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1.
Genome ; 67(4): 109-118, 2024 Apr 01.
Article in English | MEDLINE | ID: mdl-38316150

ABSTRACT

Charadriiformes, which comprises shorebirds and their relatives, is one of the most diverse avian orders, with over 390 species showing a wide range of karyotypes. Here, we isolated and characterized the whole collection of satellite DNAs (satDNAs) at both molecular and cytogenetic levels of one of its representative species, named the wattled jacana (Jacana jacana), a species that contains a typical ZZ/ZW sex chromosome system and a highly rearranged karyotype. In addition, we also investigate the in situ location of telomeric and microsatellite repeats. A small catalog of 11 satDNAs was identified that typically accumulated on microchromosomes and on the W chromosome. The latter also showed a significant accumulation of telomeric signals, being (GA)10 the only microsatellite with positive hybridization signals among all the 16 tested ones. These current findings contribute to our understanding of the genomic organization of repetitive DNAs in a bird species with high degree of chromosomal reorganization contrary to the majority of bird species that have stable karyotypes.


Subject(s)
Charadriiformes , Animals , Charadriiformes/genetics , DNA, Satellite/genetics , Heterochromatin/genetics , Repetitive Sequences, Nucleic Acid , Sex Chromosomes/genetics , Karyotype , Birds/genetics , Evolution, Molecular
2.
J Sep Sci ; 46(17): e2300214, 2023 Sep.
Article in English | MEDLINE | ID: mdl-37400419

ABSTRACT

The development of a fast, cost-effective, and efficient microextraction by packed sorbent setup was achieved by combining affordable laboratory-repackable devices of microextraction with a high-throughput cartesian robot. This setup was evaluated for the development of an analytical method to determine N-nitrosamines in losartan tablets. N-nitrosamines pose a significant concern in the pharmaceutical market due to their carcinogenic risk, necessitating their control and quantification in pharmaceutical products. The parameters influencing the performance of this sample preparation for N-nitrosamines were investigated through both univariate and multivariate experiments. Microextractions were performed using just 5.0 mg of carboxylic acid-modified polystyrene divinylbenzene copolymer as the extraction phase. Under the optimized conditions, the automated setup enabled the simultaneous treatment of six samples in less than 20 min, providing reliable analytical confidence for the proposed application. The analytical performance of the automated high-throughput microextraction by the packed sorbent method was evaluated using a matrix-matching calibration. Quantification was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry with chemical ionization at atmospheric pressure. The method exhibited limits of detection as low as 50 ng/g, good linearity, and satisfactory intra-day (1.38-18.76) and inter-day (2.66-20.08) precision. Additionally, the method showed accuracy ranging from 80% to 136% for these impurities in pharmaceutical formulations.


Subject(s)
Nitrosamines , Robotics , Nitrosamines/analysis , Losartan/analysis , Tandem Mass Spectrometry/methods , Limit of Detection , Solid Phase Microextraction/methods , Chromatography, Liquid , Chromatography, High Pressure Liquid/methods , Tablets
3.
Int J Mol Sci ; 24(14)2023 Jul 21.
Article in English | MEDLINE | ID: mdl-37511506

ABSTRACT

Although LC-MS with atmospheric pressure ionization (API) sources is the primary technique used in modern bioanalytical studies, electron ionization mass spectrometry (EI-MS) can provide some substantial advantages over it. EI-MS is a matrix effect-free technique that provides reproducible and comparable mass spectra, serving as a compound fingerprint for easy identification through automated comparison with spectral libraries. Leveraging EI-MS in biochemical studies can yield critical analytical benefits for targeted and untargeted analyses. However, to fully utilize EI-MS for heavy and non-volatile molecules, a new technology that enables the coupling of liquid chromatography with EI-MS is needed. Recent advancements in nanoLC have addressed the compatibility issues between LC and EI-MS, and innovative interfacing strategies such as Direct-EI, liquid electron ionization (LEI), and Cold-EI have extended the application of EI-MS beyond the determination of volatile organic molecules. This review provides an overview of the latest developments in nanoLC-EI-MS interfacing technologies, discussing their scope and limitations. Additionally, selected examples of nanoLC-EI-MS applications in the field of biochemical analysis are presented, highlighting the potential prospects and benefits that the establishment of this technique can bring to this field.


