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Multiple stressors are linked to declines of insects and important pollinators, such as bees. Recently, interactive effects of multiple agrochemicals on bees have been highlighted, including fungicides, which increase toxicity of neonicotinoid insecticides. Here, we use a semi-field study across two seasons in controlled foraging tunnels to test the effects of a field application of a commercial fungicide product with two active ingredients (pyraclostrobin and metconazole) applied at label rates. We also examine its interactive effects with the neonicotinoid insecticide clothianidin, at a conservative field-realistic dose of 2.23 ppb, on 48 honey bee colonies. We found combined effects of pesticide exposure, including additive 2.93-fold increases in mortality, and an additional effect of increased infestation levels of the ectoparasitic mite, Varroa destructor. Pesticide treatments also reduced colony activity, reduced colony weight, and increased sugar consumption of whole colonies. These findings indicate that typical sublethal exposure levels to common, co-occurring agrochemicals in the field significantly affect the health of whole honey bee colonies, highlighting an unintended consequence of increasing pesticide applications.
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In response to the increasing demand for spheroid-based cancer research, the importance of developing integrated platforms that can simultaneously facilitate high-throughput spheroid production and multiplexed analysis is emphasized. In addition, the understanding of how the size and cellular composition of tumors directly influence their internal structures and functionalities underlines the critical need to produce spheroids of diverse sizes and compositions on a large scale. To address this rising demand, this work presents a configurable and linkable in vitro three-dimensional (3D) cell culture kit (CLiCK) for spheroids, termed CLiCK-Spheroid. This platform consists of three primary components: a hanging drop microarray (HDMA), a concave pillar microarray (CPMA), and gradient blocks. The HDMA alone produces a homogeneous spheroid array, while its combination with the gradient block enables one-step generation of a size-gradient spheroid array. Using the size-gradient spheroid arrays, the occurrence of necrotic cores based on spheroid size is demonstrated. Additionally, spheroids in a single batch can be conveniently compartmentalized and regrouped using a CPMA, enhancing the versatility of spheroid arrays and enabling multiplexed drug treatments. By combining the different assembly methods, this work has achieved high-throughput production of cell composition-gradient spheroid arrays, with noticeable variations in morphology and vascularization based on cell compositions.
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Esferoides Celulares , Esferoides Celulares/metabolismo , Esferoides Celulares/citología , Humanos , Línea Celular Tumoral , Técnicas de Cultivo Tridimensional de Células/métodos , Técnicas de Cultivo Tridimensional de Células/instrumentación , Técnicas de Cultivo de Célula/métodos , Técnicas de Cultivo de Célula/instrumentaciónRESUMEN
Nabis stenoferus is a zoophytophagous predator that lives in grasslands around agricultural fields. It is a candidate biological control agent for use via augmentation or conservation. To find a suitable food source for mass-rearing and to better understand this predator's biology, we compared the life history characteristics of N. stenoferus under the three different diets: aphids only (Myzus persicae), moth eggs only (Ephestia kuehniella), or a mixed diet of aphids and moth eggs. Interestingly, when only aphids were supplied, N. stenoferus developed to the adult stage but lacked normal levels of fecundity. There was a significant synergism of the mixed diet on N. stenoferus fitness in both the immature and adult stages, i.e., a 13% reduction in the nymphal developmental period and an 87.3-fold increase in fecundity, compared to aphid-only diet. Furthermore, the intrinsic rate of increase was significantly higher for the mixed diet (0.139) than either aphids only (0.022) or moth eggs only (0.097). These results show that M. persicae alone is not a complete diet for the mass-rearing of N. stenoferus, whereas this aphid can be a supplementary food when combined with E. kuehniella eggs. Implications and applications of these findings for biological control are discussed.
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Áfidos , Mariposas Nocturnas , Animales , Óvulo , Heterópteros , Dieta , FemeninoRESUMEN
Profiling molecular expression in situ allows the integration of biomolecular and cellular features, enabling an in-depth understanding of biological systems. Multiplexed immunofluorescence methods can visualize tens to hundreds of proteins from individual tissue samples, but their application is usually limited to thin tissue sections. Multiplexed immunofluorescence of thick tissues or intact organs will enable high-throughput profiling of cellular protein expression within 3D tissue architectures (e.g., blood vessels, neural projections, tumors), opening a new dimension in diverse biological research and medical applications. We will review current multiplexed immunofluorescence methods and discuss possible approaches and challenges to achieve 3D multiplexed immunofluorescence.
