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1.
Heliyon ; 10(17): e37331, 2024 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-39296175

RESUMO

The elasmobranch population is declining in the Bay of Bengal of Bangladesh due to large-mesh gill net fishing, locally known as the Lakkha net, which primarily targets Indian threadfin (Leptomelanosoma indicum). This study was the first attempt to identify megafaunal bycatch in Lakkha fishing and assess its vulnerability using Productivity Susceptibility Analysis. A total of 40 elasmobranch bycatch species were identified, with sharks comprising 13 species from three families, while 27 rays belonged to six families, with the majority belonging to the Myliobatiformes order (60 %). Productivity and susceptibility scores were assigned to all identified species, with values ranging from 1.27 to 2.73 and 1.50 to 2.63, respectively. The target Lakkha fish exhibited the highest susceptibility score, followed by several pelagic sharks and eagle rays. Vulnerability assessment revealed that 31.7 % (n = 13) of species were highly vulnerable, while 43.9 % (n = 18) were classified as moderate, and 24.4 % (n = 10) were considered to have low vulnerability. All the high-risk megafauna species (n = 13) are classified as threatened by the global IUCN Red List. Sensitivity analysis highlighted susceptibility as a major contributor to species' vulnerability. Alterations in susceptibility scores led to significant changes in the vulnerability status of many species. The overall data quality assessment indicated moderate data quality across species, with variability observed between productivity (76 % of species received a poor data quality score) and susceptibility attributes. However, vulnerability of these species can be reduced through adequate gear modification, shorter net deployment periods, adoption of safe discharge techniques, identification of critical habitats, and establishment of marine protected areas within this region. This study provides valuable insights into the species composition and vulnerability of elasmobranchs in the Lakkha gill net fishery, emphasizing the need for conservation measures to mitigate bycatch impacts on threatened species.

2.
ACS Sens ; 2024 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-39289811

RESUMO

A comprehensive investigation into how nanostructures alter real-time DNA hybridization kinetics in both buffer and complex media and under a wide range of probe and target concentrations is currently lacking. In response, we use a real-time, wash-free, and in situ assay to study DNA hybridization kinetics by performing continuous electrochemical measurements in different media. We investigated the differences in hybridization kinetics under three regimes of probe density (low, medium, and high) and over three orders of magnitude of target concentrations (0.01-1 µM). Additionally, we compared the performance of planar and nanostructured electrodes in buffer, blood, urine, and saliva. Our experiments indicate that adding nanostructures to the transducer surface is only effective under a specific probe/target concentration regime. Additionally, we found that direct electrochemical readout is possible in the examined physiological media, with measurements in blood showing the highest and saliva showing the lowest signal magnitudes compared to buffer.

3.
Biol Trace Elem Res ; 2024 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-39048802

RESUMO

This study investigated heavy metal accumulations in fish, water and sediment from three rivers around a major urban area in Bangladesh, namely the Rupsha, the Atai, and the Bhairab with a view to assessing the ecological and human health risks. Samples were collected from 10 stations over two seasons (summer and winter) and concentrations of 11 metals (As, Se, Pb, Be, Cd, Co, Cr, Cu, Mn, Ni, V) were measured using ICP-MS. Heavy metals in water of these rivers were above the WHO higher thresholds. The Rupsha River, which runs close to industrially dense areas and the downstream part of the three-river network, demonstrated the highest As, Cu, and V concentrations during both seasons. On the other hand, As, Mn and Cr were highest in the Bhairab which is the upstream to the Rupsha and connected to several industrial setups which differ from Bhairab. The less anthropogenically connected Atai River only showed elevated concentrations of Cu and Se. Ecological risk indices indicated low pollution in the rivers during both seasons. In all three rivers and in nearly all fish species, the contamination was higher in winter than summer. Benthic and carnivorous fish species such as, Cynoglossus lingua, Glossogobius giuris, Pseudapocryptes elongatus showed higher metal accumulation compared to other species. Health risk indices like the target hazard quotient (THQ) and carcinogenic risk (CR) suggested low risks but pointed potential risks to human health. The outcomes of this research reports insights into metal contamination pattern in interconnected river systems.

