RESUMO
Mycobacterium tuberculosis (Mtb) defends host-mediated killing by repressing the autophagolysosome machinery. For the first time, we report NCoR1 co-repressor as a crucial host factor, controlling Mtb growth in myeloid cells by regulating both autophagosome maturation and lysosome biogenesis. We found that the dynamic expression of NCoR1 is compromised in human peripheral blood mononuclear cells (PBMCs) during active Mtb infection, which is rescued upon prolonged anti-mycobacterial therapy. In addition, a loss of function in myeloid-specific NCoR1 considerably exacerbates the growth of M. tuberculosis in vitro in THP1 differentiated macrophages, ex vivo in bone marrow-derived macrophages (BMDMs), and in vivo in NCoR1MyeKO mice. We showed that NCoR1 depletion controls the AMPK-mTOR-TFEB signalling axis by fine-tuning cellular adenosine triphosphate (ATP) homeostasis, which in turn changes the expression of proteins involved in autophagy and lysosomal biogenesis. Moreover, we also showed that the treatment of NCoR1 depleted cells by Rapamycin, Antimycin-A, or Metformin rescued the TFEB activity and LC3 levels, resulting in enhanced Mtb clearance. Similarly, expressing NCoR1 exogenously rescued the AMPK-mTOR-TFEB signalling axis and Mtb killing. Overall, our data revealed a central role of NCoR1 in Mtb pathogenesis in myeloid cells.
Assuntos
Mycobacterium tuberculosis , Correpressor 1 de Receptor Nuclear , Animais , Humanos , Camundongos , Proteínas Quinases Ativadas por AMP , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos , Leucócitos Mononucleares , Células Mieloides , Serina-Treonina Quinases TOR , Correpressor 1 de Receptor Nuclear/metabolismoRESUMO
We performed a genome-wide siRNA screen to identify host factors that regulated pathogen load in human macrophages infected with a virulent strain of Mycobacterium tuberculosis. Iterative rounds of confirmation, followed by validation, identified 275 such molecules that were all found to functionally associate with each other through a dense network of interactions. This network then yielded to a molecular description of the host cell functional modules that were both engaged and perturbed by the pathogen. Importantly, a subscreen against a panel of field isolates revealed that the molecular composition of the host interface varied with both genotype and the phenotypic properties of the pathogen. An analysis of these differences, however, permitted identification of those host factors that were invariantly involved, regardless of the diversification in adaptive mechanisms employed by the pathogen. Interestingly, these factors were found to predominantly function through the regulation of autophagy.
Assuntos
Estudo de Associação Genômica Ampla , Interações Hospedeiro-Patógeno , Macrófagos/microbiologia , Mycobacterium tuberculosis/fisiologia , Genoma Humano , Biblioteca Genômica , Humanos , Viabilidade Microbiana , Mycobacterium tuberculosis/imunologia , RNA Interferente Pequeno/genéticaRESUMO
Macromolecules such as monoclonal antibodies (mAbs) are likely to experience poor tumor penetration because of their large size, and thus low drug exposure of target cells within a tumor could contribute to suboptimal responses. Given the challenge of inadequate quantitative tools to assess mAb activity within tumors, we hypothesized that measurement of accessible target levels in tumors could elucidate the pharmacologic activity of a mAb and could be used to compare the activity of different mAbs. Using positron emission tomography (PET), we measured the pharmacodynamics of immune checkpoint protein programmed-death ligand 1 (PD-L1) to evaluate pharmacologic effects of mAbs targeting PD-L1 and its receptor programmed cell death protein 1 (PD-1). For PD-L1 quantification, we first developed a small peptide-based fluorine-18-labeled PET imaging agent, [18F]DK222, which provided high-contrast images in preclinical models. We then quantified accessible PD-L1 levels in the tumor bed during treatment with anti-PD-1 and anti-PD-L1 mAbs. Applying mixed-effects models to these data, we found subtle differences in the pharmacodynamic effects of two anti-PD-1 mAbs (nivolumab and pembrolizumab). In contrast, we observed starkly divergent target engagement with anti-PD-L1 mAbs (atezolizumab, avelumab, and durvalumab) that were administered at equivalent doses, correlating with differential effects on tumor growth. Thus, we show that measuring PD-L1 pharmacodynamics informs mechanistic understanding of therapeutic mAbs targeting PD-L1 and PD-1. These findings demonstrate the value of quantifying target pharmacodynamics to elucidate the pharmacologic activity of mAbs, independent of mAb biophysical properties and inclusive of all physiological variables, which are highly heterogeneous within and across tumors and patients.
