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Following synthesis, integral membrane proteins dwell in the endoplasmic reticulum (ER) for variable periods that are typically rate limiting for plasma membrane delivery. In neurons, the ER extends for hundreds of microns as an anastomosing network throughout highly branched dendrites. However, little is known about the mobility, spatial scales, or dynamic regulation of cargo in the dendritic ER. Here, we show that membrane proteins, including AMPA-type glutamate receptors, rapidly diffuse within the continuous network of dendritic ER but are confined by increased ER complexity at dendritic branch points and near dendritic spines. The spatial range of receptor mobility is rapidly restricted by type I mGluR signaling through a mechanism involving protein kinase C (PKC) and the ER protein CLIMP63. Moreover, local zones of ER complexity compartmentalize ER export and correspond to sites of new dendritic branches. Thus, local control of ER complexity spatially scales secretory trafficking within elaborate dendritic arbors.
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Dendritas/metabolismo , Retículo Endoplásmico/metabolismo , Proteínas de la Membrana/metabolismo , Secuencia de Aminoácidos , Animales , Embrión de Mamíferos/metabolismo , Hipocampo/citología , Hipocampo/metabolismo , Masculino , Datos de Secuencia Molecular , Proteína Quinasa C/metabolismo , Ratas , Receptores AMPA/metabolismo , Receptores de Superficie Celular/química , Receptores de Superficie Celular/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Sinapsis/metabolismoRESUMEN
PURPOSE: Hardikar syndrome (HS, MIM #301068) is a female-specific multiple congenital anomaly syndrome characterized by retinopathy, orofacial clefting, aortic coarctation, biliary dysgenesis, genitourinary malformations, and intestinal malrotation. We previously showed that heterozygous nonsense and frameshift variants in MED12 cause HS. The phenotypic spectrum of disease and the mechanism by which MED12 variants cause disease is unknown. We aim to expand the phenotypic and molecular landscape of HS and elucidate the mechanism by which MED12 variants cause disease. METHODS: We clinically assembled and molecularly characterized a cohort of 11 previously unreported individuals with HS. Additionally, we studied the effect of MED12 deficiency on ciliary biology, hedgehog, and yes-associated protein (YAP) signaling; pathways implicated in diseases with phenotypic overlap with HS. RESULTS: We report novel phenotypes associated with HS, including cardiomyopathy, arrhythmia, and vascular anomalies, and expand the molecular landscape of HS to include splice site variants. We additionally demonstrate that MED12 deficiency causes decreased cell ciliation, and impairs hedgehog and YAP signaling. CONCLUSION: Our data support updating HS standard-of-care to include regular cardiac imaging, arrhythmia screening, and vascular imaging. We further propose that dysregulation of ciliogenesis and YAP and hedgehog signaling contributes to the pathogenesis of HS.
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PURPOSE: Clinical next-generation sequencing is an effective approach for identifying pathogenic sequence variants that are medically actionable for participants and families but are not associated with the participant's primary diagnosis. These variants are called secondary findings (SFs). According to the literature, there is no report of the types and frequencies of SFs in a large pediatric cohort which includes substantial African-American participants. We sought to investigate the types (including American College of Medical Genetics and Genomics [ACMG] and non-ACMG recommended gene lists), frequencies, and rates of SFs, as well as the effects of SF disclosure on the participants and families of a large pediatric cohort at the Center for Applied Genomics at The Children's Hospital of Philadelphia (CHOP). METHODS: We systematically identified pathogenic (P) and likely pathogenic (LP) variants in established disease-causing genes, adhering to ACMG v3.2 secondary finding guidelines and beyond. For non-ACMG secondary findings, akin to incidental findings in clinical settings, we utilized a set of criteria focusing on pediatric onset, high penetrance, moderate to severe phenotypes, and the clinical actionability of the variants. This criteria-based approach was applied rather than using a fixed gene list to ensure that the variants identified are likely to impact participant health significantly. To identify and categorize these variants, we employed a clinical-grade variant classification standard per ACMG/AMP recommendations; additionally, we conducted a detailed literature search to ensure a comprehensive exploration of potential secondary findings relevant to pediatric participants. RESULTS: We report a distinctive distribution of 1,464 P/LP SF variants in 16,713 participants. There were 427 unique variants in ACMG genes and 265 in non-ACMG genes. The most frequently mutated genes among the ACMG and non-ACMG gene lists were TTR (41.6%) and CHEK2 (7.16%), respectively. Overall, variants of possible medical importance were found in 8.76% of participants in both ACMG (5.81%) and non-ACMG (2.95%) genes.