4.
Angew Chem Int Ed Engl ; 63(20): e202400413, 2024 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-38458987

RESUMO

High-precision viral detection at point of need with clinical samples plays a pivotal role in the diagnosis of infectious diseases and the control of a global pandemic. However, the complexity of clinical samples that often contain very low viral concentrations makes it a huge challenge to develop simple diagnostic devices that do not require any sample processing and yet are capable of meeting performance metrics such as very high sensitivity and specificity. Herein we describe a new single-pot and single-step electrochemical method that uses real-time kinetic profiling of the interaction between a high-affinity aptamer and an antigen on a viral surface. This method generates many data points per sample, which when combined with machine learning, can deliver highly accurate test results in a short testing time. We demonstrate this concept using both SARS-CoV-2 and Influenza A viruses as model viruses with specifically engineered high-affinity aptamers. Utilizing this technique to diagnose COVID-19 with 37 real human saliva samples results in a sensitivity and specificity of both 100 % (27 true negatives and 10 true positives, with 0 false negative and 0 false positive), which showcases the superb diagnostic precision of this method.


Assuntos
Aptâmeros de Nucleotídeos , COVID-19 , Técnicas Eletroquímicas , Aprendizado de Máquina , SARS-CoV-2 , Aptâmeros de Nucleotídeos/química , Humanos , SARS-CoV-2/isolamento & purificação , SARS-CoV-2/imunologia , Técnicas Eletroquímicas/métodos , COVID-19/diagnóstico , COVID-19/virologia , Cinética , Vírus da Influenza A , Antígenos Virais/análise , Antígenos Virais/imunologia , Técnicas Biossensoriais/métodos
5.
Heliyon ; 10(6): e27795, 2024 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-38496905

RESUMO

Bangladesh's subtropical climate with an abundance of sunlight throughout the greater portion of the year results in increased effectiveness of solar panels. Solar irradiance forecasting is an essential aspect of grid-connected photovoltaic systems to efficiently manage solar power's variation and uncertainty and to assist in balancing power supply and demand. This is why it is essential to forecast solar irradiation accurately. Many meteorological factors influence solar irradiation, which has a high degree of fluctuation and uncertainty. Predicting solar irradiance multiple steps ahead makes it difficult for forecasting models to capture long-term sequential relationships. Attention-based models are widely used in the field of Natural Language Processing for their ability to learn long-term dependencies within sequential data. In this paper, our aim is to present an attention-based model framework for multivariate time series forecasting. Using data from two different locations in Bangladesh with a resolution of 30 min, the Attention-based encoder-decoder, Transformer, and Temporal Fusion Transformer (TFT) models are trained and tested to predict over 24 steps ahead and compared with other forecasting models. According to our findings, adding the attention mechanism significantly increased prediction accuracy and TFT has shown to be more precise than the rest of the algorithms in terms of accuracy and robustness. The obtained mean square error (MSE), the mean absolute error (MAE), and the coefficient of determination (R2) values for TFT are 0.151, 0.212, and 0.815, respectively. In comparison to the benchmark and sequential models (including the Naive, MLP, and Encoder-Decoder models), TFT has a reduction in the MSE and MAE of 8.4-47.9% and 6.1-22.3%, respectively, while R2 is raised by 2.13-26.16%. The ability to incorporate long-distance dependency increases the predictive power of attention models.

