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A major objective of the Mars 2020 mission is to sample rocks in Jezero crater that may preserve organic matter for later return to Earth. Using an ultraviolet Raman and luminescence spectrometer, the Perseverance rover detected luminescence signals with maximal intensities at 330 to 350 nanometers and 270 to 290 nanometers that were initially reported as consistent with organics. Here, we test the alternative hypothesis that the 330- to 350-nanometer and 270- to 290-nanometer luminescence signals trace Ce 3+ in phosphate and silicate defects, respectively. By comparing the distributions of luminescence signals with the rover detections of x-ray fluorescence from P 2 O 5 and Si-bearing materials, we show that, while an organic origin is not excluded, the observed luminescence can be explained by purely inorganic materials. These findings highlight the importance of eventual laboratory analyses to detect and characterize organic compounds in the returned samples.

Purpose: To investigate the impact of 3D-printed temporal bone models with two different material transparencies on trainees' mastoidectomy performance., Methods: Eleven ORL residents performed two anatomical mastoidectomies with posterior tympanotomy on two 3D-printed models with different transparency and VR simulation training. Participants where divided into two groups based on their experience. Within each group participants were randomized to start with the model printed in a completely opaque material or in a material featuring some degree of transparency. After drilling on 3D-printed models, the participants performed two similar mastoidectomies on human cadavers: one on the left side of one cadaver and one on the right side of another cadaver. After drilling 3D-printed models and cadavers, the final-product performances were evaluated by two experienced raters using the 26-item modified Welling Scale. Participants also evaluated the models using a questionnaire., Results: Overall, the participants performed 25 % better on the 3D-printed models featuring transparency compared to the opaque models (18.6 points vs 14.9 points, mean difference = 3.7, 95 % CI 2.0-5.3, P < 0.001)). This difference in performance was independent of which material the participants had drilled first. In addition, the residents also subjectively rated the transparent model to be closer to cadaver dissection. The experienced group starting with the 3D-printed models scored 21.5 points (95 % CI 20.0-23.1), while the group starting with VR simulation training score 18.4 points (95 % CI 16.6-20.3)., Conclusion: We propose that material used for 3D-printing temporal bone models should feature some degree of transparency, like natural bone, for trainees to learn and exploit key visual cues during drilling., Competing Interests: Declaration of competing interest None., (Copyright © 2024. Published by Elsevier B.V.)

Hydrophobic surfaces require finely tuned process chains due to the scale, complexity, and patterning methods. For this purpose, vat photopolymerization (VPP) additive manufacturing is a promising method for surface generation; however, together with the fabrication process, the design phase needs to be optimized to achieve the desired surface property. This work presents the influence of the design features of hydrophobic surfaces through multiple studies on simple pillar structures, intrinsic single-unit geometries, and surface deposition on complex substrates. The results showed that depending on the dimensions of single pillar dimensions, wetting properties can extend between the contact angles (CA) of 83°-115.11°. The hydrophobicity was further increased by applying a re-entrant structure, reaching the CA of 115.24°. The surface deposition on the complex substrates significantly increased water droplet adhesion, preventing it from rolling off, which can be beneficial for manifold device protection from the hazardous influence of the environment. In addition, the influence of the surface on the acoustic properties was examined, which showed that the pattern application in the real-life device does not have a detrimental effect on the intrinsic functionality. This study showed that the design phase should be an essential part of the VPP process chain as it significantly influences the wetting properties of the surfaces., (Copyright 2023, Mary Ann Liebert, Inc., publishers.)

Objective: 3-D printing offers convenient and low-cost mastoidectomy training; nonetheless, training benefits using 3-D-printed temporal bones remain largely unexplored. In this study, we have collected validity evidence for a low-cost, 3-D-printed temporal bone for mastoidectomy training and established a credible pass/fail score for performance on the model., Study Design: A prospective educational study gathering validity evidence using Messick's validity framework., Setting: Seven Danish otorhinolaryngology training institutions., Participants: Eighteen otorhinolaryngology residents (novices) and 11 experienced otosurgeons (experts)., Intervention: Residents and experienced otosurgeons each performed two to three anatomical mastoidectomies on a low-cost, 3-D-printed temporal bone model produced in-house. After drilling, mastoidectomy performances were rated by three blinded experts using a 25-item modified Welling scale (WS)., Main Outcome Measure: Validity evidence using Messick's framework including reliability assessment applying both classical test theory and Generalizability theory., Results: Novices achieved a mean score of 13.9 points; experienced otosurgeons achieved 23.2 points. Using the contrasting groups method, we established a 21/25-point pass/fail level. The Generalizability coefficient was 0.91, and 75% of the score variance was attributable to participant performance, indicating a high level of assessment reliability. Subsequent D studies revealed that two raters rating one performance or one rater rating two performances were sufficiently reliable for high-stakes assessment., Conclusion: Validity evidence supports using a low-cost, 3-D-printed model for mastoidectomy training. The model can be printed in-house using consumer-grade 3-D printers and serves as an additional training tool in the temporal bone curriculum. For competency-based training, we established a cut-off score of 21 of 25 WS points using the contrasting groups method., Competing Interests: The authors disclose no conflicts of interest., (Copyright © 2023, Otology & Neurotology, Inc.)

