RESUMEN
Advancements in CRISPR technology, particularly the development of base editors, revolutionize genetic variant research. When combined with model organisms like zebrafish, base editors significantly accelerate and refine in vivo analysis of genetic variations. However, base editors are restricted by protospacer adjacent motif (PAM) sequences and specific editing windows, hindering their applicability to a broad spectrum of genetic variants. Additionally, base editors can introduce unintended mutations and often exhibit reduced efficiency in living organisms compared to cultured cell lines. Here, we engineer a suite of adenine base editors (ABEs) called ABE-Ultramax (Umax), demonstrating high editing efficiency and low rates of insertions and deletions (indels) in zebrafish. The ABE-Umax suite of editors includes ABEs with shifted, narrowed, or broadened editing windows, reduced bystander mutation frequency, and highly flexible PAM sequence requirements. These advancements have the potential to address previous challenges in disease modeling and advance gene therapy applications.
Asunto(s)
Adenina , Sistemas CRISPR-Cas , Edición Génica , Mutación INDEL , Pez Cebra , Pez Cebra/genética , Animales , Edición Génica/métodos , Adenina/metabolismo , ARN Guía de Sistemas CRISPR-Cas/genética , ARN Guía de Sistemas CRISPR-Cas/metabolismo , Animales Modificados Genéticamente , AlelosRESUMEN
The current analytical study is dedicated to the boundary layer regime where heat and mass transfer rates are ruled by natural convection. A rectangular enclosure filled with a combination of an arbitrary buoyancy ratio has an Oseen-linear solution, and the position of Beavers and Joseph's condition is employed at the porous fluid interface. Thermal radiation's interaction with a porous lining influences overall heat transfer in a system. Porous linings and radiation are employed in many applications, such as furnaces, insulation, heat exchangers, solar energy collecting and storage, and heat control in electronics. The effect of slip and radiation is to increase the flow rate because of the reduction in friction at the surface. It indicates the fact that temperature and concentration are rapidly lowering. As the slip parameter and radiation parameter increase, the heat and mass transport increase due to the rise in velocity. The Nusselt and Sherwood numbers reach their maximum when the radiation parameter, Rayleigh number, and slip parameter are increased. The findings of the Nusslet number and Sherwood numbers are related to the finite situations of the slip parameter tending to infinity, the radiation parameter going to zero and the angle 90°.
RESUMEN
The coherent perfect absorption (CPA) occurring in the graphene sheet suspended in air can be utilized to develop an ultrathin, ultra-broadband absorber working in the frequency range from a few hertz (Hz) to terahertz (THz) with perfect absorption. A graphene sheet is studied to induce the CPA to cover radio, microwave and lower THz frequency ranges. A graphene resonator able to provide the surface plasmon resonance (SPR) is combined with the graphene sheet to provide CPA at either side of a thin dielectric layer forms metamaterial structure with the cavity and enhances the absorption bandwidth in the THz region by creating a resonance near quasi-CPA frequency. A dielectric silicon resonator is embedded in the structure, which creates dipolar resonances between the resonances obtained by the formed cavity between the graphene sheet and resonator. This enhances the absorption level in the THz region. The absorption bandwidth is further enhanced to 7 THz by including a graphene disc at the top of the silicon resonator. Thus, the multiple multi-order resonances occurring in the silicon dielectric and SPR of graphene resonators are merged with the phenomena of CPA occurring in the graphene sheets to extend the CPA bandwidth in the THz regime. The doping level of graphene or its tunable Fermi energy based on the applied DC electric field provides the tunability in the total obtained absorption bandwidth. The symmetric structure provides polarization-insensitive behavior with an allowed incident angle of more than 45° with more than 90% absorption.
