13 research outputs found
Radio-synthesized protein-based nanoparticles for biomedical purposes
Protein-crosslinking whether done by enzymatic or chemically induced pathways increases the overall stability of proteins. In the continuous search for alternative routes for protein stabilization we report a novel technique - radio-induced synthesis of protein nanoparticles - to achieve size controlled particles with preserved bioactivity. Papain was used as model enzyme and the samples were irradiated at 10. kGy in a gammacell irradiator in phosphate buffer (pH=7.0) and additives such as ethanol (0-40%) and sodium chloride (0-25%). The structural rearrangement caused by irradiation under defined conditions led to an increase in papain particle size as a function of the additive and its concentration. These changes occur due to intermolecular bindings, of covalent nature, possibly involving the aromatic amino acids. Ethanol held major effects over papain particle size and particle size distribution if compared to sodium chloride. The particles presented relative retained bioactivity and the physic-chemical characterization revealed similar fluorescence spectra indicating preserved conformation. Differences in fluorescence units were observed according to the additive and its concentration, as a result of protein content changes. Therefore, under optimized conditions, the developed technique may be applied for enzyme nanoparticles formation of controllable size and preserved bioactivity.Fil: Varca, Gustavo H. C.. Comissao Nacional de Energia Nuclear. Centro de Lasers e Aplicacoes. Instituto de Pesquisas EnergĂ©ticas e Nucleares; BrasilFil: Ferraz, Caroline C.. Comissao Nacional de Energia Nuclear. Centro de Lasers e Aplicacoes. Instituto de Pesquisas EnergĂ©ticas e Nucleares; BrasilFil: Lopes, Patricia S.. Universidade de Sao Paulo; BrasilFil: Mathor, Monica Beatriz. Comissao Nacional de Energia Nuclear. Centro de Lasers e Aplicacoes. Instituto de Pesquisas EnergĂ©ticas e Nucleares; BrasilFil: Grasselli, Mariano. Consejo Nacional de Investigaciones CientĂficas y TĂ©cnicas. Centro CientĂfico TecnolĂłgico Conicet - La Plata. Instituto Multidisciplinario de BiologĂa Celular. Provincia de Buenos Aires. GobernaciĂłn. ComisiĂłn de Investigaciones CientĂficas. Instituto Multidisciplinario de BiologĂa Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de BiologĂa Celular; ArgentinaFil: LugĂŁo, Ademar B.. Comissao Nacional de Energia Nuclear. Centro de Lasers e Aplicacoes. Instituto de Pesquisas EnergĂ©ticas e Nucleares; Brasi
Determination of Organochlorines in Soil of a Suburban Area of SĂŁo Paulo Brazil
Technological advances have promoted improvements in several science fields, especially related to environmental and analytical areas with the improvement of detection and development of environmentally friendly extraction techniques. This study applied Quick, Easy, Cheap, Effective, Rugged and Safe method (QuEChERS) for soil extraction and assessed its performance through a validation study using samples from the soil of a contaminated area in Caieiras, SP, Brazil. Nine organochlorine pesticides, including the isomers alpha, beta, gamma and delta- hexachlorocyclohexane; cis- and trans-heptachlor epoxide; cis- and trans-chlordane and heptachlor were analyzed by gas chromatography coupled to electron capture detector. The method was validated according to ISO 5725-4 (2020), EURACHEM (2014) and DOQ-CGCRE-008 (2016). The limits of detection and quantification of the method for the nine organochlorines were α-HCH (1.2 and 12.6 µg kg−1), β-HCH (1.7 and 12.0 µg kg−1), γ-HCH (1.5 and 11.6 µg kg−1), δ-HCH (0.8 and 11.6 µg kg−1), heptachlor (1.0 and 10.8 µg kg−1), cis-heptachlor epoxide (0.9 and 11.5 µg kg−1), trans-heptachlor epoxide (0.9 and 11.5 µg kg−1), cis-chlordane (0.4 and 7.9 µg kg−1) and trans-chlordane (0.5 and 10.9 µg kg−1), respectively, and all of them were within the maximum limits recommended by the EPA for the compounds α-HCH (86.0 and 360.0 µg kg−1), β-HCH (300.0 and 1.3 × 103 µg kg−1), γ-HCH (570.0 and 2.5 × 103 µg kg−1), δ-HCH (not defined), heptachlor (130.0 and 630.0 µg kg−1), cis-/trans-heptachlor epoxide (7.0 and 330.0 µg kg−1), cis-/trans-chlordane (1.77 × 103 and 7.7 × 103 µg kg−1) in residential and industrial soil, respectively. Recovery results were between 65% and 105% for almost all compounds, which is an optimum result for multi-residue analytical methods, considering the complexity of the matrix used in the study. Caieiras presented contamination levels of α-HCH in the range of 2.0 to 66.0 µg g−1, which was higher than the limits established by EPA, corresponding to 0.077 µg g−1 for residential soil and 0.27 µg g−1 for industrial soil. According to the validation study, the analytical method proposed was reliable for organochlorine quantification, and the QuEChERS was considered efficient for organochlorine extraction from soil
