Brain state-dependent responses of midbrain dopaminergic neurons to footshock under urethane anaesthesia.

For a long time, it has been assumed that dopaminergic (DA) neurons in both the ventral tegmental area (VTA) and the substantia nigra pars compacta (SNc) uniformly respond to rewarding and aversive stimuli by either increasing or decreasing their activity, respectively. This response was believed to signal information about the perceived stimuli's values. The identification of VTA&SNc DA neurons that are excited by both rewarding and aversive stimuli has led to the categorisation of VTA&SNc DA neurons into two subpopulations: one signalling the value and the other signalling the salience of the stimuli. It has been shown that the general state of the brain can modulate the electrical activity of VTA&SNc DA neurons, but it remains unknown whether this factor may also influence responses to aversive stimuli, such as a footshock (FS). To address this question, we have recorded the responses of VTA&SNc DA neurons to FSs across cortical activation and slow wave activity brain states in urethane-anaesthetised rats. Adding to the knowledge of aversion signalling by midbrain DA neurons, we report that significant proportion of VTA&SNc DA neurons can change their responses to an aversive stimulus in a brain state-dependent manner. The majority of these neurons decreased their activity in response to FS during cortical activation but switched to increasing it during slow wave activity. It can be hypothesised that this subpopulation of DA neurons may be involved in the 'dual signalling' of both the value and the salience of the stimuli, depending on the general state of the brain.

When material science meets microbial ecology: Bacterial community selection on stainless steels in natural seawater.

The passive film depends on the alloy's composition and the exposure conditions. How the surface composition affects the selection of microbial biofilms though, has not been fully elucidated or incorporated into the analysis of corrosive biofilms. The degradation of stainless steel (SS) exposed to natural seawater was studied to understand how the oxide layer composition of SS could affect the selection and variability of the bacterial community. To accomplish this goal, austenitic and superferritic SS grades were exposed to natural seawater on the central coast of Chile. The deterioration of steel and qualitative description of biofilm formation was monitored at different exposure periods. Biofilms were evaluated based on massive sequencing analysis of the bacterial community and subsequent ecological studies. The results revealed that variability of the calculated corrosion rate correlated with the similarity of the bacterial community within samples from each SS and its corrosion inferred capacity. The associated bacterial families showed a higher representation in SSs with a more significant increase in the Fe/Cr ratio over the exposure time. These findings revealed that iron content in the oxide layer represents a key feature of the surface composition for selecting bacterial assemblages in marine environments.

Hybrid magnetron sputtering of ceramic superlattices for application in a next generation of combustion engines.

A hybrid magnetron sputtering process (dcMS/HiPIMS) was developed to manufacture nanostructured CrN/Cr1-xAlxN multilayers, motivated by improving the low-emission efficiency when applied on gas-nitrided diesel piston rings of a next-generation of combustion engines. In order to improve the mechanical, tribological, and corrosion behavior of the multilayers, the hybrid dcMS/HiPIMS process was designed by selecting the optimal sputtering procedure applied to AISI 440 base steel. The effect of substrate bias and carousel rotational speed on the phase composition, crystallographic texture, residual stresses, surface roughness, coating periodicity and densification, instrumented hardness, elastic modulus, as well as wear and corrosion resistance was determined. The results have demonstrated that hybrid magnetron sputtering produces multilayers with a superlattice structure, which outperforms commercial PVD coatings of CrN for diesel piston rings manufactured by cathodic arc evaporation. Also, multilayer periodicities in the range of 5 to 10 nm yield the best tribological performance under bench tests for the piston ring/cylinder liner system.

Bidirectional Communication between the Pontine Nucleus Incertus and the Medial Septum Is Carried Out by Electrophysiologically-Distinct Neuronal Populations.