Assuntos
Antineoplásicos Imunológicos/farmacologia , Antígeno B7-H1/antagonistas & inibidores , Neoplasias da Mama/tratamento farmacológico , Radioisótopos de Flúor/farmacocinética , Fragmentos de Peptídeos/farmacocinética , Tomografia por Emissão de Pósitrons/métodos , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Animais , Apoptose , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Proliferação de Células , Feminino , Humanos , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Compostos Radiofarmacêuticos/farmacocinética , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Orthopedic and dental implant failure continues to be a significant concern due to localized bacterial infections. Previous studies have attempted to improve implant surfaces by modifying their texture and roughness or coating them with antibiotics to enhance antibacterial properties for implant longevity. However, these approaches have demonstrated limited effectiveness. In this study, we attempted to engineer the titanium (Ti) alloy surface biomimetically at the nanometer scale, inspired by the cicada wing nanostructure using alkaline hydrothermal treatment (AHT) to simultaneously confer antibacterial properties and support the adhesion and proliferation of mammalian cells. The two modified Ti surfaces were developed using a 4 h and 8 h AHT process in 1 N NaOH at 230 °C, followed by a 2-hour post-calcination at 600 °C. We found that the control plates showed a relatively smooth surface, while the treatment groups (4 h & 8 h AHT) displayed nanoflower structures containing randomly distributed nano-spikes. The results demonstrated a statistically significant decrease in the contact angle of the treatment groups, which increased wettability characteristics. The 8 h AHT group exhibited the highest wettability and significant increase in roughness 0.72 ± 0.08 µm (P < 0.05), leading to more osteoblast cell attachment, reduced cytotoxicity effects, and enhanced relative survivability. The alkaline phosphatase activity measured in all different groups indicated that the 8 h AHT group exhibited the highest activity, suggesting that the surface roughness and wettability of the treatment groups may have facilitated cell adhesion and attachment and subsequently increased secretion of extracellular matrix. Overall, the findings indicate that biomimetic nanotextured surfaces created by the AHT process have the potential to be translated as implant coatings to enhance bone regeneration and implant integration.
Assuntos
Materiais Biomiméticos , Implantes Dentários , Osteoblastos , Propriedades de Superfície , Titânio , Molhabilidade , Osteoblastos/efeitos dos fármacos , Titânio/química , Animais , Materiais Biomiméticos/química , Materiais Biomiméticos/farmacologia , Adesão Celular/efeitos dos fármacos , Antibacterianos/química , Antibacterianos/farmacologia , Teste de Materiais , Biomimética , Humanos , Proliferação de Células/efeitos dos fármacos , Ligas/química , Próteses e Implantes , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Nanoestruturas/química , Sobrevivência Celular/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Hemípteros , Linhagem CelularRESUMO
Huntington's disease (HD) is an autosomal dominant ailment that affects a larger population. Due to its complex pathology operating at DNA, RNA, and protein levels, it is regarded as a protein-misfolding disease and an expansion repeat disorder. Despite the availability of early genetic diagnostics, disease-modifying treatments are still missing. Importantly, potential therapies are starting to make their way through clinical trials. Still, clinical trials are ongoing to discover potential drugs to relieve HD symptoms. However, now being aware of the root cause, the clinical studies are focused on molecular therapies to target it. The road to success has not been without bumps since a big phase III trial of tominersen was unexpectedly discontinued due to exceeding risks than drug's benefit to the patients. Although the trial's conclusion was disappointing, there is still cause to be optimistic about what this technique may achieve. We have reviewed the present disease-modifying therapies in clinical development for HD and examined the current landscape of developing clinical therapies. We further investigated the pharmaceutical development of Huntington's medicine in the pharma industries and addressed the existing challenges in their therapeutic success.