Subject(s)
Electrons , Spectrometry, Mass, Electrospray Ionization , Spectrometry, Mass, Electrospray Ionization/methods , Chromatography, Liquid/methods , Atmospheric Pressure , Technology
4.
Int J Mol Sci ; 24(10)2023 May 19.
Article in English | MEDLINE | ID: mdl-37240350

ABSTRACT

Scleropages formosus (Osteoglossiformes, Teleostei) represents one of the most valued ornamental fishes, yet it is critically endangered due to overexploitation and habitat destruction. This species encompasses three major color groups that naturally occur in allopatric populations, but the evolutionary and taxonomic relationships of S. formosus color varieties remain uncertain. Here, we utilized a range of molecular cytogenetic techniques to characterize the karyotypes of five S. formosus color phenotypes, which correspond to naturally occurring variants: the red ones (Super Red); the golden ones (Golden Crossback and Highback Golden); the green ones (Asian Green and Yellow Tail Silver). Additionally, we describe the satellitome of S. formosus (Highback Golden) by applying a high-throughput sequencing technology. All color phenotypes possessed the same karyotype structure 2n = 50 (8m/sm + 42st/a) and distribution of SatDNAs, but different chromosomal locations of rDNAs, which were involved in a chromosome size polymorphism. Our results show indications of population genetic structure and microstructure differences in karyotypes of the color phenotypes. However, the findings do not clearly back up the hypothesis that there are discrete lineages or evolutionary units among the color phenotypes of S. formosus, but another case of interspecific chromosome stasis cannot be excluded.


Subject(s)
Genome , Genomics , Animals , Fishes/genetics , Karyotype , Cytogenetic Analysis
5.
Bioinformatics ; 37(18): 3088-3090, 2021 09 29.
Article in English | MEDLINE | ID: mdl-33693580

ABSTRACT

SUMMARY: Here, we propose Fourier ring correlation-based quality estimation (FRC-QE) as a new metric for automated image quality estimation in 3D fluorescence microscopy acquisitions of cleared organoids that yields comparable measurements across experimental replicates, clearing protocols and works for different microscopy modalities. AVAILABILITY AND IMPLEMENTATION: FRC-QE is written in ImgLib2/Java and provided as an easy-to-use and macro-scriptable plugin for Fiji. Code, documentation, sample images and further information can be found under https://github.com/PreibischLab/FRC-QE. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.


Subject(s)
Imaging, Three-Dimensional , Software , Microscopy, Fluorescence
6.
Electrophoresis ; 43(15): 1587-1600, 2022 08.
Article in English | MEDLINE | ID: mdl-35531989

ABSTRACT

Mass spectrometry (MS) is a fundamental technique to identify compounds by their mass-to-charge ratio. It is known that MS can only detect target compounds when they are converted to ions in the gas phase. The ionization procedure is considered one of the most critical steps, and there are distinct techniques for it. One of them is electron ionization (EI), a widely used hard-ionization technique capable of generating several ions due to the excess energy employed. The existence of distinct ionization mechanisms turns EI capable of producing a fingerprint-like spectrum for each molecule. So, it is an essential technique for obtaining structural information. EI is often combined with chromatography to obtain a practical introduction of pretreated samples despite its excellent performance. EI-MS has been applied coupled with gas chromatography (GC) since the 1960s as both are very compatible. Currently, analytes of interest are more suitable for liquid chromatography (LC) analysis, so there are researchers dedicated to developing suitable interfaces for coupling LC and EI-MS. EI excels, as a reliable technique to fill the gap between GC and LC, possibly allowing them to coexist in a single instrument. In this work, the authors will present the fundamentals of EI-MS, emphasizing the development over the years, coupling with gas and LC, and future trends.