Theta oscillations are key brain rhythm involved in memory formation, sensorimotor integration, and control of locomotion and behavioral states. Generation and spatiotemporal synchronization of theta oscillations rely on interactions between brain nuclei forming a large neural network, which includes pontine nucleus incertus (NI). Here we identified distinct populations of NI neurons, based on the relationship of their firing to hippocampal waves, with a special focus on theta oscillations, and the direction and type of interaction with the medial septum (MS) in male, urethane-anesthetized rats. By recording NI neuronal firing and hippocampal LFP, we described NI neurons that fire action potentials in a theta phase-independent or theta phase-locked and delta wave-independent or delta wave-locked manner. Among hippocampal activity-independent NI neurons, irregular, slow-firing, and regular, fast-firing cells were observed, while hippocampal oscillation-/wave-locked NI neurons were of a bursting or nonbursting type. By projection-specific optotagging, we revealed that only fast-firing theta phase-independent NI neurons innervate the MS, rarely receiving feedback information. In contrast, the majority of theta-bursting NI neurons were inhibited by MS stimulation, and this effect was mediated by direct GABAergic input. Described NI neuronal populations differ in reciprocal connections with the septohippocampal system, plausibly forming separate neuronal loops. Our results suggest that theta phase-independent NI neurons participate in theta rhythm generation through direct innervation of the MS, while theta-bursting NI neurons further transmit the rhythmic signal received from the MS to stabilize and/or strengthen rhythmic activity in other structures.SIGNIFICANCE STATEMENT The generation and spatiotemporal synchronization of theta oscillations rely on interactions between nuclei forming a large neural network, part of which is the pontine nucleus incertus (NI). Here we describe that within NI there are populations of neurons that can be distinguished based on the relationship of their firing to hippocampal theta oscillations and delta waves. We show that these neuronal populations largely do not have reciprocal connections with the septohippocampal system, but form separate neuronal loops. Our results suggest that medial septum (MS)-projecting, fast-firing, theta phase-independent NI neurons may participate in theta rhythm generation through direct innervation of the MS, while theta-bursting NI neurons may further transmit the rhythmic signal received from the MS to other structures.

Testing the Test: A Comparative Study of Marine Microbial Corrosion under Laboratory and Field Conditions.

Microbially influenced corrosion (MIC) is an aggressive type of corrosion that occurs in aquatic environments and is sparked by the development of a complex biological matrix over a metal surface. In marine environments, MIC is exacerbated by the frequent variability in environmental conditions and the typically high diversity of microbial communities; hence, local and in situ studies are crucial to improve our understanding of biofilm composition, biological interactions among its members, MIC characteristics, and corrosivity. Typically, material performance and anticorrosion strategies are evaluated under controlled laboratory conditions, where natural fluctuations and gradients (e.g., light, temperature, and microbial composition) are not effectively replicated. To determine whether MIC development and material deterioration observed in the laboratory are comparable to those that occur under service conditions (i.e., field conditions), we used two testing setups, in the lab and in the field. Stainless steel (SS) AISI 316L coupons were exposed to southeastern Pacific seawater for 70 days using (i) acrylic tanks in a running seawater laboratory and (ii) an offshore mooring system with experimental frames immersed at two depths (5 and 15 m). Results of electrochemical evaluation, together with those of microbial community analyses and micrographs of formed biofilms, demonstrated that the laboratory setup provides critical information on the early biofilm development process (days), but the information gathered does not predict deterioration or biofouling of SS surfaces exposed to natural conditions in the field. Our results highlight the need to conduct further research efforts to understand how laboratory experiments may better reproduce field conditions where applications are to be deployed, as well as to improve our understanding of the role of eukaryotes and the flux of nutrients and oxygen in marine MIC events.

Initial adhesion suppression of biofilm-forming and copper-tolerant bacterium Variovorax sp. on laser microtextured copper surfaces.

