Совершенствование процесса формообразования в условиях плоского напряженного состояния растяжения

Thin-walled axisymmetric truncated parts made of sheet billets are actively used in rocket and aerospace engineering. Improvement to their shape formation, based on directed material thickness change will ensure the production of parts with minimum thickness variation. This will also enable aviation and space industry enterprises to attain leading positions, as well as reduce labor costs. This work studies the possibility of obtaining thin-walled axisymmetric parts of truncated tapered shape using one of the methods of sheet metal stamping under flat tensile stress conditions (flanging). The mechanism was identified and the analysis of the stress-strain state of the billet during deformation was carried out. This takes into account the minimizing of the difference between the specified and technologically possible thicknesses. A mathematical model was developed to consider the shaping method based on the process of flanging. Theoretical studies were based on the principles of the plastic deformation theory of sheet materials. This was achieved by the following factors: approximate differential equations of force equilibrium; equations of constraint; plasticity conditions; and fundamental constitutive relations under given initial and boundary conditions. The process of flanging was simulated using the LS-DYNA software package with the following initial data of a conical billet made of 12Kh18N10T steel: cone angle 16.4°, thickness Sbillet = 0.3 mm. The aim was to eliminate errors in designing a tool for future implementation of the method on a manufactured die tooling, as well as to confirm the theoretical conclusions on the selection of technological parameters and achieve minimal thickness variation. The steps of computer modeling are presented, indicating the main process parameters such as material model, mechanical characteristics of the workpiece material, type of elements, kinematic loads, conditions of contact interaction of elements with each other, etc.В ракетно-космической и авиационной технике активно применяются тонкостенные осесимметричные  детали усеченной  сужающейся  формы,  изготовленные  из  листовых  заготовок.  Совершенствование  процессов  их  формообразования, в основе которых направленное изменение толщины материала с целью получения деталей с минимальной разнотолщинностью, позволит обеспечить ведущие позиции предприятий авиационной и космической отраслей промышленности, а также гарантирует снижение трудозатрат. Данная работа посвящена исследованию возможности получения тонкостенных осесимметричных деталей усеченной сужающейся формы одним из способов листовой штамповки в условиях плоского напряженного состояния растяжения (отбортовкой). Выявлен механизм и проведен анализ напряженно-деформированного состояния заготовки в процессе формоизменения с учетом  выражения  минимизации  между  заданной  и  технологически  возможной  толщинами. Разработана математическая модель рассматриваемого способа формообразования, основанного на процессе отбортовки. Теоретические исследования основывались на положениях теории пластического деформирования листовых материалов путем совместного  решения  приближенных  дифференциальных  уравнений  равновесия  сил,  уравнений  связи,  условия  пластичности и основных определяющих соотношений при заданных начальных и граничных условиях. С целью исключения ошибок при проектировании инструмента для перспективной реализации способа на изготовленной штамповой оснастке, а также для подтверждения теоретических выводов по выбору технологических параметров и достижения минимальной разнотолщинности проведено моделирование процесса отбортовки в программном комплексе LS-DYNA с  исходными  данными  конической  заготовки из стали 12Х18Н10Т: угол конусности 16,4°, толщина Sзаг = 0,3 мм. Представлены этапы компьютерного моделирования с указанием  основных  параметров  процесса,  таких  как  модель  материала,  механические  характеристики  материала  заготовки, тип элементов, кинематические нагрузки, условия контактного взаимодействия элементов между собой и т.д

CHL1 cooperates with PAK1–3 to regulate morphological differentiation of embryonic cortical neurons

The cell adhesion molecule Close Homologue of L1 (CHL1) is important for apical dendritic projection and laminar positioning of pyramidal neurons in caudal regions of the cerebral cortex. The p21-activated kinase (PAK 1-3) subfamily of serine/threonine kinases has also been implicated in regulating cell adhesion, migration, and morphology. Immunofluorescence staining in mouse embryonic brain showed that PAK1-3 was expressed in embryonic cortex and colocalized with CHL1 during neuronal migration and differentiation. To investigate a cooperative function for CHL1 and PAK in pyramidal cell differentiation or migration, a dominant-negative PAK mutant (PAK1 AID) that inhibits PAK1-3 kinase activity while coexpressing a green fluorescent protein (GFP) reporter was electroporated into the lateral ventricles of wild type and CHL1 null mutant mouse embryos (E14.5), then brain slices were cultured and neurons analyzed for laminar positioning and morphology by confocal microscopy after 3 days in vitro. Expression of PAK1 AID in CHL1 mutant cortex inactivated PAK and caused embryonic cortical neurons to branch profusely in the intermediate zone and cortical plate. The number of nodes, terminals and length of leading processes/apical dendrites of CHL1 mutant embryos expressing PAK1 AID increased dramatically, compared to CHL1 mutants without PAK1 AID, or wild type embryos with or without PAK1 AID. These findings suggest that CHL1 and PAK1-3 kinase cooperate, most likely in independent pathways, in regulating morphological development of the leading process/apical dendrite of embryonic cortical neurons

