.
Ann Surg Oncol ; 2024 Jul 03.
Article in English | MEDLINE | ID: mdl-38958807

ABSTRACT

BACKGROUND: For women at increased risk of breast cancer, the National Comprehensive Cancer Network (NCCN) guidelines recommend clinical encounters every 6-12 months. While screening mammography has corresponded with a relative risk reduction in breast cancer mortality of approximately 20%, evidence validating clinical breast examination (CBE) as an efficacious screening modality is deficient. Our study aimed to assess the conventional merit of regular CBE for breast cancer detection among individuals at increased risk of breast cancer development. METHODS: Women > 18 years with documented high-risk encounters at Corewell Health West from 1 January 2018 to 31 December 22 were retrospectively reviewed. High-risk criteria included genetic predisposition, 5-year (> 1.7%) or lifetime (> 20%) Tyrer-Cuzick and/or Gail Model risk estimations, thoracic radiotherapy before age 30 years, lobular carcinoma in-situ, or atypical hyperplasia. Patients with a history of breast cancer or bilateral prophylactic mastectomy prior to 2018 were excluded. RESULTS: Of the 9171 cumulative high-risk encounters among 2493 women, only one breast cancer was detected by CBE. CBE resulted in 1 (0.04%) cancer diagnosis compared with 30 (1.2%) detected on screening imaging and 10 (0.4%) self-reported. Of the 30 image-detected cancers, 28 (93.3%) had no detectable clinical findings at the time of preoperative consultation. Self-reported and CBE-detected cancers were more likely to be of higher clinical stage compared with imaging-detected malignancies. CONCLUSIONS: The role of routine CBE as a cancer detection modality in the high-risk patient population appears to be limited. Telemedicine can be offered to individuals who have completed screening imaging but are unable to commit and/or are inconvenienced by in-person high-risk breast cancer assessments.

5.
Pharmaceutics ; 16(7)2024 Jun 21.
Article in English | MEDLINE | ID: mdl-39065539

ABSTRACT

The Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) causes reproductive failure and respiratory symptoms, leading to huge economic losses for the pig farming industry. Although several vaccines against PRRSV are available in the market; they show an overall low efficacy, and several countries have the need for vaccines covering the local, circulating variants. This project aims at developing a new chimeric antigen targeting specific epitopes from PRRSV and evaluating two test adjuvants to formulate a vaccine candidate. The test antigen was called LTB-PRRSV, which was produced recombinantly in Escherichia coli and consisted of the heat labile enterotoxin B subunit from E. coli (LTB) and four epitopes from PRRSV. LTB-PRRSV was rescued as inclusion bodies and methods for its solubilization, IMAC-based purification, and refolding were standardized, leading to mean yields of 18 mg of pure protein per liter culture. Layered double hydroxides (LDH) have been used as vaccine adjuvants given their biocompatibility, low cost, and positive surface charge that allows an efficient adsorption of negatively charged biomolecules. Therefore, LDH were selected as delivery vehicles of LTB-PRRSV. Pure LTB-PRRSV was adsorbed onto LDH by incubation at different LDH:LTB-PRRSV mass ratios (1:0.25, 1:0.5, 1:1, and 1:2) and at pH 9.5. The best adsorption occurred with a 1:2 mass ratio, and in a sucrose-tween solution. The conjugates obtained had a polydispersity index of 0.26, a hydrodynamic diameter of 192 nm, and a final antigen concentration of 64.2 µg/mL. An immunogenicity assessment was performed by injecting mice with LDH:LTB-PRRSV, Alum/LTB-PRRSV, or LTB-PRRSV in a scheme comprising three immunizations at two-week intervals and two dose levels (1 and 5 µg). LTB-PRRSV was capable of inducing strong humoral responses, which lasted for a longer period when LDH was used as the delivery vehicle/adjuvant. The potential of LDH to serve as an attractive carrier for veterinary vaccines is discussed.