The growth of detrimental biofilms on metal surfaces affects their structural performance and lifespan. Microtopographic texturization has emerged as an approach to suppress biofilm growth by preventing the initial stages of bacterial adhesion. This work studies the effects of linear pattern copper texturization on the initial adhesion steps of the biofilm-forming and copper-resistant bacterium Variovorax sp. Linear patterns with 4.7, 6.8, 14, and 18 µm periodicity were produced by direct laser interference patterning (DLIP) on copper coupons. Surface features were characterized by microscopic and spectroscopic techniques, and bacterial adhesion behavior was characterized by epifluorescence microscopy and functionalization of atomic force microscopy tips. We found a periodicity of 4.7 µm as the most efficient pattern to suppress Variovorax sp. initial adhesion by 31.1 % with respect to the nontextured surface. Preferential settlement in hummocks over hollows was observed for patterns with 14 and 18 µm periodicity, with adhesion events showing higher frequency in these topographies than patterns with periodicities of 4.7 and 6.8 µm. Our results highlight an understanding of the initial bacteria-copper adhesion and settlement behavior, thus contributing to the potential development of innocuous strategies for controlling biofilm growth on copper-based materials.

Nitrogen-Enriched Cr1-xAlxN Multilayer-Like Coatings Manufactured by Dynamic Glancing Angle Direct Current Magnetron Sputtering.

Multilayer-like CrN and Cr1-xAlxN coatings with different Al contents were deposited onto a stainless steel substrate using dynamic glancing angle deposition direct current magnetron sputtering (DGLAD dcMS) in a N rich atmosphere to understand the role of Al on the growth of the films and mechanical properties of the nitrides with a multilayer architecture. Chemical analysis by means of energy dispersive analysis (EDS) and glow discharge optical emission spectroscopy (GDOES) depth profiling revealed that while CrN samples were close to stoichiometric, the Cr1-xAlxN coatings presented excess N between 70 and 80% at. An expressive change in texture was observed as the CrN coating changed its preferred orientation from (111) to (200) with the addition of Al, followed by a modification in morphology from grains with faceted pyramidal tops in CrN to dome-shaped grains in Cr1-xAlxN coatings. Multilayer-like nanostructures of corrugated grains were produced with a periodicity of approximately 30 nm using dynamic glancing angle deposition. The deposition rate was drastically reduced with an increase of Al, meanwhile, the best mechanical performance was achieved for the coating with a higher content of Al, with hardness up to 27 GPa and a higher value of maximum resistance to plastic deformation.

Tribological Performance of Porous Ti-Nb-Ta-Fe-Mn Alloy in Dry Condition.

The tribological properties of a novel porous Ti-Nb-Ta-Fe-Mn alloy with 0%, 30%, and 60% porosity were evaluated for biomedical applications. The tribotesting was performed using a ball-on-disc under dry conditions, using an alumina ball and 1 N of a load. The coefficient of friction at the early stage of the porous samples was lower than that of the bulk, 0.2 and 0.7, respectively, but the samples with 30% porosity shift toward the bulk value after a variable number of cycles, while the samples with 60% remained stable after 100,000 cycles. The wear rate of the specimen with 60% porosity was twice as low as that of the bulk. The results are explained by shift in wear mechanism associated with the modified bearing ratio of the porous surface and by the accumulation of wear debris inside the pores, which prevented the development of three-body abrasion.

Electrochemical Bacterial Enrichment from Natural Seawater and Its Implications in Biocorrosion of Stainless-Steel Electrodes.

Microbial electrochemical technologies have revealed the opportunity of electrochemical enrichment for specific bacterial groups that are able to catalyze reactions of interest. However, there are unsolved challenges towards their application under aggressive environmental conditions, such as in the sea. This study demonstrates the impact of surface electrochemical potential on community composition and its corrosivity. Electrochemical bacterial enrichment was successfully carried out in natural seawater without nutrient amendments. Experiments were carried out for ten days of exposure in a closed-flow system over 316L stainless steel electrodes under three different poised potentials (-150 mV, +100 mV, and +310 mV vs. Ag/AgCl). Weight loss and atomic force microscopy showed a significant difference in corrosion when +310 mV (vs. Ag/AgCl) was applied in comparison to that produced under the other tested potentials (and an unpoised control). Bacterial community analysis conducted using 16S rRNA gene profiles showed that poised potentials are more positive as +310 mV (vs. Ag/AgCl) resulted in strong enrichment for Rhodobacteraceae and Sulfitobacter. Hence, even though significant enrichment of the known electrochemically active bacteria from the Rhodobacteraceae family was accomplished, the resultant bacterial community could accelerate pitting corrosion in 316 L stainless steel, thereby compromising the durability of the electrodes and the microbial electrochemical technologies.