The cell adhesion molecule L1 regulates the expression of choline acetyltransferase and the development of septal cholinergic neurons

Mutations in the L1 gene cause severe brain malformations and mental retardation. We investigated the potential roles of L1 in the regulation of choline acetyltransferase (ChAT) and in the development of septal cholinergic neurons, which are known to project to the hippocampus and play key roles in cognitive functions. Using stereological approaches, we detected significantly fewer ChAT-positive cholinergic neurons in the medial septum and vertical limb of the diagonal band of Broca (MS/VDB) of 2-week-old L1-deficient mice compared to wild-type littermates (1644 ± 137 vs. 2051 ± 165, P = 0.038). ChAT protein levels in the septum were 53% lower in 2-week-old L1-deficient mice compared to wild-type littermates. ChAT activity in the septum was significantly reduced in L1-deficient mice compared to wild-type littermates at 1 (34%) and 2 (40%) weeks of age. In vitro, increasing doses of L1-Fc induced ChAT activity in septal neurons with a significant linear trend (*P = 0.0065). At 4 weeks of age in the septum and at all time points investigated in the caudate-putamen (CPu), the number of ChAT-positive neurons and the levels of ChAT activity were not statistically different between L1-deficient mice and wild-type littermates. The total number of cells positive for the neuronal nuclear antigen (NeuN) in the MS/VDB and CPu was not statistically different in L1-deficient mice compared to wild-type littermates, and comparable expression of the cell cycle marker Ki67 was observed. Our results indicate that L1 is required for the timely maturation of septal cholinergic neurons and that L1 promotes the expression and activity of ChAT in septal neurons

Neural Cell Adhesion Molecule NrCAM Regulates Semaphorin 3F-Induced Dendritic Spine Remodeling

Neuron-glial related cell adhesion molecule (NrCAM) is a regulator of axon growth and repellent guidance, and has been implicated in autism spectrum disorders. Here a novel postsynaptic role for NrCAM in Semaphorin3F (Sema3F)-induced dendritic spine remodeling was identified in pyramidal neurons of the primary visual cortex (V1). NrCAM localized to dendritic spines of star pyramidal cells in postnatal V1, where it was coexpressed with Sema3F. NrCAM deletion in mice resulted in elevated spine densities on apical dendrites of star pyramidal cells at both postnatal and adult stages, and electron microscopy revealed increased numbers of asymmetric synapses in layer 4 of V1. Whole-cell recordings in cortical slices from NrCAM-null mice revealed increased frequency of mEPSCs in star pyramidal neurons. Recombinant Sema3F-Fc protein induced spine retraction on apical dendrites of wild-type, but not NrCAM-null cortical neurons in culture, while re-expression of NrCAM rescued the spine retraction response. NrCAM formed a complex in brain with Sema3F receptor subunits Neuropilin-2 (Npn-2) and PlexinA3 (PlexA3) through an Npn-2-binding sequence (TARNER) in the extracellular Ig1 domain. A trans heterozygous genetic interaction test demonstrated that Sema3F and NrCAM pathways interacted in vivo to regulate spine density in star pyramidal neurons. These findings reveal NrCAM as a novel postnatal regulator of dendritic spine density in cortical pyramidal neurons, and an integral component of the Sema3F receptor complex. The results implicate NrCAM as a contributor to excitatory/inhibitory balance in neocortical circuits

Close Homolog of L1 and Neuropilin 1 Mediate Guidance of Thalamocortical Axons at the Ventral Telencephalon

We report a cooperation between the neural adhesion molecule close homolog of L1 (CHL1) and the semaphorin 3A (Sema3A) receptor, neuropilin 1 (Npn1), important for establishment of area-specific thalamocortical projections. CHL1 deletion in mice selectively disrupted the projection of somatosensory thalamic axons from the ventrobasal (VB) nuclei, causing them to shift caudally and target the visual cortex. At the ventral telencephalon, an intermediate target with graded Sema3A expression, VB axons were caudally shifted in CHL

NrCAM Deletion Causes Topographic Mistargeting of Thalamocortical Axons to the Visual Cortex and Disrupts Visual Acuity

