RESUMO
Melanogenesis, the intricate process of melanin synthesis, is central to skin pigmentation and photoprotection and is regulated by various signaling pathways and transcription factors. To develop potential skin-whitening agents, we used B16F1 melanoma cells to investigate the inhibitory effects of anhydrous alum on melanogenesis and its underlying molecular mechanisms. Anhydrous alum (KAl(SO4)2) with high purity (>99%), which is generated through the heat-treatment of hydrated alum (KAl(SO4)2·12H2O) at 400 °C, potentiates a significant reduction in melanin content without cytotoxicity. Anhydrous alum downregulates the master regulator of melanogenesis, microphthalmia-associated transcription factor (MITF), which targets key genes involved in melanogenesis, thereby inhibiting α-melanocyte-stimulating hormone (α-MSH)-induced melanogenesis. Phosphorylation of the cAMP response element-binding protein, which acts as a co-activator of MITF gene expression, is attenuated by anhydrous alum, resulting in compromised MITF transcription. Notably, anhydrous alum promoted extracellular signal-regulated kinase phosphorylation, leading to the impaired nuclear localization of MITF. Overall, these results demonstrated the generation and mode of action of anhydrous alum in B16F1 cells, which constitutes a promising option for cosmetic or therapeutic use.
Assuntos
Melaninas , alfa-MSH , Melaninas/metabolismo , alfa-MSH/metabolismo , Monofenol Mono-Oxigenase/metabolismo , Fator de Transcrição Associado à Microftalmia/genética , Fator de Transcrição Associado à Microftalmia/metabolismo , Linhagem Celular TumoralRESUMO
Combination products (CPs) combine two or more product types such as drugs, devices, and/or biological products for increased safety and clinical effectiveness. The emergence of innovative CPs poses new challenges for regulatory agencies in assigning jurisdiction for premarket review and oversight. In US, the 1990 Safe Medical Devices Act defines and provides classification criteria for CPs, and the US government has developed a regulatory process through multiple acts, including the 21st Century Cures Act of 2016. Meanwhile, regulators in the European Union (EU) and the Republic of Korea have recently recognized the importance of premarket pathways for CPs. The European Medicines Agency (EMA) has issued guidelines and explanations on compliance issues related to drug-device CPs under MDR. EMA doesn't have the definitions of CPs, but uses the term drug-device combination products (drug-device CPs). CPs are categorized as drugs or medical devices, which follow their relevant regulatory framework. The Ministry of Food and Drug Safety (MFDS) in Korea has legal definitions of CPs under the Notice of the MFDS. CPs are designated as drugs or medical devices according to their primary mode of actions (PMOA) and follow regulatory processes through the framework of drugs or medical devices. This study aims to comprehensively summarize the regulatory oversight of CPs by analyzing the regulatory policies and legal frameworks in the US, the EU, and Korea. The regulatory challenges encountered when developing CPs are also discussed. With specific emphasis on the combination of drugs and devices, this study provides in-depth insights into the regulatory landscape, shedding light on the unique challenges associated with the development of CPs for this particular intersection of drugs and devices.
RESUMO
Human urine samples are non-invasive, readily available, and contain several components that can provide useful indicators of the health status of patients. Hence, urine is a desirable and important template to aid in the diagnosis of common clinical conditions. Conventional methods such as dipstick tests, urine culture, and urine microscopy are commonly used for urinalysis. Among them, the dipstick test is undoubtedly the most popular owing to its ease of use, low cost, and quick response. Despite these advantages, the dipstick test has limitations in terms of sensitivity, selectivity, reusability, and quantitative evaluation of diseases. Various biosensor technologies give it the potential for being developed into point-of-care (POC) applications by overcoming these limitations of the dipstick test. Here, we present a review of the biosensor technologies available to identify urine-based biomarkers that are typically detected by the dipstick test and discuss the present limitations and challenges that future development for their translation into POC applications for urinalysis.