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Proteínas , Técnica del Anticuerpo FluorescenteRESUMEN
In binocular animals that exhibit stereoscopic visual responses, the axons of retinal ganglion cells (RGCs) connect to brain areas bilaterally by forming a commissure called the optic chiasm (OC). Ventral anterior homeobox 1 (Vax1) contributes to the formation of the OC, acting endogenously in optic pathway cells and exogenously in growing RGC axons. Here, we generated Vax1AA/AA mice expressing the Vax1AA mutant, which is incapable of intercellular transfer. We found that RGC axons cannot take up Vax1AA protein from the Vax1AA/AA mouse optic stalk (OS) and grow slowly to arrive at the hypothalamus at a late stage. The RGC axons of Vax1AA/AA mice connect exclusively to ipsilateral brain areas after failing to access the midline, resulting in reduced visual acuity and abnormal oculomotor responses. Overall, our study provides physiological evidence for the necessity of intercellular transfer of Vax1 and the importance of the bilateral RGC axon projection in proper visuomotor responses.
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Neuropéptidos , Quiasma Óptico , Ratones , Animales , Quiasma Óptico/metabolismo , Células Ganglionares de la Retina , Encéfalo/metabolismo , Ratones Endogámicos C57BL , Neuropéptidos/metabolismo , Proteínas de Homeodominio/metabolismoRESUMEN
Cell-generated mechanical forces drive many cellular and tissue-level movements and rearrangements required for the tissue or organ to develop its shape1, 2, 3, 4, 5. The prevalent view of tissue morphogenesis relies on epithelial folding resulting in compressed epithelial monolayers, overlooking the involvement of stroma in morphogenesis1, 4, 6, 7. Here, we report a giant web-like network formation of stromal cells in the epithelium-stroma interface, resulting from a multi-scale mechano-reciprocity between migrating cells and their extracellular environment. In multi-layered tissues, surface wrinkles form by a stromal cell-mediated tensional force exerted at the basement membrane. The topographical cue is transmitted to the stromal cell, directing its protrusion and migration along the wrinkles. This inductive movement of the cells conveys traction forces to its surrounding extracellular matrix, remodeling the local architectures of the stroma. In this manner, stromal cells and wrinkles communicate recursively to generate the cellular network. Our observation provides a rational mechanism for network formation in living tissues and a new understanding of the role of cellular-level tensional force in morphogenesis.
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Neuronal ensembles that hold specific memory (memory engrams) have been identified in the hippocampus, amygdala, or cortex. However, it has been hypothesized that engrams of a specific memory are distributed among multiple brain regions that are functionally connected, referred to as a unified engram complex. Here, we report a partial map of the engram complex for contextual fear conditioning memory by characterizing encoding activated neuronal ensembles in 247 regions using tissue phenotyping in mice. The mapping was aided by an engram index, which identified 117 cFos+ brain regions holding engrams with high probability, and brain-wide reactivation of these neuronal ensembles by recall. Optogenetic manipulation experiments revealed engram ensembles, many of which were functionally connected to hippocampal or amygdala engrams. Simultaneous chemogenetic reactivation of multiple engram ensembles conferred a greater level of memory recall than reactivation of a single engram ensemble, reflecting the natural memory recall process. Overall, our study supports the unified engram complex hypothesis for memory storage.
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Mapeo Encefálico , Memoria , Animales , Encéfalo , Miedo/fisiología , Hipocampo/fisiología , Memoria/fisiología , RatonesRESUMEN
A handheld confocal microscope using a rapid MEMS scanning mirror facilitates real-time optical biopsy for simple cancer diagnosis. Here we report a handheld confocal microscope catheter using high definition and high frame rate (HDHF) Lissajous scanning MEMS mirror. The broad resonant frequency region of the fast axis on the MEMS mirror with a low Q-factor facilitates the flexible selection of scanning frequencies. HDHF Lissajous scanning was achieved by selecting the scanning frequencies with high greatest common divisor (GCD) and high total lobe number. The MEMS mirror was fully packaged into a handheld configuration, which was coupled to a home-built confocal imaging system. The confocal microscope catheter allows fluorescence imaging of in vivo and ex vivo mouse tissues with 30â Hz frame rate and 95.4% fill factor at 256 × 256 pixels image, where the lateral resolution is 4.35 µm and the field-of-view (FOV) is 330 µm × 330 µm. This compact confocal microscope can provide diverse handheld microscopic applications for real-time, on-demand, and in vivo optical biopsy.