Effect of added porosity on a novel porous Ti-Nb-Ta-Fe-Mn alloy exposed to simulated body fluid.

Porous titanium materials have gained interest as prosthesis materials due to their similar mechanical properties to the human bone, biocompatibility, and high corrosion resistance. The presence of pores in the metal matrix implies a decrease in the elastic modulus and an increase in the active area, perhaps improving the osseointegration. Corrosion resistance is a critical consideration as corrosion may lead not only to mechanical failure but also the release of ions and/or particles to the bloodstream. In this work, a novel Ti-Nb-Ta-Fe-Mn alloy with varying percentage of porosity (25, 31 and 37 v/v%) was exposed to simulated body fluid (SBF) at 37 °C and its corrosion resistance was investigated using electrochemical techniques and surface analysis as a function of exposure time. Open circuit potential and polarization curves revealed that the effect of porosity was mainly on the shift of the corrosion potential to more negative values with a slight increase in the anodic current. A passive range was also observed, which was not influenced either by increased exposure time or increased porosity. Therefore, a change in the surface specific area could have taken place during the exposure, which is not necessarily related to a corrosion process. Moreover, a typical porous electrode behavior was identified by electrochemical Impedance spectroscopy, without any significant change over time. No release of metal ions was detected by on line ICP-AES, either at the open circuit potential or upon polarizing the samples up to 2 V vs. SCE, whereas only traces elements (Fe and Mn 1 nmol/s cm2) were detected in the electrolyte accumulating all released ions during 30 days of exposure. Additionally, the surface analysis showed thickening of the oxide layer with exposure time. Therefore, the stability of the passive layer and low release of ions indicate that the porous alloys are suitable for further study as prosthesis materials.

On manufacturing multilayer-like nanostructures using misorientation gradients in PVD films.

Due to their applicability for manufacturing dense, hard and stable coatings, Physical Vapor Deposition (PVD) techniques, such as High Power Impulse Magnetron Sputtering (HiPIMS), are currently used to deposit transition metal nitrides for tribological applications. Cr-Al-N is one of the most promising ceramic coating systems owing to its remarkable mechanical and tribological properties along with excellent corrosion resistance and high-temperature stability. This work explores the possibility of further improving Cr-Al-N coatings by modulation of its microstructure. Multilayer-like Cr1-xAlxN single films were manufactured using the angular oscillation of the substrate surface during HiPIMS. The sputtering process was accomplished using pulse frequencies ranging from 200 to 500 Hz and the resulting films were evaluated with respect to their hardness, Young's modulus, residual stresses, deposition rate, crystallite size, crystallographic texture, coating morphology, chemical composition, and surface roughness. The multilayer-like structure, with periodicities ranging from 250 to 550 nm, were found associated with misorientation gradients and small-angle grain boundaries along the columnar grains, rather than mesoscopic chemical modulation of the microstructure. This minute modification of microstructure along with associated compressive residual stresses are concluded to explain the increased hardness ranging from 25 to 30 GPa, which is at least 20% over that expected for a film of the same chemical composition grown by a conventional PVD processing route.

Laser-Assisted Synthesis of Cu-Al-Ni Shape Memory Alloys: Effect of Inert Gas Pressure and Ni Content.