Subject(s)
Electrons , Chromatography, Liquid/methods , Gas Chromatography-Mass Spectrometry/methods , Ions , Mass Spectrometry
7.
J Toxicol Environ Health A ; 84(8): 345-355, 2021 04 18.
Article in English | MEDLINE | ID: mdl-33435828

ABSTRACT

Sida planicaulis is a weed thought to have originated in Brazil, where it is present in abundant quantities, but also this plant is also found in south-central Florida, Indian Ocean Islands, and the Pacific Islands. Sida planicaulis produces neurotoxicity that adversely affects livestock breeding with heavy animal losses and consequent negative impact on Brazil's economy. The aim of this study was to determine the chemical profile, cytotoxic and genotoxic effects of ethanolic extracts of S. planicaulis collected in winter (leaf extract) and summer (leaf extract and leaf + flower extract) using an in vitro model of human neuroblastoma cell line SH-SY5Y. Phytochemical screening demonstrated the presence of alkaloids, flavonoids, and apolar compounds. Rutin, quercetin, and swainsonine were detected by HPLC and GC/MS, respectively. Phosphorus, potassium, iron, and zinc were the inorganic elements found. Extracts produced cytotoxicity at all concentrations tested (7-4,000 µg/ml) as evidenced by the colorimetric assay [3-(4,5-dimethyl-thiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT)]. Based upon the alkaline comet assay extracts were found to induce genotoxicity at concentrations ranging from 0.437 to 7 µg/ml. DNA damage produced by extracts was affirmed using a modified comet assay with the enzymes Endo III and FPG in a concentration dependent manner. Further, enzyme-modified comet assay showed both oxidized purines and pyrimidines, and consequently oxidative stress was related to genomic instability and cell death. Data suggest that low concentrations of ethanolic extracts of S. planicaulis (different seasons) induced increased DNA damage related to oxidative stress and chemical composition.


Subject(s)
Cytotoxins/pharmacology , Mutagens/pharmacology , Plant Extracts/pharmacology , Sida Plant/chemistry , Cell Line, Tumor , Cytotoxins/chemistry , Humans , Mutagens/chemistry , Plant Extracts/chemistry , Seasons
8.
Dent Traumatol ; 37(3): 365-382, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33749978

ABSTRACT

BACKGROUND/AIM: Dr. Jens Ove Andreasen, the "father" of dental traumatology, passed away on September 26, 2020. As a tribute, the aim of this study was to catalog and analyze the top 100 most-cited papers (co)authored by Dr. Andreasen. MATERIALS AND METHODS: A bibliometric analysis was conducted on October 21, 2020, in the Web of Science Core Collection (WoS-CC) database. The numbers of citations in the WoS "All Databases" section, Scopus, and Google Scholar databases were cross-matched. Two researchers collected the following bibliometric data: title, number of citations, authors, country, year, journals, study design, and theme. The VOSviewer software was used to generate collaborative network maps for the authors and keywords. RESULTS: Of 194 papers identified in WoS-CC, a list was compiled with the 100 most-cited papers (co)authored by Dr. Andreasen. The number of citations of each paper ranged from 24 to 365 (mean: 85.4). Nine papers were cited more than 200 times. Most papers were published in Dental Traumatology (55%), between the 1990's and 2000's (60%), with a laboratory design (40%). One hundred and nine (co)authors, from 19 countries were identified in the top 100 most-cited papers. Dental traumatology was the most studied theme (52%). CONCLUSIONS: The top 100 most-cited papers (co)authored by Dr. Jens Ove Andreasen were composed mainly of laboratory and observational studies, published mostly in Dental Traumatology. The present study was a humble homage to highlight the importance of his research and work. However, his legacy goes beyond the numbers and will always remain unsurpassed.