RESUMEN
BACKGROUND: Biallelic variants in OGDHL, encoding part of the α-ketoglutarate dehydrogenase complex, have been associated with highly heterogeneous neurological and neurodevelopmental disorders. However, the validity of this association remains to be confirmed. A second OGDHL patient cohort was recruited to carefully assess the gene-disease relationship. METHODS: Using an unbiased genotype-first approach, we screened large, multiethnic aggregated sequencing datasets worldwide for biallelic OGDHL variants. We used CRISPR/Cas9 to generate zebrafish knockouts of ogdhl, ogdh paralogs, and dhtkd1 to investigate functional relationships and impact during development. Functional complementation with patient variant transcripts was conducted to systematically assess protein functionality as a readout for pathogenicity. RESULTS: A cohort of 14 individuals from 12 unrelated families exhibited highly variable clinical phenotypes, with the majority of them presenting at least one additional variant, potentially accounting for a blended phenotype and complicating phenotypic understanding. We also uncovered extreme clinical heterogeneity and high allele frequencies, occasionally incompatible with a fully penetrant recessive disorder. Human cDNA of previously described and new variants were tested in an ogdhl zebrafish knockout model, adding functional evidence for variant reclassification. We disclosed evidence of hypomorphic alleles as well as a loss-of-function variant without deleterious effects in zebrafish variant testing also showing discordant familial segregation, challenging the relationship of OGDHL as a conventional Mendelian gene. Going further, we uncovered evidence for a complex compensatory relationship among OGDH, OGDHL, and DHTKD1 isoenzymes that are associated with neurodevelopmental disorders and exhibit complex transcriptional compensation patterns with partial functional redundancy. CONCLUSIONS: Based on the results of genetic, clinical, and functional studies, we formed three hypotheses in which to frame observations: biallelic OGDHL variants lead to a highly variable monogenic disorder, variants in OGDHL are following a complex pattern of inheritance, or they may not be causative at all. Our study further highlights the continuing challenges of assessing the validity of reported disease-gene associations and effects of variants identified in these genes. This is particularly more complicated in making genetic diagnoses based on identification of variants in genes presenting a highly heterogenous phenotype such as "OGDHL-related disorders".
Asunto(s)
Proteínas , Pez Cebra , Animales , Humanos , Frecuencia de los Genes , Complejo Cetoglutarato Deshidrogenasa/genética , Complejo Cetoglutarato Deshidrogenasa/metabolismo , Fenotipo , Proteínas/genética , Pez Cebra/genéticaRESUMEN
TSPEAR variants cause autosomal recessive ectodermal dysplasia (ARED) 14. The function of TSPEAR is unknown. The clinical features, the mutation spectrum, and the underlying mechanisms of ARED14 are poorly understood. Combining data from new and previously published individuals established that ARED14 is primarily characterized by dental anomalies such as conical tooth cusps and hypodontia, like those seen in individuals with WNT10A-related odontoonychodermal dysplasia. AlphaFold-predicted structure-based analysis showed that most of the pathogenic TSPEAR missense variants likely destabilize the ß-propeller of the protein. Analysis of 100000 Genomes Project (100KGP) data revealed multiple founder TSPEAR variants across different populations. Mutational and recombination clock analyses demonstrated that non-Finnish European founder variants likely originated around the end of the last ice age, a period of major climatic transition. Analysis of gnomAD data showed that the non-Finnish European population TSPEAR gene-carrier rate is â¼1/140, making it one of the commonest AREDs. Phylogenetic and AlphaFold structural analyses showed that TSPEAR is an ortholog of drosophila Closca, an extracellular matrix-dependent signaling regulator. We, therefore, hypothesized that TSPEAR could have a role in enamel knot, a structure that coordinates patterning of developing tooth cusps. Analysis of mouse single-cell RNA sequencing (scRNA-seq) data revealed highly restricted expression of Tspear in clusters representing enamel knots. A tspeara -/-;tspearb -/- double-knockout zebrafish model recapitulated the clinical features of ARED14 and fin regeneration abnormalities of wnt10a knockout fish, thus suggesting interaction between tspear and wnt10a. In summary, we provide insights into the role of TSPEAR in ectodermal development and the evolutionary history, epidemiology, mechanisms, and consequences of its loss of function variants.
Asunto(s)
Displasia Ectodérmica , Diente , Animales , Ratones , Filogenia , Pez Cebra , Displasia Ectodérmica/epidemiología , Diente/patologíaRESUMEN
A terahertz (THz) antenna with tunable filtering is designed and numerically studied. A slotted monopole radiator with defected ground structure is used for operating with wideband response with an impedance bandwidth in the range of 3.80-11.98 THz for S 11≤-10d B. A slot is engraved in the radiator for obtaining the filtering characteristics in the antenna response. By varying its chemical potential, the frequency band 5.05-6.69 THz with graphene material can provide the tunability in the frequency and bandwidth of the filtered band in the antenna response. Furthermore, the antenna can provide radiation efficiency of more than 90% and gain with a peak value of 6.6 dBi in the passband.
