Synchronization of Isolated Downstates (K-Complexes) May Be Caused by Cortically-Induced Disruption of Thalamic Spindling

Sleep spindles and K-complexes (KCs) define stage 2 NREM sleep (N2) in humans. We recently showed that KCs are isolated downstates characterized by widespread cortical silence. We demonstrate here that KCs can be quasi-synchronous across scalp EEG and across much of the cortex using electrocorticography (ECOG) and localized transcortical recordings (bipolar SEEG). We examine the mechanism of synchronous KC production by creating the first conductance based thalamocortical network model of N2 sleep to generate both spontaneous spindles and KCs. Spontaneous KCs are only observed when the model includes diffuse projections from restricted prefrontal areas to the thalamic reticular nucleus (RE), consistent with recent anatomical findings in rhesus monkeys. Modeled KCs begin with a spontaneous focal depolarization of the prefrontal neurons, followed by depolarization of the RE. Surprisingly, the RE depolarization leads to decreased firing due to disrupted spindling, which in turn is due to depolarization-induced inactivation of the low-threshold Ca²⁺ current (I_T). Further, although the RE inhibits thalamocortical (TC) neurons, decreased RE firing causes decreased TC cell firing, again because of disrupted spindling. The resulting abrupt removal of excitatory input to cortical pyramidal neurons then leads to the downstate. Empirically, KCs may also be evoked by sensory stimuli while maintaining sleep. We reproduce this phenomenon in the model by depolarization of either the RE or the widely-projecting prefrontal neurons. Again, disruption of thalamic spindling plays a key role. Higher levels of RE stimulation also cause downstates, but by directly inhibiting the TC neurons. SEEG recordings from the thalamus and cortex in a single patient demonstrated the model prediction that thalamic spindling significantly decreases before KC onset. In conclusion, we show empirically that KCs can be widespread quasi-synchronous cortical downstates, and demonstrate with the first model of stage 2 NREM sleep a possible mechanism whereby this widespread synchrony may arise.     

Molecular analysis of the SCARECROW gene in maize reveals a common basis for radial patterning in diverse meristems

Maize and Arabidopsis root apical meristems differ in several aspects of their radial organization and ontogeny. Despite the large evolutionary distance and differences in root radial patterning, analysis of the putative maize ortholog of the Arabidopsis patterning gene SCARECROW (SCR) revealed expression localized to the endodermis, which is similar to its expression in Arabidopsis. Expression in maize extends through the quiescent center, a population of mitotically inactive cells formerly thought to be undifferentiated and to lack radial pattern information. Zea mays SCARECROW (ZmSCR), the putative maize SCR ortholog, was used as a molecular marker to investigate radial patterning during regeneration of the root tip after either whole or partial excision. Analysis of the dynamic expression pattern of ZmSCR as well as other markers indicates the involvement of positional information as a primary determinant in regeneration of the root radial pattern.     

Comparative LCA of multi-product processes with non-common products: a systematic approach applied to chlorine electrolysis technologies

Purpose

Multi-product processes are one source of multi-functionality causing widely discussed methodological problems within life cycle assessment. A multi-functionality problem exists for comparative life cycle assessment (LCA) of multi-product processes with non-common products. This work develops a systematic workflow for fixing the multi-functionality problem caused by the non-common products. A novel technology for chlor-alkali electrolysis is analyzed and compared to the industrial standard technology to illustrate the approach and to benchmark the new technology's environmental impact.

Methods

A matrix-based workflow for comparative LCA of multi-product systems is presented. Products are distinguished in main products and by-products based on the reason of process operation. We argue that only main products form the reference flows of the compared multi-product systems. Fixing the multi-functionality problem follows directly from the chosen reference flows. The framework suggests system expansion to fix the multi-functionality problem if non-common main products exist. Non-common by-products still cause a multi-functionality problem. These by-products are systematically identified and the multi-functionality problem is fixed with avoided burden and allocation. A case study applies the workflow for comparing environmental impacts of the standard chlorine electrolysis to a novel process using oxygen-depolarized cathodes. Three scenarios are derived and evaluated. The assessed impact categories are cumulative energy demand, global warming potential, acidification potential, photochemical ozone creation potential, eutrophication potential, and human toxicity potential.