6.
Pharmaceutics ; 16(7)2024 Jul 02.
Article in English | MEDLINE | ID: mdl-39065588

ABSTRACT

The COVID-19 pandemic required great efforts to develop efficient vaccines in a short period of time. However, innovative vaccines against SARS-CoV-2 virus are needed to achieve broad immune protection against variants of concern. Polymeric-based particles can lead to innovative vaccines, serving as stable, safe and immunostimulatory antigen delivery systems. In this work, polymeric-based particles called thiolated PAA/Schizo were developed. Poly (acrylic acid) (PAA) was thiolated with cysteine ethyl ester and crosslinked with a Schizochytrium sp. cell wall fraction under an inverse emulsion approach. Particles showed a hydrodynamic diameter of 313 ± 38 nm and negative Zeta potential. FT-IR spectra indicated the presence of coconut oil in thiolated PAA/Schizo particles, which, along with the microalgae, could contribute to their biocompatibility and bioactive properties. TGA analysis suggested strong interactions between the thiolated PAA/Schizo components. In vitro assessment revealed that thiolated particles have a higher mucoadhesiveness when compared with non-thiolated particles. Cell-based assays revealed that thiolated particles are not cytotoxic and, importantly, increase TNF-α secretion in murine dendritic cells. Moreover, immunization assays revealed that thiolated PAA/Schizo particles induced a humoral response with a more balanced IgG2a/IgG1 ratio. Therefore, thiolated PAA/Schizo particles are deemed a promising delivery system whose evaluation in vaccine prototypes is guaranteed.

7.
Diagnostics (Basel) ; 14(12)2024 Jun 20.
Article in English | MEDLINE | ID: mdl-38928727

ABSTRACT

Major cardiovascular events (MACEs) are a cause of major mortality worldwide. The narrowing and blockage of coronary arteries with atherosclerotic plaques are diagnosed and treated with percutaneous coronary intervention (PCI). During this procedure, coronary angiography (CAG) remains the most widely used guidance modality for the evaluation of the affected blood vessel. The measurement of the blood vessel diameter is an important factor to consider in order to decide if stent colocation is suitable for the intervention. In this regard, a small blood vessel (<2.75 mm) is majorly left without stent colocation; however, small vessel coronary artery disease (SvCAD) is a significant risk factor for the recurrence of MACEs, maybe due to the lack of a standardized treatment related to the diameter of the affected blood vessel; therefore, a more precise measurement is needed. The use of CAG for the measurement of the blood vessel diameter has some important limitations that can be improved with the use of newer techniques such as intravascular ultrasound (IVUS), although at higher costs, which might explain its underuse. To address differences in blood vessel diameter measurements and identify specific cases where IVUS might be of additional benefit for the patient, we conducted a retrospective study in patients who underwent PCI for MACEs with affection for at least one small blood vessel. We compared the measurements of the affected small blood vessels' diameter obtained by CAG and IVUS to identify cases of reclassification of the affected blood vessel; additionally, we underwent a multivariate analysis to identify risk factors associated with blood vessel reclassification. We included information from 48 patients with a mean ± SD age of 69.1 ± 11.9 years; 70.8% were men and 29.2% were women. The mean diameter with CAG and IVUS was 2.1 mm (95% CI 1.9-2.2), and 2.8 (2.8-3.0), respectively. The estimated difference was of 0.8 mm (95% CI 0.7-0.9). We found a significant positive low correlation in diameter measurements of small blood vessels obtained with CAG and IVUS (r = 0.1242 p = 0.014). In total, 37 (77%) patients had a reclassification of the affected blood vessel with IVUS. In 21 cases, the affected blood vessel changed from a small to a medium size (2.75-3.00 mm), and in 15 cases, the affected vessel changed from a small to a large size (<3.00 mm). The Bland-Altman plot was used to evaluate agreement in measurements with CAG and IVUS. The change in blood vessel classification with IVUs was important for the decision of intervention and stent collocation. The only variable associated with reclassification of blood vessels after adjustment in a multivariate analysis was T2D (type 2 diabetes) (p = 0 0.035). Our findings corroborate that blood vessels might appear smaller with CAG, especially in patients with T2D; therefore, at least in these cases, the use of IVUS is recommended over CAG.

