Pre-fusion motion state determines the heterogeneity of membrane fusion dynamics for large dense-core vesicles.

Aim: In neuroendocrine cells, large dense-core vesicles (LDCVs) undergo highly regulated pre-fusion processes before releasing hormones via membrane fusion. Significant heterogeneity has been found for LDCV population based on the dynamics of membrane fusion. However, how the pre-fusion status impacts the heterogeneity of LDCVs still remains unclear.

Identification of a Prognostic Signature for Ovarian Cancer Based on Ubiquitin-Related Genes Suggesting a Potential Role for FBXO9.

Background: Ovarian cancer (OV) is associated with high mortality and poses challenges in diagnosis and prognosis prediction. Ubiquitin-related genes (UbRGs) are involved in the initiation and progression of cancers, but have still not been utilized for diagnosis and prognosis of OV.

Methods: K48-linked ubiquitination in ovarian tissues from our OV and control cohort was assessed using immunohistochemistry.

Elastin-like recombinamer-mediated hierarchical mineralization coatings on Zr-16Nb-xTi (x = 4,16 wt%) alloy surfaces improve biocompatibility.

The biocompatibility of biomedical materials is vital to their applicability and functionality. However, modifying surfaces for enhanced biocompatibility using traditional surface treatment techniques is challenging. We employed a mineralizing elastin-like recombinamer (ELR) self-assembling platform to mediate mineralization on Zr-16Nb-xTi (x = 4,16 wt%) alloy surfaces, resulting in the modification of surface morphology and bioactivity while improving the biocompatibility of the material.

Activated or Impaired: An Overview of DNA Repair in Neurodegenerative Diseases.

As the population ages, age-related neurodegenerative diseases have become a major challenge in health science. Currently, the pathology of neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and Huntington's disease, is still not fully understood. Remarkably, emerging evidence indicates a role of genomic DNA damage and repair in various neurodegenerative disorders.

A two-subpopulation model that reflects heterogeneity of large dense core vesicles in exocytosis.

Exocytosis of large dense core vesicles is responsible for hormone secretion in neuroendocrine cells. The population of primed vesicles ready to release upon cell excitation demonstrates large heterogeneity. However, there are currently no models that clearly reflect such heterogeneity.

Lycorine hydrochloride suppresses stress-induced premature cellular senescence by stabilizing the genome of human cells.

Lycorine, a natural compound isolated from the traditional Chinese medicinal herb Lycoris radiata, exhibits multiple pharmacological effects, such as anti-inflammatory, antiviral, and anticancer effects. Accumulating evidence also indicates that lycorine might hold the potential to treat age-associated Alzheimer's disease. However, whether lycorine is involved in delaying the onset of cellular senescence and its underlying mechanisms has not been determined.

Novel Biocompatible Zr-Based Alloy with Low Young's Modulus and Magnetic Susceptibility for Biomedical Implants.

The microstructure, mechanical properties, magnetic susceptibility, electrochemical corrosion performance, in vitro cell compatibility and blood consistency of Zr-16Nb-xTi (x = 0, 4, 8, 12 and 16 wt.%) materials were investigated as potential materials for biomedical implants. X-ray diffraction (XRD) and Transmission electron microscopy (TEM) analyses revealed the secondary phase martensite α' formed during the quenching process.

Diosmetin enhances the sensitivity of radiotherapy by suppressing homologous recombination in endometrial cancer.

Radiotherapy is an essential treatment for endometrial cancer (EC), especially in advanced, metastatic, and recurrent cases. Combining radiotherapy, which mainly causes DNA double-strand breaks (DSBs), with small molecules targeting aberrantly activated homologous recombination (HR) repair pathways holds great potential for treating ECs in advanced stages. Here, we demonstrate that diosmetin (DIO), a natural flavonoid, suppresses HR, therefore inhibiting cell proliferation and enhancing the sensitivity of EC to radiotherapy.

Doc2-mediated superpriming supports synaptic augmentation.

Various forms of synaptic plasticity underlie aspects of learning and memory. Synaptic augmentation is a form of short-term plasticity characterized by synaptic enhancement that persists for seconds following specific patterns of stimulation. The mechanisms underlying this form of plasticity are unclear but are thought to involve residual presynaptic Ca Here, we report that augmentation was reduced in cultured mouse hippocampal neurons lacking the Ca sensor, Doc2; other forms of short-term enhancement were unaffected.