Selective laser sintering (SLS) is a well-established technology that is used for additive manufacturing. Significant efforts have been made to improve SLS by optimizing the powder deposition, laser beam parameters, and temperature settings. The purpose is to ensure homogeneous sintering and prevent geometric and appearance inaccuracies in the manufactured objects. We evaluated the differences in the surface roughness and grain size of curved objects manufactured by using upcoming SLS technology that features two CO laser sources. Our analysis was carried out on polyamide 11 (PA11), which is a sustainable biobased polymer that has been gaining popularity due to its high-performance properties: its low melting point, high viscosity, and excellent mechanical properties. By using a Taguchi experimental design and analysis of variance (ANOVA), we examined the influence on the surface roughness and grain size of the build setup, the presence of thin walls, and the position of the sample on the powder bed. We found significant differences in some surface roughness and grain size measurements when these parameters were changed.

Disruption of adipocyte de novo lipogenesis (DNL) by deletion of fatty acid synthase (FASN) in mice induces browning in inguinal white adipose tissue (iWAT). However, adipocyte FASN knockout (KO) increases acetyl-coenzyme A (CoA) and malonyl-CoA in addition to depletion of palmitate. We explore which of these metabolite changes triggers adipose browning by generating eight adipose-selective KO mouse models with loss of ATP-citrate lyase (ACLY), acetyl-CoA carboxylase 1 (ACC1), ACC2, malonyl-CoA decarboxylase (MCD) or FASN, or dual KOs ACLY/FASN, ACC1/FASN, and ACC2/FASN. Preventing elevation of acetyl-CoA and malonyl-CoA by depletion of adipocyte ACLY or ACC1 in combination with FASN KO does not block the browning of iWAT. Conversely, elevating malonyl-CoA levels in MCD KO mice does not induce browning. Strikingly, adipose ACC1 KO induces a strong iWAT thermogenic response similar to FASN KO while also blocking malonyl-CoA and palmitate synthesis. Thus, ACC1 and FASN are strong suppressors of adipocyte thermogenesis through promoting lipid synthesis rather than modulating the DNL intermediates acetyl-CoA or malonyl-CoA., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: 3D-printed temporal bone models can potentially provide a cost-effective alternative to cadaver surgery that can be manufactured locally at the training department. The objective of this study was to create a cost-effective 3D-printed model suitable for mastoidectomy training using entry level and commercially available print technologies, enabling individuals, without prior experience on 3D-printing, to manufacture their own models for basic temporal bone training., Methods: Expert technical professionals and an experienced otosurgeon identified the best material for replicating the temporal bone and created a cost-effective printing routine for the model using entry-level print technologies. Eleven participants at a temporal bone dissection course evaluated the model using a questionnaire., Results: The 3D-printed temporal bone model was printed using a material extrusion 3D-printer with a heat resistant filament, reducing melting during drilling. After printing, a few simple post-processing steps were designed to replicate the dura, sigmoid sinus and facial nerve. Modifying the 3D-printer by installing a direct-drive and ruby nozzle resulted in more successful prints and less need for maintenance. Upon evaluation by otorhinolaryngology trainees, unanimous feedback was that the model provided a good introduction to the mastoidectomy procedure, and supplementing practice to cadaveric temporal bones., Conclusion: In-house production of a cost-effective 3D-printed model for temporal bone training is feasible and enables training institutions to manufacture their own models. Further, this work demonstrates the feasibility of creating new temporal bone models with anatomical variation to provide ample training opportunity., (© 2023. The Author(s).)

Collocated crystal sizes and mineral identities are critical for interpreting textural relationships in rocks and testing geological hypotheses, but it has been previously impossible to unambiguously constrain these properties using in situ instruments on Mars rovers. Here, we demonstrate that diffracted and fluoresced x-rays detected by the PIXL instrument (an x-ray fluorescence microscope on the Perseverance rover) provide information about the presence or absence of coherent crystalline domains in various minerals. X-ray analysis and multispectral imaging of rocks from the Séítah formation on the floor of Jezero crater shows that they were emplaced as coarsely crystalline igneous phases. Olivine grains were then partially dissolved and filled by finely crystalline or amorphous secondary silicate, carbonate, sulfate, and chloride/oxychlorine minerals. These results support the hypothesis that Séítah formation rocks represent olivine cumulates altered by fluids far from chemical equilibrium at low water-rock ratios.

