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In 2020, the Geophysical and Astronomical Observatory of the University of Coimbra will celebrate the 95th anniversary of its first spectroheliographic observation. Keeping a daily service of solar observations since then, making almost a century, led to one of the largest continuous solar data collections in the world. This long–term solar database is essential for studies where solar activity is involved. This work reviews the development of synoptic observations made at the Observatory of Coimbra since 1925 and presents a summary of some of the principal stages of the Observatory’s history since its founding in 1772. We refer the main technical improvements and present some perspectives for the near future. One of the most significant upgrades was the installation of a CCD camera in 2007. The transition from photographic emulsion to digital recording methods allowed the development of image analysis algorithms to process solar images and improved data sharing with other institutions. This upgrade enabled also to carry–out modern climate and space weather studies. This valuable advancement makes it possible to create a new catalogue of solar observations to be published in the future.

Background: Interns experience tremendous challenges on the transition from student to new physician. There is limited literature describing curriculum that enhances medical students' preparation for internship. Description: To prepare graduating medical students at our institution for the new responsibilities and stressors that they will face as interns, an elective course, Integrative Clinical Experience, was implemented. Over a 2-week period, participating medical students rotate through 1 to 3-hr modules that cover four major domains: managing acutely ill patients, teaching, communicating, and coping with stressors. Evaluation: Participants evaluated the course qualitatively and quantitatively through verbal and written feedback. Students found the course useful and educationally valuable. They also had a statistically significant increase in perceived preparedness for internship upon completion of the course. Conclusions: A 2-week, concentrated course is a feasible and effective method for raising students' perceived preparedness for internship. [ABSTRACT FROM AUTHOR]

Prostate sarcoma is extremely rare, comprising less than 0.1 % of prostate cancers. A 61-year-old male presented to the emergency department with urinary retention and hematuria. Upon resolution of urinary retention, abdominal computed tomography showed a giant prostatic tumor, of approximately 1700 cubic centimeters, causing bilateral ureteric obstruction, and invasion of rectum and sigmoid colon. Laparotomy due to bowel obstruction showed peritoneal carcinomatosis. Palliative chemotherapy was initiated; however, he died due to hematological toxicity related to doxorubicin. Radical surgery is the ideal treatment; in cases of advanced or metastatic disease, adjuvant or palliative chemotherapy or radiotherapy withholds little or no benefit., (© 2024 The Authors.)

Background: To evaluate the anatomical and functional outcomes of high-density subthreshold micropulse laser (HSML) treatment in a cohort of patients diagnosed with chronic central serous chorioretinopathy (CSCR) whose treatment with photodynamic therapy (PDT) was delayed due to the worldwide shortage of verteporfin., Methods: Prospective interventional study which included 42 eyes of 40 patients diagnosed with chronic CSCR and on the waiting list for PDT who received rescue therapy with HSML using the Navilas® System device (OD-OS GmBH, Teltwo, Germany). Best corrected visual acuity (BCVA), subretinal fluid (SRF), and subfoveal choroidal thickness (SFCT) were measured at inclusion and during the follow-up visits at 2, 4, and 6 months., Results: The mean waiting time from the indication of PDT until treatment with HSML was 14.6 ± 9.7 months (range 5-21). There were no differences in the pre-treatment BCVA compared with the 6-month follow-up visit (67 ± 16.7 letters and 67.5 ± 8.2 letters respectively, p = 0.136). However, there was a significant decrease in the mean SFCT of -39.6 ± 37.1 μm (p = 0.030). Additionally, there was a decrease in SRF height between the pre-treatment measure (123.0 ± 49.8 μm) and the 2, 4, and 6-month follow-up visits after HSML of -58.5 ± 68.2 μm, -53.2 ± 76.3 μm, and -65.4 ± 53.6 μm respectively (p < 0.001). A complete resolution of the SRF was observed in 16/42 eyes (38.1 %) and a reduction of the SRF height in 85.7 % of the overall cohort was observed after HSML treatment., Conclusion: A significant anatomical improvement in SRF and a decrease in SFCT were observed in patients with CSCR who were previously waiting for PDT and were rescued by HSML. However, the rate of complete SRF resolution was low., Competing Interests: Declaration of competing interest None to declare, (Copyright © 2024. Published by Elsevier B.V.)