NrCAM is a neural cell adhesion molecule of the L1 family that has been linked to autism spectrum disorders (ASDs), a disease spectrum in which abnormal thalamocortical connectivity may contribute to visual processing defects. Here we show that NrCAM interaction with Neuropilin-2 (Npn-2) is critical for Semaphorin3F (Sema3F)-induced guidance of thalamocortical axon subpopulations at the ventral telencephalon (VTe), an intermediate target for thalamic axon sorting. Genetic deletion of NrCAM or Npn-2 caused contingents of embryonic thalamic axons to misproject caudally in the VTe, away from a caudal-high Sema3F gradient. The resultant thalamocortical map of NrCAM null mutants showed striking mistargeting of motor and somatosensory thalamic axon contingents to the primary visual cortex, but retino-geniculate targeting and segregation were normal. NrCAM formed a molecular complex with Npn-2 in brain and neural cells, and was required for Sema3F-induced growth cone collapse in thalamic neuron cultures, consistent with a vital function for NrCAM in Sema3F-induced axon repulsion. NrCAM null mice displayed reduced responses to visual evoked potentials (VEPs) recorded from layer IV in the binocular zone of primary visual cortex (V1), particularly when evoked from the ipsilateral eye, indicating abnormal visual acuity and ocularity. These results demonstrate that NrCAM is required for normal maturation of cortical visual acuity, and suggest that the aberrant projection of thalamic motor and somatosensory axons to the visual cortex in NrCAM null mutant mice impairs cortical functions

РАДИОНУКЛИДНАЯ ОЦЕНКА АЛЬВЕОЛЯРНО КАПИЛЛЯРНОЙ ПРОНИЦАЕМОСТИ ДЛЯ ДИФФЕРЕНЦИАЛЬНОЙ ДИАГНОСТИКИ ЛЕГОЧНОГО ИНФИЛЬТРАТА

Summary.Ventilation scintigraphy of the lungs with analysis of alveolar-capillary permeability (ACP) was performed in patients with confirmed diagnoses of community-acquired pneumonia (CAP, n = 40), infiltrative pulmonary tuberculosis (IPT, n = 20), distal pulmonary embolism (DPE, n = 11),peripheral lung carcinoma (PLC, n = 17), and in 22 healthy volunteers. ACP values were higher in affected and intact lungs in patients with CAP at 10 and 30 min of the investigation vs those with DPE and were higher at 30 min in the affected lung in patients with CAP compared to patients with IPT. Comparison of ACPs in DPE and IPT patients revealed the ACP reduction in affected and intact lungs of DPE patients at 30 min of the investigation. PLC patients demonstrated the ACP reduction in affected lung compared to contralateral lung. Therefore, ACP gave additional information for differential diagnosis of CAP, IPT, DPE and PLCРезюме. У 88 пациентов с верифицированными диагнозами: внебольничная пневмония (ВП) – 40, инфильтративный туберкулез легких(ИТЛ) – 20, тромбоэмболия дистальных ветвей легочной артерии (ТДВЛА) – 11, периферический рак легких (ПРЛ) – 17, а также у 22 здоровых добровольцев проведена вентиляционная пульмоносцинтиграфия с анализом альвеолярно-капиллярной проницаемости (АКП). При ВП, в отличие от ТДВЛА и ИТЛ, АКП была выше на 10-й и 30-й мин исследования в пораженном и интактном легких по сравнению с ТДВЛА и на 30-й мин исследования – в пораженном легком по сравнению с ИТЛ. Сопоставление значений АКП у больных ТДВЛА и ИТЛ показало снижение АКП на 30-й мин исследования у больных ТДВЛА в пораженном и интактном легких. У пациентов с ПРЛ регистрировалось снижение АКП на стороне поражения по сравнению с контралатеральным легким. Таким образом, оценка АКП позволяет расширить возможности дифференциальной диагностики ВП, ИТЛ, ТДВЛА и ПРЛ

The 14-3-3ζ Protein Binds to the Cell Adhesion Molecule L1, Promotes L1 Phosphorylation by CKII and Influences L1-Dependent Neurite Outgrowth