Assuntos
Doença de Huntington , Humanos , Doença de Huntington/tratamento farmacológico , Doença de Huntington/genética , RNA , DNA , Desenvolvimento de Medicamentos , Proteína Huntingtina/genética , Proteína Huntingtina/metabolismoRESUMO
BACKGROUND: Chimeric antigen receptor (CAR) T cell therapy is an exciting cell-based cancer immunotherapy. Unfortunately, CAR-T cell therapy is associated with serious toxicities such as cytokine release syndrome (CRS) and neurotoxicity. The mechanism of these serious adverse events (SAEs) and how homing, distribution and retention of CAR-T cells contribute to toxicities is not fully understood. Enabling in vitro methods to allow meaningful, sensitive in vivo biodistribution studies is needed to better understand CAR-T cell disposition and its relationship to both effectiveness and safety of these products. METHODS: To determine if radiolabelling of CAR-T cells could support positron emission tomography (PET)-based biodistribution studies, we labeled IL-13Rα2 targeting scFv-IL-13Rα2-CAR-T cells (CAR-T cells) with 89Zirconium-oxine (89Zr-oxine) and characterized and compared their product attributes with non-labeled CAR-T cells. The 89Zr-oxine labeling conditions were optimized for incubation time, temperature, and use of serum for labeling. In addition, T cell subtype characterization and product attributes of radiolabeled CAR-T cells were studied to assess their overall quality including cell viability, proliferation, phenotype markers of T-cell activation and exhaustion, cytolytic activity and release of interferon-γ upon co-culture with IL-13Rα2 expressing glioma cells. RESULTS: We observed that radiolabeling of CAR-T cells with 89Zr-oxine is quick, efficient, and radioactivity is retained in the cells for at least 8 days with minimal loss. Also, viability of radiolabeled CAR-T cells and subtypes such as CD4 + , CD8 + and scFV-IL-13Rα2 transgene positive T cell population were characterized and found similar to that of unlabeled cells as determined by TUNEL assay, caspase 3/7 enzyme and granzyme B activity assay. Moreover, there were no significant changes in T cell activation (CD24, CD44, CD69 and IFN-γ) or T cell exhaustion (PD-1, LAG-3 and TIM3) markers expression between radiolabeled and unlabeled CAR-T cells. In chemotaxis assays, migratory capability of radiolabeled CAR-T cells to IL-13Rα2Fc was similar to that of non-labeled cells. CONCLUSIONS: Importantly, radiolabeling has minimal impact on biological product attributes including potency of CAR-T cells towards IL-13Rα2 positive tumor cells but not IL-13Rα2 negative cells as measured by cytolytic activity and release of IFN-γ. Thus, IL-13Rα2 targeting CAR-T cells radiolabeled with 89Zr-oxine retain critical product attributes and suggest 89Zr-oxine radiolabeling of CAR-T cells may facilitate biodistribution and tissue trafficking studies in vivo using PET.
Assuntos
Imunoterapia Adotiva , Radioisótopos , Linfócitos T , Zircônio , Zircônio/farmacocinética , Radioisótopos/farmacocinética , Tomografia por Emissão de Pósitrons , Rastreamento de Células/métodos , Anticorpos de Cadeia Única , Linfócitos T/citologia , Distribuição Tecidual , Células Jurkat , Animais , Camundongos , Proliferação de Células , Sobrevivência CelularRESUMO
Tuberculosis caused by Mycobacterium tuberculosis (Mtb) is a significant public health concern, exacerbated by the emergence of drug-resistant TB. To combat the host's dynamic environment, Mtb encodes multiple DNA repair enzymes that play a critical role in maintaining genomic integrity. Mtb possesses a GC-rich genome, rendering it highly susceptible to cytosine deaminations, resulting in the occurrence of uracils in the DNA. UDGs encoded by ung and udgB initiate the repair; hence we investigated the biological impact of deleting UDGs in the adaptation of pathogen. We generated gene replacement mutants of uracil DNA glycosylases, individually (RvΔung, RvΔudgB) or together (RvΔdKO). The double KO mutant, RvΔdKO exhibited remarkably higher spontaneous mutation rate, in the presence of antibiotics. Interestingly, RvΔdKO showed higher survival rates in guinea pigs and accumulated large number of SNPs as revealed by whole-genome sequence analysis. Competition assays revealed the superior fitness of RvΔdKO over Rv, both in ex vivo and in vivo conditions. We propose that compromised DNA repair results in the accumulation of mutations, and a subset of these drives adaptation in the host. Importantly, this property allowed us to utilize RvΔdKO for the facile identification of drug targets.