The paper explores the applicability of laser-assisted synthesis for producing high density Cu-Al-Ni alloys with shape memory characteristics, that could be further developed towards a method of additive manufacturing of large size Cu-based shape memory alloys (SMA). The manufacturing approach consists in laser melting of elemental powder mixture in a controlled atmosphere of varying relative pressure of protective argon gas, producing alloys of 14.2 wt.% Al and Ni content varying between 2 and 4 wt.%. All the fabricated alloys are found to have attained martensitic microstructures capable of SMA specific phase transformations in the temperature range from 85 to 192 °C. Both gas pressure and content of Ni are found to affect the specific transformation temperatures, transformation enthalpies, and mechanical properties. In particular, increasing gas pressure suppresses the austenite to martensite transformation reducing microhardness. In conclusion, the selective laser melting (SLM) employed in this work is shown capable of producing high density Cu-Al-Ni SMA (porosity ≈ 2%).

Several behavioral traits relevant for alcoholism are controlled by ɣ2 subunit containing GABAA receptors on dopamine neurons in mice.

The risk factors for developing alcohol addiction include impulsivity, high sensitivity to the rewarding action of ethanol, and low sensitivity to its sedative and intoxicating effects. Genetic variation in GABAA receptor subunits, including the ɣ2 subunit (Gabrg2), affects the risk for developing alcoholism. Alcohol directly potentiates GABAA receptors and activates the mesolimbic dopamine system. Here, we deleted Gabrg2 selectively in dopamine cells of adult mice. The deletion resulted in elevated firing of dopamine neurons and made them less sensitive to drugs acting at GABAA receptors. At the behavioral level, the deletion increased exploratory behavior and augmented both correct and incorrect responding in the go/no-go task, a test often used to assay the response inhibition component of impulsivity. In addition, conditioned place preference to alcohol, but not to cocaine or morphine, was increased. Ethanol-induced locomotor activation was enhanced in the mice lacking Gabrg2 on dopaminergic cells, whereas the sedative effect of alcohol was reduced. Finally, the alcohol drinking, but not the alcohol preference, at a high concentration was increased in the mutant mice. In summary, deletion of Gabrg2 on dopamine cells induced several behavioral traits associated with high risk of developing alcoholism. The findings suggest that mice lacking Gabrg2 on dopaminergic cells could be used as models for individuals at high risk for developing alcoholism and that GABAA receptors on dopamine cells are protective against the development of excessive alcohol drinking.

Midbrain dopaminergic neuron activity across alternating brain states of urethane anaesthetized rat.

Midbrain dopaminergic neurons are implicated in the control of motor functions and reward-driven behaviours. The function of this neuronal population is strongly connected with distinct patterns of firing - irregular or bursting, which either maintains basal levels of dopamine (DA) or leads to phasic release, respectively. Heterogeneity of dopaminergic neurons, observed on both structural and functional levels, is also reflected in different responses of DA neurons to changes in global brain states. Preparation of urethane anaesthetized animal is a broadly used model to study brain state dependent activity of neurons. Unfortunately activity of midbrain DA neurons across urethane induced cyclic, spontaneous brain state alternations is poorly described. To fulfil this gap in our knowledge we have performed simultaneous, extracellular recordings of the firing of single putative DA neurons combined with continuous brain state monitoring. We found that during slow wave activity, the firing rate of recorded putative DA neurons was significantly higher compared to firing rates during activated state, both in ventral tegmental area (VTA) and substantia nigra pars compacta (SNc). In the presence of cortical slow waves, putative dopaminergic neurons also intensified bursting activity, but the magnitude of this phenomena differed in respect to the examined region (VTA or SNc). Our results show that activity of DA neurons under urethane anaesthesia is brain-state dependent and emphasize the importance of brain state monitoring during electrophysiological experiments.

Loss of NMDA receptors in dopamine neurons leads to the development of affective disorder-like symptoms in mice.

