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2.
Nat Mater ; 20(11): 1559-1570, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34326506

RESUMO

Flexible electronic/optoelectronic systems that can intimately integrate onto the surfaces of vital organ systems have the potential to offer revolutionary diagnostic and therapeutic capabilities relevant to a wide spectrum of diseases and disorders. The critical interfaces between such technologies and living tissues must provide soft mechanical coupling and efficient optical/electrical/chemical exchange. Here, we introduce a functional adhesive bioelectronic-tissue interface material, in the forms of mechanically compliant, electrically conductive, and optically transparent encapsulating coatings, interfacial layers or supporting matrices. These materials strongly bond both to the surfaces of the devices and to those of different internal organs, with stable adhesion for several days to months, in chemistries that can be tailored to bioresorb at controlled rates. Experimental demonstrations in live animal models include device applications that range from battery-free optoelectronic systems for deep-brain optogenetics and subdermal phototherapy to wireless millimetre-scale pacemakers and flexible multielectrode epicardial arrays. These advances have immediate applicability across nearly all types of bioelectronic/optoelectronic system currently used in animal model studies, and they also have the potential for future treatment of life-threatening diseases and disorders in humans.

3.
Pain ; 162(12): 2865-2880, 2021 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-34160168

RESUMO

ABSTRACT: Accumulating evidence suggests hippocampal impairment under the chronic pain phenotype. However, it is unknown whether neuropathic behaviors are related to dysfunction of the hippocampal circuitry. Here, we enhanced hippocampal activity by pharmacological, optogenetic, and chemogenetic techniques to determine hippocampal influence on neuropathic pain behaviors. We found that excitation of the dorsal (DH), but not the ventral (VH) hippocampus induces analgesia in 2 rodent models of neuropathic pain (SNI and SNL) and in rats and mice. Optogenetic and pharmacological manipulations of DH neurons demonstrated that DH-induced analgesia was mediated by N-Methyl-D-aspartate and µ-opioid receptors. In addition to analgesia, optogenetic stimulation of the DH in SNI mice also resulted in enhanced real-time conditioned place preference for the chamber where the DH was activated, a finding consistent with pain relief. Similar manipulations in the VH were ineffective. Using chemo-functional magnetic resonance imaging (fMRI), where awake resting-state fMRI was combined with viral vector-mediated chemogenetic activation (PSAM/PSEM89s) of DH neurons, we demonstrated changes of functional connectivity between the DH and thalamus and somatosensory regions that tracked the extent of relief from tactile allodynia. Moreover, we examined hippocampal functional connectivity in humans and observe differential reorganization of its anterior and posterior subdivisions between subacute and chronic back pain. Altogether, these results imply that downregulation of the DH circuitry during chronic neuropathic pain aggravates pain-related behaviors. Conversely, activation of the DH reverses pain-related behaviors through local excitatory and opioidergic mechanisms affecting DH functional connectivity. Thus, this study exhibits a novel causal role for the DH but not the VH in controlling neuropathic pain-related behaviors.

4.
Nat Neurosci ; 24(7): 1035-1045, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33972800

RESUMO

Advanced technologies for controlled delivery of light to targeted locations in biological tissues are essential to neuroscience research that applies optogenetics in animal models. Fully implantable, miniaturized devices with wireless control and power-harvesting strategies offer an appealing set of attributes in this context, particularly for studies that are incompatible with conventional fiber-optic approaches or battery-powered head stages. Limited programmable control and narrow options in illumination profiles constrain the use of existing devices. The results reported here overcome these drawbacks via two platforms, both with real-time user programmability over multiple independent light sources, in head-mounted and back-mounted designs. Engineering studies of the optoelectronic and thermal properties of these systems define their capabilities and key design considerations. Neuroscience applications demonstrate that induction of interbrain neuronal synchrony in the medial prefrontal cortex shapes social interaction within groups of mice, highlighting the power of real-time subject-specific programmability of the wireless optogenetic platforms introduced here.