Assuntos
Técnicas Biossensoriais , Urinálise , Humanos , Urinálise/métodos , Sistemas Automatizados de Assistência Junto ao Leito , Microscopia , BiomarcadoresRESUMO
Rapid diagnosis and case isolation are pivotal to controlling the current pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, a label-free DNA capacitive biosensor for the detection of SARS-CoV-2 that demonstrates real-time, low-cost, and high-throughput screening of nucleic acid samples is presented. Our novel biosensor composed of the interdigitated platinum/titanium electrodes on the glass substrate can detect the hybridization of analyte DNA with probe DNA. The hybridization signals of specific DNA sequences were verified through exhaustive physicochemical analytical techniques such as Fourier transform infrared (FT-IR) spectrometry, contact-angle analysis, and capacitance-frequency measurements. For a single-step hybridized reaction, the fabricated kit exhibited significant sensitivity (capacitance change, ΔC = ~2 nF) in detecting the conserved region of the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) gene with high sensitivity of 0.843 nF/nM. In addition to capacitive measurements, this selective detection was confirmed by the fluorescence image and intensity from a SARS-CoV-2 gene labeled with a fluorescent dye. We also demonstrated that the kits are recyclable by surface ozone treatment using UV irradiation. Thus, these kits could potentially be applied to various types of label-free DNA, thereby acting as rapid, cost-effective biosensors for several diseases.
Assuntos
Técnicas Biossensoriais , COVID-19 , DNA , Humanos , Sistemas Automatizados de Assistência Junto ao Leito , RNA Viral , SARS-CoV-2 , Sensibilidade e Especificidade , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Widespread neuronal cell death occurs during normal development and as a result of pathological conditions in the nervous system of many organisms. In this study, we investigated the cytotoxicity induced by H(2)O(2) in Aplysia mechanosensory neurons, which serve as a useful model in the study of learning and memory. Treatment with hydrogen peroxide (10(-2)-10 mM) for 3 h produced a nuclear DNA breakage in Aplysia sensory neurons, as revealed by TdT-mediated dUTP nick end labeling (TUNEL) staining, in a dose-dependent manner. Prolonged treatment (6-18 h) of Aplysia sensory neurons with 1 mM hydrogen peroxide produced dramatic morphological changes, such as neurite fragmentation, disintegration of the cell body, and a change in the resting membrane potential. This change in the resting potential was biphasic, and was initially hyperpolarized about 6 h after hydrogen peroxide treatment, but this later shifted to a depolarization some 13-18 h after treatment. Electron microscopic analysis also showed that this hydrogen peroxide-induced cell death was associated with apoptotic nuclear shrinkage, chromatin condensation, and necrotic swelling of organelles. Our results demonstrate that Aplysia sensory neurons show both apoptotic and necrotic characteristics as well as biphasic changes in resting potential during hydrogen peroxide-induced cell death.
Assuntos
Morte Celular/efeitos dos fármacos , Peróxido de Hidrogênio/efeitos adversos , Neurônios Aferentes/efeitos dos fármacos , Oxidantes/efeitos adversos , Animais , Aplysia , Técnicas de Cultura de Células , Eletrofisiologia , Marcação In Situ das Extremidades Cortadas , Microscopia Eletrônica , Estresse Oxidativo/efeitos dos fármacosRESUMO
In order to elucidate the structural requirements for the dual neuroprotective activity of aspirin against N-methyl-D-aspartate (NMDA) and zinc ion neurotoxicity, various aspirin analogues and derivatives, modified at the carboxylic group, the acetyl group, and the chain length between the carboxylic acid moiety and phenyl ring, were synthesized. Replacement of the carboxylic acid group with alkyl groups (compounds 2c and 2d) resulted in a dramatic increase in neuroprotective activity against NMDA neurotoxicity, while reduction of the carboxylic acid group to the alcohol (compound 2g) completely abolished this activity. In contrast to NMDA neurotoxicity, compounds that are devoid of the carboxylic acid group did not show any activity against zinc ion neurotoxicity. Replacement of the acetyl group with a propionyl (compound 5a) or butyryl group (compound 5b) did not significantly change the activity against NMDA neurotoxicity, but replacement of the acetyl group with a propionyl group (compound 5a) resulted in a slight decrease in activity against zinc ion neurotoxicity. Compound 12, which has ethylene units between the carboxylic acid moiety and phenyl ring in the structure of aspirin, exhibited greater neuroprotective activity against NMDA neurotoxicity than the compared compounds (aspirin, compound 9 and compound 17), which have different chain lengths. A similar trend was also observed in the neuroprotective activity against zinc ion neurotoxicity. These results indicate that the carboxylic acid group in aspirin is not indispensable for the inhibitory effect against NMDA neurotoxicity, but is essential for the inhibitory effect against zinc ion neurotoxicity. The acetyl group and ethylene unit's distance are favourable for the inhibitory effect against NMDA neurotoxicity as well as zinc ion neurotoxicity.