Results and discussion

The proposed workflow minimizes the methodological choices. The multi-functionality problem is systematically fixed based on the distinction between the main products and by-products. Inconsistent solutions are prevented by rigorous identification of unequal by-products within the compared systems. Selecting avoided burden processes or allocation factors is the remaining ambiguous choice common to the standard methods. The case study demonstrates the applicability of the workflow to comparative LCA of multi-product systems. The case study results show lower environmental impacts for the novel electrolysis technology in all practically relevant scenarios and impact categories.

Conclusions

The framework for comparative LCA of multi-product systems with non-common products adds systematic clarity to the general ISO standards. The approach reduces the subjective choices of LCA practitioners to the identification of reason of process operation. This reason is defined if the site-specific economic conditions are known. The matrix-based formulation allows identification of inconsistencies caused by multi-functionality. For the novel electrolysis technology, the results indicate significant potential for environmental impact reduction.     

Secretome analysis of Magnaporthe oryzae using in vitro systems

Magnaporthe oryzae is a devastating blast fungal pathogen of rice (Oryza sativa L.) that causes dramatic decreases in seed yield and quality. During the early stages of infection by this pathogen, the fungal spore senses the rice leaf surface, germinates, and penetrates the cell via an infectious structure known as an appressorium. During this process, M. oryzae secretes several proteins; however, these proteins are largely unknown mainly due to the lack of a suitable method for isolating secreted proteins during germination and appressoria formation. We examined the secretome of M. oryzae by mimicking the early stages of infection in vitro using a glass plate (GP), PVDF membrane, and liquid culture medium (LCM). Microscopic observation of M. oryzae growth revealed appressorium formation on the GP and PVDF membrane resembling natural M. oryzae-rice interactions; however, appresorium formation was not observed in the LCM. Secreted proteins were collected from the GP (3, 8, and 24 h), PVDF membrane (24 h), and LCM (48 h) and identified by two-dimensional gel electrophoresis (2DE) followed by tandem mass spectrometry. The GP, PVDF membrane, and LCM-derived 2D gels showed distinct protein patterns, indicating that they are complementary approaches. Collectively, 53 nonredundant proteins including previously known and novel secreted proteins were identified. Six biological functions were assigned to the proteins, with the predominant functional classes being cell wall modification, reactive oxygen species detoxification, lipid modification, metabolism, and protein modification. The in vitro system using GPs and PVDF membranes applied in this study to survey the M. oryzae secretome, can be used to further our understanding of the early interactions between M. oryzae and rice leaves.     

Selective Gαi subunits as novel direct activators of transient receptor potential canonical (TRPC)4 and TRPC5 channels

The ubiquitous transient receptor potential canonical (TRPC) channels function as non-selective, Ca(2+)-permeable channels and mediate numerous cellular functions. It is commonly assumed that TRPC channels are activated by stimulation of Gα(q)-PLC-coupled receptors. However, whether the Gα(q)-PLC pathway is the main regulator of TRPC4/5 channels and how other Gα proteins may regulate these channels are poorly understood. We previously reported that TRPC4/TRPC5 can be activated by Gα(i). In the current work, we found that Gα(i) subunits, rather than Gα(q), are the primary and direct activators of TRPC4 and TRPC5. We report a novel molecular mechanism in which TRPC4 is activated by several Gα(i) subunits, most prominently by Gα(i2), and TRPC5 is activated primarily by Gα(i3). Activation of Gα(i) by the muscarinic M2 receptors or expression of the constitutively active Gα(i) mutants equally and fully activates the channels. Moreover, both TRPC4 and TRPC5 are activated by direct interaction of their conserved C-terminal SESTD (SEC14-like and spectrin-type domains) with the Gα(i) subunits. Two amino acids (lysine 715 and arginine 716) of the TRPC4 C terminus were identified by structural modeling as mediating the interaction with Gα(i2). These findings indicate an essential role of Gα(i) proteins as novel activators for TRPC4/5 and reveal the molecular mechanism by which G-proteins activate the channels.     