6.
Langmuir ; 2024 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-38325360

RESUMO

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the gastrointestinal tract with a complex and multifactorial etiology, making it challenging to treat. While recent advances in immunomodulatory biologics, such as antitumor necrosis factor-α (TNF-α) antibodies, have shown moderate success, systemic administration of antibody therapeutics may lead to several adverse effects, including the risk of autoimmune disorders due to systemic cytokine depletion. Transient RNA interference using exogenous short interfering RNA (siRNA) to regulate target gene expression at the transcript level offers an alternative to systemic immunomodulation. However, siRNAs are susceptible to premature degradation and have poor cellular uptake. Graphene oxide (GO) nanoparticles have been shown to be effective nanocarriers for biologics due to their reduced cytotoxicity and enhanced bioavailability. In this study, we evaluate the therapeutic efficacy of GO mediated TNF-α_siRNA using in vitro models of chronic inflammation generated by treating murine small intestines (enteroids) and large intestines (colonoids) with inflammatory agents IL-1ß, TNF-α, and LPS. The organotypic mouse enteroids and colonoids developed an inflammatory phenotype similar to that of IBD, characterized by impaired epithelial homeostasis and an increased production of inflammatory cytokines such as TNF-α, IL-1ß, and IL-6. We assessed siRNA delivery to these inflamed organoids using three different GO formulations. Out of the three, small-sized GO with polymer and dendrimer modifications (smGO) demonstrated the highest transfection efficiency, which led to the downregulation of inflammatory cytokines, indicating an attenuation of the inflammatory phenotype. Moreover, the transfection efficiency and inflammation-ameliorating effects could be further enhanced by increasing the TNF-α_siRNA/smGO ratio from 1:1 to 3:1. Overall, the results of this study demonstrate that ex vivo organoids with disease-specific phenotypes are invaluable models for assessing the therapeutic potential of nanocarrier-mediated drug and biologic delivery systems.

7.
ACS Appl Mater Interfaces ; 15(48): 55287-55296, 2023 Dec 06.
Artigo em Inglês | MEDLINE | ID: mdl-37976404

RESUMO

Healthcare-acquired infections place a significant burden on the cost and quality of patient care in hospitals. Reducing contamination on surfaces within healthcare environments is critical for halting the spread of these infections. Herein, we report a bifunctional─repel and kill─surface developed using photoactive TiO2 nanoparticles integrated into a hierarchical scaffold (OmniKill). To quantify the repellency of OmniKill, we developed a touch-based assay, capable of simulating the transfer of individual pathogens, multiple pathogens, or pathogen-latent fecal matter from hands to surfaces. OmniKill repels bacterial pathogens by at least 2.77-log (99.8%). The photoactive material within OmniKill further reduces the viability of transferred pathogens on the surface by an additional 2.43-log (99.6%) after 1 h of light exposure. The antipathogenic effects─repel and kill─remain robust under complex biological contaminates such as feces. These findings show the potential use of OmniKill in reducing the physical transmission of bacterial pathogens in healthcare settings.


Assuntos
Anti-Infecciosos , Humanos , Bactérias , Propriedades de Superfície
8.
Biosens Bioelectron ; 235: 115359, 2023 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-37187062

RESUMO

Photoelectrochemical (PEC) signal transduction is of great interest for ultrasensitive biosensing; however, signal-on PEC assays that do not require target labeling remain elusive. In this work, we developed a signal-on biosensor that uses nucleic acids to modulate PEC currents upon target capture. Target presence removes a biorecognition probe from a DNA duplex carrying a gold nanoparticle, bringing the gold nanoparticle in direct contact to the photoelectrode and increasing the PEC current. This assay was used to develop a universal bacterial detector by targeting peptidoglycan using an aptamer, demonstrating a limit-of-detection of 82 pg/mL (13 pM) in buffer and 239 pg/mL (37 pM) in urine for peptidoglycan and 1913 CFU/mL forEscherichia coliin urine. When challenged with a panel of unknown targets, the sensor identified samples with bacterial contamination versus fungi. The versatility of the assay was further demonstrated by analyzing DNA targets, which yielded a limit-of-detection of 372 fM.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Técnicas Eletroquímicas , Ouro , Peptidoglicano , DNA , Oligonucleotídeos
9.
Adv Sci (Weinh) ; 10(19): e2207223, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37088731