Mucoadhesive Polymers and Their Applications in Drug Delivery Systems for the Treatment of Bladder Cancer
Bladder cancer (BC) is the tenth most common type of cancer worldwide, affecting up to four times more men than women. Depending on the stage of the tumor, different therapy protocols are applied. Non-muscle-invasive cancer englobes around 70% of the cases and is usually treated using the transurethral resection of bladder tumor (TURBIT) followed by the instillation of chemotherapy or immunotherapy. However, due to bladder anatomy and physiology, current intravesical therapies present limitations concerning permeation and time of residence. Furthermore, they require several frequent catheter insertions with a reduced interval between doses, which is highly demotivating for the patient. This scenario has encouraged several pieces of research focusing on the development of drug delivery systems (DDS) to improve drug time residence, permeation capacity, and target release. In this review, the current situation of BC is described concerning the disease and available treatments, followed by a report on the main DDS developed in the past few years, focusing on those based on mucoadhesive polymers as a strategy. A brief review of methods to evaluate mucoadhesion properties is also presented; lastly, different polymers suitable for this application are discussed
Radiation-grafting of thermo- and pH-responsive poly (N-vinylcaprolactam-co-acrylic acid) onto silicone rubber and polypropylene films for biomedical purposes
This work focuses on the effects of gamma-ray irradiation conditions on the stimuli-responsiveness of polypropylene (PP) films and silicone (SR) rubber substrates grafted with N-vinylcaprolactam (NVCL) and acrylic acid (AAc). PP films and SR rubber were modified by simultaneous polymerization and grafting of NVCL and AAc, using pre-irradiation oxidative method at a dose rate of 1223 kGy h(-1) and doses ranging from 5 to 70 kGy. NVCL and AAc solutions (1/1, v/v) at 50% monomer concentration (v/v) in toluene were added to the sample substrates, degassed, sealed and heated at 60 and 70 degrees C for 12 h. After grafting, the samples were soaked in ethanol and distilled water for 24 h successively, followed by drying under vacuum. Samples were characterized by FTIR-ATR, DSC and swelling measurements. Critical points (pH critical or LCST) of grafts were obtained in a pH-environment (pH ranges from 2.2 to 9) and in a thermo-environment (temperature ranges from 22 to 50 C). Cytotoxicity evaluation was performed using fibroblast BALB/c 3T3 cells. the relationship between NVCL-co-AAc grafting and radiation dose was different for each substrate, PP and SR. At 50% NVCL/AAc concentration in toluene, grafting values were higher for SR than for PP. Despite the fact that PP-g-(NVCL-co-AAc) membrane presented a cytotoxic profile at the highest experimental concentration assayed, cytotoxicity evaluation revealed noncytotoxic profiles for the membranes synthesized highlighting their applications for biomedical purposes. (C) 2013 Elsevier B.V. All rights reserved.DGAPA-UNAMConselho Nacional de Desenvolvimento CientĂfico e TecnolĂłgico (CNPq)Ibero-American Programme for Science, and Technology and Development of CYTEDInst Pesquisas Energet & Nucl IPEN CNEN SP, BR-05508000 SĂŁo Paulo, BrazilUniv Nacl Autonoma Mexico, Inst Ciencias Nucl, Dept Quim Radiac & Radioquim, Mexico City 04510, DF, MexicoUniversidade Federal de SĂŁo Paulo UNIFESP, BR-09913030 Diadema, SP, BrazilUniversidade Federal de SĂŁo Paulo UNIFESP, BR-09913030 Diadema, SP, BrazilDGAPA-UNAM: IN202311CNPq: 174378Web of Scienc
The molecular structure of β-alanine is resistant to sterilising doses of gamma radiation.