Subject(s)
Bibliometrics , Traumatology , Research Design
9.
Am J Med Genet A ; 182(12): 2891-2901, 2020 12.
Article in English | MEDLINE | ID: mdl-32946203

ABSTRACT

The aim of this study was to evaluate the oral, dental, and craniofacial features of individuals affected by the chronic forms of acid sphingomyelinase deficiency (ASMD). This study comprised a sample of adult and pediatric patients (n = 8) with chronic ASMD. The individuals underwent oral examinations to evaluate the occurrence of caries, as well as full-mouth periodontal examinations, to assess the occurrence and severity of periodontal diseases. Panoramic and profile radiographs were obtained to analyze dental conditions and craniofacial parameters. Participants also answered questionnaires to identify systemic impairment, parafunctional habits, and bruxism. Dental anomalies of size, shape, and number were found, with agenesis and microdontia being the predominant findings. The average of caries experience was 11.75 (±8.1). Only one patient had periodontal health and all adult individuals had periodontitis at different stages and degrees. Bruxism was found in 87.5% of the sample. The convex profile and maxillary and mandibular retrusion were the most relevant findings in the cephalometric analysis. It is concluded that individuals with chronic ASMD, in addition to several systemic manifestations, present significant modifications in their oral health, from a greater occurrence of dental anomalies, caries, periodontal disease, in addition to skeletal changes.


Subject(s)
Bruxism/pathology , Craniofacial Abnormalities/pathology , Mouth Diseases/pathology , Niemann-Pick Disease, Type B/complications , Periodontal Diseases/pathology , Sphingomyelin Phosphodiesterase/deficiency , Tooth Abnormalities/pathology , Adolescent , Adult , Bruxism/etiology , Child , Craniofacial Abnormalities/etiology , Female , Humans , Male , Middle Aged , Mouth Diseases/etiology , Niemann-Pick Disease, Type B/enzymology , Periodontal Diseases/etiology , Prognosis , Tooth Abnormalities/etiology , Young Adult
10.
Anal Bioanal Chem ; 412(28): 7789-7797, 2020 Nov.
Article in English | MEDLINE | ID: mdl-32929570

ABSTRACT

Cachaça or "sugarcane spirit" is a Brazilian beverage considered the third most consumed beverage worldwide. Sugarcane, its raw material, is one of the main crops developed in the country, placing Brazil as the largest producer of this commodity on a global scale. Considering the growth in sugarcane production, many farmers use pesticides in their crops. However, excess pesticides can be accumulated in products derived from sugarcane, creating an environmental and public health concern. In this context, the development of analytical methods capable of identifying residues of pesticides in cachaças and other sugarcane-derived products is essential to ensure the beverage's quality. This work presents a method to quantify multiclass pesticides in Brazilian sugarcane spirits (cachaças) through an automated multidimensional system. The first dimension consists of an extraction column packed with a graphene-silica phase, followed by a capillary liquid chromatography-tandem mass spectrometry system as the second dimension. The method was optimized by an experimental design, in which the influence of three variables was evaluated on the extraction process: percentage of acetonitrile, loading flow, and loading time. Afterward, twenty-two cachaças were analyzed to ascertain the applicability of the proposed method. The analyses reported five samples containing clomazone (a type of herbicide widely used in sugarcane production). The method showed good linearity under optimized conditions, with correlation coefficients greater than 0.981, and limits of detection and quantification of 5 µg L-1 and 10 µg L-1, respectively. The herein discussed results suggest that the proposed method could be a practical option for identifying pesticides in beverages. Graphical Abstract.


Subject(s)
Beverages/analysis , Chromatography, Liquid/methods , Pesticide Residues/analysis , Saccharum/chemistry , Tandem Mass Spectrometry/methods , Crops, Agricultural/chemistry , Limit of Detection , Reference Standards , Reproducibility of Results
11.
J Fish Biol ; 97(3): 776-784, 2020 Sep.
Article in English | MEDLINE | ID: mdl-32535919