8.
Vaccine ; 42(17): 3674-3683, 2024 Jun 20.
Article in English | MEDLINE | ID: mdl-38749821

ABSTRACT

The Zika virus (ZIKV) is considered a public health problem worldwide due to its association with the development of microcephaly and the Guillain-Barré syndrome. Currently, there is no specific treatment or vaccine approved to combat this disease, and thus, developing safe and effective vaccines is a relevant goal. In this study, a multi-epitope protein called rpZDIII was designed based on a series of ZIKV antigenic sequences, a bacterial carrier, and linkers. The analysis of the predicted 3D structure of the rpZDIII chimeric antigen was performed on the AlphaFold 2 server, and it was produced in E. coli and purified from inclusion bodies, followed by solubilization and refolding processes. The yield achieved for rpZDIII was 11 mg/L in terms of pure soluble recombinant protein per liter of fermentation. rpZDIII was deemed immunogenic since it induced serum IgG and IgM responses in mice upon subcutaneous immunization in a three-dose scheme. Moreover, sera from mice immunized with rpZDIII showed neutralizing activity against ZIKV. Therefore, this study reveals rpZDIII as a promising immunogen for the development of a rationally designed multi-epitope vaccine against ZIKV, and completion of its preclinical evaluation is guaranteed.


Subject(s)
Antibodies, Neutralizing , Antibodies, Viral , Antigens, Viral , Zika Virus Infection , Zika Virus , Animals , Zika Virus/immunology , Antibodies, Neutralizing/immunology , Antibodies, Neutralizing/blood , Mice , Antibodies, Viral/immunology , Antibodies, Viral/blood , Zika Virus Infection/prevention & control , Zika Virus Infection/immunology , Antigens, Viral/immunology , Antigens, Viral/genetics , Viral Vaccines/immunology , Viral Vaccines/administration & dosage , Epitopes/immunology , Immunoglobulin G/blood , Immunoglobulin G/immunology , Female , Escherichia coli/genetics , Escherichia coli/metabolism , Immunoglobulin M/immunology , Immunoglobulin M/blood , Mice, Inbred BALB C
9.
Planta Med ; 90(10): 744-756, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38698590

ABSTRACT

The carrot-made LTB-Syn antigen (cLTB-Syn) is a vaccine candidate against synucleinopathies based on carrot cells expressing the target antigen LTB and syn epitopes. Therefore, the development of an efficient production process is required with media culture optimization to increase the production yields as the main goal. In this study, the effect of two nitrogen sources (urea and glutamate) on callus cultures producing cLTB-Syn was studied, observing that the addition of 17 mM urea to MS medium favored the biomass yield. To optimize the MS media composition, the influence of seven medium components on biomass and cLTB-Syn production was first evaluated by a Plackett-Burman design (PBD). Then, three factors were further analyzed using a central composite design (CCD) and response surface methodology (RSM). The results showed a 1.2-fold improvement in biomass, and a 4.5-fold improvement in cLTB-Syn production was achieved at the shake-flask scale. At the bioreactor scale, there was a 1.5-fold increase in biomass and a 2.8-fold increase in cLTB-Syn yield compared with the standard MS medium. Moreover, the cLTB-Syn vaccine induced humoral responses in BALB/c mice subjected to either oral or subcutaneous immunization. Therefore, cLTB-Syn is a promising vaccine candidate that will aid in developing immunotherapeutic strategies to combat PD and other neurodegenerative diseases without the need for cold storage, making it a financially viable option for massive immunization.


Subject(s)
Daucus carota , Mice, Inbred BALB C , Animals , Mice , Vaccines , Cell Culture Techniques , Female , Biomass , Culture Media
10.
Braz J Microbiol ; 55(2): 1151-1166, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38472698