The role of changes in dopamine neuronal activity during the development of symptoms in affective disorders remains controversial. Here, we show that inactivation of NMDA receptors on dopaminergic neurons in adult mice led to the development of affective disorder-like symptoms. The loss of NMDA receptors altered activity and caused complete NMDA-insensitivity in dopamine-like neurons. Mutant mice exhibited increased immobility in the forced swim test and a decrease in social interactions. Mutation also led to reduced saccharin intake, however the preference of sweet taste was not significantly decreased. Additionally, we found that while mutant mice were slower to learn instrumental tasks, they were able to reach the same performance levels, had normal sensitivity to feedback and showed similar motivation to exert effort as control animals. Taken together these results show that inducing the loss of NMDA receptor-dependent activity in dopamine neurons is associated with development of affective disorder-like symptoms.

Uterus banding with the Osada method effectively reduces intraoperative blood loss during myomectomy.

INTRODUCTION: The surgery of fibroid removal and reconstruction of the uterus is associated with increased blood loss. That is a significant limitation of surgical myomectomy. There are many methods to decrease blood loss during myomectomy. However, in women planning to conceive their reversibility is important. The procedure of uterus banding with the Osada method meets this condition. The objective of this study was a comparison of intraoperative blood loss during the myomectomy with banding according to the Osada technique with blood loss during a classic myomectomy with the Martin method. MATERIAL AND METHODS: The study group consisted of 140 women with myomatous uterus. In 70 patients myomectomy was performed with the Osada uterus banding method, for the remaining 70 patients the Martin method was applied. RESULTS: Myomectomy with banding according to the Osada method versus myomectomy with the Martin method: intraoperative blood loss (ml): 56 ± 23 vs 378 ± 186, p < 0.05; a drop in hematocrit levels over 24 hours postoperatively (%): 0.32 ± 0.12 vs 3.42 ± 2.54, p < 0.05; a drop in hemoglobin concentration over 24 hours postoperatively (g/dl): 0.13 ± 0.04 vs 0.79 ± 0.38, p < 0.05. the need for blood transfusion (% of women): 0 vs 4.28, p < 0.05. CONCLUSION: Myomectomy performed according to the Osada method of uterus banding is associated with less intraoperative blood loss.

Local mechanical response of cells to the controlled rotation of magnetic nanorods.

The mechanical response of the cytoplasm was investigated by the intracellular implantation of magnetic nanorods and exposure to low-frequency rotatory magnetic fields. Nanorods (Pt-Ni, ∼200 nm diameter) fabricated by electrodeposition in templates of porous alumina with lengths of approximately 2 and 5 µm were inserted into NIH/3T3 fibroblasts and manipulated with a rotational magnetic field. Nanorod rotation was observed only for torques greater than 3.0 × 10(-16) Nm, suggesting a Bingham-type behavior of the cytoplasm. Higher torques produced considerable deformation of the intracellular material. The cell nucleus and cell membrane were significantly deformed by nanorods actuated by 4.5 × 10(-15) Nm torques. Our results demonstrate that nanorods under magnetic fields are an effective tool to mechanically probe the intracellular environment. We envision that our findings may contribute to the noninvasive and direct mechanical characterization of the cytoplasm.

Multi-technique approach to assess the effects of microbial biofilms involved in copper plumbing corrosion.

Microbially influenced corrosion (MIC) is recognized as an unusual and severe type of corrosion that causes costly failures around the world. A microbial biofilm could enhance the copper release from copper plumbing into the water by forming a reactive interface. The biofilm increases the corrosion rate, the mobility of labile copper from its matrix and the detachment of particles enriched with copper under variable shear stress due to flow conditions. MIC is currently considered as a series of interdependent processes occurring at the metal-liquid interface. The presence of a biofilm results in the following effects: (a) the formation of localized microenvironments with distinct pH, dissolved oxygen concentrations, and redox conditions; (b) sorption and desorption of labile copper bonded to organic compounds under changing water chemistry conditions; (c) change in morphology by deposition of solid corrosion by-products; (d) diffusive transport of reactive chemical species from or towards the metal surface; and (e) detachment of scale particles under flow conditions. Using a multi-technique approach that combines pipe and coupon experiments this paper reviews the effects of microbial biofilms on the corrosion of copper plumbing systems, and proposes an integrated conceptual model for this phenomenon supported by new experimental data.