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PURPOSE: RNF213, encoding a giant E3 ubiquitin ligase, has been recognized for its role as a key susceptibility gene for moyamoya disease. Case reports have also implicated specific variants in RNF213 with an early-onset form of moyamoya disease with full penetrance. We aimed to expand the phenotypic spectrum of monogenic RNF213-related disease and to evaluate genotype-phenotype correlations. METHODS: Patients were identified through reanalysis of exome sequencing data of an unselected cohort of unsolved pediatric cases and through GeneMatcher or ClinVar. Functional characterization was done by proteomics analysis and oxidative phosphorylation enzyme activities using patient-derived fibroblasts. RESULTS: We identified 14 individuals from 13 unrelated families with (de novo) missense variants in RNF213 clustering within or around the Really Interesting New Gene (RING) domain. Individuals presented either with early-onset stroke (n = 11) or with Leigh syndrome (n = 3). No genotype-phenotype correlation could be established. Proteomics using patient-derived fibroblasts revealed no significant differences between clinical subgroups. 3D modeling revealed a clustering of missense variants in the tertiary structure of RNF213 potentially affecting zinc-binding suggesting a gain-of-function or dominant negative effect. CONCLUSION: De novo missense variants in RNF213 clustering in the E3 RING or other regions affecting zinc-binding lead to an early-onset syndrome characterized by stroke or Leigh syndrome.
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Enfermedad de Leigh , Enfermedad de Moyamoya , Accidente Cerebrovascular , Humanos , Niño , Enfermedad de Moyamoya/genética , Enfermedad de Leigh/complicaciones , Factores de Transcripción/genética , Ubiquitina-Proteína Ligasas/genética , Zinc , Predisposición Genética a la Enfermedad , Adenosina Trifosfatasas/genéticaRESUMEN
Children with birth defects (BD) express distinct clinical features that often have various medical consequences, one of which is predisposition to the development of cancers. Identification of the underlying genetic mechanisms related to the development of cancer in BD patients would allow for preventive measures. We performed a whole genome sequencing (WGS) study on blood-derived DNA samples from 1566 individuals without chromosomal anomalies, including 454 BD probands with at least one type of malignant tumors, 767 cancer-free BD probands, and 345 healthy individuals. Exclusive recurrent variants were identified in BD-cancer and BD-only patients and mapped to their corresponding genomic regions. We observed statistically significant overlaps for protein-coding/ncRNA with exclusive variants in exons, introns, ncRNAs, and 3'UTR regions. Exclusive exonic variants, especially synonymous variants, tend to occur in prior exons locus in BD-cancer children. Intronic variants close to splicing site (< 500 bp from exon) have little overlaps in BD-cancer and BD-only patients. Exonic variants in non-coding RNA (ncRNA) tend to occur in different ncRNAs exons regardless of the overlaps. Notably, genes with 5' UTR variants are almost mutually exclusive between the two phenotypes. In conclusion, we conducted the first genomic study to explore the impact of recurrent variants exclusive to the two distinguished clinical phenotypes under study, BD with or without cancer, demonstrating enrichment of selective protein-coding/ncRNAs differentially expressed between these two phenotypes, suggesting that selective genetic factors may underlie the molecular processes of pediatric cancer development in BD children.
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Neoplasias , Empalme del ARN , Humanos , Mutación , Exones , Genómica , Neoplasias/genética , IntronesRESUMEN
AMOTL1 encodes angiomotin-like protein 1, an actin-binding protein that regulates cell polarity, adhesion, and migration. The role of AMOTL1 in human disease is equivocal. We report a large cohort of individuals harboring heterozygous AMOTL1 variants and define a core phenotype of orofacial clefting, congenital heart disease, tall stature, auricular anomalies, and gastrointestinal manifestations in individuals with variants in AMOTL1 affecting amino acids 157-161, a functionally undefined but highly conserved region. Three individuals with AMOTL1 variants outside this region are also described who had variable presentations with orofacial clefting and multi-organ disease. Our case cohort suggests that heterozygous missense variants in AMOTL1, most commonly affecting amino acid residues 157-161, define a new orofacial clefting syndrome, and indicates an important functional role for this undefined region.
