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Neurodegeneration is a hallmark of many diseases and disorders of the central nervous system (CNS). High levels of neuroinflammation are often associated with irreparable damage to CNS cells due to the dysregulation of signaling cascades that are unable to restore a homeostatic balance. Due to the inherent complexity of the CNS, development of CNS-related therapeutics has met limited success. While stem cell therapy has been evaluated in the context of CNS repair, the mechanisms responsible for their functional properties have not been clearly defined. In recent years, there has been growing interest in the use of stem cell extracellular vesicles (EVs) for the treatment of various CNS pathologies as these vesicles are believed to mediate many of the functional effects associated with their donor stem cells. The potency of stem cell EVs is believed to be largely driven by their biological cargo which includes various types of RNAs, proteins, and cytokines. In this review, we describe the characteristic properties of stem cell EVs and summarize their reported neuroprotective and immunomodulatory functions. A special emphasis is placed on the identification of specific biological cargo, including proteins and non-coding RNA molecules, that have been found to be associated with stem cell EVs. Collectively, this review highlights the potential of stem cell EVs as an alternative to traditional stem cell therapy for the repair of cellular damage associated with diverse CNS pathologies.
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Soaring demand for technology metals (e.g., Cd, Ni) and its ever-depleting primary resources ask for alternative recovery from secondary sources. Ni-Cd battery is one such source that can abridge the gap between demand and supply of such metals. Biogenic recovery, being environmentally benign, is explored for Cd and Ni recovery to manage the menace of spent Ni-Cd battery. Studies with 20, 40 and 60 mg/L Cd2+ initial concentrations in batch mode (in triplicates) at pH 7.0 ± 0.2, 30 ± 0.5 °C and 120 rpm were conducted using sulfate-reducing bacteria for 10 days. Analysis of extracellular polymeric substance revealed that protein secretion was enhanced, thereby forming Cd-EPS binding. Biosolids were collected and freeze-dried for morphological analysis viz. FESEM/EDX, PXRD and TEM, which revealed the formation of CdS nanoparticles (JCPDS card #00-042-1411) in range of 2-6 nm. Similarly, combined effect with 5, 10 and 20 mg/L Ni2+ at 20 mg/L Cd2+ were also investigated. Furthermore, to test the efficacy for real field application, spent Ni-Cd battery was dismantled and its powder was characterized, digested with concentrated HCl at 70 °C and was fed in batch mode after cooling, wherein nanoparticles of Ni and Cd sulfides were formed that has potential as semi-conducting material.
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Cádmio , Nanopartículas , Matriz Extracelular de Substâncias Poliméricas , Metais , Níquel , SulfetosRESUMO
Vermicomposting is a traditional technology that produces the best quality of compost, but factors such as maturity, presence of heavy metals, etc. need to be tackled prior to agrarian application. The present study investigates the influence of varying biochar dose (2.5, 5, and 10% on a weight basis) on the maturity of compost and heavy metals during vermicomposting of vegetable waste using epigeic earthworm. Biochar amendment notably enhanced the electrical conductivity (up to 2.7â¯mS/cm), nitrogen content (up to 3.1%), NO3-N (up to 630â¯mg/kg) and nutritional value. The heavy metals, oxygen uptake rate (below 0.96â¯mg/g VS/day) and CO2 evolution rate (below 1â¯mg/g VS/day) were attenuated along with degradation of complex organic crystals as observed in powder X-Ray Diffraction (PXRD) spectra. Furthermore, biochar aid in reducing pathogens (below 1.1â¯×â¯103 MPN/g dry weight) as inferred from the Most Probable Number (MPN) results as well as degrading the complex organics into simpler compounds as revealed from the Fourier-transform infrared spectroscopy (FTIR) spectra. The present study inferred that the vegetable waste was biologically stabilized through biochar amendment during vermicomposting process with improved nutritional and physico-chemical properties.
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Carvão Vegetal , Compostagem/métodos , Metais Pesados , Oligoquetos , Poluentes do Solo , Animais , Verduras , ResíduosRESUMO
Neurological diseases and disorders are leading causes of death and disability worldwide. Many of these pathologies are associated with high levels of neuroinflammation and irreparable tissue damage. As the global burden of these pathologies continues to rise there is a significant need for the development of novel therapeutics. Due to their multipotent properties, stem cells have broad applications for tissue repair; additionally, stem cells have been shown to possess both immunomodulatory and neuroprotective properties. It is now believed that paracrine factors, such as extracellular vesicles (EVs), play a critical role in the functionality associated with stem cells. The diverse biological cargo contained within EVs are proposed to mediate these effects and, to date, the reparative and regenerative effects of stem cell EVs have been demonstrated in a wide range of cell types. While a high potential for their therapeutic use exists, there is a gap of knowledge surrounding their characterization, mechanisms of action, and how they may regulate cells of the CNS. Here, we report the isolation, characterization, and functional assessment of EVs from two sources of human stem cells, mesenchymal stem cells and induced pluripotent stem cells. We demonstrate the ability of these EVs to enhance the processes of cellular migration and angiogenesis, which are critical for both normal cellular development as well as cellular repair. Furthermore, we investigate their reparative effects on damaged cells, specifically those with relevance to the central nervous system. Collectively, our data highlight the similarities and differences among these EV populations and support the view that stem cells EV can be used to repair or partially reverse cellular damage. Graphical Abstract Stem cell-derived Extracellular Vesicles (EVs) for repair of damaged cells. EVs isolated from human induced pluripotent stem cells and mesenchymal stem cells contribute to the partial reversal of phenotypes induced by different sources of cellular damage.