Contaminant concentrations in filter-feeding shellfish may indicate the health of coastal waters and consumption risks. Widespread expansion of the Pacific oyster (Crassostrea gigas) and its popularity as food make it a useful sentinel. We surveyed intertidal Pacific oysters in San Diego Bay, California for contaminants during summer 2018 and winter 2019. We compared contaminants in Pacific oyster to California mussel from California's State Mussel Watch Program (1993-2003) and human consumption thresholds. Contaminants such as neonicotinoid and chlorinated pesticides, selenium, and several metals were higher in Pacific oysters in summer, while PBDEs, benzylbutyl phthalate, and plastics were higher in winter. Contaminant levels were generally lower in Pacific oyster than mussel except for copper and zinc. Bay-wide PCB concentrations in oysters exceeded thresholds but individual samples (locations) also met or surpassed chlordane, PCB and PAH thresholds. Monitoring and risk assessments that consider species' biology, season, location, effects of multiple contaminants, and human consumption patterns will contribute to more effective consumption guidelines., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objective: Mastoidectomy is a cornerstone in the surgical management of middle and inner ear diseases. Unfortunately, training is challenged by insufficient access to human cadavers. Three-dimensional (3D) printing of temporal bones could alleviate this problem, but evidence on their educational effectiveness is lacking. It is largely unknown whether training on 3D-printed temporal bones improves mastoidectomy performance, including on cadavers, and how this training compares with virtual reality (VR) simulation. To address this knowledge gap, this study investigated whether training on 3D-printed temporal bones improves cadaveric dissection performance, and it compared this training with the already-established VR simulation., Study Design: Prospective cohort study of an educational intervention., Setting: Tertiary university hospital, cadaver dissection laboratory, and simulation center in Copenhagen, Denmark., Methods: Eighteen otorhinolaryngology residents (intervention) attending the national temporal bone dissection course received 3 hours of mastoidectomy training on 3D-printed temporal bones. Posttraining cadaver mastoidectomy performances were rated by 3 experts using a validated assessment tool and compared with those of 66 previous course participants (control) who had received time-equivalent VR training prior to dissection., Results: The intervention cohort outperformed the controls during cadaver dissection by 29% ( P < .001); their performances were largely similar across training modalities but remained at a modest level (~50% of the maximum score)., Conclusion: Mastoidectomy skills improved from training on 3D-printed temporal bone and seemingly more so than on time-equivalent VR simulation. Importantly, these skills transferred to cadaveric dissection. Training on 3D-printed temporal bones can effectively supplement cadaver training when learning mastoidectomy., (© The Authors 2021.)

Objective: 3D-printed models hold great potential for temporal bone surgical training as a supplement to cadaveric dissection. Nevertheless, critical knowledge on manufacturing remains scattered, and little is known about whether use of these models improves surgical performance. This systematic review aims to explore (1) methods used for manufacturing and (2) how educational evidence supports using 3D-printed temporal bone models., Data Sources: PubMed, Embase, the Cochrane Library, and Web of Science., Review Methods: Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, relevant studies were identified and data on manufacturing and validation and/or training extracted by 2 reviewers. Quality assessment was performed using the Medical Education Research Study Quality Instrument tool; educational outcomes were determined according to Kirkpatrick's model., Results: The search yielded 595 studies; 36 studies were found eligible and included for analysis. The described 3D-printed models were based on computed tomography scans from patients or cadavers. Processing included manual segmentation of key structures such as the facial nerve; postprocessing, for example, consisted of removal of print material inside the model. Overall, educational quality was low, and most studies evaluated their models using only expert and/or trainee opinion (ie, Kirkpatrick level 1). Most studies reported positive attitudes toward the models and their potential for training., Conclusion: Manufacturing and use of 3D-printed temporal bones for surgical training are widely reported in the literature. However, evidence to support their use and knowledge about both manufacturing and the effects on subsequent surgical performance are currently lacking. Therefore, stronger educational evidence and manufacturing knowhow are needed for widespread implementation of 3D-printed temporal bones in surgical curricula.

The manufacturing of inserts for micro injection moulding made of mortar material is presented in this work. The fabrication of the mortar insert described in this publication relied on a versatile and relatively fast rapid prototyping process based on soft tooling. The mortar insert has a QR code with micro features on its surface, which was replicated in acrylonitrile butadiene styrene (ABS) polymer by the micro injection moulding process. With this approach, it is possible to fabricate hard inserts for micro injection moulding purposes that are able to compete with conventional-made inserts made of tool steel.

Metal-supported oxide cells (MSCs) are considered as the third-generation solid oxide cells (SOCs) succeeding electrolyte-supported (first generation) and anode-supported (second generation) cells, which have gained much attention and progress in the past decade. The use of metal supports and advanced technical methods (such as infiltrated electrodes) has vastly improved cell performance, especially with its rapid startup ability and power density, showing a significant decrease in raw materials cost. However, new degradation mechanisms appeared, limiting the further improvement of the performance and lifetime. This review encapsulates the degradation mechanisms and countermeasures in the field of MSCs, reviewing the challenges and recommendations for future development.