RESUMO

There is a need for point-of-care bacterial sensing and identification technologies that are rapid and simple to operate. Technologies that do not rely on growth cultures, nucleic acid amplification, step-wise reagent addition, and complex sample processing are the key for meeting this need. Herein, multiple materials technologies are integrated for overcoming the obstacles in creating rapid and one-pot bacterial sensing platforms. Liquid-infused nanoelectrodes are developed for reducing nonspecific binding on the transducer surface; bacterium-specific RNA-cleaving DNAzymes are used for bacterial identification; and redox DNA barcodes embedded into DNAzymes are used for binding-induced electrochemical signal transduction. The resultant single-step and one-pot assay demonstrates a limit-of-detection of 102 CFU mL-1 , with high specificity in identifying Escherichia coli amongst other Gram positive and negative bacteria including Klebsiella pneumoniae, Staphylococcus aureus, and Bacillus subtilis. Additionally, this assay is evaluated for analyzing 31 clinically obtained urine samples, demonstrating a clinical sensitivity of 100% and specify of 100%. When challenging this assay with nine clinical blood cultures, E. coli-positive and E. coli-negative samples can be distinguished with a probability of p < 0.001.


Assuntos
DNA Catalítico , Escherichia coli , Escherichia coli/genética , Sensibilidade e Especificidade , Bactérias , DNA
10.
iScience ; 26(2): 105945, 2023 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-36866046

RESUMO

The bendability of genomic DNA impacts chromatin packaging and protein-DNA binding. However, we do not have a comprehensive understanding of the motifs influencing DNA bendability. Recent high-throughput technologies such as Loop-Seq offer an opportunity to address this gap but the lack of accurate and interpretable machine learning models still remains. Here we introduce DeepBend, a convolutional neural network model with convolutions designed to directly capture the motifs underlying DNA bendability and their periodic occurrences or relative arrangements that modulate bendability. DeepBend consistently performs on par with alternative models while giving an extra edge through mechanistic interpretations. Besides confirming the known motifs of DNA bendability, DeepBend also revealed several novel motifs and showed how the spatial patterns of motif occurrences influence bendability. DeepBend's genome-wide prediction of bendability further showed how bendability is linked to chromatin conformation and revealed the motifs controlling the bendability of topologically associated domains and their boundaries.

11.
J Cachexia Sarcopenia Muscle ; 13(6): 3106-3121, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36254806

RESUMO

BACKGROUND: Human pluripotent stem cell-derived muscle models show great potential for translational research. Here, we describe developmentally inspired methods for the derivation of skeletal muscle cells and their utility in skeletal muscle tissue engineering with the aim to model skeletal muscle regeneration and dystrophy in vitro. METHODS: Key steps include the directed differentiation of human pluripotent stem cells to embryonic muscle progenitors followed by primary and secondary foetal myogenesis into three-dimensional muscle. To simulate Duchenne muscular dystrophy (DMD), a patient-specific induced pluripotent stem cell line was compared to a CRISPR/Cas9-edited isogenic control line. RESULTS: The established skeletal muscle differentiation protocol robustly and faithfully recapitulates critical steps of embryonic myogenesis in two-dimensional and three-dimensional cultures, resulting in functional human skeletal muscle organoids (SMOs) and engineered skeletal muscles (ESMs) with a regeneration-competent satellite-like cell pool. Tissue-engineered muscle exhibits organotypic maturation and function (up to 5.7 ± 0.5 mN tetanic twitch tension at 100 Hz in ESM). Contractile performance could be further enhanced by timed thyroid hormone treatment, increasing the speed of contraction (time to peak contraction) as well as relaxation (time to 50% relaxation) of single twitches from 107 ± 2 to 75 ± 4 ms (P < 0.05) and from 146 ± 6 to 100 ± 6 ms (P < 0.05), respectively. Satellite-like cells could be documented as largely quiescent PAX7+ cells (75 ± 6% Ki67- ) located adjacent to muscle fibres confined under a laminin-containing basal membrane. Activation of the engineered satellite-like cell niche was documented in a cardiotoxin injury model with marked recovery of contractility to 57 ± 8% of the pre-injury force 21 days post-injury (P < 0.05 compared to Day 2 post-injury), which was completely blocked by preceding irradiation. Absence of dystrophin in DMD ESM caused a marked reduction of contractile force (-35 ± 7%, P < 0.05) and impaired expression of fast myosin isoforms resulting in prolonged contraction (175 ± 14 ms, P < 0.05 vs. gene-edited control) and relaxation (238 ± 22 ms, P < 0.05 vs. gene-edited control) times. Restoration of dystrophin levels by gene editing rescued the DMD phenotype in ESM. CONCLUSIONS: We introduce human muscle models with canonical properties of bona fide skeletal muscle in vivo to study muscle development, maturation, disease and repair.