Kidney proteome responses in the teleost fish Paralichthys olivaceus indicate a putative immune response against Streptococcus parauberis

The proteomic response to bacterial infection in a teleost fish (Paralichthys olivaceus) infected with Streptococcus parauberis was analyzed using label-free protein quantitation coupled with LC-MS(E) tandem mass spectrometry. A total of 82 proteins from whole kidney, a major lymphoid organ in this fish, were found to be differentially expressed between healthy and diseased fish analyzed 6, 24, 72 and 120 h post-infection. Among the differentially expressed proteins, those involved in mediating immune responses (e.g., heat shock proteins, cathepsins, goose-type lysozyme and complement components) were most significantly up-regulated by infection. In addition, cell division cycle 48 (CDC48) and calreticulin, which are associated with cellular recovery and glycoprotein synthesis, were up-regulated in the universal protein group, whereas the other proteins in that group were down-regulated. There was continuous activation of expression of immune-associated proteins during infection, but there was also loss of expression of proteins not involved in immune function. We expect that our findings regarding immune response at the protein level would offer new insight into the systemic response to bacterial infection of a major immune organ in teleost fish.     

Cigarette Smoking Increases Abdominal and Visceral Obesity but Not Overall Fatness: An Observational Study

Background

Cigarette smoking and obesity are leading public health concerns. Both increase the risk for cardiovascular disease, cancer, and metabolic abnormalities. This study was conducted to assess the association between cigarette smoking and different types of obesity.

Methodology/Principal Findings

Two hundred eighty-three visitors to university hospitals located in four main provinces of South Korea were participated. All participants were classified as either current/past or never smokers and were divided into quartiles according to the total pack-years. Body mass index, waist circumference, total body fat percentage, and area of visceral and abdominal subcutaneous fat were measured. These results of each groups were compared. Waist circumference, and visceral fat area showed a J- or U-shaped association with total smoking amount during a lifetime. After restricting the analyses to past/current smokers, we found significant dose-dependent associations of smoking pack-years with abdominal and visceral obesity. Overall obesity measured by body mass index and total body fat percentage did not show such associations. Although current smokers clearly showed significant associations, we could not demonstrate these in past smokers, possibly because of the limited sample size.

Conclusions/Significance

Although smokers did not show significant difference in mean body mass index than those who never smoked, they showed more metabolically adverse fat distributions with increasing smoking amounts. This finding suggests that smoking is not beneficial for weight control. Therefore, smoking cessation and avoidance of smoking commencement should be addressed as important public health issues in preventing obesity and related complications.     

Remediation of Gulf War Oil Spill Contaminated Soil by a Subcritical Water Extraction Process: Oil Removal,Recovery, and Degradation

Due to the unique properties of subcritical water (marked change in water's dielectric constant and viscosity), the extraction by subcritical water offers a great opportunity to remediate soil contaminated with organic pollutants as an alternative and green remediation method. In this study, subcritical water extraction is proposed as an efficient remediation technique for the Gulf War oil spill contaminated soil. The subcritical water extraction experiment was carried out in a lab-scale continuous flow apparatus. The three major operating factors, temperature, time and water flow rate, were evaluated in terms of optimum removal efficiency. The results show that crude oil removal depended largely on water temperature, whereas an extraction run time higher than 1 h and a water flow rate higher than 1.5 mL/min marginally or negatively affected removal efficiency. During subcritical water treatment at 300°C for 1 h at a flow rate of 1.5 mL/min, removal efficiency was almost 95%. Under these operating conditions, the subcritical water treatment demonstrated a similar removal efficiency to those of organic solvents like acetone. In contrast, the efficiency of oil recovery decreased with an increase in extraction temperature, due to degradation by a water self-oxidizing agent. Several degradation products identified in the treated soil and in the effluent sample (which initially were absent in the contaminated soil) were oxygen-containing aromatic compounds, confirming the oxidation-degradation.