ABSTRACT

Developing efficient microbiological methods to convert polysaccharide-rich materials into fermentable sugars, particularly monosaccharides, is vital for advancing the bioeconomy and producing renewable chemicals and energy sources. This study focused on optimizing the production conditions of an enzyme cocktail from Aspergillus niger ATCC 9642 using solid-state fermentation (SSF) and assessing its effectiveness in saccharifying mango peels through a simple, rapid, and efficient one-step process. A rotatable central composite design was employed to determine optimal conditions of moisture, time, and pH for enzyme production in SSF medium. The optimized enzyme cocktail exhibited cellulase activity (CMCase) at 6.28 U/g, filter paper activity (FPase) at 3.29 U/g, and pectinase activity at 117.02 U/g. These optimal activities were achieved with an SSF duration of 81 h, pH of 4.66, and a moisture content of 59%. The optimized enzyme cocktail effectively saccharified the mango peels without the need for chemical agents. The maximum saccharification yield reached approximately 81%, indicating efficient conversion of mango peels into sugars. The enzyme cocktail displayed consistent thermal stability within the tested temperature range of 30-60°C. Notably, the highest sugar release occurred within 36 h, with glucose, arabinose, galactose, and xylose being the primary monosaccharides released during saccharification. This study highlights the potential application of Aspergillus niger ATCC 9642 and SSF for enzymatic production, offering a simple and high-performance process for monosaccharide production. The optimized enzyme cocktail obtained through solid-state fermentation demonstrated efficient saccharification of mango peels, suggesting its suitability for industrial-scale applications.


Subject(s)
Aspergillus niger , Fermentation , Mangifera , Aspergillus niger/enzymology , Aspergillus niger/metabolism , Mangifera/microbiology , Mangifera/chemistry , Hydrogen-Ion Concentration , Cellulase/metabolism , Cellulase/chemistry , Temperature , Polygalacturonase/metabolism , Enzyme Stability , Hydrolysis , Fungal Proteins/metabolism
11.
J Gastrointest Oncol ; 15(1): 478-484, 2024 Feb 29.
Article in English | MEDLINE | ID: mdl-38482230

ABSTRACT

Background: Small molecule fibroblast growth factor receptor (FGFR) inhibitors, such as pemigatinib, have been developed for the treatment of cholangiocarcinoma (CCA) with rearrangements or fusions in the FGFR2. FGFR inhibitors (FGFRis) have dermatologic side effects such as dry skin or nail bed damage. However, in very rare instances, a life-threatening vascular calcification disease known as calciphylaxis has been linked to these therapies. Case Description: We report a patient with metastatic CCA, who developed calciphylaxis following the start of their pemigatinib treatment. Calciphylaxis is associated with skin lesions and affects the dermal microvasculature in addition to the vascular calcification. This case focuses on the management strategy used for this rare adverse event (AE) as well as the pathology and complicated mechanism of calciphylaxis. We highlight the unclear pathophysiology behind this disease by identifying key players in the signaling and molecular pathways in the microenvironment that are needed to trigger this pathology. Conclusions: Calciphylaxis is normally associated with advanced renal failure in the setting of high phosphate and calcium. However, the patient we present here did not have advanced renal failure or high calcium levels and calcium dysregulation. As FGFRi use becomes more widespread, the more important it becomes to identify and have a treatment strategy for this rare AE.

12.
Pharmaceuticals (Basel) ; 17(3)2024 Feb 27.
Article in English | MEDLINE | ID: mdl-38543088

ABSTRACT

Subunit vaccines stand as a leading approach to expanding the current portfolio of vaccines to fight against COVID-19, seeking not only to lower costs but to achieve long-term immunity against variants of concern and have the main attributes that could overcome the limitations of the current vaccines. Herein a chimeric protein targeting S1 and S2 epitopes, called LTp50, was designed as a convenient approach to induce humoral responses against SARS-CoV-2. LTp50 was produced in recombinant Escherichia coli using a conventional pET vector, recovering the expected antigen in the insoluble fraction. LTp50 was purified by chromatography (purity > 90%). The solubilization and refolding stages helped to obtain a stable protein amenable for vaccine formulation. LTp50 was adsorbed onto alum, resulting in a stable formulation whose immunogenic properties were assessed in BALB/c mice. Significant humoral responses against the S protein (BA.5 variant) were detected in mice subjected to three subcutaneous doses (10 µg) of the LTp50/alum formulation. This study opens the path for the vaccine formulation optimization using additional adjuvants to advance in the development of a highly effective anti-COVID-19 vaccine directed against the antigenic regions of the S protein, which are less prone to mutations.