Different states of synaptotagmin regulate evoked versus spontaneous release.

The tandem C2-domains of synaptotagmin 1 (syt) function as Ca(2+)-binding modules that trigger exocytosis; in the absence of Ca(2+), syt inhibits spontaneous release. Here, we used proline linkers to constrain and alter the relative orientation of these C2-domains. Short poly-proline helices have a period of three, so large changes in the relative disposition of the C2-domains result from changing the length of the poly-proline linker by a single residue.

Structural elements that underlie Doc2β function during asynchronous synaptic transmission.

Double C2-like domain-containing proteins alpha and beta (Doc2α and Doc2β) are tandem C2-domain proteins proposed to function as Ca(2+) sensors for asynchronous neurotransmitter release. Here, we systematically analyze each of the negatively charged residues that mediate binding of Ca(2+) to the β isoform. The Ca(2+) ligands in the C2A domain were dispensable for Ca(2+)-dependent translocation to the plasma membrane, with one exception: neutralization of D220 resulted in constitutive translocation.

Linker mutations reveal the complexity of synaptotagmin 1 action during synaptic transmission.

The Ca(2+) sensor for rapid synaptic vesicle exocytosis, synaptotagmin 1 (syt), is largely composed of two Ca(2+)-sensing C2 domains, C2A and C2B. We investigated the apparent synergy between the tandem C2 domains by altering the length and rigidity of the linker that connects them. The behavior of the linker mutants revealed a correlation between the ability of the C2 domains to penetrate membranes in response to Ca(2+) and to drive evoked neurotransmitter release in cultured mouse neurons, uncovering a step in excitation-secretion coupling.

Mutations that disrupt Ca²⁺-binding activity endow Doc2β with novel functional properties during synaptic transmission.

Double C2-domain protein (Doc2) is a Ca(2+)-binding protein implicated in asynchronous and spontaneous neurotransmitter release. Here we demonstrate that each of its C2 domains senses Ca(2+); moreover, the tethered tandem C2 domains display properties distinct from the isolated domains. We confirm that overexpression of a mutant form of Doc2β, in which two acidic Ca(2+) ligands in the C2A domain and two in the C2B domain have been neutralized, results in markedly enhanced asynchronous release in synaptotagmin 1-knockout neurons.

The crosstalks between adipokines and catecholamines.

Adipocytes, which secrete a spectrum of adipokines, play an integral role in metabolism via communications with other endocrine cells. In the present work, we have studied the interplays between adipokines and catecholamines, using 3T3-L1 adipocytes and PC12 cells as the cell models and an integrative experimental platform. We demonstrate that all catecholamines inhibit vesicle trafficking and secretion of leptin and resistin through β-adrenergic receptors, while leptin and resistin enhance the vesicle trafficking and secretion of catecholamines through PKC, PKA, MAPK kinase and Ca(2+) dependent pathways.

Roles of cholesterol in vesicle fusion and motion.

Although it is well established that exocytosis of neurotransmitters and hormones is highly regulated by numerous secretory proteins, such as SNARE proteins, there is an increasing appreciation of the importance of the chemophysical properties and organization of membrane lipids to various aspects of the exocytotic program. Based on amperometric recordings by carbon fiber microelectrodes, we show that deprivation of membrane cholesterol by methyl-beta-cyclodextrin not only inhibited the extent of membrane depolarization-induced exocytosis, it also adversely affected the kinetics and quantal size of vesicle fusion in neuroendocrine PC12 cells. In addition, total internal fluorescence microscopy studies revealed that cholesterol depletion impaired vesicle docking and trafficking, which are believed to correlate with the dynamics of exocytosis.

PKC epsilon facilitates recovery of exocytosis after an exhausting stimulation.

It has been well documented that protein kinase Cs (PKCs) play multifaceted roles in regulating exocytosis of neurotransmitters and hormones. But the isoform-specific PKC effects are still poorly elucidated mainly because of the large variety of PKC isoforms and the dubious specificity of the commonly used pharmacological agents. In the present study, based on overexpression of wild-type or dominant negative PKC epsilon, we demonstrate in neuroendocrine PC12 cells that PKC epsilon, but not PKC alpha, facilitates recovery of exocytosis after an exhausting stimulation.