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Labio Leporino , Fisura del Paladar , Cardiopatías Congénitas , Humanos , Fisura del Paladar/diagnóstico , Fisura del Paladar/genética , Labio Leporino/diagnóstico , Labio Leporino/genética , Mutación , Mutación Missense/genética , Cardiopatías Congénitas/diagnóstico , Cardiopatías Congénitas/genética , AngiomotinasRESUMEN
Age-related effects on motor asymmetry provide insight into changes in cortical activation during aging. To investigate potential changes in manual performance associated with aging, we conducted the Jamar hand function test and the Purdue Pegboard test on young and older adults. All tests indicated reduced motor asymmetry in the older group. Further analysis suggested that a significant decline in dominant (right) hand function resulted in less asymmetric performance in older adults. The finding is inconsistent with the application of the HAROLD model in the motor domain, which assumes improved performance in the non-dominant hand, leading to a reduction of motor asymmetry in older adults. Based on the manual performance in young and older adults, it is suggested that aging reduces manual asymmetry in both force production and manual dexterity due to the reduced performance of the dominant hand.
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Rezafungin is a novel echinocandin being developed for treatment of candidemia and invasive candidiasis and for prevention of invasive fungal disease caused by Candida, Aspergillus, and Pneumocystis spp. in recipients of blood and marrow transplantation. Studies using [14C]-radiolabeled rezafungin were conducted in rats, monkeys, and humans to characterize the mass balance, excretion, and pharmacokinetics of [14C]-rezafungin and to evaluate relative amounts of rezafungin metabolites compared with parent drug. Fecal excretion was the main route of elimination in rats, monkeys, and humans. Radioactivity was primarily excreted as unchanged drug, with ≥95% average total recovery in rats (through 336 h) and monkeys (through 720 h). In humans, cumulative recovery of radioactivity through the first 17 days was 52% (38% in feces, 14% in urine) with estimated mean overall recovery through day 60 of 88.3% (73% in feces, 27% in urine). The clinical pharmacokinetics of rezafungin following a single 400-mg intravenous infusion (200 µCi of [14C]-rezafungin) were similar in plasma, plasma total radioactivity, and whole blood total radioactivity. Unchanged rezafungin represented the majority of total radioactivity in plasma, and the partitioning of total radioactivity into red blood cells was negligible. Across species, rezafungin was primarily metabolized by hydroxylation of the terphenyl, pentyl ether side chain. In these excretion/mass balance, metabolism, and PK studies, clinical observations were consistent with findings in the rat and monkey demonstrating the minimal metabolism and slow elimination of rezafungin after intravenous administration, with fecal excretion as the major route of elimination.
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Antifúngicos , Candidiasis Invasiva , Administración Oral , Animales , Antifúngicos/uso terapéutico , Candida , Candidiasis Invasiva/tratamiento farmacológico , Equinocandinas/uso terapéutico , Heces/química , Humanos , RatasRESUMEN
TBX6 encodes transcription-factor box 6, a transcription factor critical to paraxial mesoderm segmentation and somitogenesis during embryonic development. TBX6 haploinsufficiency is believed to drive the skeletal and kidney phenotypes associated with the 16p11.2 deletion syndrome. Heterozygous and biallelic variants in TBX6 are associated with vertebral and rib malformations (TBX6-associated congenital scoliosis) and spondylocostal dysostosis, and heterozygous TBX6 variants are associated with increased risk of genitourinary tract malformations. Combined skeletal and kidney phenotypes in individuals harboring heterozygous or biallelic TBX6 variants are rare. Here, we present seven individuals with vertebral and rib malformations and structural kidney differences associated with heterozygous TBX6 gene deletion in trans with a hypomorphic TBX6 allele or biallelic TBX6 variants. Our case series highlights the association between TBX6 and both skeletal and kidney disease.