Assuntos
Adaptação Fisiológica/genética , Reparo do DNA/fisiologia , Especificidade de Hospedeiro/genética , Mycobacterium tuberculosis/genética , Animais , Cobaias , CamundongosRESUMO
We describe a case of a ruptured sinus of Valsalva with severe aortic regurgitation treated by transcatheter device closure. The regurgitation was purely due to leaflet entrapment because of the hemodynamic Venturi effect which was effectively treated.
Assuntos
Ruptura Aórtica , Insuficiência da Valva Aórtica , Seio Aórtico , Humanos , Insuficiência da Valva Aórtica/diagnóstico por imagem , Insuficiência da Valva Aórtica/etiologia , Insuficiência da Valva Aórtica/cirurgia , Ruptura Aórtica/diagnóstico por imagem , Ruptura Aórtica/etiologia , Ruptura Aórtica/cirurgia , Seio Aórtico/diagnóstico por imagem , Resultado do Tratamento , HemodinâmicaRESUMO
We propose a photonic crystal ring resonator for the enhancement of quality factor that supports two-dimensionally bounded topological edge states. Crystal parameters are obtained through finite-difference time-domain numerical simulation to get the enhanced quality factor using the topological properties of the photonic crystal. Topological edge states are created when two regions with dissimilar band topologies come together at an interface and are contained within a slab of dielectric material. These edge states can move along sharp edges without backscattering. The transmission dropout issue arises whenever the quality factor is enhanced in a conventional photonic system and is eliminated remarkably by employing the present approach. Such nanoscale photonic crystal structures promote robust interactions between quantum emitters and photonic edge states.
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The uptake and storage of extracellular orthophosphate (Pi) by polyphosphate (polyP) accumulating bacteria may contribute to mineral dissolution in the oral cavity. To test the effect of potential inhibitors of polyP kinases on Rothia dentocariosa, gallein (0, 25, 50, and 100 µM) and fluoride (0, 50, and 100 ppm) were added to R. dentocariosa cultures grown in brain-heart infusion broth. At a late log growth phase (8 h), extracellular Pi was measured using an ascorbic acid assay, and polyP was isolated from bacterial cells treated with RNA/DNAases using a neutral phenol/chloroform extraction. Extracts were hydrolyzed and quantified as above. Gallein and fluoride had minor effects on bacterial growth with NaF having a direct effect on media pH. Gallein (≥25 µM) and fluoride (≥50 ppm) attenuated the bacterial drawdown of extracellular Pi by 56.7% (P < 0.05) and 37.3% (P < 0.01). There was a corresponding polyP synthesis decrease of 73.2% (P < 0.0001) from gallein and 83.1% (P < 0.0001) from fluoride. Attenuated total reflectance-Fourier-transform infrared spectroscopy validated the presence of polyP and its reduced concentration in R. dentocariosa bacterial cells following gallein and fluoride treatment. Rothia dentocariosa can directly change extracellular Pi and accumulate intracellular polyP, but the mechanism is attenuated by gallein and NaF.
Assuntos
Actinomycetales , Fluoretos , Polifosfatos , Boca/microbiologiaRESUMO
A radiochemical synthesis of [18 F]DK222, a peptide binder of programmed death ligand 1 protein, suitable for human PET studies is described, and results from validation productions are presented. The high specific activity radiotracer product is prepared as a sterile, apyrogenic solution that conforms to current Good Manufacturing Practice (cGMP) requirements. In addition, the production is extended to use a commercial synthesizer platform (General Electric FASTlab 2).
Assuntos
Antígeno B7-H1 , Tomografia por Emissão de Pósitrons , Humanos , Tomografia por Emissão de Pósitrons/métodos , Radioisótopos de Flúor , Compostos Radiofarmacêuticos , Radioquímica/métodosRESUMO
Accumulating research suggests that the tumor immune microenvironment (TIME) plays an essential role in regulation of tumor growth and metastasis. The cellular and molecular nature of the TIME influences cancer progression and metastasis by altering the ratio of immune- suppressive versus cytotoxic responses in the vicinity of the tumor. Targeting or activating the TIME components show a promising therapeutic avenue to combat cancer. The success of immunotherapy is both astounding and unsatisfactory in the clinic. Advancements in RNA-based technology have improved understanding of the complexity and diversity of the TIME and its effects on therapy. TIME-related RNA or RNA regulators could be promising targets for anticancer immunotherapy. In this review, we discuss the available RNA-based cancer immunotherapies targeting the TIME. More importantly, we summarize the potential of various RNA-based therapeutics clinically available for cancer treatment. RNA-dependent targeting of the TIME, as monotherapy or combined with other evolving therapeutics, might be beneficial for cancer patients' treatment in the near future.