Care for the older patient living with a chronic wound comes with challenges not seen in younger patients. The aging skin, impacted by the environment and intrinsic physiologic changes, makes it susceptible to injury and poor healing. Likewise, older adults' goals with regards to wound healing may vary depending on their functional abilities and quality of life. The clinician must pay attention to these nuances and collaborate with the older patient in developing a treatment plan. Careful systematic description, documentation, and communication with the patient/caregiver aids the clinician in tracking the treatment goals and potentially reducing medical liability risk., Competing Interests: Disclosure The authors have no financial affiliations or conflicts of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Carbohydrates, in particular the d-enantiomers of ribose, 2-deoxyribose, and glucose, are essential to life's informational biopolymers (RNA/DNA) and for supplying energy to living cells through glycolysis. Considered to be potential biosignatures in the search of past or present life, our capacity to detect and quantify these essential sugars is crucial for future space missions to the Moon, Mars or Titan as well as for sample-return missions. However, the enantioselective analysis of carbohydrates is challenging and both research and routine applications, are lacking efficient methods that combine highly sensitive and reproducible detection with baseline enantioselective resolution and reliable enantiomeric excess (ee) measurements. Here, we present four different derivatization strategies in combination with multidimensional gas chromatography coupled to a reflectron time-of-flight mass spectrometer (GC×GC-TOF-MS) for the enantioselective resolution of C3 to C6 carbohydrates potentially suitable for sample-return analyses. Full mass spectral interpretation and calibration curves for one single-step (cyclic boronate derivatives) and three two-step derivatization protocols (aldononitrile-acetate, hemiacetalization-trifluoroacetylation, and hemiacetalization-permethylation) are presented for concentrations ranging from 1 to 50 pmol μL⁻ 1 with correlation coefficients R 2  > 0.94. We compared several analytical parameters including reproducibility, sensitivity (LOD and LOQ), overall separation, chiral resolution (R S ), mass spectrum selectivity, stability during long term storage, and reliability of ee measurements to guide the application-dependent selection of optimal separation and quantification performance., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Cornelia Meinert reports financial support was provided by European Research Council. Jana Bocková reports financial support was provided by French Space Agency. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Glycerophospholipid membranes are one of the key cellular components. Still, their species-dependent composition and homochirality remain an elusive subject. In the context of the astrophysical circularly polarized light scenario likely involved in the generation of a chiral bias in meteoritic amino and sugar acids in space, and consequently in the origin of life's homochirality on Earth, this study reports the first measurements of circular dichroism and anisotropy spectra of a selection of glycerophospholipids, their chiral backbones and their analogs. The rather low asymmetry in the interaction of UV/VUV circularly polarized light with sn-glycerol-1/3-phosphate indicates that chiral photons would have been unlikely to directly induce symmetry breaking to membrane lipids. In contrast, the anisotropy spectra of d-3-phosphoglyceric acid and d-glyceraldehyde-3-phosphate unveil up to 20 and 100 times higher maximum anisotropy factor values, respectively. This first experimental report, targeted on investigating the origins of phospholipid symmetry breaking, opens up new avenues of research to explore alternative mechanisms leading to membrane lipid homochirality, while providing important clues for the search for chiral biosignatures of extant and/or extinct life in space, in particular for the ExoMars 2028 mission., (© 2024 The Authors. Chirality published by Wiley Periodicals LLC.)

Purpose: The timely delivery of health care is an important quality indicator that has been shown to correlate with outcomes for cancer patients. We present our single-institution experience with the implementation of a same-day-access scheduling initiative in outpatient radiation oncology., Methods and Materials: From March 2021 to March 2023, a total of 4301 consecutive new patients referred for radiation oncology consultation were offered same-day appointments as part of a prospective pilot initiative conducted at a tertiary-based academic medical center. Descriptive statistics were used to study the effect of this initiative on access-related benchmarks compared with historical control patients referred during a preceding 24-month period., Results: Among the 3414 patients scheduled, 477 (14%) opted for same-day appointments. Black, Latino, and Asian patients were significantly more likely to use same-day access versus Caucasian patients (P < .001). The same-day-access initiative increased the proportion of patients seen within 5 days from referral from 22% to 61% (P < .001). The median time from referral to consult was 12 days (range, 0-149 days) before the implementation of the same-day-access initiative compared with 3 days (range, 0-101 days) after (P < .001). The no-show rate was reduced from 15% to 7% with the initiative (P < .001). All patients who requested a same-day appointment were successfully accommodated., Conclusions: The implementation of this same-day-access initiative enhanced operational efficiency and reduced barriers to care in the outpatient setting., Competing Interests: Disclosures Allen M. Chen holds volunteer leadership positions within the American Society of Radiation Oncology and the American Board of Radiology. Maheswari Senthil holds volunteer leadership positions with the Society of Surgical Oncology, American College of Surgeons, and California Institute of Regenerative Medicine., (Copyright © 2023 American Society for Radiation Oncology. Published by Elsevier Inc. All rights reserved.)

