Morphometric and immunohistochemical studies of the suprachiasmatic nucleus in the hereditary microphthalmic rat]

Morphometric and immunohistochemical analyses of the suprachiasmatic nucleus (SCN) were performed on hereditary microphthalmic rats. In normal rats, the number of cells and the volume of the SCN were 11, 631 and 6.7 x 10(-2) mm3 (an average taken from 12 SCNs). However, the neuronal population and volume of the SCN in hereditary microphthalmic rats were 7,450 and 4.5 x 10(-2) mm3 (an average taken from 14 SCNs), respectively. There were no significant differences in the size of neurons between normal and microphthalmic SCN neurons. Immunohistochemical studies showed that a considerable number of antivasopressin positive neurons were present in microphthalmic rats, despite their lack of the optic nerve. However, further detailed studies revealed that the number of antivasopressin positive neurons present in microphthalmic rats was only 68% of those found in normal rats. These findings suggest that the complete development of the SCN and vasopressin neurons depends on the visual input.     

Evaluation of Poly(2-Ethyl-2-Oxazoline) Containing Copolymer Networks of Varied Composition as Sustained Metoprolol Tartrate Delivery Systems

Segmented copolymer networks (SCN) based on poly(2-ethyl-2-oxazoline) and containing 2-hydroxyethyl methacrylate, 2-hydroxypropyl acrylate, and/or methyl methacrylate segments have been evaluated as potential sustained release systems of the water soluble cardioselective β-blocker metoprolol tartrate. The structure and properties of the drug carriers were investigated by differential scanning calorimetry, attenuated total reflectance Fourier transform infrared spectroscopy, scanning electron microscopy, and atomic force microscopy. Swelling kinetics of SCNs in various media was followed, and the conditions for effective MT loading were specified. MT-loaded SCNs with drug content up to 80 wt.% were produced. The release kinetics of metoprolol tartrate from the systems was studied and it was shown that the conetworks of different structure and composition are able to sustain the metoprolol tartrate release without additional excipients.     

Properties of the serotonergic cerebral ganglion neurons of the gastropod mollusc, Philine aperta

The characteristics of a pair of identified neurons found in the cerebral ganglia of the gastropod mollusc Philine aperta have been examined. Because they appear to contain serotonin, and since they probably also use serotonin as a neurotransmitter, these neurons were named the serotonergic cerebral neurons (SCNs). Each SCN sent an axon out of the ipsilateral cerebro-buccal connective to the buccal ganglia. The SCNs also had extensive projections to all the ipsilateral, and most of the contralateral, buccal nerve trunks. Stimulating the SCNs produced the hyperpolarization of a pair of identified buccal ganglion mechano-sensory neurons (the S-cells), and had an excitatory action on the electrical activity of acinar cells of the salivary glands. A comparison of the properties of the Philine SCNs with those of similar serotonin-containing cerebral ganglion neurons in other gastropod molluscs provides evidence of homology with these neurons.     

Cellular Requirements of Suprachiasmatic Nucleus Transplants for Restoration of Orcadian Rhythm

Fetal neurografts containing the suprachiasmatic nucleus (SCN) can restore the circadian locomotor and drinking rhythm of SCN-lesioned (SCNX) rat and hamster. This functional outcome finally proves that the endogenous biological clock autonomously resides in the SCN. Observations on the cellular requirements of the “new” SCN for restoration of the arrhythmic SCNX animals have led to some new insights and confirmed findings from other studies. A critical mass of SCN neurons appeared necessary for functional effects, whereas the temporal profile of reinstatement of rhythm correlated with the delayed maturation of the grafted SCN. Cytoarchitectoni-cally, the grafted SCN does not seem to develop normally for all anatomical aspects. Complementary clusters of vasoactive intestinal polypeptide(VIP)-and vasopressin(VP)ergic neurons are formed, but somatostatin(SOM)ergic neurons do not always “join” this group, as is normally seen in situ. Nevertheless, these new SCNs can restore the ablated functions. As the period length of restored rhythms tends to vary, it might be that the grafted SCN underwent an altered or impaired maturation that resulted in a different setting of its clock mechanism. A prominent role of VIPergic neurons seems indicated by their presence in all functional grafts, but, although they may be required, these cells do not appear to be a sufficient condition for restoration of rhythm. Many grafts exhibit the presence of VIPergic cells without counteracting the arrhythmia, whereas VP- and SOMergic SCN neurons are usually present as well. Findings with VP-deficient Brattleboro rat grafts indicated that VP is not the primary obligatory signal of circadian activity. It is argued that perhaps the role of SOMergic neurons in the clock function of the (grafted) SCN has been insufficiently considered. However, one should keep in mind that the peptides of the various types of SCN neurons may function only as cofactors, mutually modulating molecular or bioelectrical cellular activities within the nucleus or the message of the main transmitter γ-aminobutyric acid.     