RESUMEN
Objective: When there is a complete slippage of facet joints of C1 over C2 such that there is no contact between the articulating surfaces of C1 and C2, the condition is known as atlantoaxial spondyloptosis (AAS). AAS represents an extremely rare manifestation of atlantoaxial instability. This study was performed to highlight the presentation, radiological features, and management of unilateral AAS in pediatric patients. Material and Methods: We retrospectively identified four pediatric patients with AAS from our hospital records in the last 6 years (2014-2019). Results: Among the four patients with unilateral AAS, three were posttraumic and one was diagnosed with craniovertebral junction tuberculosis (CVJ TB). All the patients had a varying degree of spastic quadriparesis on presentation. One patient with CVJ TB presented with neck tilt. All patients with traumatic unilateral AAS were associated with an odontoid fracture. These patients underwent C1-C2 fixation with complete reduction of spondyloptosis using the techniques of joint manipulation and joint remodeling with a posterior only approach. Complete reduction of AAS in patients with trauma was also associated with the realignment of the odontoid fracture. All patients improved neurologically after surgery and achieved excellent correction of the deformity on a follow-up imaging. Conclusion: Pediatric unilateral AAS is an extremely rare phenomenon. A single-stage posterior approach with C1-C2 fixation is a feasible technique for the treatment of this seemingly difficult to correct deformity in pediatric patients and the clinical outcomes are excellent.
Asunto(s)
Articulación Atlantoaxoidea , Apófisis Odontoides , Fracturas de la Columna Vertebral , Espondilolistesis , Humanos , Niño , Apófisis Odontoides/diagnóstico por imagen , Apófisis Odontoides/cirugía , Articulación Atlantoaxoidea/diagnóstico por imagen , Articulación Atlantoaxoidea/cirugía , Articulación Atlantoaxoidea/lesiones , Estudios Retrospectivos , Espondilolistesis/cirugía , Fracturas de la Columna Vertebral/diagnóstico por imagen , Fracturas de la Columna Vertebral/cirugía , Fracturas de la Columna Vertebral/complicacionesAsunto(s)
Edición Génica , Pez Cebra , Animales , Humanos , Pez Cebra/genética , Sistemas CRISPR-Cas/genéticaRESUMEN
PURPOSE: The mediator (MED) multisubunit-complex modulates the activity of the transcriptional machinery, and genetic defects in different MED subunits (17, 20, 27) have been implicated in neurologic diseases. In this study, we identified a recurrent homozygous variant in MED11 (c.325C>T; p.Arg109Ter) in 7 affected individuals from 5 unrelated families. METHODS: To investigate the genetic cause of the disease, exome or genome sequencing were performed in 5 unrelated families identified via different research networks and Matchmaker Exchange. Deep clinical and brain imaging evaluations were performed by clinical pediatric neurologists and neuroradiologists. The functional effect of the candidate variant on both MED11 RNA and protein was assessed using reverse transcriptase polymerase chain reaction and western blotting using fibroblast cell lines derived from 1 affected individual and controls and through computational approaches. Knockouts in zebrafish were generated using clustered regularly interspaced short palindromic repeats/Cas9. RESULTS: The disease was characterized by microcephaly, profound neurodevelopmental impairment, exaggerated startle response, myoclonic seizures, progressive widespread neurodegeneration, and premature death. Functional studies on patient-derived fibroblasts did not show a loss of protein function but rather disruption of the C-terminal of MED11, likely impairing binding to other MED subunits. A zebrafish knockout model recapitulates key clinical phenotypes. CONCLUSION: Loss of the C-terminal of MED subunit 11 may affect its binding efficiency to other MED subunits, thus implicating the MED-complex stability in brain development and neurodegeneration.