Introduction: The differential diagnosis for serous SRF can involve diseases with widely different pathogenic mechanisms that can range from vascular ocular diseases to ocular tumours and paraneoplastic syndromes. Recently, van Dijk et al. have described in three patients a new entity which they have called serous maculopathy with an absence of retinal pigment epithelium (SMARPE). We hereby describe a case of this infrequent macular disease and report its characteristic findings on multimodal imaging., Case Description: We present the case of a 65-year-old hyperopic woman with a three-year history of visual acuity (VA) loss in her left eye. Prior optical coherence tomography (OCT) had revealed the presence of serous subretinal fluid that had shown no response to treatment with intravitreal injections. On swept source OCT angiography scan, no macular alterations in the retinal vascular plexus structure were noted and there was no evidence of choroidal neovascularization. Ultra-widefield fluorescence angiography of the left eye revealed an early hyperfluorescent macular spot corresponding to the area of absent RPE and late fluorescein pooling. On ultra-widefield indocyanine green angiography there were no central or peripheral abnormalities of choroidal vascularization., Conclusion: This recently described entity should be considered as a differential diagnosis in persistent serous subretinal fluid. Multimodal imaging helps differentiate SMARPE from its main differential diagnoses , and care should be taken to identify and differentiate it from similar conditions to avoid unnecessary treatment with its possible side effects and complications., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Purpose: We present our single-institution experience with the development of a same day access scheduling initiative for an outpatient radiation oncology unit, focusing on its potential influence on ameliorating racial disparities., Methods and Materials: From March 2021 to August 2022, a pilot initiative was conducted such that all new patients referred to a tertiary care-based radiation oncology department were offered the ability to be seen as a same day consultation. The timespan of this analysis was categorized into 2 distinct successive periods over 36 months-a 18-month pre-initiative period (September 2019 to February 2021) and another subsequent one (March 2021 to August 2022). Descriptive statistics were used to study the impact of this initiative on access-related benchmarks., Results: A total of 2897 patients were referred. Among the 2107 patients scheduled, three hundred and sixteen (15 %) opted for same day appointments. Black, Latino, and Asian patients were significantly more likely to use the same day access initiative versus Caucasian patients (p = 0.01). The same day access initiative increased the proportion of patients seen within 5 days from referral from 8 % to 34 % for Blacks, 12-57 % for Latinos, and 18-67 % for Asians, compared to 39-55 % for Caucasians (p < 0.001). The no-show rate was reduced from 20 % to 7 % and 14-5 %, for Black and Latino patients, respectively (p < 0.001)., Conclusions: The implementation of a same day access initiative narrowed disparities with respect to access-related benchmarks., Competing Interests: Declaration of Competing Interest The authors have no disclosures or conflict of interest to report., (Copyright © 2023. Published by Elsevier Ltd.)

Fungal infections are a serious problem affecting many people worldwide, creating critical economic and medical consequences. Fungi are ubiquitous and can cause invasive diseases in individuals mostly living in developing countries or with weakened immune systems, and antifungal drugs currently available have important limitations in tolerability and efficacy. In an effort to counteract the high morbidity and mortality rates associated with invasive fungal infections, various approaches are being utilized to discover and develop new antifungal agents. This review discusses the challenges posed by fungal infections, outlines different methods for developing antifungal drugs and reports on the status of drugs currently in clinical trials, which offer hope for combating this serious global problem.