Assuntos
Distrofia Muscular de Duchenne , Células Satélites de Músculo Esquelético , Humanos , Distrofia Muscular de Duchenne/genética , Músculo Esquelético/metabolismo , Desenvolvimento Muscular/genética , Células Satélites de Músculo Esquelético/metabolismo , Fibras Musculares Esqueléticas/metabolismo
12.
Molecules ; 27(16)2022 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-36014550

RESUMO

This investigation is motivated by increasing interest in the development of magnetically ordered pseudocapacitors (MOPC), which exhibit interesting magnetocapacitive effects. Here, advanced pseudocapacitive properties of magnetic CuFe2O4 nanoparticles in negative potential range are reported, suggesting that CuFe2O4 is a promising MOPC and advanced negative electrode material for supercapacitors. A high capacitance of 2.76 F cm-2 is achieved at a low electrode resistance in a relatively large potential window of 0.8 V. The cyclic voltammograms and galvanostatic charge-discharge data show nearly ideal pseudocapacitive behavior. Good electrochemical performance is achieved at a high active mass loading due to the use of chelating molecules of ammonium salt of purpuric acid (ASPA) as a co-dispersant for CuFe2O4 nanoparticles and conductive multiwalled carbon nanotube (MCNT) additives. The adsorption of ASPA on different materials is linked to structural features of ASPA, which allows for different interaction and adsorption mechanisms. The combination of advanced magnetic and pseudocapacitive properties in a negative potential range in a single MOPC material provides a platform for various effects related to the influence of pseudocapacitive/magnetic properties on magnetic/pseudocapacitive behavior.

13.
Cells ; 11(13)2022 06 27.
Artigo em Inglês | MEDLINE | ID: mdl-35805117

RESUMO

Understanding changes in the expression of genes involved in regulating various components of the neural extracellular matrix (ECM) during aging can provide an insight into aging-associated decline in synaptic and cognitive functions. Hence, in this study, we compared the expression levels of ECM-related genes in the hippocampus of young, aged and very aged mice. ECM gene expression was downregulated, despite the accumulation of ECM proteoglycans during aging. The most robustly downregulated gene was carbohydrate sulfotransferase 3 (Chst3), the enzyme responsible for the chondroitin 6-sulfation (C6S) of proteoglycans. Further analysis of epigenetic mechanisms revealed a decrease in H3K4me3, three methyl groups at the lysine 4 on the histone H3 proteins, associated with the promoter region of the Chst3 gene, resulting in the downregulation of Chst3 expression in non-neuronal cells. Cluster analysis revealed that the expression of lecticans-substrates of CHST3-is tightly co-regulated with this enzyme. These changes in ECM-related genes were accompanied by an age-confounded decline in cognitive performance. Despite the co-directional impairment in cognitive function and average Chst3 expression in the studied age groups, at the individual level we found a negative correlation between mRNA levels of Chst3 and cognitive performance within the very aged group. An analysis of correlations between the expression of ECM-related genes and cognitive performance in novel object versus novel location recognition tasks revealed an apparent trade-off in the positive gene effects in one task at the expense of another. Further analysis revealed that, despite the reduction in the Chst3 mRNA, the expression of CHST3 protein is increased in glial cells but not in neurons, which, however, does not lead to changes in the absolute level of C6S and even results in the decrease in C6S in perineuronal, perisynaptic and periaxonal ECM relative to the elevated expression of its protein carrier versican.