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Osteocondrodisplasias , Escoliosis , Humanos , Proteínas de Dominio T Box/genética , Escoliosis/genética , Columna Vertebral/diagnóstico por imagen , Columna Vertebral/anomalías , Fenotipo , Factores de Transcripción/genética , Túbulos Renales ProximalesRESUMEN
Ichthyosis follicularis, atrichia, and photophobia syndrome (IFAP syndrome) is a rare, X-linked disorder caused by pathogenic variants in membrane-bound transcription factor protease, site 2 (MBTPS2). Pathogenic MBTPS2 variants also cause BRESHECK syndrome, characterized by the IFAP triad plus intellectual disability and multiple congenital anomalies. Here we present a patient with ichthyosis, sparse hair, pulmonic stenosis, kidney dysplasia, hypospadias, growth failure, thrombocytopenia, anemia, bone marrow fibrosis, and chronic diarrhea found by research-based exome sequencing to harbor a novel, maternally inherited MBTPS2 missense variant (c.766 G>A; (p.Val256Leu)). In vitro modeling supports variant pathogenicity, with impaired cell growth in cholesterol-depleted media, attenuated activation of the sterol regulatory element-binding protein pathway, and failure to activate the endoplasmic reticulum stress response pathway. Our case expands both the genetic and phenotypic spectrum of BRESHECK syndrome to include a novel MBTPS2 variant and cytopenias, bone marrow fibrosis, and chronic diarrhea.
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Discapacidad Intelectual , Alopecia/genética , Encéfalo/anomalías , Anomalías Congénitas , Oído/anomalías , Displasia Ectodérmica , Estrés del Retículo Endoplásmico/genética , Enfermedades Genéticas Ligadas al Cromosoma X , Enfermedad de Hirschsprung , Humanos , Discapacidad Intelectual/genética , Riñón/anomalías , Masculino , Metaloendopeptidasas/genética , Péptido Hidrolasas , Esteroles , Factores de TranscripciónRESUMEN
Animals with natural protections against diabetes complications may provide clues to improve human health. Birds are unique in their ability to avoid hyperglycemia-associated complications (e.g., glycation and oxidative stress) despite having naturally high blood glucose (BG) concentrations. This makes them useful models to elucidate strategies to prevent and/or treat diabetes-related complications in mammals. As diet plays a key role in BG concentration and diabetes risk, this systematic review aimed to summarize the effects of macro and micronutrient manipulation on avian BG. Three databases were searched (PubMed, SCOPUS, and Web of Science) for articles that met inclusion criteria: altered at least one nutrient and measured BG in at least one avian species. The search yielded 91 articles that produced 128 datasets (i.e., one nutrient manipulation in one sample). Across all macronutrient manipulations (n = 69 datasets), 62% reported no change in BG and 23% measured an increase (p < 0.001). Within the macronutrient groups (carbohydrate, lipid, protein, and mixed) most datasets showed no change in BG (67%, 62%, 52%, and 86%, respectively). Across micronutrient manipulations (n = 59 datasets), 51% demonstrated no change and 41% decreased BG (p < 0.001). While manipulations that altered vitamin intake largely produced no change in BG (62%), 48% of datasets examining altered mineral intake found no change and 46% decreased BG. Chromium was the most studied micronutrient (n = 24 datasets), where 67% of datasets reported a decrease in BG. These results suggest birds are largely able to maintain blood glucose homeostasis in response to altered nutrient intake indicative of dietary flexibility.
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Hiperglucemia , Oligoelementos , Animales , Aves/metabolismo , Glucemia/metabolismo , Dieta/veterinaria , Ingestión de Energía , Humanos , Mamíferos/metabolismo , MicronutrientesRESUMEN
Oral-facial-digital syndromes (OFDS) are a heterogeneous and rare group of Mendelian disorders characterized by developmental abnormalities of the oral cavity, face, and digits caused by dysfunction of the primary cilium, a mechanosensory organelle that exists atop most cell types that facilitates organ patterning and growth. OFDS is inherited both in an X-linked dominant, X-linked recessive, and autosomal recessive manner. Importantly, though many of the causal genes for OFDS have been identified, up to 40% of OFD syndromes are of unknown genetic basis. Here we describe three children with classical presentations of OFDS including lingual hamartomas, polydactyly, and characteristic facial features found by exome sequencing to harbor variants in causal genes not previously associated with OFDS. We describe a female with hypothalamic hamartoma, urogenital sinus, polysyndactyly, and multiple lingual hamartomas consistent with OFDVI with biallelic pathogenic variants in CEP164, a gene associated with ciliopathy-spectrum disease, but never before with OFDS. We additionally describe two unrelated probands with postaxial polydactyly, multiple lingual hamartomas, and dysmorphic features both found to be homozygous for an identical TOPORS missense variant, c.29 C>A; (p.Pro10Gln). Heterozygous TOPORS pathogenic gene variants are associated with autosomal dominant retinitis pigmentosa, but never before with syndromic ciliopathy. Of note, both probands are of Dominican ancestry, suggesting a possible founder allele.