Introduction: Circulating immature precursor cells indicate malignant diseases like acute myeloid or lymphoid leukemia, and blast cells are key finds for disease management. Automatized cell counters are an essential contemporary appliance for blast detection, but false-positive samples remain challenging in terms of time and resources. To reduce this issue, the White Precursor Channel (WPC) was introduced to Sysmex XN series; however, sensitivity may reduce when accommodating low specificity. Therefore, our aim was to evaluate WPC blast alarm flag performance with regard to detecting blast cells., Methods: At two major Danish hospitals, random blood samples were collected from the routine setting   in a four-week period and analyzed on WPC XN20 (Sysmex, Japan). Results were compared with manual differential white blood cell count (Manual WBCC) assisted by CellaVisionDM96., Results: In 117 samples, we found 0.2 to 34.4% blasts, WPC blast flag specificity = 82% and a low sensitivity = 40%. However, other XN alarm flags forwarded samples to Manual WBCC, so blast cells were detected despite missing a specific blast flag: With all alarm flags, combined sensitivity increased to 88%. Overall, the WPC application stopped 18% of the 117 samples going to Manual WBCC (three false negatives). Q values are arbitrary probability measurements for the blast flag, and in five samples (0.5 to 47.3% blasts) imprecision ranged from 5.3 to 122 CV%., Conclusions: WPC blast alarm flags are imprecise and inaccurate, especially when blast counts are low. However, the XN20 will alarm samples with other flags so that most samples containing blast cells will be manually reviewed after all. Hence, the presented flag types should not bias the decisions of manual reviewers., (© 2020 John Wiley & Sons Ltd.)

Advanced microbiology technologies are rapidly changing our ability to diagnose infections, improve patient care, and enhance clinical workflow. These tools are increasing the breadth, depth, and speed of diagnostic data generated per patient, and testing is being moved closer to the patient through rapid diagnostic technologies, including point-of-care (POC) technologies. While select stakeholders have an appreciation of the value/importance of improvements in the microbial diagnostic field, there remains a disconnect between clinicians and some payers and hospital administrators in terms of understanding the potential clinical utility of these novel technologies. Therefore, a key challenge for the clinical microbiology community is to clearly articulate the value proposition of these technologies to encourage payers to cover and hospitals to adopt advanced microbiology tests. Specific guidance on how to define and demonstrate clinical utility would be valuable. Addressing this challenge will require alignment on this topic, not just by microbiologists but also by primary care and emergency room (ER) physicians, infectious disease specialists, pharmacists, hospital administrators, and government entities with an interest in public health. In this article, we discuss how to best conduct clinical studies to demonstrate and communicate clinical utility to payers and to set reasonable expectations for what diagnostic manufacturers should be required to demonstrate to support reimbursement from commercial payers and utilization by hospital systems., (Copyright © 2019 Miller et al.)

Objective: Crosstalk between adipocytes and local neurons may be an important regulatory mechanism to control energy homeostasis. We previously reported that perturbation of adipocyte de novo lipogenesis (DNL) by deletion of fatty acid synthase (FASN) expands sympathetic neurons within white adipose tissue (WAT) and stimulates the appearance of "beige" adipocytes. Here we tested whether WAT DNL activity can also influence neuronal regulation and thermogenesis in brown adipose tissue (BAT)., Methods and Results: Induced deletion of FASN in all adipocytes in mature mice (iAdFASNKO) enhanced sympathetic innervation and neuronal activity as well as UCP1 expression in both WAT and BAT. This increased sympathetic innervation could be observed at both 22 °C and 30 °C, indicating it is not a response to heat loss but rather adipocyte signaling. In contrast, selective ablation of FASN in brown adipocytes of mice (iUCP1FASNKO) failed to modulate sympathetic innervation and the thermogenic program in BAT. Surprisingly, DNL in brown adipocytes was also dispensable in maintaining euthermia when UCP1FASNKO mice were cold-exposed., Conclusion: These results indicate that DNL in white adipocytes influences long distance signaling to BAT, which can modify BAT sympathetic innervation and expression of genes involved in thermogenesis., (Copyright © 2018 The Authors. Published by Elsevier GmbH.. All rights reserved.)

The purpose of this paper is to present the method of a soft tooling process chain employing Additive Manufacturing (AM) for fabrication of injection molding inserts with micro surface features. The Soft Tooling inserts are manufactured by Digital Light Processing (vat photo polymerization) using a photopolymer that can withstand relatively high temperaturea. The part manufactured here has four tines with an angle of 60°. Micro pillars (Ø200 µm, aspect ratio of 1) are arranged on the surfaces by two rows. Polyethylene (PE) injection molding with the soft tooling inserts is used to fabricate the final parts. This method demonstrates that it is feasible to obtain injection-molded parts with microstructures on complex geometry by additive manufactured inserts. The machining time and cost is reduced significantly compared to conventional tooling processes based on computer numerical control (CNC) machining. The dimensions of the micro features are influenced by the applied additive manufacturing process. The lifetime of the inserts determines that this process is more suitable for pilot production. The precision of the inserts production is limited by the additive manufacturing process as well.