Gas chromatography (GC) is a separation technique commonly developed for targeted in situ analyses in planetary space missions. It is coupled with low-resolution mass spectrometry to obtain additional structural information and allow compound identification. However, ground-based analyses of extraterrestrial samples have shown the presence of large molecular diversities. For future targeted in situ analyses, it is therefore essential to develop new technologies. High resolution mass spectrometry (HRMS) is currently being spatialized using FT-orbitrap-MS technology. In this contribution, the coupling of gas chromatography with FT-orbitrap-MS is studied for targeted amino acid analyses. The method for enantioselective separation of amino acids was optimized on a standard mixture comprising 47 amino acid enantiomers. Different ionization modes were optimized, chemical ionization with three different reactive gasses (NH 3 , CH 4 and NH 3 /CH 4 ) and electron impact ionization at different electron energies. Single ion and full scan monitoring modes were compared, and detection and quantification limits were estimated by internal calibration under the optimized conditions. The GC-FT-orbitrap-MS demonstrated its ability to separate 47 amino acid enantiomers with minimal co-elution. Furthermore, due to the high mass resolution and accuracy of FT-orbitrap-MS, with mass extraction, the S/N is close to zero, allowing average LOD values of 10⁻ 7  M, orders of magnitude lower than conventional GC-MS techniques. Finally, these conditions were tested for enantioselective analysis of amino acids on an analog of a pre-cometary organic material showing similarities to that of extraterrestrial materials., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Introduction Benign prostatic hyperplasia (BPH) is one of the most common causes of lower urinary tract symptoms in men. Holmium Laser Enucleation of the Prostate (HoLEP) has been recommended by international guidelines as an alternative to transurethral resection of the prostate (TURP). HoLEP's learning curve and the lack of adequate mentorship remains an obstacle for the worldwide adoption of this technique. Objective To report the first-year learning curve of a newly established mentorship program in young urologists without any previous HoLEP experience. Methods We report a cohort of patients with BPH, with prostate size ≥70 grams, treated with HoLEP, analyzed for perioperative data and complications, and short-term postoperative complications and functional outcomes, at three and six months after surgery. Results A total of 47 patients were managed with HoLEP. Mean total operative time was 149.8 ± 42.9 minutes. We experienced five (10.6%) intraoperative complications, including one intravesical resection of the prostate with bipolar energy, three conversions to TURP and one conversion to open prostatectomy. We experienced four postoperative complications, all of them Clavien-Dindo ≤2. Median International Prostate Symptom Score (IPSS) decrease at six months was -17 points from baseline. Mean post-void residual volume and prostate-specific antigen significantly decreased by the third postoperative month. Multiple linear regression showed that prostate size is directly related to increased surgical time during the early learning curve. Discussion Our experience adequately reflects the importance of HoLEP mentorship in young urologists seeking training in this technique. Both surgeons had perioperative and postoperative outcomes deemed satisfactory based on previously published learning curves. Conclusions HoLEP is a technically difficult procedure, however, adequate mentorship leads to satisfying short-term outcomes since the early stages of the learning curve in young urologists with no previous training on this technique., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Martinez-Salas et al.)

The Venezuelan equine encephalitis virus (VEEV) nonstructural protein 2 (nsP2) cysteine protease (EC 3.4.22.B79) is essential for viral replication. High throughput in silico/in vitro screening using a focused set of known cysteine protease inhibitors identified two epoxysuccinyl prodrugs, E64d and CA074 methyl ester (CA074me) and a reversible oxindole inhibitor. Here, we determined the X-ray crystal structure of the CA074-inhibited nsP2 protease and compared it with our E64d-inhibited structure. We found that the two inhibitors occupy different locations in the protease. We designed hybrid inhibitors with improved potency. Virus yield reduction assays confirmed that the viral titer was reduced by >5 logs with CA074me. Cell-based assays showed reductions in viral replication for CHIKV, VEEV, and WEEV, and weaker inhibition of EEEV by the hybrid inhibitors. The most potent was NCGC00488909-01 which had an EC 50 of 1.76 µM in VEEV-Trd-infected cells; the second most potent was NCGC00484087 with an EC 50 = 7.90 µM. Other compounds from the NCATS libraries such as the H1 antihistamine oxatomide (>5-log reduction), emetine, amsacrine an intercalator (NCGC0015113), MLS003116111-01, NCGC00247785-13, and MLS00699295-01 were found to effectively reduce VEEV viral replication in plaque assays. Kinetic methods demonstrated time-dependent inhibition by the hybrid inhibitors of the protease with NCGC00488909-01 (K i = 3 µM) and NCGC00484087 (K i = 5 µM). Rates of inactivation by CA074 in the presence of 6 mM CaCl 2 , MnCl 2 , or MgCl 2 were measured with varying concentrations of inhibitor, Mg 2+ and Mn 2+ slightly enhanced inhibitor binding (3 to 6-fold). CA074 inhibited not only the VEEV nsP2 protease but also that of CHIKV and WEEV.