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Alelos , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Variación Genética , Síndromes Orofaciodigitales/diagnóstico , Síndromes Orofaciodigitales/genética , Femenino , Pruebas Genéticas , Genotipo , Humanos , Lactante , Recién Nacido , Imagen por Resonancia Magnética , Masculino , Mutación , Proteínas de Neoplasias/genética , Proteínas Nucleares/genética , Fenotipo , Ubiquitina-Proteína Ligasas/genética , Secuenciación del ExomaRESUMEN
Ciliopathy syndromes are a diverse spectrum of disease characterized by a combination of cystic kidney disease, hepatobiliary disease, retinopathy, skeletal dysplasia, developmental delay, and brain malformations. Though generally divided into distinct disease categories based on the pattern of system involvement, ciliopathy syndromes are known to display certain phenotypic overlap. We performed next-generation sequencing panel testing, clinical exome sequencing, and research-based exome sequencing reanalysis on patients with suspected ciliopathy syndromes with additional features. We identified biallelic pathogenic variants in BBS1 in a child with features of cranioectodermal dysplasia, and biallelic variants in BBS12 in a child with the clinical stigmata of Bardet-Biedl syndrome, but also with anal atresia. We additionally identified biallelic pathogenic variants in WDR35 and DYNC2H1 in children with predominant liver disease and ductal plate malformation without skeletal dysplasia. Our study highlights the phenotypic and genetic diversity of ciliopathy syndromes, the importance of considering ciliopathy syndromes as a disease-spectrum and screening for all associated complications in all patients, and describes exclusive extra-skeletal manifestations in two classical skeletal dysplasia syndromes.
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Anomalías Múltiples/patología , Chaperoninas/genética , Ciliopatías/patología , Dineínas Citoplasmáticas/genética , Proteínas del Citoesqueleto/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Proteínas Asociadas a Microtúbulos/genética , Mutación , Anomalías Múltiples/genética , Adulto , Niño , Preescolar , Ciliopatías/genética , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Fenotipo , PronósticoRESUMEN
Ring-finger protein 213 (RNF213) encodes a protein of unknown function believed to play a role in cellular metabolism and angiogenesis. Gene variants are associated with susceptibility to moyamoya disease. Here, we describe two children with moyamoya disease who also demonstrated kidney disease, elevated aminotransferases, and recurrent skin lesions found by exome sequencing to have de novo missense variants in RNF213. These cases highlight the ability of RNF213 to cause Mendelian moyamoya disease in addition to acting as a genetic susceptibility locus. The cases also suggest a new, multi-organ RNF213-spectrum disease characterized by liver, skin, and kidney pathology in addition to severe moyamoya disease caused by heterozygous, de novo C-terminal RNF213 missense variants.