ABSTRACT

Resource partitioning has an essential role in interspecific relations, especially in congener species, which share many morphological traits. In some places, small characids coexist through resource partitioning, which may reduce their interspecific competition. Astyanax species (e.g., Astyanax minor, Astyanax gymnodontus and Astyanax bifasciatus), for example, coexist in different water bodies from the Iguaçu River basin. These species have high phenotypic plasticity and many morphologic specializations that allow them to live in different habitats. Based on evidences that these species modified their feeding habits because of changes in resource availability in Iguaçu River, this study tested two hypotheses: (a) there are differences in head morphology, number of teeth and number of gill rakers among the species of Astyanax; and (b) there are differences in gill arch and gill raker morphology among the species of Astyanax, which may favour their trophic resource partitioning in Iguaçu River. The head morphological traits and quantitative morphological characters were summarized in a principal coordinates analysis (PCoA), and the analysis of similarities (ANOSIM) showed significant differences among species. Gill morphological measurements were analysed through analysis of covariance (ANCOVA), and it also showed significant differences in gill arch and gill raker morphology among species. Therefore, the analysis of ecomorphological traits related to trophic habits revealed some differences that may suggest a tendency of reducing competition for trophic resources in the Iguaçu River basin.


Subject(s)
Characidae/anatomy & histology , Food Chain , Rivers , Animals , Brazil , Gills/anatomy & histology , Nutritional Status , Species Specificity , Tropical Climate
12.
J Cell Physiol ; 234(2): 1398-1415, 2019 02.
Article in English | MEDLINE | ID: mdl-30078202

ABSTRACT

The mechanisms of cancer involve changes in multiple biological pathways. Multitarget molecules, which are components of animal venoms, are therefore a potential strategy for treating tumors. The objective of this study was to screen the effects of Phoneutria nigriventer spider venom (PnV) on tumor cell lines. Cultured human glioma (NG97), glioblastoma (U-251) and cervix adenocarcinoma (HeLa) cells, and nontumor mouse fibroblasts (L929) were treated with low (14 µg/ml) and high (280 µg/ml) concentrations of PnV, and analyzed through assays for cell viability (thiazolyl blue tetrazolium blue), proliferation (carboxyfluorescein succinimidyl ester), death (annexin V/propidium iodide [Pi]), the cell cycle (Pi), and migration (wound healing and transwell assay). The venom decreased the viability of U-251 cells, primarily by inducing cell death, and reduced the viability of NG97 cells, primarily by inhibiting the cell cycle. The migration of all the tumor cell lines was delayed when treated with venom. The venom significantly affected all the tumor cell lines studied, with no cytotoxic effect on normal cells (L929), although the nonglial tumor cell (HeLa) was less sensitive to PnV. The results of the current study suggest that PnV may be composed of peptides that are highly specific for the multiple targets involved in the hallmarks of cancer. Experiments are underway to identify these molecules.


Subject(s)
Adenocarcinoma/drug therapy , Antineoplastic Agents/pharmacology , Brain Neoplasms/drug therapy , Cell Cycle Checkpoints/drug effects , Cell Movement/drug effects , Glioma/drug therapy , Spider Venoms/pharmacology , Uterine Cervical Neoplasms/drug therapy , Adenocarcinoma/pathology , Animals , Apoptosis/drug effects , Brain Neoplasms/pathology , Cell Proliferation/drug effects , Cell Survival/drug effects , Female , Glioma/pathology , HeLa Cells , Humans , Male , Mice , Necrosis , Neoplasm Invasiveness , Uterine Cervical Neoplasms/pathology
13.
Int Rev Psychiatry ; 31(7-8): 603-607, 2019.
Article in English | MEDLINE | ID: mdl-31612743

ABSTRACT

In line with studies from around the world, several studies have shown equal or higher rates of anxiety, depression, alcohol and other drug use, and burnout in medical students in Brazil. The aim of the present study was to identify difficulties Brazilian medical students face regarding stressors, psychological distress, and psychiatric morbidity using standardized reliable and valid instruments. In addition, this study also investigated differences associated with sex. Students (n = 129) were asked to complete a one-off survey from January to June 2019, including demographics, previous mental health issues, CAGE, GHQ-12, and OLBI. A higher prevalence of previous mental disorders was found in comparison to during the medical course. This may suggest that perhaps students are under more stress or that they form a more vulnerable sub-population in medical school in comparison with the general population. However, the types of mental disorders before and during the undergraduate course appear to vary. During the course, they show higher levels of anxiety rather than that of depression. Before commencing the course, they showed higher levels of depression rather than anxiety. Cannabis and ecstasy experimentation levels were higher than those of students from other courses in previous studies. The opposite was true for stimulants. Just over a quarter reported alcohol abuse above the CAGE cut-off point. The levels of psychological distress and burnout were extremely high, and women had even higher levels. Male respondents were more likely to have a history of mental ill-health, but female subjects scored higher on burnout. Overall over 80% students reported burnout. These findings are discussed in the context of Brazilian culture.