Assuntos
Optogenética/instrumentação , Comportamento Social , Tecnologia sem Fio/instrumentação , Animais , Camundongos
5.
Int J Mol Sci ; 22(8)2021 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-33923724

RESUMO

Coxsackievirus A16 (CA16) is one of the major causative agents of hand, foot, and mouth disease (HFMD). Children aged <5 years are the most affected by CA16 HFMD globally. Although clinical symptoms of CA16 infections are usually mild, severe complications, such as aseptic meningitis or even death, have been recorded. Currently, no vaccine or antiviral therapy for CA16 infection exists. Single-chain variable fragment (scFv) antibodies significantly inhibit viral infection and could be a potential treatment for controlling the infection. In this study, scFv phage display libraries were constructed from splenocytes of a laying hen immunized with CA16-infected lysate. The pComb3X vector containing the scFv genes was introduced into ER2738 Escherichia coli and rescued by helper phages to express scFv molecules. After screening with five cycles of bio-panning, an effective scFv antibody showing favorable binding activity to proteins in CA16-infected lysate on ELISA plates was selected. Importantly, the selected scFv clone showed a neutralizing capability against the CA16 virus and cross-reacted with viral proteins in EV71-infected lysate. Intriguingly, polyclonal IgY antibody not only showed binding specificity against proteins in CA16-infected lysate but also showed significant neutralization activities. Nevertheless, IgY-binding protein did not cross-react with proteins in EV71-infected lysate. These results suggest that the IgY- and scFv-binding protein antibodies provide protection against CA16 viral infection in in vitro assays and may be potential candidates for treating CA16 infection in vulnerable young children.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Galinhas/imunologia , Enterovirus/imunologia , Animais , Especificidade de Anticorpos , Linhagem Celular Tumoral , Humanos , Anticorpos de Cadeia Única/imunologia , Vacinas Virais/imunologia
7.
J Med Imaging (Bellingham) ; 8(1): 014004, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33634205

RESUMO

Purpose: Deep learning is a promising technique for spleen segmentation. Our study aims to validate the reproducibility of deep learning-based spleen volume estimation by performing spleen segmentation on clinically acquired computed tomography (CT) scans from patients with myeloproliferative neoplasms. Approach: As approved by the institutional review board, we obtained 138 de-identified abdominal CT scans. A sum of voxel volume on an expert annotator's segmentations establishes the ground truth (estimation 1). We used our deep convolutional neural network (estimation 2) alongside traditional linear estimations (estimation 3 and 4) to estimate spleen volumes independently. Dice coefficient, Hausdorff distance, R 2 coefficient, Pearson R coefficient, the absolute difference in volume, and the relative difference in volume were calculated for 2 to 4 against the ground truth to compare and assess methods' performances. We re-labeled on scan-rescan on a subset of 40 studies to evaluate method reproducibility. Results: Calculated against the ground truth, the R 2 coefficients for our method (estimation 2) and linear method (estimation 3 and 4) are 0.998, 0.954, and 0.973, respectively. The Pearson R coefficients for the estimations against the ground truth are 0.999, 0.963, and 0.978, respectively (paired t -tests produced p < 0.05 between 2 and 3, and 2 and 4). Conclusion: The deep convolutional neural network algorithm shows excellent potential in rendering more precise spleen volume estimations. Our computer-aided segmentation exhibits reasonable improvements in splenic volume estimation accuracy.

8.
Arch Biochem Biophys ; 701: 108776, 2021 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-33515532

RESUMO

Cancer is a global health issue that origins thousands of deaths annually worldwide. Cyclic peptides are polypeptide chains which are formed by cyclic sequence of amide bonds between proteinogenic or non-proteinogenic amino acids. Numerous evidences indicate that cyclic peptides are implicated with the occurrence and development of cancer. This review presents the current knowledge about the role of cyclic peptides in cancer, such as liver cancer, colorectal cancer, ovarian cancer, breast cancer as well as prostate cancer. Specifically, the precise molecular mechanisms between cyclic peptides and cancer are elaborated. Some cyclic peptides from nature and synthesis prevent the occurrence and development of cancer. However, some other cyclic peptides including endothelin-1, urotensinⅡand melanin-concentrating hormone deteriorate the pathogenesis of cancer. Given the pleiotropic actions of cyclic peptides, the identification and development of cyclic peptides and their derivates as drug may be a potent therapeutic strategy for cancer.