β-alanine is the rate-limiting point for the endogenous synthesis of carnosine in skeletal muscle. Carnosine has a wide range of implications for health, normal function and exercise performance. Whilst the physiological relevance of carnosine to different tissues remains enigmatic, β-alanine administration is a useful strategy to investigate the physiological roles of carnosine in humans. Intravenous administration of β-alanine is an interesting approach to study carnosine metabolism. However, sterilisation is mandatory due to the nature of the administration route. We evaluated whether sterilising doses of gamma radiation damages the molecular structure and leads to the loss of functional characteristics of β-alanine. Pure β-alanine was sterilised by gamma radiation in sealed glass vials using a 60Co multipurpose irradiator at a dose rate of 8.5 kGy.hour-1 totalising 10, 20, 25 30 and 40 kGy. The molecular integrity was assessed by X-ray Diffraction and changes in content were determined by High Performance Liquid Chromatography (UV-HPLC) and Triple Quadrupole Mass Spectrometer (HPLC/MS-MS). Sterility assurance was evaluated by inoculation assay. To examine whether functional properties were preserved, β-alanine was infused in one participant, who rated the level of paraesthesia on the skin using a 0-3 scale. Urinary β-alanine was quantified before and 24-h following β-alanine infusion using HPLC-ESI+-MS/MS. Irradiation resulted in no change in the crystal structure of β-alanine, no degradation, and no new peaks were identified in the dose range assayed. The inoculation assay showed the absence of viable microorganisms in all β-alanine samples, including those that did not undergo irradiation. Intravenous infusion of β-alanine resulted in paraesthesia and it detected in the urine as per normal. We conclude that gamma radiation is a suitable technique for the sterilisation of β-alanine. It does not lead to degradation, damage to the β-alanine structure, content or loss of function within the evaluated irradiation conditions
Phenobarbital pharmacological findings on the nerve-muscle basis
Phenobarbital and carbamazepine are antiepileptic drugs that act at the nervous central system by different mechanisms of action. In this work we investigated the pharmacological effects of these drugs on mouse phrenic nerve-diaphragm preparations through the myographic technique. Carbamazepine (0.105, 1.05, 2.1 and 4.2 mM, n = 8, 6, 6 and 6, respectively), induced a dose-dependent neuromuscular blockade, under indirect or direct muscle stimulation and the neurotransmission was reestablished after washing. Conversely, phenobarbital caused an unexpected facilitatory effect, under several formulations, such as the acid-extracted commercial tablets (1.05, 2.1 and 4.2 mM, n = 7, 6 and 7, respectively), commercial phenobarbital solution (4.2 mM, n = 7) or its correspondent pure active ingredient (4.2 and 2.1 mM, n = 6 each). Only at a higher concentration the acid-extracted phenobarbital performed a neuromuscular blockade (8.4 mM, n = 10). The different responses between carbamazepine (paralysis) and phenobarbital (facilitatory effect) evidentiated a new effect for phenobarbital until now concealed at the neuromuscular junction and may involve the glutamatergic regulation, since its role as an acetylcholine co-transmitter in motoneurons was already established.Colegio de Farmacéuticos de la Provincia de Buenos Aire