Subject(s)
Anxiety/epidemiology , Depression/epidemiology , Mental Health , Stress, Psychological/psychology , Students, Medical/statistics & numerical data , Alcoholism/psychology , Brazil/epidemiology , Female , Humans , Male , Prevalence , Schools, Medical/statistics & numerical data , Sex Factors , Students, Medical/psychology , Surveys and Questionnaires
14.
Clin Oral Investig ; 23(10): 3713-3720, 2019 Oct.
Article in English | MEDLINE | ID: mdl-30623309

ABSTRACT

OBJECTIVES: The aim of this study was to investigate the effect of a dentifrice that contains calcium silicate, sodium phosphate, and fluoride on erosive-abrasive enamel wear. MATERIAL AND METHODS: This randomized, single-blind in situ/ex vivo study was conducted with four crossover phases of 5 days (one group tested per phase). Bovine enamel blocks (n = 256) were allocated to 16 volunteers and 8 groups. The groups under study were test dentifrice, with calcium silicate, sodium phosphate, and 1450 ppm sodium monofluorophosphate; tin dentifrice, with 3500 ppm stannous chloride, 700 ppm amine fluoride, and 700 ppm sodium fluoride; conventional dentifrice, with 1450 ppm sodium monofluorophosphate; and control (deionized water). Half of the enamel blocks were subjected to erosion and the other half to erosion plus abrasion. The daily extraoral protocol consisted in four citric acid exposures (2 min) and two applications of dentifrice slurry on all blocks for 30 s; after, half of the blocks were brushed for 15 s. The response variable was enamel loss. Data were analyzed by two-way ANOVA and Fisher's test (p < 0.05). RESULTS: For erosion, the test dentifrice promoted less enamel loss than water (4.7 ± 3.1 and 5.8 ± 2.5 µm, respectively, p < 0.05), and did not differ from tin (4.8 ± 2.5 µm) and conventional (4.8 ± 1.4 µm) dentifrices (p > 0.05). However, the test dentifrice (7.7 ± 3.8 µm) promoted higher wear after erosive plus abrasive procedures than tin (5.4 ± 1.5 µm) and conventional (6.2 ± 1.7 µm, p < 0.05) dentifrices, and did not differ from water (6.9 ± 2.0 µm). CONCLUSIONS: The investigated dentifrice reduced enamel loss against acid challenge but had no effect against acid and brushing challenge. CLINICAL RELEVANCE: Little is known regarding the preventive effect of dentifrices indicated for dental erosion. The tested anti-erosive dentifrice was unable to protect enamel when erosion was associated to toothbrushing abrasion.


Subject(s)
Calcium Compounds/chemistry , Dentifrices/chemistry , Phosphates/chemistry , Silicates/chemistry , Sodium Fluoride/chemistry , Tooth Erosion/prevention & control , Animals , Cattle , Cross-Over Studies , Dental Enamel , Humans , Single-Blind Method , Toothbrushing
15.
Drug Dev Ind Pharm ; 44(5): 723-728, 2018 May.
Article in English | MEDLINE | ID: mdl-29192518

ABSTRACT

OBJECTIVE: This study aimed to develop and validate an in vitro dissolution method based on in silico-in vivo data to determine whether an in vitro-in vivo relationship could be established for rivaroxaban in immediate-release tablets. SIGNIFICANCE: Oral drugs with high permeability but poorly soluble in aqueous media, such as the anticoagulant rivaroxaban, have a major potential to reach a high level of in vitro-in vivo relationship. Currently, there is no study on scientific literature approaching the development of RIV dissolution profile based on its in vivo performance. METHODS AND RESULTS: Drug plasma concentration values were modeled using computer simulation with adjustment of pharmacokinetic properties. Those values were converted into drug fractions absorbed by the Wagner-Nelson deconvolution approach. Gradual and continuous dissolution of RIV tablets was obtained with a 30 rpm basket on 50 mM sodium acetate +0.2% SDS, pH 6.5 medium. Dissolution was conducted for up to 180 min. The fraction absorbed was plotted against the drug fraction dissolved, and a linear point-to-point regression (R2 = 0.9961) obtained. CONCLUSION: The in vitro dissolution method designed promoted a more convenient dissolution profile of RIV tablets, whereas it suggests a better relationship with in vivo performance.