Estimation of the mean waiting time of a customer subject to balking: a simulation study

To a customer, the waiting time for order processing for a product or service is important information for order placement. If the time foreseen for order fulfillment is long, the order might be lost to a competitor. In particular, modern principles of supply chain management highly suggest information sharing between entities in the chain and information technology has enabled customers to conveniently consider the waiting time for a potential balking decision. To help determine the design and operation of a manufacturing or service system in which a customer may balk based on the foreseen waiting time, this paper develops procedures to estimate the average waiting time of an order. Either the procedures allow the maximum waiting time for a balking decision to be random or do not require knowledge of the arrival process of customers before balking if the balking limit is known. For generality of the model, this paper considers general inter-arrival and service time distributions, and uses the simulation and regression approach.     

Resting-state brain organization revealed by functional covariance networks

Background

Brain network studies using techniques of intrinsic connectivity network based on fMRI time series (TS-ICN) and structural covariance network (SCN) have mapped out functional and structural organization of human brain at respective time scales. However, there lacks a meso-time-scale network to bridge the ICN and SCN and get insights of brain functional organization.

Methodology and Principal Findings

We proposed a functional covariance network (FCN) method by measuring the covariance of amplitude of low-frequency fluctuations (ALFF) in BOLD signals across subjects, and compared the patterns of ALFF-FCNs with the TS-ICNs and SCNs by mapping the brain networks of default network, task-positive network and sensory networks. We demonstrated large overlap among FCNs, ICNs and SCNs and modular nature in FCNs and ICNs by using conjunctional analysis. Most interestingly, FCN analysis showed a network dichotomy consisting of anti-correlated high-level cognitive system and low-level perceptive system, which is a novel finding different from the ICN dichotomy consisting of the default-mode network and the task-positive network.

Conclusion

The current study proposed an ALFF-FCN approach to measure the interregional correlation of brain activity responding to short periods of state, and revealed novel organization patterns of resting-state brain activity from an intermediate time scale.     

Fractal Patterns of Neural Activity Exist within the Suprachiasmatic Nucleus and Require Extrinsic Network Interactions

The mammalian central circadian pacemaker (the suprachiasmatic nucleus, SCN) contains thousands of neurons that are coupled through a complex network of interactions. In addition to the established role of the SCN in generating rhythms of ∼24 hours in many physiological functions, the SCN was recently shown to be necessary for normal self-similar/fractal organization of motor activity and heart rate over a wide range of time scales—from minutes to 24 hours. To test whether the neural network within the SCN is sufficient to generate such fractal patterns, we studied multi-unit neural activity of in vivo and in vitro SCNs in rodents. In vivo SCN-neural activity exhibited fractal patterns that are virtually identical in mice and rats and are similar to those in motor activity at time scales from minutes up to 10 hours. In addition, these patterns remained unchanged when the main afferent signal to the SCN, namely light, was removed. However, the fractal patterns of SCN-neural activity are not autonomous within the SCN as these patterns completely broke down in the isolated in vitro SCN despite persistence of circadian rhythmicity. Thus, SCN-neural activity is fractal in the intact organism and these fractal patterns require network interactions between the SCN and extra-SCN nodes. Such a fractal control network could underlie the fractal regulation observed in many physiological functions that involve the SCN, including motor control and heart rate regulation.     

Cytological features of serotonin-containing neurons and their processes in the retina of the carp (Cyprinus carpio). An immunohistochemical study using flat-mount preparations

The morphological features of serotonin-containing neurons (SCNs) and their processes in the retina of the carp (Cyprinus carpio) were immunohistochemically studied by applying a modified peroxidase-antiperoxidase technique using flat-mount preparations. The somata of immunoreactive SCNs were mostly located in the innermost part of the inner nuclear layer (INL). These cells were distributed at a density of 64.8 cells/mm2, this being similar to the density of dopamine-containing interplexiform cells. The processes of the SCNs ramified successively into two finer branches, eventually forming a broad, extensive network in the thin layer just subjacent to the plane of the somata of the SCNs. Processes originating from neighboring SCNs exhibited cytoplasmatic continuity with each other at the lightmicroscope level. Due to their location and cytological features, these SCNs appeared to correspond to amacrine cells.     

