Reconstruction and Simulation of Neocortical Microcircuitry.

We present a first-draft digital reconstruction of the microcircuitry of somatosensory cortex of juvenile rat. The reconstruction uses cellular and synaptic organizing principles to algorithmically reconstruct detailed anatomy and physiology from sparse experimental data. An objective anatomical method defines a neocortical volume of 0.29 ± 0.01 mm(3) containing ~31,000 neurons, and patch-clamp studies identify 55 layer-specific morphological and 207 morpho-electrical neuron subtypes. When digitally reconstructed neurons are positioned in the volume and synapse formation is restricted to biological bouton densities and numbers of synapses per connection, their overlapping arbors form ~8 million connections with ~37 million synapses. Simulations reproduce an array of in vitro and in vivo experiments without parameter tuning. Additionally, we find a spectrum of network states with a sharp transition from synchronous to asynchronous activity, modulated by physiological mechanisms. The spectrum of network states, dynamically reconfigured around this transition, supports diverse information processing strategies. PAPERCLIP: VIDEO ABSTRACT.

Charcot-Marie-Tooth: From Molecules to Therapy.

Charcot-Marie-Tooth (CMT) is the most prevalent category of inherited neuropathy. The most common inheritance pattern is autosomal dominant, though there also are X-linked and autosomal recessive subtypes. In addition to a variety of inheritance patterns, there are a myriad of genes associated with CMT, reflecting the heterogeneity of this disorder. Next generation sequencing (NGS) has expanded and simplified the diagnostic yield of genes/molecules underlying and/or associated with CMT, which is of paramount importance in providing a substrate for current and future targeted disease-modifying treatment options. Considerable research attention for disease-modifying therapy has been geared towards the most commonly encountered genetic mutations (PMP22, GJB1, MPZ, and MFN2). In this review, we highlight the clinical background, molecular understanding, and therapeutic investigations of these CMT subtypes, while also discussing therapeutic research pertinent to the remaining less common CMT subtypes.

Adsorption and oxidation of ciprofloxacin in a fixed bed column using activated sludge derived activated carbon.

In this study, the performance of activated sludge derived granular activated carbon (SGAC) was investigated for ciprofloxacin (CPX) removal from synthetic and simulated wastewaters in a fixed-bed adsorption column operated in continuous mode. The adsorbent was synthesized using chemical activation using ZnCl2 as activating agent. Its surface area and pore volume were found comparable to that of the commercial granular activated carbon (CGAC). The maximum saturation adsorption capacities for CPX were ~16 mg/g and ~14 mg/g, respectively, with SGAC column under identical operating conditions (CPX concentration = 50 mg/L, bed height = 4 cm and wastewater flow rate = 1.5 mL/min) for synthetic and simulated wastewaters. The presence of other organics reduced CPX adsorption capacity of SGAC. The breakthrough curve data for both wastewaters could be adequately fit in Thomas and Yoon-Nelson kinetic models. The addition of H2O2 in wastewater showed no considerable improvement in CPX removal. However, H2O2 oxidation of spent adsorbent exhibited better results compared to thermal treatment for adsorbent regeneration. The results showed that sewage sludge can be recycled as an efficient adsorbent for the removal of recalcitrant organic pollutants from wastewater.

Disruption of the histidine triad nucleotide-binding hint2 gene in mice affects glycemic control and mitochondrial function.