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Amblyseius eharai is a generalist predatory mite that consumes spider mites, rust mites, thrips, and pollen, with a high adaptability to various plants. To better understand ecological and behavioral aspects of this species, we investigated its functional response to different stages of two-spotted spider mite, Tetranychus urticae. Furthermore, we compared its environmental adaptability with that of other referenced phytoseiids using a temperature-dependent model of the intrinsic rate of increase. We were able to calculate the functional response parameters of both sexes of A. eharai when preying on eggs or larvae of T. urticae and, for females only, when preying on the deutonymph of T. urticae. Among the various combinations tested herein, A. eharai females preying on T. urticae larvae had the highest attack rate and shortest handling time. For eggs of T. urticae, A. eharai showed a lower attack rate; however, its handling time for eggs was significantly shorter compared to other phytoseiids. Using T. urticae larva as a prey, the attack rate of female A. eharai was higher and the handling time of both sexes of this species was shorter than those of other phytoseiid mites. Amblyseius eharai populations can show maximum performance quickly due to this species' lower optimal temperature for population growth (28.1°C) compared to other phytoseiid mites. Thus, we provided evidence that this predatory mite has the potential to be a new, effective biological control agent of greenhouse pests such as T. urticae due to its high predation capacity and low optimal temperature.
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Conducta Alimentaria , Ácaros/fisiología , Control Biológico de Vectores/métodos , Conducta Predatoria , Temperatura , Tetranychidae/fisiología , Animales , Femenino , MasculinoRESUMEN
An essential step toward understanding brain function is to establish a structural framework with cellular resolution on which multi-scale datasets spanning molecules, cells, circuits and systems can be integrated and interpreted1. Here, as part of the collaborative Brain Initiative Cell Census Network (BICCN), we derive a comprehensive cell type-based anatomical description of one exemplar brain structure, the mouse primary motor cortex, upper limb area (MOp-ul). Using genetic and viral labelling, barcoded anatomy resolved by sequencing, single-neuron reconstruction, whole-brain imaging and cloud-based neuroinformatics tools, we delineated the MOp-ul in 3D and refined its sublaminar organization. We defined around two dozen projection neuron types in the MOp-ul and derived an input-output wiring diagram, which will facilitate future analyses of motor control circuitry across molecular, cellular and system levels. This work provides a roadmap towards a comprehensive cellular-resolution description of mammalian brain architecture.
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Corteza Motora/anatomía & histología , Corteza Motora/citología , Neuronas/clasificación , Animales , Atlas como Asunto , Femenino , Neuronas GABAérgicas/citología , Neuronas GABAérgicas/metabolismo , Glutamatos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Neuroimagen , Neuronas/citología , Neuronas/metabolismo , Especificidad de Órganos , Análisis de Secuencia de ARN , Análisis de la Célula IndividualRESUMEN
Bemisia tabaci Mediterranean (Gennadius) (Hemiptera: Aleyrodidae) is an economically important insect pest worldwide. Previously, we have reported that most B. tabaci Mediterranean (MED) populations occurring in greenhouse tomatoes in Korea have been displaced from well-differentiated two genetic clusters (C1 and C2) to one (C2) during one-year period. To elucidate factors responsible for this phenomenon, we compared life history parameters of these two different genetic clusters through single and cross mating experiments on two different host plants, cucumber and tobacco, at 26°C. Intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (Ro) were significantly higher in the dominating cluster (C2) (0.247, 1.280, and 192.402, respectively on cucumber; 0.226, 1.253, and 133.792, respectively on tobacco) than in the other cluster (C1) (0.149, 1.161, and 50.539, respectively on cucumber; 0.145, 1.156, and 53.332, respectively on tobacco). Overall performances of cross mating groups, C2fC1m (C2 female × C1 male) and C1fC2m (C1 female × C2 male), were in-between those of C2 and C1, with C2fC1m performing better than C1fC2m. Thus, maternal inheritance appeared to be significantly associated with their life history parameters, with partial involvement of paternal inheritance. Our results demonstrated that the rapid displacement of genetic clusters of B. tabaci MED populations was clearly associated with differences in their life history parameters.