Assuntos
Antineoplásicos/uso terapêutico , Neoplasias/tratamento farmacológico , Peptídeos Cíclicos/uso terapêutico , Antineoplásicos/química , Humanos , Neoplasias/metabolismo , Neoplasias/patologia , Peptídeos Cíclicos/química
9.
Clin Toxicol (Phila) ; 59(1): 28-37, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32400229

RESUMO

INTRODUCTION: Protobothrops mucrosquamatus bite induces wound necrosis, coagulopathy, thrombocytopenia, rhabdomyolysis, and acute renal failure. The severity of the hematological derangements and associated factors for wound necrosis and subsequent surgery and the appropriate management of these conditions have not been well characterized. Although severe renal failure requiring hemodialysis has been reported following P. mucrosquamatus bite, the culprit snake may be erroneously classified. MATERIALS AND METHODS: A total of 186 patients with P. mucrosquamatus bites were retrospectively evaluated. They were categorized into group 1 (patients receiving debridement or finger/toe amputation) and group 2 (all other patients) to identify the associated factors for surgery. Characteristic data were compared between groups 1 and 2 and between definite and suspected cases. RESULTS: No differences were observed between definite and suspected cases in terms of symptomatology and management. Of the 186 patients, 7 (3.8%) were asymptomatic, 179 (96.2%) experienced tissue swelling and pain, and 107 (57.5%) had local ecchymosis. Coagulopathy, thrombocytopenia, and renal impairment were found in 13 (7%), 19 (10.2%), and 7 (3.8%) patients, respectively. None of the patients required transfusion therapy or hemodialysis. Furthermore, no systemic bleeding or death occurred. Antivenom was administered to all 179 envenomed patients at a median of 1.5 h post-bite. The median total dose of the specific antivenom was 5.5 vials. In multivariate logistic regression analysis, finger as the bite site, bullae and blister formation, and wound infection were significantly associated with wound necrosis; whereas finger as the bite site and bullae and blister formation were related to debridement or finger/toe amputation. DISCUSSION AND CONCLUSIONS: Protobothrops mucrosquamatus envenomation mainly exerts effects on local tissue. Systemic effects are uncommon and generally nonsevere and transient after the treatment with the specific antivenom. We speculated that severe renal failure requiring hemodialysis is not a typical finding of P. mucrosquamatus envenomation. Patients with finger as the bite site and bullae or blister formation should be carefully examined for wound necrosis, secondary infection, and subsequent surgery. Further evaluations of the efficacy of antivenom against local tissue effects and the effect of selective antibiotics in the management of bite wound infection are urgently required. Although the antivenom manufacturer suggested a skin test prior to use, we believed that it could be omitted because it does not accurately predict the allergic responses.


Assuntos
Amputação , Antivenenos/uso terapêutico , Venenos de Crotalídeos/antagonistas & inibidores , Desbridamento , Dedos/cirurgia , Mordeduras de Serpentes/terapia , Dedos do Pé/cirurgia , Trimeresurus , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Antibacterianos/uso terapêutico , Antivenenos/efeitos adversos , Criança , Pré-Escolar , Protocolos Clínicos , Venenos de Crotalídeos/metabolismo , Feminino , Dedos/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Necrose , Diálise Renal , Insuficiência Renal/etiologia , Insuficiência Renal/terapia , Estudos Retrospectivos , Mordeduras de Serpentes/sangue , Mordeduras de Serpentes/complicações , Mordeduras de Serpentes/patologia , Taiwan , Dedos do Pé/patologia , Trimeresurus/metabolismo , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/terapia , Adulto Jovem
10.
J Venom Anim Toxins Incl Trop Dis ; 26: e20200056, 2020 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-33281887