UNLABELLED: The histidine triad nucleotide-binding (HINT2) protein is a mitochondrial adenosine phosphoramidase expressed in the liver and pancreas. Its physiological function is unknown. To elucidate the role of HINT2 in liver physiology, the mouse Hint2 gene was deleted. Hint2(-/-) and Hint2(+/+) mice were generated in a mixed C57Bl6/J × 129Sv background. At 20 weeks, the phenotypic changes in Hint2(-/-) relative to Hint2(+/+) mice were an accumulation of hepatic triglycerides, decreased tolerance to glucose, a defective counter-regulatory response to insulin-provoked hypoglycemia, and an increase in plasma interprandial insulin but a decrease in glucose-stimulated insulin secretion and defective thermoregulation upon fasting. Leptin messenger RNA (mRNA) in adipose tissue and plasma leptin were elevated. In mitochondria from Hint2(-/-) hepatocytes, state 3 respiration was decreased, a finding confirmed in HepG2 cells where HINT2 mRNA was silenced. The linked complex II-III electron transfer was decreased in Hint2(-/-) mitochondria, which was accompanied by a lower content of coenzyme Q. Hypoxia-inducible factor-2α expression and the generation of reactive oxygen species were increased. Electron microscopy of mitochondria in Hint2(-/-) mice aged 12 months revealed clustered, fused organelles. The hepatic activities of 3-hydroxyacyl-coenzyme A dehydrogenase short chain and glutamate dehydrogenase (GDH) were decreased by 68% and 60%, respectively, without a change in protein expression. GDH activity was similarly decreased in HINT2-silenced HepG2 cells. When measured in the presence of purified sirtuin 3, latent GDH activity was recovered (126% in Hint2(-/-) versus 83% in Hint2(+/+) ). This suggests a greater extent of acetylation in Hint2(-/-) than in Hint2(+/+) . CONCLUSION: Hint2/HINT2 positively regulates mitochondrial lipid metabolism and respiration and glucose homeostasis. The absence of Hint2 provokes mitochondrial deformities and a change in the pattern of acetylation of selected proteins.

Marked reduction in paralytic attacks in a patient with Andersen-Tawil syndrome switched from acetazolamide to dichlorphenamide.

Andersen-Tawil syndrome is a rare, autosomal dominant, multisystem disorder for which the majority of cases are caused by pathogenic variants in the KCNJ2 gene. The syndrome is characterized by the clinical triad of episodic paralysis, cardiac conduction abnormalities, and dysmorphic facial and skeletal features. Treatment of Andersen-Tawil syndrome is primarily focused on management of cardiac arrhythmias and preventive management of paralytic attacks. Dichlorphenamide is approved by the US Food and Drug Administration for use in primary periodic paralysis based on several randomized, controlled trials but has not been studied in patients with Andersen-Tawil syndrome. Here, we report a case of the syndrome caused by a de novo pathogenic variant in the KCNJ2 gene (c.95_98del). The paralytic attack rate for this patient was better controlled with dichlorphenamide compared with acetazolamide, further supporting the use of dichlorphenamide in patients with Andersen-Tawil syndrome.

Magnetic properties of biogenic selenium nanomaterials.

Bioreduction of selenium oxyanions to elemental selenium is ubiquitous; elucidating the properties of this biogenic elemental selenium (BioSe) is thus important to understand its environmental fate. In this study, the magnetic properties of biogenic elemental selenium nanospheres (BioSe-Nanospheres) and nanorods (BioSe-Nanorods) obtained via the reduction of selenium(IV) using anaerobic granular sludge taken from an upflow anaerobic sludge blanket (UASB) reactor treating paper and pulp wastewater were investigated. The study indicated that the BioSe nanomaterials have a strong paramagnetic contribution with some ferromagnetic component due to the incorporation of Fe(III) (high-spin and low-spin species) as indicated by electron paramagnetic resonance (EPR). The paramagnetism did not saturate up to 50,000 Oe at 5 K, and the hysteresis curve showed the coercivity of 100 Oe and magnetic moment saturation around 10 emu. X-ray photoelectron spectroscopy (XPS) and EPR evidenced the presence of Fe(III) in the nanomaterial. Signals for Fe(II) were observed neither in EPR nor in XPS ruling out its presence in the BioSe nanoparticles. Fe(III) being abundantly present in the sludge likely got entrapped in the extracellular polymeric substances (EPS) coating the biogenic nanomaterials. The presence of Fe(III) in BioSe nanomaterial increases the mobility of Fe(III) and may have an effect on phytoplankton growth in the environment. Furthermore, as supported by the literature, there is a potential to exploit the magnetic properties of BioSe nanomaterials in drug delivery systems as well as in space refrigeration.

Degradation of ciprofloxacin using Fenton's oxidation: Effect of operating parameters, identification of oxidized by-products and toxicity assessment.