Assuntos
Antineoplásicos , Neoplasias , Antineoplásicos/farmacologia , Humanos , Imunoterapia , Neoplasias/tratamento farmacológico , Neoplasias/terapia , RNA , Microambiente TumoralRESUMO
BACKGROUND: Cancer stem cells (CSCs) play crucial role in tumor progression, drug resistance and relapse in various cancers. CSC niche is comprised of various stromal cell types including Tumor-associated macrophages (TAMs). Extrinsic ques derived from these cells help in maintenance of CSC phenotype. TAMs have versatile roles in tumor progression however their function in enrichment of CSC is poorly explored. METHODS: Mouse macrophages (RAW264.7) cells were activated by interaction with conditioned media (CM) of murine breast cancer cells (4T1) into TAMs and the effect of activated macrophage (TAM) derived factors was examined on enrichment of cancer stem cells (CSCs) and tumor growth using in vitro and in vivo models. RESULTS: In this study, we report that macrophages upon interaction with breast cancer cells activate tumor promoting function and exhibit differential expression of various proteins as shown by secretome analysis using proteomics studies. Based on secretome data, we found that Interleukin-6 (IL-6) is one of the up-regulated genes expressed in activated macrophages. Further, we confirm that TAMs produce high levels of IL-6 and breast cancer cell derived factors induce IL-6 production in activated macrophages via p38-MAPK pathway. Furthermore, we demonstrate that tumor activated macrophages induce enrichment of CSCs and expression of CSC specific transcription factors such as Sox-2, Oct-3/4 and Nanog in breast cancer cells. We further prove that TAM derived IL-6 plays a key role in TAM mediated CSC enrichment through activation of Signal transducer and activator of transcription 3 (STAT-3) signaling. TAM derived IL-6 influences breast cancer cell migration and angiogenesis. Moreover, our in vivo findings indicated that TAM derived IL-6 induces CSC population and resulting tumor growth in breast cancer. CONCLUSION: These finding provide evidence that TAM derived IL-6 plays a major role in CSC enrichment and tumor progression in breast cancer and IL-6 and its regulated signalling network may act as potential therapeutic target for management of breast cancer.
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Iron is an essential element for Mycobacterium tuberculosis; it has at least 40 enzymes that require iron as a cofactor. Accessibility of iron at the phagosomal surface inside macrophage is crucial for survival and virulence of M. tuberculosis ESAT-6, a 6-kDa-secreted protein of region of difference 1, is known to play a crucial role in virulence and pathogenesis of M. tuberculosis In our earlier study, we demonstrated that ESAT-6 protein interacts with ß-2-microglobulin (ß2M) and affects class I Ag presentation through sequestration of ß2M inside endoplasmic reticulum, which contributes toward inhibition of MHC class I:ß2M:peptide complex formation. The 6 aa at C-terminal region of ESAT-6 are essential for ESAT6:ß2M interaction. ß2M is essential for proper folding of HFE, CD1, and MHC class I and their surface expression. It is known that M. tuberculosis recruit holotransferrin at the surface of the phagosome. But the upstream mechanism by which it modulates holotransferrin-mediated iron uptake at the surface of macrophage is not well understood. In the current study, we report that interaction of the ESAT-6 protein with ß2M causes downregulation of surface HFE, a protein regulating iron homeostasis via interacting with transferrin receptor 1 (TFR1). We found that ESAT-6:ß2M interaction leads to sequestration of HFE in endoplasmic reticulum, causing poorer surface expression of HFE and HFE:TFR1 complex (nonfunctional TFR1) in peritoneal macrophages from C57BL/6 mice, resulting in increased holotransferrin-mediated iron uptake in these macrophages. These studies suggest that M. tuberculosis probably targets the ESAT-6 protein to increase iron uptake.