Assuntos
Epigênese Genética , Proteoglicanas , Envelhecimento/genética , Animais , Cognição , Camundongos , RNA Mensageiro , Sulfotransferases , Carboidrato Sulfotransferases
14.
Front Endocrinol (Lausanne) ; 13: 892342, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35757431

RESUMO

An in vitro system to study testicular maturation in rats, an important model organism for reproductive toxicity, could serve as a platform for high-throughput drug and toxicity screening in a tissue specific context. In vitro maturation of somatic cells and spermatogonia in organ culture systems has been reported. However, this has been a challenge for organoids derived from dissociated testicular cells. Here, we report generation and maintenance of rat testicular organoids in microwell culture for 28 days. We find that rat organoids can be maintained in vitro only at lower than ambient O2 tension of 15% and organoids cultured at 34°C have higher somatic cell maturation and spermatogonial differentiation potential compared to cultures in 37°C. Upon exposure to known toxicants, phthalic acid mono-2-ethylhexyl ester and cadmium chloride, the organoids displayed loss of tight-junction protein Claudin 11 and altered transcription levels of somatic cell markers that are consistent with previous reports in animal models. Therefore, the microwell-derived rat testicular organoids described here can serve as a novel platform for the study of testicular cell maturation and reproductive toxicity in vitro.


Assuntos
Organoides , Espermatogônias , Animais , Diferenciação Celular , Masculino , Ratos , Espermatogônias/metabolismo , Testículo/metabolismo
15.
Small ; 18(15): e2108112, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35224860

RESUMO

The surface fouling of biomedical devices has been an ongoing issue in healthcare. Bacterial and blood adhesion in particular, severely impede the performance of such tools, leading to poor patient outcomes. Various structural and chemical modifications have been shown to reduce fouling, but all existing strategies lack the combination of physical, chemical, and economic traits necessary for widespread use. Herein, a lubricant infused, hierarchically micro- and nanostructured polydimethylsiloxane surface is presented. The surface is easy to produce and exhibits the high flexibility and optical transparency necessary for incorporation into various biomedical tools. Tests involving two clinically relevant, priority pathogens show up to a 98.5% reduction in the biofilm formation of methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa. With blood, the surface reduces staining by 95% and suppresses thrombin generation to background levels. Furthermore, the surface shows applicability within applications such as catheters, extracorporeal circuits, and microfluidic devices, through its effectiveness in dynamic conditions. The perfusion of bacterial media shows up to 96.5% reduction in bacterial adhesion. Similarly, a 95.8% reduction in fibrin networks is observed following whole blood perfusion. This substrate stands to hold high applicability within biomedical systems as a means to prevent fouling, thus improving performance.


Assuntos
Staphylococcus aureus Resistente à Meticilina , Trombose , Aderência Bacteriana , Biofilmes , Dimetilpolisiloxanos , Humanos , Propriedades de Superfície
16.
Int J Mol Sci ; 22(24)2021 Dec 17.
Artigo em Inglês | MEDLINE | ID: mdl-34948348