Ciprofloxacin (CPX), a fluoroquinolone antibiotic, is found in the bulk effluents emerged from pharmaceutical industries. Its presence in the effluent may lead to toxicity to the aquatic life and antimicrobial resistance. Hence, the oxidative degradation of CPX by classical Fenton's process was investigated for the present study. The maximum CPX and total organic carbon (TOC) removal from the synthetic wastewater (initial CPX concentration = 100 mg L-1) were 70% and 55%, respectively, under the best reaction conditions ([H2O2]:[Fe2+] = 10, stoichiometric H2O2 concentration = 14.2 mM and initial wastewater pH = 3.0). Five major degradation products could be identified by high resolution liquid chromatography-mass spectrometry (HR LC-MS) which could have resulted by the occurrence of decarboxylation, defluorination, hydroxylation and cleavage of the piperazine ring. The time based detection of ferrous and ferric ions in aqueous phase confirmed the CPX degradation by hydroxyl radicals. A pathway has been proposed for CPX degradation by Fenton's process based on the kinetic decay and/or evolution profiles of CPX and intermediates as well as the release of inorganic ions into the aqueous solution. It was indicated from the rapid respirometric test that the wastewater toxicity was lowered after Fenton's treatment.

Effects of Urea and TMAO on Lipid Self-Assembly under Osmotic Stress Conditions.

Most land-living organisms regularly experience dehydration. In nature, one commonly applied strategy to protect against this osmotic stress is to introduce small polar molecules with low vapor pressure, commonly called osmolytes. Two examples of naturally occurring small polar compounds are urea and trimethylamine N-oxide (TMAO), which are known to have counteracting effects on protein stability. In this work, we investigate the effects of urea and TMAO on lipid self-assembly at varying water contents, focusing on dehydrated conditions. By using complementary experimental techniques, including sorption microcalorimetry, NMR, and X-ray scattering, together with molecular dynamics simulations in model systems composed of phosphatidylcholine lipids, water, and solute, we characterize interactions and self-assembly over a large range of hydration conditions. It is shown that urea and TMAO show qualitatively similar effects on lipid self-assembly at high water contents, whereas they have clearly different effects in dehydrated conditions. The latter can be explained by differences in the molecular interactions between the solutes and the lipid headgroups. TMAO is repelled from the bilayer interface, and it is thereby expelled from lipid lamellar systems with low water contents and narrow inter-bilayer regions. In these conditions, TMAO shows no effect on the lipid phase behavior. Urea, on the other hand, shows a slight affinity for the lipid headgroup layer, and it is present in the lipid lamellar system at all water contents. As a result, urea may exchange with water in dry conditions and thereby prevent dehydration-induced phase transitions. In nature, urea and TMAO are sometimes found together in the same organisms and it is possible that their combined effect is to both protect lipid membranes against dehydration and still avoid denaturation of proteins.

Stereotypy in neocortical microcircuits.

A central debate regarding neocortical function concerns the degree to which the underlying microcircuitry is stereotypically organized. Stereotypy reflects invariance in structure and function, as a result of common genetic templates and environmental conditions, whereas uniqueness can be caused by genetic variations, differences in environmental conditions as well as random processes. Stereotypy is an appealing concept because it provides strong support for determinism in the formation of neuronal microcircuits and in the relationship between their specific structure and function.

Utilisation of sewage sludge derived adsorbents for the removal of recalcitrant compounds from wastewater: Mechanistic aspects, isotherms, kinetics and thermodynamics.

In the present study, the performance of sewage sludge based adsorbents was examined for the removal of two recalcitrant pollutants (i.e. lignin and amoxicillin) from synthetic wastewater solutions (adsorbate concentration=50-250 mg/l). The effect of various reaction parameters such as wastewater pH, adsorbent dosage and temperature was studied. Possible mechanisms for the adsorption process have been proposed which depends upon the behaviour of adsorbent surface and adsorbate molecules under specific reaction conditions. Three-parameter Redlich-Peterson isotherm model was found the best fit to the equilibrium data. Pseudo first and second order models validated the kinetic data for lignin and amoxicillin adsorption systems, respectively and the corresponding activation energy was 3.5-4.5 and 12-22 kJ/mol. The nature of adsorption was elucidated from the thermodynamic parameters.

Interneurons of the neocortical inhibitory system.

Mammals adapt to a rapidly changing world because of the sophisticated cognitive functions that are supported by the neocortex. The neocortex, which forms almost 80% of the human brain, seems to have arisen from repeated duplication of a stereotypical microcircuit template with subtle specializations for different brain regions and species. The quest to unravel the blueprint of this template started more than a century ago and has revealed an immensely intricate design. The largest obstacle is the daunting variety of inhibitory interneurons that are found in the circuit. This review focuses on the organizing principles that govern the diversity of inhibitory interneurons and their circuits.