Assuntos
Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Regulação para Baixo/fisiologia , Proteína da Hemocromatose/metabolismo , Macrófagos Peritoneais/metabolismo , Mycobacterium tuberculosis/metabolismo , Transferrina/metabolismo , Animais , Transporte Biológico/fisiologia , Retículo Endoplasmático/metabolismo , Ferro/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Receptores da Transferrina/metabolismo , Virulência/fisiologia , Microglobulina beta-2/metabolismoRESUMO
BACKGROUND: Preliminary data highlights the importance of anticoagulation therapy in the prevention and treatment of thromboembolism in SARS CoV-2 infection. There is insufficient data comparing the safety and efficacy of direct oral anticoagulants (DOACs) and subcutaneous enoxaparin in the prophylactic management of COVID-19 associated thromboembolic disease, particularly in mild to moderate cases of COVID-19 infection. OBJECTIVES: The study was designed to investigate the efficacy of oral rivaroxaban as a prophylactic anticoagulant in mild to moderate SARS CoV-2 infection. METHODS: In this randomized, open-label, prospective superiority trial involving hospitalized patients with confirmed mild or moderate COVID-19 disease without known thromboembolism, we assigned 230 patients to receive either once-daily oral rivaroxaban (10mg or 15mg) or once-daily subcutaneous enoxaparin (40mg or 60mg) for a median duration of 8 days. The primary outcome was a composite of all major, clinically relevant haemorrhagic and thrombotic events. RESULTS: The primary efficacy outcome occurred in 4 of 115 patients in the rivaroxaban group (3.5%) versus 16 of 113 patients in the enoxaparin group (14.2%) (hazard ratio 0.207, 95% confidence interval [CI], 0.069 to 0.621, P=0.005). Adverse events developed in 4.3% of patients in the study group and 12.4% in the enoxaparin group (hazard ratio 0.328; 95% CI, 0.118 to 0.910; P=0.032). Major bleeding was seen in 1 patient (0.9%) in the rivaroxaban group and 3 patients (2.7%) in the enoxaparin group. CONCLUSIONS: Rivaroxaban alone was superior to enoxaparin for the prophylactic management of coagulopathy associated with mild to moderate SARS CoV-2 infection.
Assuntos
COVID-19 , Tromboembolia , Anticoagulantes/efeitos adversos , Enoxaparina/efeitos adversos , Hemorragia/induzido quimicamente , Humanos , Estudos Prospectivos , Rivaroxabana/efeitos adversosRESUMO
Disease models have proven useful tools for gaining deeper mechanistic insights into neurodegenerative diseases. In this context, stem cell technology is effective, especially induced pluripotent stem cell (iPSC)-derived brain organoids and cell replacement/restoration which can be used for personalized medicine, allowing physicians to test the efficacy of drugs in vitro before delivering them to patients, enabling more precise and personalized treatment. Nonetheless, it offers the potential to minimize (or even eliminate) the use of animals, provides important clues for disease processes, and accelerates therapeutic strategies. Perhaps in the not-too-distant future, organoid models of the human brain will be able to link blood-brain barrier cultures with other liver cultures, simulating blood flow across organs and as a method of testing medicines, giving crucial pharmacokinetics and pharmacodynamics data. Simultaneously, stem cell interventions for cell replacements or restoration therapy would enable us to realize efficacious and realistic therapeutic options for Neurodegenerative diseases.
Assuntos
Barreira Hematoencefálica/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Descoberta de Drogas/métodos , Doenças Neurodegenerativas/terapia , Organoides/efeitos dos fármacos , Animais , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Encéfalo/patologia , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Células-Tronco Pluripotentes Induzidas/transplante , Organoides/citologia , Organoides/metabolismo , Medicina de Precisão/métodos , Transplante de Células-Tronco/métodosRESUMO
Identifying reliable biomarkers and ultra-sensitive techniques are crucial for the early detection of neurodegenerative disorders (NDDs) to improve the clinical diagnosis and development of effective disease-modifying treatments. Here, we discussed recent technological advancements that enabled scientists to monitor brain health by detecting biological molecules even at lower levels. These technologies enabled the detection of neurological biomarkers in blood, revolutionizing the diagnosis and prognosis of NDDs. Moreover, it provided a better understanding of disease pathology's long-term effects, resulting in fewer invasive tests, early diagnosis, faster drug development, and possibly more effective therapies as possible outcomes.