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Cruzamientos Genéticos , Hemípteros/crecimiento & desarrollo , Hemípteros/genética , Estadios del Ciclo de Vida/genética , Familia de Multigenes , Conducta Sexual Animal/fisiología , Animales , Teorema de Bayes , Peso Corporal , Femenino , Fertilidad , Hemípteros/anatomía & histología , Masculino , Oviposición , Razón de Masculinidad , Análisis de SupervivenciaRESUMEN
The basal ganglia regulate a wide range of behaviors, including motor control and cognitive functions, and are profoundly affected in Parkinson's disease (PD). However, the functional organization of different basal ganglia nuclei has not been fully elucidated at the circuit level. In this study, we investigated the functional roles of distinct parvalbumin-expressing neuronal populations in the external globus pallidus (GPe-PV) and their contributions to different PD-related behaviors. We demonstrate that substantia nigra pars reticulata (SNr)-projecting GPe-PV neurons and parafascicular thalamus (PF)-projecting GPe-PV neurons are associated with locomotion and reversal learning, respectively. In a mouse model of PD, we found that selective manipulation of the SNr-projecting GPe-PV neurons alleviated locomotor deficit, whereas manipulation of the PF-projecting GPe-PV neurons rescued the impaired reversal learning. Our findings establish the behavioral importance of two distinct GPe-PV neuronal populations and, thereby, provide a new framework for understanding the circuit basis of different behavioral deficits in the Parkinsonian state.
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Globo Pálido/fisiopatología , Vías Nerviosas/fisiopatología , Neuronas/fisiología , Trastornos Parkinsonianos/fisiopatología , Animales , Femenino , Trastornos Neurológicos de la Marcha/fisiopatología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Parvalbúminas , Aprendizaje Inverso/fisiologíaRESUMEN
The zoophytophagous mirid Nesidiocoris tenuis (Hemiptera: Miridae) is one of the biological control agents against the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae), a major pest of greenhouse crops. The successful establishment of a biological control agent and its co-occurrence with the target pests increases the efficacy of biological control programs in greenhouses. In this study, we explored the effects of different wavelengths of LED light on establishment of N. tenuis in laboratory condition, with the goal of enhancing the biological control of B. tabaci in greenhouse crops. Nesidiocoris tenuis was most strongly attracted by LED light at a wavelength of 385 nm. This same wavelength was also highly attractive to B. tabaci in Y-tube experiments with lights of specific wavelengths provided is each arm of the apparatus. In trials in growth chambers, we verified the attraction of N. tenuis to 385 nm wavelength. When LED light at a wavelength of 385 nm was used in a growth chamber for 6 hours out of 24 hours, it significantly increased the remaining number of N. tenuis in growth chamber and level of predation compared to treatment with white LED light or without LED light. In conclusion, UV-LED light at a wavelength of 385 nm attracts both B. tabaci and N. tenuis. Thus, it would be used for enhancing early establishment of this mirid bug, better spatial congruence of both mirid bug and whitefly, and better control of the whitefly.
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Hemípteros , Control Biológico de Vectores , Rayos Ultravioleta , AnimalesRESUMEN
Understanding complex biological systems requires the system-wide characterization of both molecular and cellular features. Existing methods for spatial mapping of biomolecules in intact tissues suffer from information loss caused by degradation and tissue damage. We report a tissue transformation strategy named stabilization under harsh conditions via intramolecular epoxide linkages to prevent degradation (SHIELD), which uses a flexible polyepoxide to form controlled intra- and intermolecular cross-link with biomolecules. SHIELD preserves protein fluorescence and antigenicity, transcripts and tissue architecture under a wide range of harsh conditions. We applied SHIELD to interrogate system-level wiring, synaptic architecture, and molecular features of virally labeled neurons and their targets in mouse at single-cell resolution. We also demonstrated rapid three-dimensional phenotyping of core needle biopsies and human brain cells. SHIELD enables rapid, multiscale, integrated molecular phenotyping of both animal and clinical tissues.