Anatomical, physiological, molecular and circuit properties of nest basket cells in the developing somatosensory cortex.

Anatomical, electrophysiological and molecular diversity of basket cell-like interneurons in layers II-IV of rat somatosensory cortex were studied using patch-clamp electrodes filled with biocytin. This multiparametric study shows that neocortical basket cells (BCs) are composed of three distinct subclasses: classical large (LBC) and small (SBC) basket cells and a third subclass, the nest basket cell (NBC). Anatomically, NBCs were distinct from LBCs and SBCs in that they formed simpler dendritic arbors and an axonal plexus of inter-mediate density, composed of a few long, smooth axonal branches. Electrophysiologically, NBCs exhibited diverse discharge responses to depolarizing current injections including accommodation, non-accommodation and stuttering. Single-cell multiplex RT-PCR revealed distinct mRNA expression patterns for the calcium binding proteins parvalbumin (PV), calbindin (CB) and calretinin (CR), and the neuropeptides somatostatin (SOM), vasoactive intestinal peptide (VIP), cholecystokinin (CCK) and neuropeptide Y (NPY) for each BC-subclass. SBCs lacked NPY expression but invariably expressed VIP, whereas neither VIP, CR nor SOM expression was detected in LBCs, and VIP and CR expression was absent in NBCs. Electro-physiologically distinct types of NBCs formed GABAergic synapses with specific dynamics onto pyramidal cells (PCs) and received either strongly facilitating or depressing synaptic inputs from PCs. Finally, NBCs were found to be the most common basket cell in layers II/III, while LBCs were the most common in layer IV. These data provide multiparametric distinguishing features of three major subclasses of basket cells and indicate that NBCs are powerful interneurons that provide most of the (peri-)somatic inhibition in the supragranular layers.

Anatomical, physiological and molecular properties of Martinotti cells in the somatosensory cortex of the juvenile rat.

Whole-cell patch-clamp recordings followed by histochemical staining and single-cell RT-PCR were obtained from 180 Martinotti interneurones located in layers II to VI of the somatosensory cortex of Wistar rats (P13-P16) in order to examine their anatomical, electrophysiological and molecular properties. Martinotti cells (MCs) mostly displayed ovoid-shaped somata, bitufted dendritic morphologies, and axons with characteristic spiny boutons projecting to layer I and spreading horizontally across neighbouring columns more than 1 mm. Electron microscopic examination of MC boutons revealed that all synapses were symmetrical and most synapses (71%) were formed onto dendritic shafts. MCs were found to contact tuft, apical and basal dendrites in multiple neocortical layers: layer II/III MCs targeted mostly layer I and to a lesser degree layer II/III; layer IV MCs targeted mostly layer IV and to a lesser degree layer I; layer V and VI MCs targeted mostly layer IV and layer I and to a lesser degree the layer in which their somata was located. MCs typically displayed spike train accommodation (90%; n = 127) in response to depolarizing somatic current injections, but some displayed non-accommodating (8%) and a few displayed irregular spiking responses (2%). Some accommodating and irregular spiking MCs also responded initially with bursts (17%). Accommodating responses were found in all layers, non-accommodating mostly in upper layers and bursting mostly in layer V. Single-cell multiplex RT-PCR performed on 63 MCs located throughout layers II-VI, revealed that all MCs were somatostatin (SOM) positive, and negative for parvalbumin (PV) as well as vasoactive intestinal peptide (VIP). Calbindin (CB), calretinin (CR), neuropeptide Y (NPY) and cholecystokinin (CCK) were co- expressed with SOM in some MCs. Some layer-specific trends seem to exist. Finally, 24 accommodating MCs were examined for the expression of 26 ion channel genes. The ion channels with the highest expression in these MCs were (from highest to lowest); Cabeta1, Kv3.3, HCN4, Cabeta4, Kv3.2, Kv3.1, Kv2.1, HCN3, Caalpha1G, Kv3.4, Kv4.2, Kv1.1 and HCN2. In summary, this study provides the first detailed analysis of the anatomical, electrophysiological and molecular properties of Martinotti cells located in different neocortical layers. It is proposed that MCs are crucial interneurones for feedback inhibition in and between neocortical layers and columns.