13.
Int J Biol Macromol ; 254(Pt 1): 127691, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37898249

ABSTRACT

Among the conditions caused by diabetes, the diabetic foot is a significant public health problem due to its delayed healing process. That makes it essential to design, manufacture, and apply auxiliary dressings during healing. In this work, chitosan sponges were developed and evaluated as wound dressings. Metformin, fucoidan, and exopolysaccharide from Porphyridium purpureum algae were loaded into the sponges and studied as healing promoters. The composite sponges were physicochemically, morphologically, and thermally characterized, allowing us to determine the chemical mechanisms involved in the sponge formation. The mechanical analysis demonstrated that sponge composites have shape memory and good mechanical performance under compression stress, showing a compressive strength above 30 kPa. These results correlated with the materials' porosity, influencing the swelling capacity that reached a maximum of 70 %. The morphology of materials was observed by SEM, resulting in folded films with surface porosity. The results of the biocompatibility tests confirmed that the materials are not cytotoxic or hemolytic and have good antibacterial activity. In vivo wound healing evaluation showed that metformin-loaded chitosan sponges regenerated skin tissue after 21 days of treatment, highlighting the rate of healing provided when exopolysaccharide was added to promote tissue regeneration, which can be corroborated by histological analysis. These results make chitosan sponge compounds promising dressings for diabetic foot wound treatment.


Subject(s)
Chitosan , Diabetes Mellitus , Diabetic Foot , Microalgae , Humans , Chitosan/chemistry , Wound Healing , Anti-Bacterial Agents/pharmacology , Bandages
14.
Planta Med ; 90(1): 63-72, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37852270

ABSTRACT

The development of virus-free, oral vaccines against poliovirus capable of inducing mucosal protective immunity is needed to safely combat this pathogen. In the present study, a carrot cell line expressing the poliovirus VP2 antigen was established at the level of callus and cell suspensions, exploring the effects of culture media (MS and B5), supplementation with urea, phytoregulators (2,4-D : KIN), and light conditions (continuous light, photoperiod, and total darkness). The best callus growth was obtained on B5 medium supplemented with 2 mg/L of 2,4-D + 2 mg/L kinetin and 0.0136 g/L of urea and in continuous light conditions. Suspension cultures of the SMC-1 line in 250 mL Erlenmeyer flasks had a maximum growth of 16.07 ± 0.03 g/L DW on day 12 with a growth rate of µ=0.3/d and a doubling time of 2.3 days. In a 2 L airlift bioreactor, the biomass yield achieved was 25.6 ± 0.05 g/L DW at day 10 with a growth rate of µ= 0.58/d and doubling time of 1.38 d. Cell growth was 1.5 times higher in bioreactors than in shake flasks, highlighting that both systems resulted in the accumulation of VP2 throughout the time in culture. The maximum VP2 yield in flasks was 387.8 µg/g DW at day 21, while in the reactor it was 550.2 µg/g DW at day 18. In conclusion, bioreactor-based production of the VP2 protein by the SMC-1 suspension cell line offers a higher productivity when compared to flask cultures, offering a key perspective to produce low-cost vaccines against poliomyelitis.


Subject(s)
Daucus carota , Poliovirus Vaccines , Poliovirus , Cell Line , Urea , 2,4-Dichlorophenoxyacetic Acid
15.
Microb Pathog ; 186: 106488, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38061668

ABSTRACT

Trypanosoma cruzi parasite - causal Chagas disease agent - affects about 7 million people; no vaccine is available, and current medications have not been entirely effective. Multidisciplinary efforts are necessary for developing clinical vaccine prototypes. Thus, this research study aims to assess the expressed and whole-cell administration protection of the oral vaccine prototype Tc24:Co1 using Schizochytrium sp. microalga. High recombinant protein expression yields (675 µg/L) of algal culture were obtained. Additionally, Schizochytrium sp.-Tc24:Co1 resulted stable at 4 °C for up to six months and at 25 °C for three months. After receiving four oral doses of the vaccine, the mice showed a significant humoral immune response and a parasitemia reduction associated with a lack of heart inflammatory damage compared with the unvaccinated controls. The Schizochytrium sp.-Tc24:Co1 vaccine demonstrates to be promising as a prototype for further development showing protective effects against a T. cruzi challenge in a mouse model.