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CRM197, which retains the same inflammatory and immune-stimulant properties as diphtheria toxin but with reduced toxicity, has been used as a safe carrier in conjugated vaccines. Expression of recombinant CRM197 in E. coli is limited due to formation of inclusion bodies. Soluble expression attempts in Bacillus subtilis, P. fluorescens, Pichia pastoris, and E. coli were partially unsuccessful or did not generate yields sufficient for industrial scale production. Multiple approaches have been attempted to produce CRM197 in E. coli, which has attractive features such as high yield, simplicity, fast growth, etc., including expression of oxidative host, concurrent expression of chaperones, or periplasmic export. Recently, alternative methods for recovery of insoluble proteins expressed in E. coli were reported. Compared to traditional denaturation/refolding, these methods used the non-denaturing solubilization agent, N-lauroylsarkosine to obtain higher recovery yields of native proteins. Based on this work, here, we focused on solubilization of CRM197 from E. coli inclusion bodies. First, CRM197 was expressed as inclusion bodies by high-level expression of recombinant CRM197 in E. coli (126.8 mg/g dcw). Then bioactive CRM197 was isolated from these inclusion bodies with high yield (108.1 mg/g dcw) through solubilization with N-lauroylsarkosine including Triton X-100 and CHAPS, and purified by Ni-affinity chromatography and size-exclusion chromatography. In this study, we present a cost-effective alternative for the production of bioactive CRM197 and compare our recovery yield with yields in other production processes.
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Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Escherichia coli/citología , Escherichia coli/genética , Cuerpos de Inclusión/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Bacterianas/química , Clonación Molecular , Expresión Génica , Proteínas Recombinantes/química , SolubilidadRESUMEN
BACKGROUND: Lactulose, a synthetic disaccharide, has received increasing interest due to its role as a prebiotic, specifically proliferating Bifidobacilli and Lactobacilli and enhancing absorption of calcium and magnesium. The use of cellobiose 2-epimerase (CE) is considered an interesting alternative for industrial production of lactulose. CE reversibly converts D-glucose residues into D-mannose residues at the reducing end of unmodified ß-1,4-linked oligosaccharides, including ß-1,4-mannobiose, cellobiose, and lactose. Recently, a few CE 3D structure were reported, revealing mechanistic details. Using this information, we redesigned the substrate binding site of CE to extend its activity from epimerization to isomerization. RESULTS: Using superimposition with 3 known CE structure models, we identified 2 residues (Tyr114, Asn184) that appeared to play an important role in binding epilactose. We modified these residues, which interact with C2 of the mannose moiety, to prevent epimerization to epilactose. We found a Y114E mutation led to increased release of a by-product, lactulose, at 65 °C, while its activity was low at 37 °C. Notably, this phenomenon was observed only at high temperature and more reliably when the substrate was increased. Using Y114E, isomerization of lactose to lactulose was investigated under optimized conditions, resulting in 86.9 g/l of lactulose and 4.6 g/l of epilactose for 2 h when 200 g/l of lactose was used. CONCLUSION: These results showed that the Y114E mutation increased isomerization of lactose, while decreasing the epimerization of lactose. Thus, a subtle modification of the active site pocket could extend its native activity from epimerization to isomerization without significantly impairing substrate binding. While additional studies are required to scale this to an industrial process, we demonstrated the potential of engineering this enzyme based on structural analysis.
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Carbohidrato Epimerasas/química , Carbohidrato Epimerasas/metabolismo , Celobiosa/química , Celobiosa/metabolismo , Bacterias Grampositivas/enzimología , Ingeniería de Proteínas/métodos , Proteínas Bacterianas/metabolismo , Sitios de Unión , Bacterias Grampositivas/genética , Bacterias Grampositivas/metabolismo , Calor , Microbiología Industrial/métodos , Isomerismo , Lactosa/genética , Lactosa/metabolismo , Lactulosa/biosíntesis , Lactulosa/química , Lactulosa/metabolismo , Manosa/metabolismo , Oligosacáridos/metabolismo , Prebióticos , Dominios Proteicos , Especificidad por SustratoRESUMEN
The biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. We use off-the-shelf antibodies for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, and our experiments demonstrate a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.