Background: The de novo biosynthesis of fatty acids (DNL) through fatty acid synthase (FASN) in adipocytes is exquisitely regulated by nutrients, hormones, fasting, and obesity in mice and humans. However, the functions of DNL in adipocyte biology and in the regulation of systemic glucose homeostasis are not fully understood., Methods & Results: Here we show adipocyte DNL controls crosstalk to localized sympathetic neurons that mediate expansion of beige/brite adipocytes within inguinal white adipose tissue (iWAT). Induced deletion of FASN in white and brown adipocytes of mature mice (iAdFASNKO mice) enhanced glucose tolerance, UCP1 expression, and cAMP signaling in iWAT. Consistent with induction of adipose sympathetic nerve activity, iAdFASNKO mice displayed markedly increased neuronal tyrosine hydroxylase (TH) and neuropeptide Y (NPY) content in iWAT. In contrast, brown adipose tissue (BAT) of iAdFASNKO mice showed no increase in TH or NPY, nor did FASN deletion selectively in brown adipocytes (UCP1-FASNKO mice) cause these effects in iWAT., Conclusions: These results demonstrate that downregulation of fatty acid synthesis via FASN depletion in white adipocytes of mature mice can stimulate neuronal signaling to control thermogenic programming in iWAT.

Obesity promotes insulin resistance associated with liver inflammation, elevated glucose production, and type 2 diabetes. Although insulin resistance is attenuated in genetic mouse models that suppress systemic inflammation, it is not clear whether local resident macrophages in liver, denoted Kupffer cells (KCs), directly contribute to this syndrome. We addressed this question by selectively silencing the expression of the master regulator of inflammation, NF-κB, in KCs in obese mice. We used glucan-encapsulated small interfering RNA particles (GeRPs) that selectively silence gene expression in macrophages in vivo. Following intravenous injections, GeRPs containing siRNA against p65 of the NF-κB complex caused loss of NF-κB p65 expression in KCs without disrupting NF-κB in hepatocytes or macrophages in other tissues. Silencing of NF-κB expression in KCs in obese mice decreased cytokine secretion and improved insulin sensitivity and glucose tolerance without affecting hepatic lipid accumulation. Importantly, GeRPs had no detectable toxic effect. Thus, KCs are key contributors to hepatic insulin resistance in obesity and a potential therapeutic target for metabolic disease., (© FASEB.)

The liver is a major site of glucose, fatty acid, and triglyceride (TG) synthesis and serves as a major regulator of whole body nutrient homeostasis. Chronic exposure of humans or rodents to high-calorie diets promotes non-alcoholic fatty liver disease, characterized by neutral lipid accumulation in lipid droplets (LD) of hepatocytes. Here we show that the LD protein hypoxia-inducible gene 2 (Hig2/Hilpda) functions to enhance lipid accumulation in hepatocytes by attenuating TG hydrolysis. Hig2 expression increased in livers of mice on a high-fat diet and during fasting, two states associated with enhanced hepatic TG content. Hig2 expressed in primary mouse hepatocytes localized to LDs and promoted LD TG deposition in the presence of oleate. Conversely, tamoxifen-inducible Hig2 deletion reduced both TG content and LD size in primary hepatocytes from mice harboring floxed alleles of Hig2 and a cre/ERT2 transgene controlled by the ubiquitin C promoter. Hepatic TG was also decreased by liver-specific deletion of Hig2 in mice with floxed Hig2 expressing cre controlled by the albumin promoter. Importantly, we demonstrate that Hig2-deficient hepatocytes exhibit increased TG lipolysis, TG turnover, and fatty acid oxidation as compared with controls. Interestingly, mice with liver-specific Hig2 deletion also display improved glucose tolerance. Taken together, these data indicate that Hig2 plays a major role in promoting lipid sequestration within LDs in mouse hepatocytes through a mechanism that impairs TG degradation., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)

Objective: Adipose tissue (AT) inflammation is associated with systemic insulin resistance and hyperinsulinemia in obese rodents and humans. A longstanding concept is that hyperinsulinemia may promote systemic insulin resistance through downregulation of its receptor on target tissues. Here we tested the novel hypothesis that insulin also impairs systemic insulin sensitivity by specifically enhancing adipose inflammation., Methods: Circulating insulin levels were reduced by about 50% in diet-induced and genetically obese mice by treatments with diazoxide or streptozotocin, respectively. We then examined AT crown-like structures, macrophage markers and pro-inflammatory cytokine expression in AT. AT lipogenesis and systemic insulin sensitivity was also monitored. Conversely, insulin was infused into lean mice to determine its affects on the above parameters., Results: Lowering circulating insulin levels in obese mice by streptozotocin treatment decreased macrophage content in AT, enhancing insulin stimulated Akt phosphorylation and de novo lipogenesis (DNL). Moreover, responsiveness of blood glucose levels to injected insulin was improved by streptozotocin and diazoxide treatments of obese mice without changes in body weight. Remarkably, even in lean mice, infusion of insulin under constant euglycemic conditions stimulated expression of cytokines in AT. Consistent with these findings, insulin treatment of 3T3-L1 adipocytes caused a 10-fold increase in CCL2 mRNA levels within 6 h, which was blocked by the ERK inhibitor PD98059., Conclusion: Taken together, these results indicate that obesity-associated hyperinsulinemia unexpectedly drives AT inflammation in obese mice, which in turn contributes to factors that suppress insulin-stimulated adipocyte DNL and systemic insulin sensitivity.