Involvement of PKC alpha in PMA-induced facilitation of exocytosis and vesicle fusion in PC12 cells.

Phorbol-12-myristate-13-acetate, a stable analog of the important signaling membrane lipid diacylglycerol (DAG), is known to potentiate exocytosis and modulate vesicle fusion kinetics in neurons and endocrine cells. The exact mechanisms underlying the actions of PMA, however, is often not clear, largely because of the diversity of the DAG/PMA receptors involved in the exocytotic process, which include, most notably, various isoforms of protein kinase C (PKC). In this study, the roles of PKC alpha in PMA-mediated regulation of exocytosis were investigated by over-expressing wild-type PKC alpha (wt-PKC alpha) or dominant negative PKC alpha (dn-PKC alpha).

Label-free detection of ATP release from living astrocytes with high temporal resolution using carbon nanotube network.

Owing to its unique combination of electrical, physiochemical, and one-dimension structural properties, single-walled carbon nanotube (SWNT) has recently emerged as a novel nanoelectronic biosensor for biomolecular detection with extraordinary sensitivity and simple detection scheme. All the realizations so far, however, are limited to static in vitro measurement. Dynamic detection of biomolecule release from living cells which may occur in millisecond timescale has yet to be demonstrated.

Effects of phorbol ester on vesicle dynamics as revealed by total internal reflection fluorescence microscopy.

Exocytosis of neurotransmitter or hormone-filled vesicles is a highly dynamic process regulated by various proteins and lipids. As mainly revealed indirectly by the electrophysiological methods, exocytosis is believed to involve multiple kinetic steps in which vesicles transit from one state to another. Using total internal reflection fluorescence microscopy which enables direct visualization of individual vesicles, we developed an analytical framework to track and analyze vesicle dynamics.

Highly sensitive and selective method to detect dopamine in the presence of ascorbic acid by a new polymeric composite film.

A new approach was applied to modify gold electrode with a unique polymer composite for selectively detecting dopamine (DA), a neurotransmitter, in the presence of an electroactive species of ascorbic acid (AA). After self-assembly of 11-mercaptoundecanoic acid (MUA) monolayer on gold surface, polyethylene glycol (PEG) was used to perform electrochemical esterification with MUA. In general, AA is the main interference of DA detection in a biological system.

Chronic daily administration of ethyl docosahexaenoate protects against gerbil brain ischemic damage through reduction of arachidonic acid liberation and accumulation.

Recently, we reported that dietary ethyl docosahexaenoate (Et-DHA) intake decreases the level of membrane arachidonic acid (AA), which reduces the generation of AA metabolites in ischemic gerbil brain. As an extended study, we further investigated the influence of the chronic administration of Et-DHA on free AA levels after ischemia. In addition, Na,K-ATPase activity, cation content, cerebral edema and brain damage were also evaluated.

Chronic administration of ethyl docosahexaenoate decreases mortality and cerebral edema in ischemic gerbils.

Dietary docosahexaenoic acid (DHA) intake can decrease the level of membrane arachidonic acid (AA), which is liberated during cerebral ischemia and implicated in the pathogenesis of brain damage. Therefore, in the present study, we investigated the effects of chronic ethyl docosahexaenoate (E-DHA) administration on mortality and cerebral edema induced by transient forebrain ischemia in gerbils. Male Mongolian gerbils were orally pretreated with either E-DHA (100, 150 mg/kg) or vehicle, once a day, for 4 weeks and were subjected to transient forebrain ischemia by bilateral common carotid occlusion for 30 min.

Effects of docosahexaenoic acid on the survival and neurite outgrowth of rat cortical neurons in primary cultures.

Effects of docosahexaenoic acid (DHA) on survival and neurite outgrowth were investigated in primary cultures of rat cortical neurons. Cell cultures were prepared from cortex on embryonic day 18 (E-18) for treatment with a series of DHA concentrations (12.5, 25, 50, 75, 100 and 200 microM).