医师定期考核制度操作流程的再设计

针对目前医师定期考核制度操作流程中的8种缺陷,提出制度操作流程改造的策略,并明确流程改造后应该建立《执业医师专业守则》,接轨继续医学教育制度,完善医师重新注册制度。     

Cytological features of serotonin-containing neurons and their processes in the retina of the carp (Cyprinus carpio)

Summary The morphological features of serotonin-containing neurons (SCNs) and their processes in the retina of the carp (Cyprinus carpio) were immunohistochemically studied by applying a modified peroxidase-antiperoxidase technique using flat-mount preparations. The somata of immunoreactive SCNs were mostly located in the innermost part of the inner nuclear layer (INL). These cells were distributed at a density of 64.8 cells/mm², this being similar to the density of dopamine-containing interplexiform cells. The processes of the SCNs ramified successively into two finer branches, eventually, forming a broad, extensive network in the thin layer just subjacent to the plane of the somata of the SCNs. Processes originating from neighboring SCNs exhibited cytoplasmatic continuity with each other at the lightmicroscope level. Due to their location and cytological features these SCNs appeared to correspond to amacrine cells.In honour of Prof. P. van Duijn     

The conservation value of tree decay processes as a key driver structuring tree cavity nest webs in South American temperate rainforests

South American temperate rainforests, a global biodiversity hotspot, have been reduced to nearly 30% of their original extent and most remaining stands are being degraded. Cavity-nesting vertebrate communities are dependent on cavity-bearing trees and hierarchically structured within nest webs. Evaluating the actual degree of cavity dependence (obligate, non-obligate) and the preferred attributes of trees by cavity nesters is critical to design conservation strategies in areas undergoing habitat loss. During three breeding seasons (2010–2013), we studied the cavity-nesting bird community in temperate rainforests of Chile. We found the highest reported proportion of tree cavity nesters (n = 29 species; 57%) compared to non-cavity-using birds for any forest system. Four species were excavators and 25 were secondary cavity nesters (SCNs). Among SCNs, ten species were obligate and 15 were non-obligate cavity nesters. Seventy-five percent of nests of SCNs were located in cavities produced by tree decay processes and the remaining 25% were in cavities excavated mainly by Pygarrhichas albogularis and Campephilus magellanicus. Nest web structure had a low dominance and evenness, with most network interactions occurring between SCNs and large decaying trees. Tree diameter at breast height (DBH) was larger in nest-trees (57.3 cm) than in available trees (26.1 cm). Cavity nesters showed a strong preference for dead trees, both standing and fallen (58% of nests). Our results stress that retaining large decaying and standing dead trees (DBH > 57 cm), and large fallen trees, should be a priority for retention in forest management plans in this globally threatened ecosystem.     

Mild Androgen Insensitivity Syndrome: The Current Landscape

ObjectiveMild androgen insensitivity syndrome (MAIS) belongs to the androgen insensitivity syndrome (AIS) spectrum, an X-linked genetic disease that is the most common cause of differences in sex development. Unfortunately, AIS studies mainly focus on the partial and complete phenotypes, and the mild phenotype (MAIS) has been barely reported. Our purpose is to explore the MAIS facets, clinical features, and molecular aspects.MethodsWe collected all reported MAIS cases in the medical literature and presented them based on the phenotype and molecular diagnosis.ResultsWe identified 49 different androgen receptor (AR) mutations in 69 individuals in the literature. We compared the AR mutations presented in individuals with MAIS with AR mutations previously reported in other AIS phenotypes (partial and complete) regarding the type, location, genotype-phenotype correlation, and functional studies.ConclusionThis review provides a landscape of the mild phenotype of AIS. Most patients with MAIS present with male factor infertility. Therefore, AR gene sequencing should be considered during male factor infertility investigation, even in males with typically male external genitalia. In addition, MAIS can be part of other medical conditions, such as X-linked spinal and bulbar muscular atrophy (Kennedy disease).     