Asunto(s)
Complejo Mediador , Microcefalia , Enfermedades Neurodegenerativas , Animales , Humanos , Homocigoto , Complejo Mediador/genética , Microcefalia/genética , Enfermedades Neurodegenerativas/genética , ARN , Pez Cebra/genéticaRESUMEN
Aminoacyl-tRNA synthetases (ARSs) are essential enzymes for faithful assignment of amino acids to their cognate tRNA. Variants in ARS genes are frequently associated with clinically heterogeneous phenotypes in humans and follow both autosomal dominant or recessive inheritance patterns in many instances. Variants in tryptophanyl-tRNA synthetase 1 (WARS1) cause autosomal dominantly inherited distal hereditary motor neuropathy and Charcot-Marie-Tooth disease. Presently, only one family with biallelic WARS1 variants has been described. We present three affected individuals from two families with biallelic variants (p.Met1? and p.(Asp419Asn)) in WARS1, showing varying severities of developmental delay and intellectual disability. Hearing impairment and microcephaly, as well as abnormalities of the brain, skeletal system, movement/gait, and behavior were variable features. Phenotyping of knocked down wars-1 in a Caenorhabditis elegans model showed depletion is associated with defects in germ cell development. A wars1 knockout vertebrate model recapitulates the human clinical phenotypes, confirms variant pathogenicity, and uncovers evidence implicating the p.Met1? variant as potentially impacting an exon critical for normal hearing. Together, our findings provide consolidating evidence for biallelic disruption of WARS1 as causal for an autosomal recessive neurodevelopmental syndrome and present a vertebrate model that recapitulates key phenotypes observed in patients.
Asunto(s)
Aminoacil-ARNt Sintetasas , Enfermedad de Charcot-Marie-Tooth , Triptófano-ARNt Ligasa , Aminoacil-ARNt Sintetasas/genética , Enfermedad de Charcot-Marie-Tooth/genética , Exones , Humanos , Mutación , Linaje , ARN de Transferencia/genética , Síndrome , Triptófano-ARNt Ligasa/genéticaRESUMEN
Aminoacylation of transfer RNA (tRNA) is a key step in protein biosynthesis, carried out by highly specific aminoacyl-tRNA synthetases (ARSs). ARSs have been implicated in autosomal dominant and autosomal recessive human disorders. Autosomal dominant variants in tryptophanyl-tRNA synthetase 1 (WARS1) are known to cause distal hereditary motor neuropathy and Charcot-Marie-Tooth disease, but a recessively inherited phenotype is yet to be clearly defined. Seryl-tRNA synthetase 1 (SARS1) has rarely been implicated in an autosomal recessive developmental disorder. Here, we report five individuals with biallelic missense variants in WARS1 or SARS1, who presented with an overlapping phenotype of microcephaly, developmental delay, intellectual disability, and brain anomalies. Structural mapping showed that the SARS1 variant is located directly within the enzyme's active site, most likely diminishing activity, while the WARS1 variant is located in the N-terminal domain. We further characterize the identified WARS1 variant by showing that it negatively impacts protein abundance and is unable to rescue the phenotype of a CRISPR/Cas9 wars1 knockout zebrafish model. In summary, we describe two overlapping autosomal recessive syndromes caused by variants in WARS1 and SARS1, present functional insights into the pathogenesis of the WARS1-related syndrome and define an emerging disease spectrum: ARS-related developmental disorders with or without microcephaly.
Asunto(s)
Aminoacil-ARNt Sintetasas , Enfermedad de Charcot-Marie-Tooth , Microcefalia , Triptófano-ARNt Ligasa , Animales , Humanos , Aminoacil-ARNt Sintetasas/genética , Enfermedad de Charcot-Marie-Tooth/genética , Ligasas , Microcefalia/genética , Microcefalia/patología , ARN de Transferencia , Triptófano-ARNt Ligasa/genética , Pez Cebra/genéticaRESUMEN
A circularly polarized (CP) multi-input multioutput (MIMO) dielectric resonator (DR) antenna (DRA) with compact size and four ports is implemented. CP radiation was achieved using the deformed DR geometry excited with aperture coupled feeding. A CPDRA with a single and two ports is investigated. The defected ground structure (DGS) was incorporated into the antenna for improving the isolation between the ports. The DGS was incorporated in such a way that the required phase difference between the generated orthogonal degenerate modes is preserved. This concept could be utilized in implementing a compact four-port CP antenna. The MIMO antenna provides a 10 dB impedance bandwidth of 38% (8.5-12.5 GHz) and a 3 dB AR bandwidth of 9.32% (9.2-10.1 GHz). The gain of the implemented antenna was around 6 dBi in the band where CP radiation was achieved. The MIMO performance parameters were calculated, and their values remained within the acceptable limits. The implemented antenna could suitably be used in X-band applications.