RESUMO

Male survivors of childhood cancer are at risk of suffering from infertility in adulthood because of gonadotoxic chemotherapies. For adult men, sperm collection and preservation are routine procedures prior to treatment; however, this is not an option for pre-pubertal children. From young boys, a small biopsy may be taken before chemotherapy, and spermatogonia may be propagated in vitro for future transplantation to restore fertility. A robust system that allows for scalable expansion of spermatogonia within a controlled environment is therefore required. Stirred suspension culture has been applied to different types of stem cells but has so far not been explored for spermatogonia. Here, we report that pre-pubertal porcine spermatogonia proliferate more in bioreactor suspension culture, compared with static culture. Interestingly, oxygen tension provides an avenue to modulate spermatogonia status, with culture under 10% oxygen retaining a more undifferentiated state and reducing proliferation in comparison with the conventional approach of culturing under ambient oxygen levels. Spermatogonia grown in bioreactors upregulate the Wnt/ ß-catenin pathway, which, along with enhanced gas and nutrient exchange observed in bioreactor culture, may synergistically account for higher spermatogonia proliferation. Therefore, stirred suspension bioreactors provide novel platforms to culture spermatogonia in a scalable manner and with minimal handling.


Assuntos
Reatores Biológicos , Técnicas de Cultura de Células/métodos , Proliferação de Células , Espermatogônias/fisiologia , Suspensões , Via de Sinalização Wnt , Animais , Masculino , Espermatogônias/metabolismo , Sus scrofa
17.
ACS Appl Mater Interfaces ; 13(43): 50851-50861, 2021 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-34664926

RESUMO

There is an increasing interest in using photoelectrochemistry for enhancing the signal-to-noise ratio and sensitivity of electrochemical biosensors. Nevertheless, it remains challenging to create photoelectrochemical biosensors founded on stable material systems that are also easily biofunctionalized for sensing applications. Herein, a photoelectrochemical immunosensor is reported, in which the concentration of the target protein directly correlates to a change in the measured photocurrent. The material system for the photoelectrode signal transducer involves using catecholate ligands to modify the properties of TiO2 nanostructures in a three-pronged approach of morphology tuning, photoabsorption enhancement, and facilitating bioconjugation. The catecholate-modified TiO2 photoelectrode is combined with a signal-off direct immunoassay to detect interleukin-6 (IL-6), a key biomarker for diagnosing and monitoring various diseases. Catecholate ligands are added during hydrothermal synthesis of TiO2 to enable the growth of three-dimensional nanostructures to form highly porous photoelectrodes that provide a three-dimensional scaffold for immobilizing capture antibodies. Surface modification by catecholate ligands greatly enhances photocurrent generation of the TiO2 photoelectrodes by improving photoabsorption in the visible range. Additionally, catecholate molecules facilitate bioconjugation and probe immobilization by forming a Schiff-base between their -COH group and the -NH2 group of the capture antibodies. The highest photocurrent achieved herein is 8.89 µA cm-2, which represents an enhancement by a factor of 87 from unmodified TiO2. The fabricated immunosensor shows a limit-of-detection of 3.6 pg mL-1 and a log-linear dynamic range of 2-2000 pg mL-1 for IL-6 in human blood plasma.


Assuntos
Técnicas Biossensoriais , Catecóis/química , Técnicas Eletroquímicas , Imunoensaio , Interleucina-6/sangue , Titânio/química , Humanos , Estrutura Molecular , Processos Fotoquímicos
18.
Molecules ; 26(7)2021 Mar 31.
Artigo em Inglês | MEDLINE | ID: mdl-33807421