We have previously shown that deletion of protein kinase C epsilon (PKCε) in mice results in protection against glucose intolerance caused by a high fat diet. This was in part due to reduced insulin uptake by hepatocytes and insulin clearance, which enhanced insulin availability. Here we employed mouse embryonic fibroblasts (MEFs) derived from wildtype (WT) and PKCε-deficient (PKCε(-/-)) mice to examine this mechanistically. PKCε(-/-) MEFs exhibited reduced insulin uptake which was associated with decreased insulin receptor phosphorylation, while downstream signalling through IRS-1 and Akt was unaffected. Cellular fractionation demonstrated that PKCε deletion changed the localization of the insulin receptor, a greater proportion of which co-fractionated with flotillin-1, a marker of membrane microdomains. Insulin stimulation resulted in redistribution of the receptor in WT cells, while this was markedly reduced in PKCε(-/-) cells. These alterations in insulin receptor trafficking were associated with reduced expression of CEACAM1, a receptor substrate previously shown to modulate insulin clearance. Virally-mediated reconstitution of PKCε in MEFs increased CEACAM1 expression and partly restored the sensitivity of the receptor to insulin-stimulated redistribution. These data indicate that PKCε can affect insulin uptake in MEFs through promotion of receptor-mediated endocytosis, and that this may be mediated by regulation of CEACAM1 expression.

Electrochemical cycling of silver surfaces in the presence of the organophosphonates paraoxon and malathion leads to changes in the electrochemical response of silver and the formation of silver nanostructures. Adsorption of the organophosphonates onto the silver surfaces causes a significant reduction in the observed current response due to an increase in the charge transfer resistance. Surface enhanced Raman spectroscopy (SERS) measurements indicate that paraoxon adsorbs with no structural changes, while malathion decomposes and a thiophosphonate interacts with the surface. The SERS study of these adsorbates was carried out by changing the electrochemical conditions and the concentration of the organophosphonates. The size of the nanostructures greatly influences the SERS signal and it is observed that the strongest enhancement is observed for mid-sized nanostructures with a uniform thickness on the surface. The limit of detection was shown to be in the range of 10 nM to 10 pM for paraoxon and malathion, respectively.

A simple electrochemical method has been proposed for the preparation of silver nanoparticles that can be used for the detection of cyanide. Both the electrochemical behavior and morphology of the Ag nanoparticles have been characterized in the presence of KCN or diethyl cyanophosphonate (DECP) as well as in alkaline media. These were investigated by cyclic voltammetry and scanning electron microscopy (SEM). DECP is a simulant of the chemical warfare agent tabun., (© 2011 American Chemical Society)

The electrochemistry of gas-generated naked Ag nanoparticles deposited on indium-tin oxide covered glass plates is compared to bulk polycrystalline Ag. The nano-specific electrochemistry has been identified and includes the preferential formation of beta-oxides. In 100 mM KOH supporting electrolyte, disruption of beta-oxide formation is exploited to test for the presence of diethyl cyanophosphonate. While in 8.0 M KOH, beta-oxide formation is enhanced leading to testing capabilities for 2-chloroethyl ethyl sulfide. These non-redox active compounds are effective simulants for the chemical warfare agents Tabun nerve agent and sulfur mustard, respectively.

Fiber Bragg Gratings (FBGs) are known to be sensitive acoustic transducers and have previously been used for the photoacoustic detection of small solid samples. Here, we demonstrate the use of an FBG as an on-line detector for liquid chromatography. The FBG was inserted into a silica capillary and the photoacoustic response from the effluent was generated by a 10 ns pulsed laser. The acoustic pulse was quantified by the FBG through a characteristic change in the reflection spectrum. Good repeatability and linear response were obtained over three orders of magnitude (R(2) > 0.99), and the limit of detection of Coumarin 440 was determined to be 5 microM. The technique was successfully coupled to high performance liquid chromatography and applied to on-line analysis of a three-compound solution. Photoacoustic detection in liquid chromatography using FBGs is a label-free method, which can be applied to the detection of any chromogenic compound irrespective of its fluorogenic properties. It is a simple, inexpensive, and inherently micron-sized technique, insensitive to electromagnetic interference.