Asunto(s)
Tecnología Inalámbrica , Impedancia Eléctrica , Diseño de EquipoRESUMEN
Chondroitin/dermatan sulfate (CS/DS) proteoglycans are indispensable for animal development and homeostasis but the large number of enzymes involved in their biosynthesis have made CS/DS function a challenging problem to study genetically. In our study, we generated loss-of-function alleles in zebrafish genes encoding CS/DS biosynthetic enzymes and characterized the effect on development in single and double mutants. Homozygous mutants in chsy1, csgalnact1a, csgalnat2, chpfa, ust and chst7, respectively, develop to adults. However, csgalnact1a-/- fish develop distinct craniofacial defects while the chsy1-/- skeletal phenotype is milder and the remaining mutants display no gross morphological abnormalities. These results suggest a high redundancy for the CS/DS biosynthetic enzymes and to further reduce CS/DS biosynthesis we combined mutant alleles. The craniofacial phenotype is further enhanced in csgalnact1a-/-;chsy1-/- adults and csgalnact1a-/-;csgalnact2-/- larvae. While csgalnact1a-/-;csgalnact2-/- was the most affected allele combination in our study, CS/DS is still not completely abolished. Transcriptome analysis of chsy1-/-, csgalnact1a-/- and csgalnact1a-/-;csgalnact2-/- larvae revealed that the expression had changed in a similar way in the three mutant lines but no differential expression was found in any of fifty GAG biosynthesis enzymes identified. Thus, zebrafish larvae do not increase transcription of GAG biosynthesis genes as a consequence of decreased CS/DS biosynthesis. The new zebrafish lines develop phenotypes similar to clinical characteristics of several human congenital disorders making the mutants potentially useful to study disease mechanisms and treatment.
Asunto(s)
Dermatán Sulfato , Pez Cebra , Animales , Sulfatos de Condroitina/metabolismo , Dermatán Sulfato/genética , Dermatán Sulfato/metabolismo , Glicosiltransferasas/genética , Fenotipo , Pez Cebra/genética , Pez Cebra/metabolismoRESUMEN
A technique is implemented for altering the multimodal resonance generated in an ultrathin silicon ring resonator-based terahertz (THz) absorber. The absorber provides the dual-band resonance with the excitation of magnetic and electric dipole in the lower and upper band, respectively. The field of magnetic and electric dipoles is altered using a non-resonant graphene ring placed in the center of the generated dipolar arrangement and the tunability and perfect absorption is achieved. A circuit model is prepared using transmission line method and absorber operation is verified. The proposed absorber can be utilized as a biosensor for the detection of malaria virus and glucose percentage in water. The sensor offers highest sensitivity as 0.29 and 0.27 THz per thickness unit change and quality factor as 117.53 and 245 in the lower and upper band, respectively during the sensing of analyte thickness. Also, it offers the sensitivity as 0.20 and 0.10 THz per refractive index unit change and quality factor as 105.28 and 211.84 in the lower and upper band, respectively during refractive index sensing. Moreover, the structure remains insensitive to polarization angle of the incident electromagnetic wave.
Asunto(s)
Refractometría , Silicio , Luz , Dispersión de Radiación , Radiación TerahertzRESUMEN
A systematic knowledge of the roles of DNA repair genes at the level of the organism has been limited due to the lack of appropriate experimental approaches using animal model systems. Zebrafish has become a powerful vertebrate genetic model system with availability due to the ease of genome editing and large-scale phenotype screening. Here, we generated zebrafish mutants for 32 DNA repair and replication genes through multiplexed CRISPR/Cas9-mediated mutagenesis. Large-scale phenotypic characterization of our mutant collection revealed that three genes (atad5a, ddb1, pcna) are essential for proper embryonic development and hematopoiesis; seven genes (apex1, atrip, ino80, mre11a, shfm1, telo2, wrn) are required for growth and development during juvenile stage and six genes (blm, brca2, fanci, rad51, rad54l, rtel1) play critical roles in sex development. Furthermore, mutation in six genes (atad5a, brca2, polk, rad51, shfm1, xrcc1) displayed hypersensitivity to DNA damage agents. Our zebrafish mutant collection provides a unique resource for understanding of the roles of DNA repair genes at the organismal level.