Subject(s)
Chagas Disease , Protozoan Vaccines , Trypanosoma cruzi , Humans , Animals , Mice , Chagas Disease/drug therapy , Recombinant Proteins , Disease Models, Animal
16.
Biotechnol Prog ; 39(6): e3390, 2023.
Article in English | MEDLINE | ID: mdl-37702113

ABSTRACT

The aggregation and spread of alpha-synuclein (αSyn) is associated with several pathogenic pathways that lead to neurodegeneration and, ultimately, to synucleinopathies development. Hence, the establishment of a safe and effective disease-modifying therapy that limits or prevents the spread of toxic αSyn aggregation could lead to positive clinical outcomes. A rational vaccine design can be focused on the selection of specific epitopes able to induce the immune response desired, for example, antibodies able to mediate the clearance of αSyn aggregates without the induction of inflammatory responses. To develop a rapid system for the evaluation of a vaccine candidate against synucleinopathies, rLTB-Syn (an antigen based on three B cell epitopes from αSyn and the B subunit of the heat-labile Escherichia coli enterotoxin [LTB] as adjuvant/carrier) was produced using recombinant E. coli (Rosetta DE3) as the expression host. The bacterial version of rLTB-Syn was produced as soluble protein at yields up to 1.72 mg/g biomass. A method for the purification of rLTB-Syn (~18 kDa) was developed based on ion exchange chromatography, reaching purity >93% with a final concentration of 82.6 µg/mL. Furthermore, the purified soluble rLTB-Syn retained GM1 binding activity, suggesting proper folding and pentameric structure. The results from this study establish a fast and effective method to obtain rLTB-Syn, making it useful in the design of novel vaccine formulations targeting synucleinopathies.


Subject(s)
Bacterial Toxins , Escherichia coli Proteins , Synucleinopathies , Vaccines , Humans , Escherichia coli/genetics , Escherichia coli/metabolism , Epitopes , Recombinant Proteins/metabolism , Immunotherapy , Recombinant Fusion Proteins/genetics
17.
Sci Total Environ ; 905: 167426, 2023 Dec 20.
Article in English | MEDLINE | ID: mdl-37774864

ABSTRACT

Emerging pollutants (EPs) emerged as a group of new compounds whose presence in the environment has been widely detected in Mexico. In this country, different concentrations of pharmaceutical compounds, pesticides, dyes, and microplastics have been reported, which vary depending on the region and the analyzed matrix (i.e., wastewater, surface water, groundwater). The evidence of the EPs' presence focuses on the detection of them, but there is a gap in information regarding is biomonitoring and their effects in health in Mexico. The presence of these pollutants in the country associated with lack of proper regulations in the discharge and disposal of EPs. Therefore, this review aims to provide a comprehensive view of the current environmental status, policies, and frameworks regarding Mexico's situation. The review also highlights the lack of information about biomonitoring since EPs are present in water even after their treatment, leading to a critical situation, which is high exposure to humans and animals. Although, technologies to efficiently eliminate EPs are available, their application has been reported only at a laboratory scale thus far. Here, an overview of health and environmental impacts and a summary of the research works reported in Mexico from 2014 to 2023 were presented. This review concludes with a concrete point of view and perspective on the status of the EPs' research in Mexico as an alert for government entities about the necessity of measures to control the EPs disposal and treatment.