Subject(s)
Rivaroxaban/chemistry , Solubility , Tablets/chemistry , Computer Simulation , In Vitro Techniques , Linear Models , Permeability
16.
Sci Rep ; 14(1): 308, 2024 01 03.
Article in English | MEDLINE | ID: mdl-38172290

ABSTRACT

Alzheimer's disease (AD) is considered the leading cause of dementia in the elderly worldwide. It results in progressive memory loss and impairment of cognitive and motor skills, leading to a high degree of disability and dependence. The development of AD is associated with the accumulation of senile plaques in the brain, caused by the amyloidogenic pathway of the disease. Several genetic and biochemical events are linked to AD development, with oxidative stress being one of them. Due to the scarcity of drugs aimed at treating AD, antioxidant compounds are increasingly studied as therapeutic targets for the disease. In this study, we investigate the antioxidant and anti-Alzheimer potential of the Tetragonisca angustula (Jataí) pollen extract in a Drosophila melanogaster Alzheimer's model. For this purpose, we utilized a D. melanogaster AD-like model, which expresses genes related to the amyloidogenic pathway of Alzheimer's disease. We explored the floral origin of the collected pollen, conducted phytochemical prospecting, and evaluated its antioxidant capacity in vitro. In vivo experiments involved assessing the survival and climbing ability of the D. melanogaster AD-like model with various concentrations of the pollen extract. Our findings revealed that the pollen extract of Tetragonisca angustula exhibits a significant antioxidant response and high concentrations of important phytochemicals, such as flavonoids and polyphenols. Furthermore, it enhanced the survival rate of D. melanogaster, and across all concentrations tested, it improved the climbing ability of the flies after 15 days of treatment with methanolic pollen extract. Additionally, the pollen extract reduced the neurodegeneration index in histopathological analysis. Thus, our study demonstrates the potential of Tetragonisca angustula pollen as an important subject for further investigation, aiming to isolate molecules that could potentially serve as therapeutic targets for Alzheimer's disease.


Subject(s)
Alzheimer Disease , Antioxidants , Humans , Bees , Animals , Aged , Antioxidants/pharmacology , Antioxidants/therapeutic use , Alzheimer Disease/metabolism , Drosophila melanogaster , Pollen/metabolism , Plant Extracts/pharmacology , Plant Extracts/therapeutic use
17.
Biotechnol Prog ; 40(3): e3419, 2024.
Article in English | MEDLINE | ID: mdl-38247123

ABSTRACT

Extracellular vesicles (EVs) derived from mesenchymal stromal cells (MSCs) have shown increasing therapeutic potential in the last years. However, large production of EV is required for therapeutic purposes. Thereby, scaling up MSC cultivation in bioreactors is essential to allow culture parameters monitoring. In this study, we reported the establishment of a scalable bioprocess to produce MSC-EV in suspension cultures using spinner flasks and human collagen-coated microcarriers (3D culture system). We compared the EV production in this 3D culture system with the standard static culture using T-flasks (2D culture system). The EV produced in both systems were characterized and quantify by western blotting and nanoparticle tracking analysis. The presence of the typical protein markers CD9, CD63, and CD81 was confirmed by western blotting analyses for EV produced in both culture systems. The cell fold-increase was 5.7-fold for the 3D culture system and 4.6-fold for the 2D culture system, signifying a fold-change of 1.2 (calculated as the ratio of fold-increase 3D to fold-increase 2D). Furthermore, it should be noted that the total cell production in the spinner flask cultures was 4.8 times higher than that in T-flask cultures. The total cell production in the spinner flask cultures was 5.2-fold higher than that in T-flask cultures. While the EV specific production (particles/cell) in T-flask cultures (4.40 ± 1.21 × 108 particles/mL, p < 0.05) was higher compared to spinner flask cultures (2.10 ± 0.04 × 108 particles/mL, p < 0.05), the spinner flask culture system offers scalability, making it capable of producing enough MSC-EV at a large scale for clinical applications. Therefore, we concluded that 3D culture system evaluated here serves as an efficient transitional platform that enables the scaling up of MSC-EV production for therapeutic purposes by utilizing stirred tank bioreactors and maintaining xeno-free conditions.