RESUMO

Background: The venom of bamboo vipers (Trimeresurus stejnegeri - TS), commonly found in Taiwan, contains deadly hemotoxins that cause severe envenomation. Equine-derived antivenom is a specific treatment against snakebites, but its production costs are high and there are some inevitable side effects. The aim of the present work is to help in the development of an affordable and more endurable therapeutic strategy for snakebites. Methods: T. stejnegeri venom proteins were inactivated by glutaraldehyde in order to immunize hens for polyclonal immunoglobulin (IgY) antibodies production. After IgY binding assays, two antibody libraries were constructed expressing single-chain variable fragment (scFv) antibodies joined by the short or long linker for use in phage display antibody technology. Four rounds of biopanning were carried out. The selected scFv antibodies were then further tested for their binding activities and neutralization assays to TS proteins. Results: Purified IgY from egg yolk showed the specific binding ability to TS proteins. The dimensions of these two libraries contain 2.4 × 107 and 6.8 × 107 antibody clones, respectively. An increase in the titers of eluted phage indicated anti-TS clones remarkably enriched after 2nd panning. The analysis based on the nucleotide sequences of selected scFv clones indicated that seven groups of short linkers and four groups of long linkers were identified. The recombinant scFvs showed significant reactivity to TS venom proteins and a cross-reaction to Trimeresurus mucrosquamatus venom proteins. In in vivo studies, the data demonstrated that anti-TS IgY provided 100% protective effects while combined scFvs augmented partial survival time of mice injected with a lethal amount of TS proteins. Conclusion: Chickens were excellent hosts for the production of neutralization antibodies at low cost. Phage display technology is available for generation of monoclonal antibodies against snake venom proteins. These antibodies could be applied in the development of diagnostic kits or as an alternative for snakebite envenomation treatment in the near future.

11.
Quant Imaging Med Surg ; 10(12): 2297-2306, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33269228

RESUMO

Background: The underestimation of renal depth by Tønnesen formula in Gates' method, which has been confirmed by many scholars, leads to the underestimation of both separate glomerular filtration rate (gSGFR) and total glomerular filtration rate (gTGFR). This study aimed to establish the normal reference ranges of renal depth-calibrated gTGFR and gSGFR in Chinese healthy adults, and to analyze the influencing factors. Methods: Renal depth was measured by CT scan followed by technetium 99m-diethylene triamine pentaacetic acid (99mTc-DTPA) renal dynamic imaging by single-photon emission computed tomography/computed tomography (SPECT/CT) in 329 living kidney donors. The renal depth-calibrated gTGFR and gSGFR were calculated by Gates' method with renal depth measured by CT instead of being calculated by the Tønnesen formula. A general linear model based on age, gender, body height, body weight, and BMI was used to analyze factors influencing gSGFR (L), gSGFR (R) and gTGFR. Results: The average gSGFR (L), gSGFR (R), and gTGFR for patients aged 23-64 years old were 49.3±10.1, 49.9±10.4, and 99.1±18.7 mL/min/1.73 m2, respectively. The gSGFR (L), gSGFR (R) and gTGFR for patients aged 41-50 years old were 26.9-69.3, 27.7-68.8, and 57.5-135.3 mL/min/1.73 m2, respectively, and those for patients aged 51-60 years old were 31.0-61.5, 29.5-63.3, and 64.6-120.7 mL/min/1.73 m2, respectively. gSGFR (L), gSGFR (R) and gTGFR had statistical significance with body height and age (P<0.05); however, there was no significant difference with gender, body weight, and BMI (P>0.05). For each 1 year increase in age, the gSGFR (L), gSGFR (R), and gTGFR decreased by 0.17, 0.28, and 0.44 mL/min/1.73 m2, respectively, while for every 1 cm increase in body height, the gSGFR (L), gSGFR (R), and gTGFR decreased by 0.37, 0.36, and 0.74 mL/min/1.73 m2, respectively. Conclusions: Normal reference ranges for renal depth-calibrated gSGFR (L), gSGFR (R), and gTGFR were established in healthy Chinese adults aged 23-64 years, and gSGFR (L), gSGFR (R), and gTGFR decreased with age and body height.

12.
Nat Neurosci ; 23(12): 1522-1536, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33199897

RESUMO

Interest in deciphering the fundamental mechanisms and processes of the human mind represents a central driving force in modern neuroscience research. Activities in support of this goal rely on advanced methodologies and engineering systems that are capable of interrogating and stimulating neural pathways, from single cells in small networks to interconnections that span the entire brain. Recent research establishes the foundations for a broad range of creative neurotechnologies that enable unique modes of operation in this context. This review focuses on those systems with proven utility in animal model studies and with levels of technical maturity that suggest a potential for broad deployment to the neuroscience community in the relatively near future. We include a brief summary of existing and emerging neuroscience techniques, as background for a primary focus on device technologies that address associated opportunities in electrical, optical and microfluidic neural interfaces, some with multimodal capabilities. Examples of the use of these technologies in recent neuroscience studies illustrate their practical value. The vibrancy of the engineering science associated with these platforms, the interdisciplinary nature of this field of research and its relevance to grand challenges in the treatment of neurological disorders motivate continued growth of this area of study.