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Vesículas Extracelulares , Células-Tronco Pluripotentes Induzidas/ultraestrutura , Doenças do Sistema Nervoso/terapia , Células A549 , Linhagem Celular , Movimento Celular , Sobrevivência Celular , Citocinas/biossíntese , Vesículas Extracelulares/genética , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/efeitos da radiação , Humanos , Imunidade Inata , Células-Tronco Pluripotentes Induzidas/efeitos da radiação , Neovascularização Patológica/terapia , Doenças do Sistema Nervoso/patologia , Proteômica , RNA/genética , Radiação IonizanteRESUMO
One of the major hurdles in the field of extracellular vesicle (EV) research today is the ability to achieve purified EV preparations in a viral infection setting. The presented method is meant to isolate EVs away from virions (i.e., HIV-1), allowing for a higher efficiency and yield compared to conventional ultracentrifugation methods. Our protocol contains three steps: EV precipitation, density gradient separation, and particle capture. Downstream assays (i.e., Western blot, and PCR) can be run directly following particle capture. This method is advantageous over other isolation methods (i.e., ultracentrifugation) as it allows for the use of minimal starting volumes. Furthermore, it is more user friendly than alternative EV isolation methods requiring multiple ultracentrifugation steps. However, the presented method is limited in its scope of functional EV assays as it is difficult to elute intact EVs from our particles. Furthermore, this method is tailored towards a strictly research-based setting and would not be commercially viable.
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Vesículas Extracelulares , Vírion , Western Blotting , Humanos , Ultracentrifugação/métodosRESUMO
Biochar, a recalcitrant carbon, is known to enhance organic matter degradation and improve physical properties. The objective of the study is to examine the probable effect of biochar addition during composting of a fibrous aquatic waste, i.e., water hyacinth though degradation kinetics and spectroscopic (FTIR and PXRD) analysis. Four dosages of biochar (0, 2.5, 5, and 10% w/w) were mixed to a mixture of water hyacinth, cow-dung and saw-dust comprising a total weight of 150â¯kg and composted using rotary drum composter for 20 days in batch mode. The study outcomes indicated that the amendment of biochar prolonged the duration of the thermophilic temperatures, reduced salinity, and promoted nutritional quality of compost. Moreover, biochar amendment enhanced the organic matter degradation with a rate constant of 0.029 day-1 and increased the total Kjeldahl nitrogen content up to 1.75% from an initial value of 1.10% in the reactor with 2.5% biochar amendment. Concurrently, biochar amendment aided in reducing Cu and Cr in the final product inferring 2.5% biochar is best suited for composting of water hyacinth. However, future studies are encouraged to decipher the microbial shifts and bioavailability of metals due to biochar dosage during composting for mitigating and managing the menace of such fibrous waste like water hyacinth by converting it to a soil conditioner.
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Compostagem , Animais , Carbono , Bovinos , Feminino , Cinética , Nitrogênio , SoloRESUMO
Sewage sludge produced is either applied to land or used as fertilizer for crops or disposed of in landfills, causing several environmental problems. Recent studies revealed that composting is a proven technology in reducing organic content, heavy metals, and harmful pathogens, improving the nutritional value of sewage sludge, which is useful for crops. But studies on variation in physical properties are rare. Composting physics or physical properties during composting plays a vital role from handling, management, and utilization of end product, i.e., compost. This study mainly deals with the detailed information on physics involved during the degradation process, which is crucial for land and geotechnical applications. In the present study, sewage sludge was used as a composting substrate in 550â¯L in-vessel rotary drum composter. Emphasis was given in deciphering the changes in physical parameters such as bulk density, porosity, and air-filled porosity and few chemical parameters during the composting process. Besides, a relationship between different physical properties during rotary drum composting was investigated statistically. Bulk density was observed to have increased from 643 to 707â¯kgâ¯m-3 as a result of volume reduction of compost matrix. Moreover, the gravimetric moisture content was found to be less than 45% in the end product, which is recommended for compost.