Subject(s)
Cell Culture Techniques , Extracellular Vesicles , Mesenchymal Stem Cells , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Extracellular Vesicles/metabolism , Extracellular Vesicles/chemistry , Humans , Cell Culture Techniques/methods , Bioreactors , Cells, Cultured
18.
Adv Rheumatol ; 64(1): 2, 2024 01 02.
Article in English | MEDLINE | ID: mdl-38167239

ABSTRACT

Systemic lupus erythematosus (SLE) is an autoimmune disease that can affect several organs and systems. The central and/or peripheral nervous system can suffer from complications known as neuropsychiatric lupus (NPSLE). Studies have associated the manifestations of SLE or NPSLE with vitamin D deficiency. It has been shown that hypovitaminosis D can lead to cognition deficits and cerebral hypoperfusion in patients with NPSLE. In this review article, we will address the main features related to vitamin D supplementation or serum vitamin D levels with neuropsychiatric manifestations, either in patients or in animal models of NPSLE.


Subject(s)
Lupus Erythematosus, Systemic , Lupus Vasculitis, Central Nervous System , Animals , Humans , Lupus Vasculitis, Central Nervous System/drug therapy , Lupus Vasculitis, Central Nervous System/complications , Vitamin D/therapeutic use , Lupus Erythematosus, Systemic/complications , Lupus Erythematosus, Systemic/drug therapy
19.
Genes (Basel) ; 15(2)2024 02 19.
Article in English | MEDLINE | ID: mdl-38397247

ABSTRACT

Vanellus (Charadriidae; Charadriiformes) comprises around 20 species commonly referred to as lapwings. In this study, by integrating cytogenetic and genomic approaches, we assessed the satellite DNA (satDNA) composition of one typical species, Vanellus chilensis, with a highly conserved karyotype. We additionally underlined its role in the evolution, structure, and differentiation process of the present ZW sex chromosome system. Seven distinct satellite DNA families were identified within its genome, accumulating on the centromeres, microchromosomes, and the W chromosome. However, these identified satellite DNA families were not found in two other Charadriiformes members, namely Jacana jacana and Calidris canutus. The hybridization of microsatellite sequences revealed the presence of a few repetitive sequences in V. chilensis, with only two out of sixteen displaying positive hybridization signals. Overall, our results contribute to understanding the genomic organization and satDNA evolution in Charadriiform birds.


Subject(s)
Charadriiformes , Animals , Charadriiformes/genetics , DNA, Satellite/genetics , Birds/genetics , Sex Chromosomes , Repetitive Sequences, Nucleic Acid
20.
Discov Nano ; 19(1): 92, 2024 May 27.
Article in English | MEDLINE | ID: mdl-38801473

ABSTRACT

This study explores the green synthesis of silver nanoparticles (AgNPs) using a methanolic extract of fermented pollen from Tetragonisca angustula, a species of stingless bees. The AgNPs exhibit spherical morphology, low charge values, and suspension stability, with their unique composition attributed to elements from the pollen extract. Antioxidant assays show comparable activity between the pollen extract and AgNPs, emphasizing the retention of antioxidant effects. The synthesized AgNPs demonstrate antimicrobial activity against multidrug-resistant bacteria, highlighting their potential in combating bacterial resistance. The AgNPs exhibit no toxic effects on Drosophila melanogaster and even enhance the hatching rate of eggs. The study underscores the innovative use of stingless bee pollen extract in green synthesis, offering insights into the varied applications of AgNPs in biomedicine.

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