Assuntos
Neurociências/tendências , Tecnologia/tendências , Animais , Encéfalo/fisiologia , Estimulação Elétrica/instrumentação , Estimulação Elétrica/métodos , Humanos , Neurociências/métodos , Optogenética/tendências , Farmacologia/métodos , Pesquisa
13.
J Venom Anim Toxins Incl Trop Dis ; 26: e20200043, 2020 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-32983233

RESUMO

Background: Trimeresurus stejnegeri stejnegeri bite induces tissue swelling, pain, thrombocytopenia, rhabdomyolysis, and acute renal failure. However, the incidence of coagulopathy, factors associated with wound necrosis, and the appropriate management of this condition have not been well characterized yet. Materials: This study included patients bitten by T. s. stejnegeri that were admitted to the study hospitals from 2001 to 2016. Patient characteristics, laboratory data, and management approaches were compared in victims with and without wound necrosis. Results: A total of 185 patients were evaluated: three patients (1.6%) were asymptomatic; whereas tissue swelling and pain, local ecchymosis, wound necrosis, coagulopathy, thrombocytopenia, rhabdomyolysis, and renal impairment were present in 182, 53, 13, 15, 10, 1, and 3 patients, respectively. One patient died from coagulopathy and hemorrhagic shock. Antivenom was administered to all envenomed patients at a median time of 1.8 h after the bite. The median total dose of antivenom was five vials. Chi-square analysis showed that bitten fingers, using cold packs during first aid, presence of bullae or blisters, lymphangitis or lymphadenitis, local numbness and suspected infection to be significantly associated with wound necrosis. After adjustment using a multivariate logistic regression model, only cold packs as first aid, bulla or blister formation, and wound infection remained significant. Conclusions: The main effects of T. s. stejnegeri envenomation are tissue swelling, pain, and local ecchymosis. We do not recommend the use of cold packs during first aid to reduce wound pain, as this may be a risk factor for wound necrosis. In addition, patients with bulla or blister formation should be carefully examined for subsequent wound necrosis. Antiplatelet use may worsen systemic bleeding. No severe rhabdomyolysis or renal failure was observed in this large case series, we therefore considered that they were not prominent effects of T. s. stejnegeri bite.

15.
Int J Mol Sci ; 21(8)2020 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-32326294

RESUMO

Candida albicans (C. albicans) is an opportunistic human pathogen responsible for approximately a half of clinical candidemia. The emerging Candida spp. with resistance to azoles is a major challenge in clinic, suggesting an urgent demand for new drugs and therapeutic strategies. Alpha-enolase (Eno1) is a multifunctional protein and represents an important marker for invasive candidiasis. Thus, C. albicans Eno1 (CaEno1) is believed to be an important target for the development of therapeutic agents and antibody drugs. Recombinant CaEno1 (rCaEno1) was first used to immunize chickens. Subsequently, we used phage display technology to construct two single chain variable fragment (scFv) antibody libraries. A novel biopanning procedure was carried out to screen anti-rCaEno1 scFv antibodies, whose specificities were further characterized. The polyclonal IgY antibodies showed binding to rCaEno1 and native CaEno1. A dominant scFv (CaS1) and its properties were further characterized. CaS1 attenuated the growth of C. albicans and inhibited the binding of CaEno1 to plasminogen. Animal studies showed that CaS1 prolonged the survival rate of mice and zebrafish with candidiasis. The fungal burden in kidney and spleen, as well as level of inflammatory cytokines were significantly reduced in CaS1-treated mice. These results suggest CaS1 has potential of being immunotherapeutic drug against C. albicans infections.


Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Candida albicans/enzimologia , Inibidores Enzimáticos/farmacologia , Fosfopiruvato Hidratase/antagonistas & inibidores , Anticorpos de Cadeia Única/farmacologia , Animais , Avaliação Pré-Clínica de Medicamentos , Camundongos , Ligação Proteica , Peixe-Zebra
16.
Clin Chim Acta ; 504: 125-137, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32017925