RESUMO

A dip-coating technique is designed for deposition of poly(methyl methacrylate) (PMMA) from water/2-propanol mixture, avoiding the use of traditional toxic solvents. Solutions of PMMA macromolecules with high molecular weight (MW) are obtained for a water/2-propanol ratio of 0.15-0.33 and the solubilization mechanism is discussed. The ability to use concentrated PMMA solutions and high MW of the polymer are the key factors for the successful dip coating deposition. The coating mass for 10 g L-1 polymer solutions shows a maximum at a water/2-propanol ratio of 0.25. The deposition yield increases with the polymer concentration increase and with an increasing number of the deposited layers. PMMA deposits protect stainless steel from aqueous corrosion. The coating technique allows for the fabrication of composite coatings, containing flame-retardant materials (FRMs), such as commercial halloysite, huntite, hydrotalcite, and synthesized Al(OH)3, in the PMMA matrix. The FRM content in the coatings is modified by variation of the FRM content in colloidal suspensions. A fundamentally new method is developed, which is based on the salting out aided dispersive extraction of Al(OH)3 from the aqueous synthesis medium to 2-propanol. It is based on the use of hexadecylphosphonic acid molecules as extractors. The method offers advantages of reduced agglomeration.

19.
Endocrinology ; 162(4)2021 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-33570577

RESUMO

Organoids are 3-dimensional (3D) structures grown in vitro that emulate the cytoarchitecture and functions of true organs. Therefore, testicular organoids arise as an important model for research on male reproductive biology. These organoids can be generated from different sources of testicular cells, but most studies to date have used immature primary cells for this purpose. The complexity of the mammalian testicular cytoarchitecture and regulation poses a challenge for working with testicular organoids, because, ideally, these 3D models should mimic the organization observed in vivo. In this review, we explore the characteristics of the most important cell types present in the testicular organoid models reported to date and discuss how different factors influence the regulation of these cells inside the organoids and their outcomes. Factors such as the developmental or maturational stage of the Sertoli cells, for example, influence organoid generation and structure, which affect the use of these 3D models for research. Spermatogonial stem cells have been a focus recently, especially in regard to male fertility preservation. The regulation of the spermatogonial stem cell niche inside testicular organoids is discussed in the present review, as this research area may be positively affected by recent progress in organoid generation and tissue engineering. Therefore, the testicular organoid approach is a very promising model for male reproductive biology research, but more studies and improvements are necessary to achieve its full potential.


Assuntos
Organoides/citologia , Testículo/citologia , Animais , Diferenciação Celular , Humanos , Masculino , Organoides/crescimento & desenvolvimento , Células de Sertoli/citologia , Espermatogônias/citologia , Testículo/crescimento & desenvolvimento
20.
Development ; 147(13)2020 07 13.
Artigo em Inglês | MEDLINE | ID: mdl-32554530

RESUMO

In mammalian testis, contractile peritubular myoid cells (PMCs) regulate the transport of sperm and luminal fluid, while secreting growth factors and extracellular matrix proteins to support the spermatogonial stem cell niche. However, little is known about the role of testicular smooth muscle cells during postnatal testicular development. Here we report age-dependent expression of hypermethylated in cancer 1 (Hic1; also known as ZBTB29) in testicular smooth muscle cells, including PMCs and vascular smooth muscle cells, in the mouse. Postnatal deletion of Hic1 in smooth muscle cells led to their increased proliferation and resulted in dilatation of seminiferous tubules, with increased numbers of PMCs. These seminiferous tubules contained fewer Sertoli cells and more spermatogonia, and fibronectin was not detected in their basement membrane. The expression levels of genes encoding smooth muscle contractile proteins, Acta2 and Cnn1, were downregulated in the smooth muscle cells lacking Hic1, and the seminiferous tubules appeared to have reduced contractility. These data imply a role for Hic1 in determining the size of seminiferous tubules by regulating postnatal smooth muscle cell proliferation, subsequently affecting spermatogenesis in adulthood.


Assuntos
Fibronectinas/metabolismo , Miócitos de Músculo Liso/metabolismo , Testículo/metabolismo , Actinas/genética , Actinas/metabolismo , Animais , Fibronectinas/genética , Fatores de Transcrição Kruppel-Like/genética , Fatores de Transcrição Kruppel-Like/metabolismo , Masculino , Proteínas dos Microfilamentos/genética , Proteínas dos Microfilamentos/metabolismo
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