Asunto(s)
Edición Génica , Animales , Pez CebraRESUMEN
The role of the cannabinoid receptor 2 (CNR2) is still poorly described in sensory epithelia. We found strong cnr2 expression in hair cells (HCs) of the inner ear and the lateral line (LL), a superficial sensory structure in fish. Next, we demonstrated that sensory synapses in HCs were severely perturbed in larvae lacking cnr2. Appearance and distribution of presynaptic ribbons and calcium channels (Cav1.3) were profoundly altered in mutant animals. Clustering of membrane-associated guanylate kinase (MAGUK) in post-synaptic densities (PSDs) was also heavily affected, suggesting a role for cnr2 for maintaining the sensory synapse. Furthermore, vesicular trafficking in HCs was strongly perturbed suggesting a retrograde action of the endocannabinoid system (ECs) via cnr2 that was modulating HC mechanotransduction. We found similar perturbations in retinal ribbon synapses. Finally, we showed that larval swimming behaviors after sound and light stimulations were significantly different in mutant animals. Thus, we propose that cnr2 is critical for the processing of sensory information in the developing larva.
RESUMEN
OBJECTIVE: In a recent genome-wide association study, a significant genetic association between rs34330 of CDKN1B and risk of systemic lupus erythematosus (SLE) in Han Chinese was identified. This study was undertaken to validate the reported association and elucidate the biochemical mechanisms underlying the effect of the variant. METHODS: We performed an allelic association analysis in patients with SLE, followed by a meta-analysis assessing genome-wide association data across 11 independent cohorts (n = 28,872). In silico bioinformatics analysis and experimental validation in SLE-relevant cell lines were applied to determine the functional consequences of rs34330. RESULTS: We replicated a genetic association between SLE and rs34330 (meta-analysis P = 5.29 × 10-22 , odds ratio 0.84 [95% confidence interval 0.81-0.87]). Follow-up bioinformatics and expression quantitative trait locus analysis suggested that rs34330 is located in active chromatin and potentially regulates several target genes. Using luciferase and chromatin immunoprecipitation-real-time quantitative polymerase chain reaction, we demonstrated substantial allele-specific promoter and enhancer activity, and allele-specific binding of 3 histone marks (H3K27ac, H3K4me3, and H3K4me1), RNA polymerase II (Pol II), CCCTC-binding factor, and a critical immune transcription factor (interferon regulatory factor 1 [IRF-1]). Chromosome conformation capture revealed long-range chromatin interactions between rs34330 and the promoters of neighboring genes APOLD1 and DDX47, and effects on CDKN1B and the other target genes were directly validated by clustered regularly interspaced short palindromic repeat (CRISPR)-based genome editing. Finally, CRISPR/dead CRISPR-associated protein 9-based epigenetic activation/silencing confirmed these results. Gene-edited cell lines also showed higher levels of proliferation and apoptosis. CONCLUSION: Collectively, these findings suggest a mechanism whereby the rs34330 risk allele (C) influences the presence of histone marks, RNA Pol II, and IRF-1 transcription factor to regulate expression of several target genes linked to proliferation and apoptosis. This process could potentially underlie the association of rs34330 with SLE.