Subject(s)
Environmental Pollutants , Water Pollutants, Chemical , Animals , Humans , Wastewater , Water , Mexico , Plastics , Water Pollutants, Chemical/analysis , Policy , Environmental Monitoring
18.
Materials (Basel) ; 16(15)2023 Aug 04.
Article in English | MEDLINE | ID: mdl-37570172

ABSTRACT

The discovery and validation of new adjuvants are critical areas for vaccinology. Mineral materials (e.g., alum microparticles) have been used for a long time as adjuvants in human vaccine formulations. Nonetheless, the use of nanosized materials is a promising approach to diversify the properties of adjuvants. Nanoclays are potential adjuvants proposed by some research groups. However, their adjuvant mechanisms and safety have not been fully elucidated. Herein, we aimed at expanding the knowledge on the potential adjuvanticity of layered double hydroxide (LDH) nanoparticles by reporting a detailed method for the synthesis and characterization of LDHs and the adsorption of a model antigen (bovine serum albumin, BSA). LDHs varying in diameter (from 56 to 88 nm) were obtained, and an in vitro evaluation revealed that the LDHs are not inherently toxic. BSA was passively adsorbed onto the LDHs, and the immunogenicity in mice of the conjugates obtained was compared to that of free BSA and BSA co-administered with alum (Alum-BSA). The LDH-BSA conjugates induced a higher humoral response that lasted for a longer period compared with that of free BSA and Alum-BSA, confirming that LDH exerts adjuvant effects. The 56 nm LDH particles were deemed as the more efficient carrier since they induced a higher and more balanced Th1/Th2 response than the 88 nm particles. This study is a contribution toward expanding the characterization and use of nanoclays in vaccinology and justifies further studies with pathogen-specific antigens.

19.
Int J Biol Macromol ; 250: 126007, 2023 Oct 01.
Article in English | MEDLINE | ID: mdl-37524277

ABSTRACT

Cellulose nanomaterials (CNs) are promising green materials due to their unique properties as well as their environmental benefits. Among these materials, cellulose nanofibrils (CNFs) and nanocrystals (CNCs) are the most extensively researched types of CNs. While they share some fundamental properties like low density, biodegradability, biocompatibility, and low toxicity, they also possess unique differentiating characteristics such as morphology, rheology, aspect ratio, crystallinity, mechanical and optical properties. Therefore, numerous comparative studies have been conducted, and recently, various studies have reported the synergetic advantages resulting from combining CNF and CNC. In this review, we initiate by addressing the terminology used to describe combinations of these and other types of CNs, proposing "hybrid cellulose nanomaterials" (HCNs) as the standardized classifictation for these materials. Subsequently, we briefly cover aspects of properties-driven applications and the performance of CNs, from both an individual and comparative perspective. Next, we comprehensively examine the potential of HCN-based materials, highlighting their performance for various applications. In conclusion, HCNs have demonstraded remarkable success in diverse areas, such as food packaging, electronic devices, 3D printing, biomedical and other fields, resulting in materials with superior performance when compared to neat CNF or CNC. Therefore, HCNs exhibit great potential for the development of environmentally friendly materials with enhanced properties.

20.
Int J Biol Macromol ; 246: 125655, 2023 Aug 15.
Article in English | MEDLINE | ID: mdl-37399864

ABSTRACT

Advanced materials used in the biomedicine field comprises a diverse group of organic molecules, including polymers, polysaccharides, and proteins. A significant trend in this area is the design of new micro/nano gels whose small size, physical stability, biocompatibility, and bioactivity could lead to new applications. Herein a new synthesis route is described to obtain core-shell microgels based on chitosan and Porphyridium exopolysaccharides (EPS) crosslinked with sodium tripolyphosphate (TPP). First, the synthesis of EPS-chitosan gels through ionic interactions was explored, leading to the formation of unstable gels. Alternatively, the use of TTP as crosslinker agent led to stable core-shell structures. The influence of reaction temperature, sonication time, and exopolysaccharide concentration, pH and TPP concentration were determined as a function of particle size and polydispersity index (PDI). The obtained EPS-chitosan gels were characterized by TEM, TGA, and FTIR; followed by the assessment of protein load capacity, stability upon freezing, cytotoxicity, and mucoadhesivity. Experimentation revealed that the core-shell particles size ranges 100-300 nm, have a 52 % loading capacity for BSA and a < 90 % mucoadhesivity, and no toxic effects in mammalian cell cultures. The potential application of the obtained microgels in the biomedical field is discussed.


Subject(s)
Chitosan , Microgels , Porphyridium , Animals , Chitosan/chemistry , Gels/chemistry , Ions , Particle Size , Mammals
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