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Encéfalo/metabolismo , Imagenología Tridimensional/métodos , Imagen Molecular/métodos , Proteoma/metabolismo , Sinapsis/metabolismo , Sinapsis/ultraestructura , Animales , Encéfalo/ultraestructura , Femenino , Perfilación de la Expresión Génica/métodos , Aumento de la Imagen/métodos , Interpretación de Imagen Asistida por Computador/métodos , Inmunoensayo/métodos , Masculino , Ratones , Proteínas del Tejido Nervioso/metabolismo , Proteoma/ultraestructura , Distribución TisularRESUMEN
Combined measurement of diverse molecular and anatomical traits that span multiple levels remains a major challenge in biology. Here, we introduce a simple method that enables proteomic imaging for scalable, integrated, high-dimensional phenotyping of both animal tissues and human clinical samples. This method, termed SWITCH, uniformly secures tissue architecture, native biomolecules, and antigenicity across an entire system by synchronizing the tissue preservation reaction. The heat- and chemical-resistant nature of the resulting framework permits multiple rounds (>20) of relabeling. We have performed 22 rounds of labeling of a single tissue with precise co-registration of multiple datasets. Furthermore, SWITCH synchronizes labeling reactions to improve probe penetration depth and uniformity of staining. With SWITCH, we performed combinatorial protein expression profiling of the human cortex and also interrogated the geometric structure of the fiber pathways in mouse brains. Such integrated high-dimensional information may accelerate our understanding of biological systems at multiple levels.
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Imagen Molecular/métodos , Conservación de Tejido/métodos , Algoritmos , Animales , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Fibras Nerviosas Mielínicas/química , Proteómica , Sustancias Reductoras , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
When subjected to stress, some individuals develop maladaptive symptoms whereas others retain normal behavior. The medial prefrontal cortex (mPFC) is known to control these adaptive responses to stress. Here, we show that mPFC neurons in the left hemisphere control stress effects on social behavior. Mice made socially avoidant by the stress of chronic social defeats showed depressed neural activity in the left mPFC. Photoactivation of these neurons reversed social avoidance and restored social activity. Despite social defeats, resilient mice with normal sociability showed normal firing rates in the left mPFC; however, photoinhibition of these neurons induced social avoidance. The same photomodulation administered to the right mPFC caused no significant effects. These results explain how stressed individuals develop maladaptive behaviors through left cortical depression, as reported in mood and anxiety disorders.
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Dominancia Cerebral , Red Nerviosa/fisiopatología , Corteza Prefrontal/fisiopatología , Trastorno de la Conducta Social/fisiopatología , Conducta Social , Estrés Psicológico/fisiopatología , Adaptación Fisiológica , Animales , Reacción de Prevención , Conducta Animal , Masculino , Ratones , Ratones Endogámicos C57BL , Trastorno de la Conducta Social/complicaciones , Estrés Psicológico/complicacionesRESUMEN
This study investigated the effects of youngiaside A (YA), youngiaside C (YC), and Youngia denticulatum extract (YDE) on extrinsic aging and assessed its molecular mechanisms in UVB-irradiated HaCaT keratinocytes and human dermal fibroblasts (HDFs). The results showed that YA, YC, and YDE decreased matrix metalloproteinase (MMP) expression and production in HaCaT cell and HDFs and increased collagen expression and production in HDFs. In addition, YA, YC, and YDE significantly increased antioxidant enzyme expression, thereby down-regulating UVB-induced reactive oxygen species (ROS) production and ROS-induced mitogen-activated protein kinase (MAPK) and activator protein-1 (AP-1) signaling in HaCaT cells. Furthermore, YA, YC, and YDE reduced phosphorylation of IκBα and IKKα/ß, blocked nuclear factor-κB (NF-κB) p65 nuclear translocation, and strongly suppressed pro-inflammatory mediators. Finally, YA, YC, and YDE augmented UVB-induced adenosine monophosphate activated protein kinase (AMPK) phosphorylation and YA and YC did not inhibit MMP-1 production in AMPK inhibitor or nuclear factor-erythroid 2-related factor-2 (Nrf2) siRNA-treated HaCaT cells. The results suggest that these compounds could be potential therapeutic agents for prevention and treatment of skin photoaging.