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Adenosina Trifosfatasas/genética , Enfermedades Renales/genética , Enfermedad de Moyamoya/genética , Enfermedades de la Piel/genética , Ubiquitina-Proteína Ligasas/genética , Femenino , Predisposición Genética a la Enfermedad , Humanos , Lactante , Recién Nacido , Enfermedades Renales/complicaciones , Enfermedades Renales/patología , Masculino , Enfermedad de Moyamoya/complicaciones , Enfermedad de Moyamoya/patología , Neovascularización Fisiológica/genética , Enfermedades de la Piel/complicaciones , Enfermedades de la Piel/patología , Transaminasas/genética , Secuenciación del ExomaRESUMEN
Summary: Enhanced Recovery After Surgery (ERAS) is a model of care that was introduced in the late 1990s by a group of surgeons in Europe. The model consists of a number of evidence-based principles that support better outcomes for surgical patients, including improved patient experience, reduced length of stay in hospital, decreased complication rates and fewer hospital readmissions. A number of Canadian surgical care teams have already adopted ERAS principles and have reported positive outcomes. Arising from the Canadian Patient Safety Institute's Integrated Patient Safety Action Plan for Surgical Care Safety, and with support from numerous partner organizations from across the country, Enhanced Recovery Canada is leading the drive to improve surgical safety across the country and help disseminate these ERAS principles. We discuss the development of a multidisciplinary clinical pathway for elective colorectal surgery to help guide Canadian clinicians.
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Cirugía Colorrectal/normas , Vías Clínicas/normas , Recuperación Mejorada Después de la Cirugía/normas , Grupo de Atención al Paciente/normas , Seguridad del Paciente/normas , Canadá , Medicina Basada en la Evidencia/normas , HumanosRESUMEN
Despite the ubiquitous nature of scar tissue, there is not a single, reliable strategy to prevent or treat excessive scarring. The difficulty in arriving at a universally accepted form of management is multifaceted: there is an incomplete understanding of the complex pathophysiology of scar formation; a lack of common metrics hampers the accurate description of scar quality and characteristics; model systems do not exist for proper investigation in the controlled environment of a laboratory; and there is only limited data from prospective, randomized controlled clinical trials. Accordingly, the management of cutaneous scars is typically based upon the experience from practitioners rather than from evidence-based data. This article will review the pathophysiology of excessive scar formation, define the most common scars-hypertrophic scars and keloids-and discuss the evidence to support the current nonsurgical therapies in use to both prevent and treat excessive scars.
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Cicatriz Hipertrófica , Queloide , Humanos , Estudios ProspectivosRESUMEN
Hippocampal long-term potentiation (LTP) is a cellular memory mechanism. For LTP to endure, new protein synthesis is required immediately after induction and some of these proteins must be delivered to specific, presumably potentiated, synapses. Local synthesis in dendrites could rapidly provide new proteins to synapses, but the spatial distribution of translation following induction of LTP is not known. Here, we quantified polyribosomes, the sites of local protein synthesis, in CA1 stratum radiatum dendrites and spines from postnatal day 15 rats. Hippocampal slices were rapidly fixed at 5, 30, or 120 min after LTP induction by theta-burst stimulation (TBS). Dendrites were reconstructed through serial section electron microscopy from comparable regions near the TBS or control electrodes in the same slice, and in unstimulated hippocampus that was perfusion-fixed in vivo. At 5 min after induction of LTP, polyribosomes were elevated in dendritic shafts and spines, especially near spine bases and in spine heads. At 30 min, polyribosomes remained elevated only in spine bases. At 120 min, both spine bases and spine necks had elevated polyribosomes. Polyribosomes accumulated in spines with larger synapses at 5 and 30 min, but not at 120 min. Small spines, meanwhile, proliferated dramatically by 120 min, but these largely lacked polyribosomes. The number of ribosomes per polyribosome is variable and may reflect differences in translation regulation. In dendritic spines, but not shafts, there were fewer ribosomes per polyribosome in the slice conditions relative to in vivo, but this recovered transiently in the 5 min LTP condition. Overall, our data show that LTP induces a rapid, transient upregulation of large polyribosomes in larger spines, and a persistent upregulation of small polyribosomes in the bases and necks of small spines. This is consistent with local translation supporting enlargement of potentiated synapses within minutes of LTP induction.