We determined the effect of coingestion of caffeine (Caff) with carbohydrate (CHO) on rates of muscle glycogen resynthesis during recovery from exhaustive exercise in seven trained subjects who completed two experimental trials in a randomized, double-blind crossover design. The evening before an experiment subjects performed intermittent exhaustive cycling and then consumed a low-CHO meal. The next morning subjects rode until volitional fatigue. On completion of this ride subjects consumed either CHO [4 g/kg body mass (BM)] or the same amount of CHO + Caff (8 mg/kg BM) during 4 h of passive recovery. Muscle biopsies and blood samples were taken at regular intervals throughout recovery. Muscle glycogen levels were similar at exhaustion [ approximately 75 mmol/kg dry wt (dw)] and increased by a similar amount ( approximately 80%) after 1 h of recovery (133 +/- 37.8 vs. 149 +/- 48 mmol/kg dw for CHO and Caff, respectively). After 4 h of recovery Caff resulted in higher glycogen accumulation (313 +/- 69 vs. 234 +/- 50 mmol/kg dw, P < 0.001). Accordingly, the overall rate of resynthesis for the 4-h recovery period was 66% higher in Caff compared with CHO (57.7 +/- 18.5 vs. 38.0 +/- 7.7 mmol x kg dw(-1) x h(-1), P < 0.05). After 1 h of recovery plasma Caff levels had increased to 31 +/- 11 microM (P < 0.001) and at the end of the recovery reached 77 +/- 11 microM (P < 0.001) with Caff. Phosphorylation of CaMK(Thr286) was similar after exercise and after 1 h of recovery, but after 4 h CaMK(Thr286) phosphorylation was higher in Caff than CHO (P < 0.05). Phosphorylation of AMP-activated protein kinase (AMPK)(Thr172) and Akt(Ser473) was similar for both treatments at all time points. We provide the first evidence that in trained subjects coingestion of large amounts of Caff (8 mg/kg BM) with CHO has an additive effect on rates of postexercise muscle glycogen accumulation compared with consumption of CHO alone.

In type 2 diabetes, pancreatic beta cells fail to secrete sufficient insulin to overcome peripheral insulin resistance. Intracellular lipid accumulation contributes to beta cell failure through poorly defined mechanisms. Here we report a role for the lipid-regulated protein kinase C isoform PKCepsilon in beta cell dysfunction. Deletion of PKCepsilon augmented insulin secretion and prevented glucose intolerance in fat-fed mice. Importantly, a PKCepsilon-inhibitory peptide improved insulin availability and glucose tolerance in db/db mice with preexisting diabetes. Functional ablation of PKCepsilon selectively enhanced insulin release ex vivo from diabetic or lipid-pretreated islets and optimized the glucose-regulated lipid partitioning that amplifies the secretory response. Independently, PKCepsilon deletion also augmented insulin availability by reducing both whole-body insulin clearance and insulin uptake by hepatocytes. Our findings implicate PKCepsilon in the etiology of beta cell dysfunction and highlight that enhancement of insulin availability, through separate effects on liver and beta cells, provides a rationale for inhibiting PKCepsilon to treat type 2 diabetes.

To determine whether preexercise muscle glycogen content influences the transcription of several early-response genes involved in the regulation of muscle growth, seven male strength-trained subjects performed one-legged cycling exercise to exhaustion to lower muscle glycogen levels (Low) in one leg compared with the leg with normal muscle glycogen (Norm) and then the following day completed a unilateral bout of resistance training (RT). Muscle biopsies from both legs were taken at rest, immediately after RT, and after 3 h of recovery. Resting glycogen content was higher in the control leg (Norm leg) than in the Low leg (435 +/- 87 vs. 193 +/- 29 mmol/kg dry wt; P < 0.01). RT decreased glycogen content in both legs (P < 0.05), but postexercise values remained significantly higher in the Norm than the Low leg (312 +/- 129 vs. 102 +/- 34 mmol/kg dry wt; P < 0.01). GLUT4 (3-fold; P < 0.01) and glycogenin mRNA abundance (2.5-fold; not significant) were elevated at rest in the Norm leg, but such differences were abolished after exercise. Preexercise mRNA abundance of atrogenes was also higher in the Norm compared with the Low leg [atrogin: approximately 14-fold, P < 0.01; RING (really interesting novel gene) finger: approximately 3-fold, P < 0.05] but decreased for atrogin in Norm following RT (P < 0.05). There were no differences in the mRNA abundance of myogenic regulatory factors and IGF-I in the Norm compared with the Low leg. Our results demonstrate that 1) low muscle glycogen content has variable effects on the basal transcription of select metabolic and myogenic genes at rest, and 2) any differences in basal transcription are completely abolished after a single bout of heavy resistance training. We conclude that commencing resistance exercise with low muscle glycogen does not enhance the activity of genes implicated in promoting hypertrophy.