BACKGROUND: The cell adhesion molecule L1 is crucial for mammalian nervous system development. L1 acts as a mediator of signaling events through its intracellular domain, which comprises a putative binding site for 14-3-3 proteins. These regulators of diverse cellular processes are abundant in the brain and preferentially expressed by neurons. In this study, we investigated whether L1 interacts with 14-3-3 proteins, how this interaction is mediated, and whether 14-3-3 proteins influence the function of L1. METHODOLOGY/PRINCIPAL FINDINGS: By immunoprecipitation, we demonstrated that 14-3-3 proteins are associated with L1 in mouse brain. The site of 14-3-3 interaction in the L1 intracellular domain (L1ICD), which was identified by site-directed mutagenesis and direct binding assays, is phosphorylated by casein kinase II (CKII), and CKII phosphorylation of the L1ICD enhances binding of the 14-3-3 zeta isoform (14-3-3ζ). Interestingly, in an in vitro phosphorylation assay, 14-3-3ζ promoted CKII-dependent phosphorylation of the L1ICD. Given that L1 phosphorylation by CKII has been implicated in L1-triggered axonal elongation, we investigated the influence of 14-3-3ζ on L1-dependent neurite outgrowth. We found that expression of a mutated form of 14-3-3ζ, which impairs interactions of 14-3-3ζ with its binding partners, stimulated neurite elongation from cultured rat hippocampal neurons, supporting a functional connection between L1 and 14-3-3ζ. CONCLUSIONS/SIGNIFICANCE: Our results suggest that 14-3-3ζ, a novel direct binding partner of the L1ICD, promotes L1 phosphorylation by CKII in the central nervous system, and regulates neurite outgrowth, an important biological process triggered by L1

Using C. elegans to decipher the cellular and molecular mechanisms underlying neurodevelopmental disorders

Prova tipográfica (uncorrected proof)Neurodevelopmental disorders such as epilepsy, intellectual disability (ID), and autism spectrum disorders (ASDs) occur in over 2 % of the population, as the result of genetic mutations, environmental factors, or combination of both. In the last years, use of large-scale genomic techniques allowed important advances in the identification of genes/loci associated with these disorders. Nevertheless, following association of novel genes with a given disease, interpretation of findings is often difficult due to lack of information on gene function and effect of a given mutation in the corresponding protein. This brings the need to validate genetic associations from a functional perspective in model systems in a relatively fast but effective manner. In this context, the small nematode, Caenorhabditis elegans, presents a good compromise between the simplicity of cell models and the complexity of rodent nervous systems. In this article, we review the features that make C. elegans a good model for the study of neurodevelopmental diseases. We discuss its nervous system architecture and function as well as the molecular basis of behaviors that seem important in the context of different neurodevelopmental disorders. We review methodologies used to assess memory, learning, and social behavior as well as susceptibility to seizures in this organism. We will also discuss technological progresses applied in C. elegans neurobiology research, such as use of microfluidics and optogenetic tools. Finally, we will present some interesting examples of the functional analysis of genes associated with human neurodevelopmental disorders and how we can move from genes to therapies using this simple model organism.The authors would like to acknowledge Fundação para a Ciência e Tecnologia (FCT) (PTDC/SAU-GMG/112577/2009). AJR and CB are recipients of FCT fellowships: SFRH/BPD/33611/2009 and SFRH/BPD/74452/2010, respectively

Perinatal asphyxia: current status and approaches towards neuroprotective strategies, with focus on sentinel proteins

Delivery is a stressful and risky event menacing the newborn. The mother-dependent respiration has to be replaced by autonomous pulmonary breathing immediately after delivery. If delayed, it may lead to deficient oxygen supply compromising survival and development of the central nervous system. Lack of oxygen availability gives rise to depletion of NAD+ tissue stores, decrease of ATP formation, weakening of the electron transport pump and anaerobic metabolism and acidosis, leading necessarily to death if oxygenation is not promptly re-established. Re-oxygenation triggers a cascade of compensatory biochemical events to restore function, which may be accompanied by improper homeostasis and oxidative stress. Consequences may be incomplete recovery, or excess reactions that worsen the biological outcome by disturbed metabolism and/or imbalance produced by over-expression of alternative metabolic pathways. Perinatal asphyxia has been associated with severe neurological and psychiatric sequelae with delayed clinical onset. No specific treatments have yet been established. In the clinical setting, after resuscitation of an infant with birth asphyxia, the emphasis is on supportive therapy. Several interventions have been proposed to attenuate secondary neuronal injuries elicited by asphyxia, including hypothermia. Although promising, the clinical efficacy of hypothermia has not been fully demonstrated. It is evident that new approaches are warranted. The purpose of this review is to discuss the concept of sentinel proteins as targets for neuroprotection. Several sentinel proteins have been described to protect the integrity of the genome (e.g. PARP-1; XRCC1; DNA ligase IIIα; DNA polymerase β, ERCC2, DNA-dependent protein kinases). They act by eliciting metabolic cascades leading to (i) activation of cell survival and neurotrophic pathways; (ii) early and delayed programmed cell death, and (iii) promotion of cell proliferation, differentiation, neuritogenesis and synaptogenesis. It is proposed that sentinel proteins can be used as markers for characterising long-term effects of perinatal asphyxia, and as targets for novel therapeutic development and innovative strategies for neonatal care