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Biomarcadores/metabolismo , Diagnóstico Precoce , Imunoensaio/métodos , Doenças Neurodegenerativas/diagnóstico , Doenças Neurodegenerativas/metabolismo , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/metabolismo , Humanos , Doença de Parkinson/diagnóstico , Doença de Parkinson/metabolismo , Sensibilidade e Especificidade , Ubiquitina-Proteína Ligases/metabolismoRESUMO
ESAT-6 is a small secreted protein of Mycobacterium tuberculosis involved in the ESAT-6 secretion system (ESX-1)-mediated virulence and pathogenesis. The protein interacts with ß2M, causing downregulation of MHC class I Ag presentation, which could be one of the mechanisms by which it favors increased survival of the bacilli inside the host. In an earlier study, we have shown that the C-terminal region of ESAT-6 is crucial for its interaction with ß2M. However, the interface of ß2M involved in interaction with ESAT-6 and detailed physicochemical changes associated with ESAT-6:ß2M complexation are not fully defined. In this study, using computational and site-directed mutagenesis studies, we demonstrate the presence of strong noncovalent hydrophobic interactions between ESAT-6 and ß2M in addition to the vital hydrogen bonding between the aspartate residue (Asp53) of ß2M and methionine (Met93) of ESAT-6. Docking-based high-throughput virtual screening followed by 16-point screening on microscale thermophoresis resulted in the identification of two potent inhibitors (SM09 and SM15) that mask the critical Met93 residue of ESAT-6 that is required for ESAT-6:ß2M interaction and could rescue cell surface expression of ß2M and HLA in human macrophages as well as MHC class I Ag presentation suppressed by ESAT-6 in peritoneal macrophages isolated from C57BL/6 mice. Both SM09 and SM15 significantly inhibited intracellular survival of M. tuberculosis in human macrophages. Further, we characterized the physicochemical properties involved in the ESAT-6:ß2M complexation, which may help in understanding host-pathogen interactions.
Assuntos
Antígenos de Bactérias/química , Proteínas de Bactérias/química , Simulação de Acoplamento Molecular , Mycobacterium tuberculosis/química , Microglobulina beta-2/química , Substituição de Aminoácidos , Animais , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Ácido Aspártico/química , Ácido Aspártico/genética , Ácido Aspártico/imunologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Interações Hospedeiro-Patógeno/imunologia , Humanos , Macrófagos Peritoneais/química , Macrófagos Peritoneais/imunologia , Camundongos , Mutagênese Sítio-Dirigida , Mutação de Sentido Incorreto , Mycobacterium tuberculosis/fisiologia , Estrutura Quaternária de Proteína , Microglobulina beta-2/genética , Microglobulina beta-2/imunologiaRESUMO
OBJECTIVES: To assess the prevalence of Iron Deficiency and impact of Parenteral Iron therapy in patients with Symptomatic Heart failure, the role of exercise capacity and serial Echocardiography in assessing treatment response. METHODS: Study was performed in a Government Hospital for 24 months, December 2017 to December 2019.120 participants were recruited. Patients with symptomatic heart failure and Serum Ferritin <100 mg/dl were recruited and those with diagnosed Ischemic Cardiomyopathy or unwilling to give consent were excluded. They underwent a functional assessment and 2D Echo at baseline, after 30 and 90 days of IV Carboxymaltose. The data was analysed represented in appropriate figures. A P value <0.05 was considered significant. RESULTS: Of 120 patients recruited, 28 were male and 92 were female. The mean age of presentation was 44 +/- 5.4 years. The Mean baseline Haemoglobin was 11.7 +/- 0.38 gm/dl.The baseline Ferritin levels were 16.69 +/-2.9 ug/L. HFpEF was predominant with 65% cases. The NYHA status and 6min HWT tests showed a statistically significant improvement and Echocardiography findings showed a statistically insignificant improvement after Parenteral Iron. CONCLUSION: Iron Deficiency is a major risk factor in Heart Failure including HFpEF and prevails in the younger population.Parenteral Iron Carboxymaltose followed by oral iron supplementation is effective in Heart Failure patients, especially in HFpEF. Functional capacity and NYHA status appear to be the time tested markers for Iron repletion.
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Anemia Ferropriva , Insuficiência Cardíaca , Adulto , Ecocardiografia , Feminino , Insuficiência Cardíaca/tratamento farmacológico , Insuficiência Cardíaca/epidemiologia , Humanos , Ferro , Masculino , Pessoa de Meia-Idade , Volume SistólicoRESUMO
[This corrects the article DOI: 10.1371/journal.ppat.1006236.].