We report on the size and charge dependence of the C-O stretching frequency, nu(CO), in complexes of CO with gas phase anionic, neutral, and cationic cobalt clusters (Co(n)CO(-0+)), anionic, neutral, and cationic rhodium clusters (Rh(n)CO(-0+)), and cationic nickel clusters (Ni(n)CO(+)) for n up to 37. We develop models, based on the established vibrational spectroscopy of organometallic carbonyl compounds, to understand how cluster size and charge relate to nu(CO) in these complexes. The dominating factor is the available electron density for backdonation from the metal to the CO pi* orbital. Electrostatic effects play a significant but minor role. For the charged clusters, the size trends are related to the dilution of the charge density at the binding site on the cluster as n increases. At large n, nu(CO) approaches asymptotes that are not the same as found for nu(CO) on the single crystal metal surfaces, reflecting differences between binding sites on medium sized clusters and the more highly coordinated metal surface sites.

The adsorption of chloroethyl ethyl sulfide and 2,2'-dichloro ethyl sulfide to gold nanoparticles has been examined in the gas phase using surface-plasmon resonance spectroscopy. The equilibrium constants for the gas-phase adsorption reactions are 14 +/- 4 and 25 +/- 5 atm(-1), respectively. The bond energy of the Au-chloroethyl ethyl sulfide complex is 44 +/- 9 kJ mol(-1). The bond energy of Au-2,2'-dichloro ethyl sulfide is estimated to be 47 kJ mol(-1). Halogenation of the dialkyl side chains is found to have little impact on the chemistry, and only physisorption processes occur, analogous to dialkyl sulfide adsorption on Au surfaces.

We report on the interaction of carbon monoxide with cationic gold clusters in the gas phase. Successive adsorption of CO molecules on the Au(n)(+) clusters proceeds until a cluster size specific saturation coverage is reached. Structural information for the bare gold clusters is obtained by comparing the saturation stoichiometry with the number of available equivalent sites presented by candidate structures of Au(n)(+). Our findings are in agreement with the planar structures of the Au(n)(+) cluster cations with n < or = 7 that are suggested by ion mobility experiments [Gilb, S.; Weis, P.; Furche, F.; Ahlrichs, R.; Kappes, M. M. J. Chem. Phys. 2001, 116, 4094]. By inference we also establish the structure of the saturated Au(n)(CO)(m)(+) complexes. In certain cases we find evidence suggesting that successive adsorption of CO can distort the metal cluster framework. In addition, the vibrational spectra of the Au(n)(CO)(m)(+) complexes in both the CO stretching region and in the region of the Au-C stretch and the Au-C-O bend are measured using infrared photodepletion spectroscopy. The spectra further aid in the structure determination of Au(n)(+), provide information on the structure of the Au(n)(+)-CO complexes, and can be compared with spectra of CO adsorbates on deposited clusters or surfaces.

The photoionization efficiency spectra of gas-phase Nb 3 CO and Nb 3 (CO) 2 have been acquired using a laser-ablation, photoionization mass spectrometer. The adiabatic ionization energies of the species are 5.82 ± 0.02 and 5.85 ± 0.02 eV, respectively, where error bars reflect precision, the values are accurate to within 0.1 eV. From these data and literature values of cation bond strengths, the bond energies of Nb 3 -CO and Nb 3 CO-CO are found to lie within the 2.8-3.4 and 1.7-1.8 eV ranges, respectively. The former is most consistent with dissociatively adsorbed CO, while the latter may correspond to molecularly or dissociatively adsorbed CO. These conclusions are supported by density-functional theory calculations. In addition to properties of Nb 3 CO, the properties of the transition state to dissociative adsorption of CO have been calculated. It is found that the transition state lies lower in energy than the separated reagents. Spontaneous dissociation is expected to follow molecular adsorption of CO on Nb 3 , accordingly. Over the 1540-2220 cm -1 range, no infrared multiphoton dissociation of Nb 3 CO or Nb 3 (CO) 2 is observed, consistent with CO being dissociatively adsorbed for both species.

An Al-cytosine association complex has been generated via laser ablation of a mixture of aluminum and cytosine powders that were pressed into a rod form. The ionization energy of the complex is found to be 5.16 +/- 0.01 eV. The photoionization efficiency spectrum of Al-cytosine has also been collected. DFT calculations indicate that binding of Al to cytosine manifests a significant weakening of the N-H bond, predicted to have a strength of 1.5 eV in the complex, and a significant stabilization of the oxo tautomeric form relative to the hydroxy forms. The predicted ionization energy of 5.2 eV agrees well with the experimental value. The threshold for dehydrogenation/ionization of Al-cytosine, forming (Al-cytosine-H)+, is found to occur at photoexcitation energies between 11.4 and 12.8 eV. This is a two-photon process that is proposed to occur via photoinduced electron transfer from Al to an antibonding (sigma) orbital localized on N-H. In the context of this mechanism, this work constitutes the first time charge transfer between a metal and DNA base has been photoinitiated in the gas phase.