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Región CA1 Hipocampal/metabolismo , Potenciación a Largo Plazo/fisiología , Polirribosomas/ultraestructura , Biosíntesis de Proteínas/fisiología , Sinapsis/metabolismo , Animales , Región CA1 Hipocampal/ultraestructura , Espinas Dendríticas/metabolismo , Espinas Dendríticas/ultraestructura , Masculino , Ratas , Ratas Long-Evans , Sinapsis/ultraestructuraRESUMEN
The composition dependence of the short-range order (SRO) structure in highly modified mixed glass former sodium thiosilicophosphate glasses, yNa2S + (1 - y)[xSiS2 + (1 - x)PS5/2], were investigated using infrared (IR), Raman, and 29Si and 31P magic angle spinning nuclear magnetic resonance (MAS NMR) spectroscopies. Both the y = 0.5 and 0.67 glasses undergo disproportionation reactions among the Si and P SRO structures, which lead to various and complex SRO structural units for the Si and P, as shown via the spectra used to characterize the glasses. In the y = 0.5 series, the compositionally expected and experimentally observed SRO units are the P1 and Si2 units in the two binary end-member glasses, where the superscript is the number of bridging sulfur atoms on the P or Si units. However, in the ternary mixed glasses, 0 < x < 1, these units were found to react to form P0 (more highly modified, y = 0.60) and Si3 (less highly modified, y = 0.33) units, indicating preferential association of Na+ ions with the P SRO structures. The Raman spectra were used to resolve the heretofore incompletely studied Si3 SRO unit, which was otherwise difficult to elucidate using 29Si MAS NMR alone. In the y = 0.67 series glasses, the expected P0 and Si0 SRO units were observed for the end-member binary glasses. Like in the y = 0.5 series, the 29Si MAS NMR showed that edge-sharing Si2 (ESi2, y = 0.5) structures were also present in these highly modified glasses, which meant Na2S was not completely incorporated in the network. Evidence of this was shown in the Raman spectra in the form of polysulfide structures Na2Sx (x = 2, 4).
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In adult hippocampus, long-term potentiation (LTP) produces synapse enlargement while preventing the formation of new small dendritic spines. Here, we tested how LTP affects structural synaptic plasticity in hippocampal area CA1 of Long-Evans rats at postnatal day 15 (P15). P15 is an age of robust synaptogenesis when less than 35% of dendritic spines have formed. We hypothesized that LTP might therefore have a different effect on synapse structure than in adults. Theta-burst stimulation (TBS) was used to induce LTP at one site and control stimulation was delivered at an independent site, both within s. radiatum of the same hippocampal slice. Slices were rapidly fixed at 5, 30, and 120 min after TBS, and processed for analysis by three-dimensional reconstruction from serial section electron microscopy (3DEM). All findings were compared to hippocampus that was perfusion-fixed (PF) in vivo at P15. Excitatory and inhibitory synapses on dendritic spines and shafts were distinguished from synaptic precursors, including filopodia and surface specializations. The potentiated response plateaued between 5 and 30 min and remained potentiated prior to fixation. TBS resulted in more small spines relative to PF by 30 min. This TBS-related spine increase lasted 120 min, hence, there were substantially more small spines with LTP than in the control or PF conditions. In contrast, control test pulses resulted in spine loss relative to PF by 120 min, but not earlier. The findings provide accurate new measurements of spine and synapse densities and sizes. The added or lost spines had small synapses, took time to form or disappear, and did not result in elevated potentiation or depression at 120 min. Thus, at P15 the spines formed following TBS, or lost with control stimulation, appear to be functionally silent. With TBS, existing synapses were awakened and then new spines formed as potential substrates for subsequent plasticity.
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Hipocampo/crecimiento & desarrollo , Hipocampo/fisiología , Potenciación a Largo Plazo/fisiología , Neurogénesis/fisiología , Sinapsis/fisiología , Análisis de Varianza , Animales , Animales Recién Nacidos , Biofisica , Espinas Dendríticas/fisiología , Espinas Dendríticas/ultraestructura , Estimulación Eléctrica , Imagenología Tridimensional , Técnicas In Vitro , Microscopía Electrónica , Técnicas de Placa-Clamp , Ratas , Ratas Long-Evans , Sinapsis/ultraestructuraRESUMEN
Volume loss due to facial aging can be restored by facial volumization using a variety of materials. Volumization can be performed in isolation or concurrent with other facial rejuvenation procedures to obtain an optimal aesthetic result. There is a myriad of manufactured products available for volumization. The use of autologous fat as facial filler has been adopted more recently and possesses certain advantages; however, the ideal filler is still lacking. Tissue engineering may offer a solution. This technology would provide autologous soft-tissue components for use in facial volumization. The use of stem cells may enable customization of the engineered product for the specific needs of each patient.