RESUMO

Endoplasmic reticulum (ER) is an intracellular membranous organelle involved in the synthesis, folding, maturation and post-translation modification of secretory and transmembrane proteins. Therefore, ER is closely related to the maintenance of intracellular homeostasis and the good balance between health and diseases. Endoplasmic reticulum stress (ERS) occurs when unfolded/misfolded proteins accumulate after disturbance of ER environment. In response to ERS, cells trigger an adaptive response called the Unfolded protein response (UPR), which helps cells cope with the stress. In recent years, a large number of studies show that ERS can aggravate cardiovascular diseases. ERS-related proteins expression in cardiovascular diseases is on the rise. Therefore, down-regulation of ERS is critical for alleviating symptoms of cardiovascular diseases, which may be used in the near future to treat cardiovascular diseases. This article reviews the relationship between ERS and cardiovascular diseases and drugs that inhibit ERS. Furthermore, we detail the role of ERS inhibitors in the treatment of cardiovascular disease. Drugs that inhibit ERS are considered as promising strategies for the treatment of cardiovascular diseases.


Assuntos
Doenças Cardiovasculares , Estresse do Retículo Endoplasmático , Doenças Cardiovasculares/tratamento farmacológico , Doenças Cardiovasculares/metabolismo , Descoberta de Drogas , Retículo Endoplasmático/metabolismo , Humanos , Resposta a Proteínas não Dobradas
17.
Adv Mater ; 32(14): e1908424, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32100406

RESUMO

Deterministic transformations of 2D patterns of materials into well-controlled 3D mesostructures serve as the basis for manufacturing methods that can bypass limitations of conventional 3D micro/nanofabrication. Here, guided mechanical buckling processes provide access to a rich range of complex 3D mesostructures in high-performance materials, from inorganic and organic semiconductors, metals and dielectrics, to ceramics and even 2D materials (e.g., graphene, MoS2 ). Previous studies demonstrate that iterative computational procedures can define design parameters for certain targeted 3D configurations, but without the ability to address complex shapes. A technical need is in efficient, generalized inverse design algorithms that directly yield sets of optimized parameters. Here, such schemes are introduced, where the distributions of thicknesses across arrays of separated or interconnected ribbons provide scalable routes to 3D surfaces with a broad range of targeted shapes. Specifically, discretizing desired shapes into 2D ribbon components allows for analytic solutions to the inverse design of centrally symmetric and even general surfaces, in an approximate manner. Combined theoretical, numerical, and experimental studies of ≈20 different 3D structures with characteristic sizes (e.g., ribbon width) ranging from ≈200 µm to ≈2 cm and with geometries that resemble hemispheres, fire balloons, flowers, concave lenses, saddle surfaces, waterdrops, and rodents, illustrate the essential ideas.

18.
Int J Mol Sci ; 21(2)2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31940993

RESUMO

Zika virus (ZIKV) is a new and emerging virus that has caused outbreaks worldwide. The virus has been linked to congenital neurological malformations in neonates and Guillain-Barré syndrome in adults. Currently there are no effective vaccines available. As a result, there is a great need for ZIKV treatment. In this study, we developed single chain variable fragment (scFv) antibodies that target the ZIKV envelope protein using phage display technology. We first induced an immune response in white leghorn laying hens against the ZIKV envelope (E) protein. Chickens were immunized and polyclonal immunoglobulin yolk (IgY) antibodies were extracted from egg yolks. A high-level titer of anti-ZIKV_E IgY antibodies was detected using enzyme-linked immunosorbent assay (ELISA) after the third immunization. The titer persisted for at least 9 weeks. We constructed two antibody libraries that contained 5.3 × 106 and 4.5 × 106 transformants. After biopanning, an ELISA phage assay confirmed the enrichment of specific clones. We randomly selected 26 clones that expressed ZIKV scFv antibodies and classified them into two groups, short-linker and long-linker. Of these, four showed specific binding activities toward ZIKV_E proteins. These data suggest that the polyclonal and monoclonal scFv antibodies have the diagnostic or therapeutic potential for ZIKV.