Importance of being Nernst: Synaptic activity and functional relevance in stem cell-derived neurons

Functional synaptogenesis and network emergence are signature endpoints of neurogenesis. These behaviors provide higher-order confirmation that biochemical and cellular processes necessary for neurotransmitter release, post-synaptic detection and network propagation of neuronal activity have been properly expressed and coordinated among cells. The development of synaptic neurotransmission can therefore be considered a defining property of neurons. Although dissociated primary neuron cultures readily form functioning synapses and network behaviors in vitro, continuously cultured neurogenic cell lines have historically failed to meet these criteria. Therefore, in vitro-derived neuron models that develop synaptic transmission are critically needed for a wide array of studies, including molecular neuroscience, developmental neurogenesis, disease research and neurotoxicology. Over the last decade, neurons derived from various stem cell lines have shown varying ability to develop into functionally mature neurons. In this review, we will discuss the neurogenic potential of various stem cells populations, addressing strengths and weaknesses of each, with particular attention to the emergence of functional behaviors. We will propose methods to functionally characterize new stem cell-derived neuron (SCN) platforms to improve their reliability as physiological relevant models. Finally, we will review how synaptically active SCNs can be applied to accelerate research in a variety of areas. Ultimately, emphasizing the critical importance of synaptic activity and network responses as a marker of neuronal maturation is anticipated to result in in vitro findings that better translate to efficacious clinical treatments.     

Electrical and pharmacological properties of the suprachiasmatic nuclei

The suprachiasmatic nuclei (SCN) of the mammalian hypothalamus are in important circadian pacemaker. The electrical activity of these nuclei exhibits an intrinsic circadian rhythm. The rhythmicity of the SCN is also reflected in cyclic glucose consumption and serotonin metabolism. These rhythms are entrained to the light-dark cycle via the retinohypothalamic projection. This pathway, possibly together with a visual projection via the ventral lateral geniculate nuclei, innervates light-responsive SCN cells, which exhibit the functional properties of luminance detectors. The SCN contain various peptides, acetylcholine, and serotonin either intrinsically or in terminals of afferent projections. For acetylcholine it has been demonstrated that the SCN mediate the process of photic entrainment and light suppression of pineal synthetic activity. In the case of serotonin and vasopressin it seems certain that the SCN do not depend on their presence for generating circadian rhythms or for entrainment. Both substances may modulate the intrinsic pacemaker frequency through mechanisms that remain to be established.     

Enhanced stability of core-surface cross-linked micelles fabricated from amphiphilic brush copolymers

"Stealth" nanoparticles made from polymer micelles have been widely explored as drug carriers for targeted drug delivery. High stability (i.e., low critical micelle concentration (CMC)) is required for their intravenous applications. Herein, we present a "core-surface cross-linking" concept to greatly enhance nanoparticle's stability: amphiphilic brush copolymers form core-surface cross-linked micelles (nanoparticles) (SCNs). The amphiphilic brush copolymers consisted of hydrophobic poly(epsilon-caprolactone) (PCL) and hydrophilic poly(ethylene glycol) (PEG) or poly(2-(N,N-dimethylamino)ethyl methacrylate) (PDMA) chains were synthesized by macromonomer copolymerization method and used to demonstrate this concept. The resulting SCNs were about 100 times more stable than micelles from corresponding amphiphilic block copolymers. The size and surface properties of the SCNs could be easily tailored by the copolymer's compositions.     

A combined experimental and quantum chemical study on the putative protonophoric activity of thiocyanate

Inhibition of gastric acid secretion by thiocyanate is explained by a protonophoric mechanism assuming that thiocyanate induces a H(+) back flux from the acidic gastric lumen into the parietal cells of gastric mucosa. Protonophoric activity of thiocyanate was examined by swelling measurements using rat liver mitochondria and theoretically by quantum chemical methods. Mitochondria suspended in K-thiocyanate medium plus nigericin (an H/K-exchanger) swelled when the medium pH was acidic, indicating that SCN(-) initiates a transfer of H(+) across the inner membrane. To rationalize the protonophoric activity of thiocyanate, we considered the dehydration of SCN(-) to be critical for transmembranal H(+) transfer. For modeling this process, various hydrate clusters of SCN(-) and Cl(-) were generated and optimized by density functional theory (DFT) at the B3-LYP/6-311++G(d,p) level. The cluster hydration energy was lower for SCN(-) than for Cl(-). The total Gibbs free energies of hydration of the ions were estimated by a hybrid supermolecule-continuum approach based on DFT. The calculated hydration energies also led to the conclusion that SCN(-) is less efficiently solvated than Cl(-). Due to the easier removal of the hydration shell of SCN(-) relative to Cl(-), SCN(-) is favored in going across the membrane, giving rise to the protonophoric activity.