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Compostagem , Metais Pesados , Esgotos , SoloRESUMO
Co-occurrence of arsenic and iron along with nitrate in groundwater makes the trio an onerous combination both in terms of potability and treatment. To meet drinking water guidelines, batch and column laboratory trials were conducted on simulated and bore-well water for attenuation of arsenic (1000 µg/L), iron (5 mg/L) and nitrate (150 mg/L). Increment in sulphate showed a direct individual impact on iron removal, meeting WHO guidelines. The bio-kinetic parameters were in the range of: µmax = 0.079-0.551/d, Ks = 116.18-645.19 mg/L, Kd = 0.0009-0.0077/d and Y = 0.034-0.094 mg MLVSS/mg COD. Transmission electron microscopy (TEM) and X-ray diffraction (XRD) confirmed orpiment precipitation and/or co-precipitation with mackinawite are the key mechanisms for arsenic and iron attenuation. Column experiments were conducted by charging simulated groundwater containing arsenic (500 µg/L), nitrate (50 mg/L), sulphate (25 mg/L) and iron (3 mg/L) in an acetate (105 mg/L as COD) fed flow-through bioreactor at constant empty bed contact time of 60 min. Profile sampling illustrated segregation of different terminal electron accepting zones following thermodynamic yield for sequential removal of different oxyanions. This study showed the importance of considering microbially mediated terminal electron-accepting processes (TEAP) for multi-oxyanion removal in engineered systems.
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Papillomaviruses, of the family Papillomaviridae, are epitheliotropic, nonenveloped, circular, double-stranded DNA viruses that contribute to benign and malignant tumors in humans and animals. We report here the whole-genome sequence of canine papillomavirus type 12, found at a pigmented plaque located on the skin of a mixed-breed bloodhound.
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In vitro evolution of nucleic acids and proteins is a powerful strategy to optimize their biological and physical properties. To select proteins with the desired phenotype from large gene libraries, the proteins need to be linked to the gene they are encoded by. To facilitate selection of the desired phenotype and isolation of the encoding DNA, a novel bead display approach was developed, in which each member of a library of beads is first linked to multiple copies of a clonal gene variant by emulsion polymerase chain reaction. Beads are transferred to a second emulsion for an in vitro transcription-translation reaction, in which the protein encoded by each bead's amplicon covalently binds to the bead present in the same picoliter reactor. The beads then contain multiple copies of a clonal gene variant and multiple molecules of the protein encoded by the bead's gene variant and serve as the unit of selection. As a proof of concept, we screened a randomized library of the T7 promoter for high expression levels by flow cytometry and identified a T7 promoter variant with an ~10-fold higher in vitro transcriptional activity, confirming that the multi-copy bead display approach can be efficiently applied to in vitro evolution.
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Evolução Molecular Direcionada/métodos , Regiões Promotoras Genéticas , Proteínas Recombinantes de Fusão/genética , Transcrição Gênica , Bacteriófago T7/genética , DNA , Citometria de Fluxo , Mutação , Reação em Cadeia da Polimerase , Biossíntese de Proteínas , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/químicaRESUMO
Tat, an important regulatory protein of HIV-1, has been implicated in HIV-related pathogenesis. Immune responses to Tat, although underrepresented, confer protection against disease progression, in natural infection and experimental immunization, making Tat an attractive vaccine candidate. Information on immune responses to Tat from India which has the second largest HIV incidence has been lacking. Here we report a cross-sectional study evaluating the humoral response to Tat from a large number of samples from two southern states of India. 14% of the seropositive (63/447) and 4.6% of seronegative samples (7/150) harbored Tat-reactive antibodies. A significant number of the seropositive samples contained high levels of anti-Tat antibodies (31/447) which demonstrated class-switch to IgG1 and bound to Tat with high avidity. Cross-reactivity analysis showed that these antibodies interacted with Tat from different clades with variable degree with the highest interaction with subtype-AE and the least with subtype-B Tat. Importantly, a B-cell epitope in the cysteine-rich domain was found to be the most immunodominant one and antibodies interacting with this epitope blocked extracellular Tat efficiently. To the best of our knowledge this is the first report on immune responses to Tat from Indian populations and the data presented here could significantly contribute to HIV Tat vaccine design.
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Epitopos de Linfócito B/imunologia , Anticorpos Anti-HIV/sangue , Infecções por HIV/imunologia , Imunoglobulina G/sangue , Produtos do Gene tat do Vírus da Imunodeficiência Humana/imunologia , Adolescente , Adulto , Afinidade de Anticorpos , Reações Cruzadas , Estudos Transversais , Mapeamento de Epitopos , Feminino , Anticorpos Anti-HIV/imunologia , Soropositividade para HIV , HIV-1/imunologia , Humanos , Imunidade Humoral , Epitopos Imunodominantes , Switching de Imunoglobulina , Imunoglobulina G/imunologia , Índia , Masculino , Pessoa de Meia-Idade , Testes de Neutralização , Adulto JovemRESUMO
The People's Republic of Bangladesh is located in South Asia. The total land area of Bangladesh is 147 570 km2. Its total population in 2001 was about 123 million. The population growth rate is 1.47%; of the total population, 75% live in rural areas and 25% in urban areas (Bangladesh Bureau of Statistics, 2000).