Assuntos
Anticorpos Antivirais , Proteínas Aviárias , Galinhas , Anticorpos de Cadeia Única , Proteínas do Envelope Viral/imunologia , Zika virus/imunologia , Animais , Anticorpos Antivirais/química , Anticorpos Antivirais/genética , Anticorpos Antivirais/imunologia , Anticorpos Antivirais/isolamento & purificação , Proteínas Aviárias/química , Proteínas Aviárias/genética , Proteínas Aviárias/imunologia , Proteínas Aviárias/isolamento & purificação , Galinhas/genética , Galinhas/imunologia , Expressão Gênica , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Anticorpos de Cadeia Única/química , Anticorpos de Cadeia Única/genética , Anticorpos de Cadeia Única/imunologia , Anticorpos de Cadeia Única/isolamento & purificação
19.
Artigo em Inglês | MEDLINE | ID: mdl-34040274

RESUMO

Deep learning has achieved many successes in medical imaging, including lung nodule segmentation and lung cancer prediction on computed tomography (CT). Recently, multi-task networks have shown to both offer additional estimation capabilities, and, perhaps more importantly, increased performance over single-task networks on a "main/primary" task. However, balancing the optimization criteria of multi-task networks across different tasks is an area of active exploration. Here, we extend a previously proposed 3D attention-based network with four additional multi-task subnetworks for the detection of lung cancer and four auxiliary tasks (diagnosis of asthma, chronic bronchitis, chronic obstructive pulmonary disease, and emphysema). We introduce and evaluate a learning policy, Periodic Focusing Learning Policy (PFLP), that alternates the dominance of tasks throughout the training. To improve performance on the primary task, we propose an Internal-Transfer Weighting (ITW) strategy to suppress the loss functions on auxiliary tasks for the final stages of training. To evaluate this approach, we examined 3386 patients (single scan per patient) from the National Lung Screening Trial (NLST) and de-identified data from the Vanderbilt Lung Screening Program, with a 2517/277/592 (scans) split for training, validation, and testing. Baseline networks include a single-task strategy and a multi-task strategy without adaptive weights (PFLP/ITW), while primary experiments are multi-task trials with either PFLP or ITW or both. On the test set for lung cancer prediction, the baseline single-task network achieved prediction AUC of 0.8080 and multi-task baseline failed to converge (AUC 0.6720). However, applying PFLP helped multi-task network clarify and achieved test set lung cancer prediction AUC of 0.8402. Furthermore, our ITW technique boosted the PFLP enabled multi-task network and achieved an AUC of 0.8462 (McNemar test, p < 0.01). In conclusion, adaptive consideration of multi-task learning weights is important, and PFLP and ITW are promising strategies.

20.
J. venom. anim. toxins incl. trop. dis ; 26: e20200056, 2020. tab, graf
Artigo em Inglês | ID: biblio-1135145

RESUMO

The venom of bamboo vipers (Trimeresurus stejnegeri - TS), commonly found in Taiwan, contains deadly hemotoxins that cause severe envenomation. Equine-derived antivenom is a specific treatment against snakebites, but its production costs are high and there are some inevitable side effects. The aim of the present work is to help in the development of an affordable and more endurable therapeutic strategy for snakebites. Methods: T. stejnegeri venom proteins were inactivated by glutaraldehyde in order to immunize hens for polyclonal immunoglobulin (IgY) antibodies production. After IgY binding assays, two antibody libraries were constructed expressing single-chain variable fragment (scFv) antibodies joined by the short or long linker for use in phage display antibody technology. Four rounds of biopanning were carried out. The selected scFv antibodies were then further tested for their binding activities and neutralization assays to TS proteins. Results: Purified IgY from egg yolk showed the specific binding ability to TS proteins. The dimensions of these two libraries contain 2.4 × 107 and 6.8 × 107 antibody clones, respectively. An increase in the titers of eluted phage indicated anti-TS clones remarkably enriched after 2nd panning. The analysis based on the nucleotide sequences of selected scFv clones indicated that seven groups of short linkers and four groups of long linkers were identified. The recombinant scFvs showed significant reactivity to TS venom proteins and a cross-reaction to Trimeresurus mucrosquamatus venom proteins. In in vivo studies, the data demonstrated that anti-TS IgY provided 100% protective effects while combined scFvs augmented partial survival time of mice injected with a lethal amount of TS proteins. Conclusion: Chickens were excellent hosts for the production of neutralization antibodies at low cost. Phage display technology is available for generation of monoclonal antibodies against snake venom proteins. These antibodies could be applied in the development of diagnostic kits or as an alternative for snakebite envenomation treatment in the near future.(AU)


Assuntos
Animais , Venenos de Serpentes , Antivenenos , Galinhas , Trimeresurus , Anticorpos , Bacteriófagos
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