Asunto(s)
Apoptosis/genética , Proliferación Celular/genética , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/genética , Lupus Eritematoso Sistémico/genética , Polimorfismo de Nucleótido Simple , Alelos , Biología Computacional , Predisposición Genética a la Enfermedad , Estudio de Asociación del Genoma Completo , Humanos , Regiones Promotoras GenéticasRESUMEN
Deafness, the most frequent sensory deficit in humans, is extremely heterogeneous with hundreds of genes involved. Clinical and genetic analyses of an extended consanguineous family with pre-lingual, moderate-to-profound autosomal recessive sensorineural hearing loss, allowed us to identify CLRN2, encoding a tetraspan protein, as a new deafness gene. Homozygosity mapping followed by exome sequencing identified a 14.96 Mb locus on chromosome 4p15.32p15.1 containing a likely pathogenic missense variant in CLRN2 (c.494C > A, NM_001079827.2) segregating with the disease. Using in vitro RNA splicing analysis, we show that the CLRN2 c.494C > A variant leads to two events: (1) the substitution of a highly conserved threonine (uncharged amino acid) to lysine (charged amino acid) at position 165, p.(Thr165Lys), and (2) aberrant splicing, with the retention of intron 2 resulting in a stop codon after 26 additional amino acids, p.(Gly146Lysfs*26). Expression studies and phenotyping of newly produced zebrafish and mouse models deficient for clarin 2 further confirm that clarin 2, expressed in the inner ear hair cells, is essential for normal organization and maintenance of the auditory hair bundles, and for hearing function. Together, our findings identify CLRN2 as a new deafness gene, which will impact future diagnosis and treatment for deaf patients.
Asunto(s)
Sustitución de Aminoácidos , Cromosomas Humanos Par 4/química , Células Ciliadas Auditivas Internas/metabolismo , Pérdida Auditiva Sensorineural/genética , Proteínas de la Membrana/genética , Mutación Puntual , Tetraspaninas/genética , Adulto , Alelos , Animales , Secuencia de Bases , Mapeo Cromosómico , Consanguinidad , Femenino , Expresión Génica , Genes Recesivos , Células Ciliadas Auditivas Internas/patología , Pérdida Auditiva Sensorineural/metabolismo , Pérdida Auditiva Sensorineural/patología , Humanos , Masculino , Proteínas de la Membrana/deficiencia , Ratones , Linaje , Tetraspaninas/deficiencia , Secuenciación del Exoma , Pez CebraRESUMEN
Surface plasmons generated at the graphene dielectric interface can be altered by trapping the electric charge. A technique is implemented for trapping the bipolar electric charge on the graphene surface and arranged in a desired way to enhance the performance of a monolayer graphene metamaterial based tunable, ultrathin, dual narrow band terahertz (THz) absorber. A monolayer graphene sheet placed on the dielectric substrate can provide dual-band resonance by utilizing the surface plasmons of the fundamental and third order mode index and an absorption of more than 99% and 50% can be obtained in the lower and upper band, respectively. The absorption is further enhanced to the level of perfect-absorption by utilizing the charge trapping mechanism on the graphene and generating bipolar charged nodes to create higher order surface plasmons. The multiple interference and reflection theory proves that the destructive interference in the dielectric layer is the cause of perfect absorption. The applied technique in the dual-band absorber configuration provides a tunable response which remains insensitive to the polarization and incident angle of the electromagnetic wave. The proposed perfect absorber can be utilized as a biosensor for refractive index sensing and the detection of glucose in water and the malaria virus in blood. It can provide an ultrahigh sensitivity of 14.88 THz RIU-1 with FOM as 53.09 RIU-1 with the variation in the chemical potential of graphene and 12.7 THz RIU-1 and FOM as 41.1 RIU-1 during glucose detection in water.
RESUMEN
Anaplastic lymphoma kinase (Alk) is a receptor tyrosine kinase of the insulin receptor super-family that functions as oncogenic driver in a range of human cancers such as neuroblastoma. In order to investigate mechanisms underlying Alk oncogenic signaling, we conducted a genetic suppressor screen in Drosophila melanogaster. Our screen identified multiple loci important for Alk signaling, including members of Ras/Raf/ERK-, Pi3K-, and STAT-pathways as well as tailless (tll) and foxo whose orthologues NR2E1/TLX and FOXO3 are transcription factors implicated in human neuroblastoma. Many of the identified suppressors were also able to modulate signaling output from activated oncogenic variants of human ALK, suggesting that our screen identified targets likely relevant in a wide range of contexts. Interestingly, two misexpression alleles of wallenda (wnd, encoding a leucine zipper bearing kinase similar to human DLK and LZK) were among the strongest suppressors. We show that Alk expression leads to a growth advantage and induces cell death in surrounding cells. Our results suggest that Alk activity conveys a competitive advantage to cells, which can be reversed by over-expression of the JNK kinase kinase Wnd.