Kuiper Belt objects exhibit a wider color range than any other solar system population. The origin of this color diversity is unknown, but likely the result of the prolonged irradiation of organic materials by galactic cosmic rays (GCRs). Here, we combine ultrahigh-vacuum irradiation experiments with comprehensive spectroscopic analyses to examine the color evolution during GCR processing methane and acetylene under Kuiper Belt conditions. This study replicates the colors of a population of Kuiper Belt objects such as Makemake, Orcus, and Salacia. Aromatic structural units carrying up to three rings as in phenanthrene (C 14 H 10 ), phenalene (C 9 H 10 ), and acenaphthylene (C 12 H 8 ), of which some carry structural motives of DNA and RNA connected via unsaturated linkers, were found to play a key role in producing the reddish colors. These studies demonstrate the level of molecular complexity synthesized of GCR processing hydrocarbon and hint at the role played by irradiated ice in the early production of biological precursor molecules.

Background: Critically ill pediatric patients can have difficulty with establishing and maintaining stable vascular access. A long-dwelling peripheral intravenous catheter placement decreases the need for additional vascular interventions., Aim: The study sought to compare longevity, catheter-associated complications, and the need for additional vascular interventions when using ultrasound-guided longer peripheral intravenous catheters comparing to a traditional approach using standard-sized peripheral intravenous catheters in pediatric critically ill patients with difficult vascular access., Methods: This single-center retrospective cohort study included children 0-18 years of age with difficult vascular access admitted to the pediatric intensive care unit between 01/01/2018-06/01/2021., Results: One hundred and eighty seven placements were included in the study, with 99 ultrasound-guided long intravenous catheters placed and 88 traditionally placed standard-sized intravenous catheters. In the univariate analysis, patients in the traditional approach were at a higher risk of intravenous failure compared to those in the ultrasound-guided approach (HR = 2.20, 95% CI [1.45-3.34], p = .001), with median intravenous survival times of 108 and 219 h, respectively. Adjusting for age, patients in the traditional approach remained at higher risk of intravenous failure (HR = 1.99, 95% CI: [1.28-3.08], p = .002). Adjusting for hospital length of stay, patients in the ultrasound-guided approach were less likely to have additional peripheral intravenous access placed during hospitalization (OR = 0.39, 95% CI [0.18-0.85] p = .017)., Conclusion: In critically ill pediatric patients with difficult vascular access, ultrasound-guided long peripheral intravenous catheters provide an alternative to traditional approach standard-sized intravenous catheters with improved longevity, lower failure rates, and reduced need for additional vascular interventions., (© 2023 The Authors. Pediatric Anesthesia published by John Wiley & Sons Ltd.)

Biofilm formation is important for microbial survival in hostile environments and a phenotype that provides microorganisms with antimicrobial resistance. Zinc oxide (ZnO) and Zinc sulfide (ZnS) nanoparticles (NPs) present potential antimicrobial properties for biomedical and food industry applications. Here, we aimed to analyze, for the first time, the bactericidal and antibiofilm activity of ZnS NPs against Staphylococcus aureus, Klebsiella oxytoca, and Pseudomonas aeruginosa, all medically important bacteria in developed countries. We compared ZnS NPs antimicrobial activity to ZnO NPs, which have been extensively studied. Using the colorimetric XTT reduction assay to observe the metabolic activity of bacterial cells and the crystal violet assay to measure biofilm mass, we demonstrated that ZnS and ZnO had similar efficacy in killing planktonic bacterial cells and reducing biofilm formation, with S. aureus being more susceptible to both therapeutics than K. oxytoca and P. aeruginosa. Crystal violet staining and confocal microscopy validated that Zn NPs inhibit biofilm formation and cause architectural damage. Our findings provide proof of principle that ZnS NPs have antibiofilm activity, and can be potentially used in medical and food industry applications, such as treatment of wound infections or package coating for food preservation. IMPORTANCE Zinc (Zn)-based nanoparticles (NPs) can be potentially used in medical and food preservation applications. As proof of principle, we investigated the bactericidal and antibiofilm activity of zinc oxide (ZnO) and zinc sulfide (ZnS) NPs against medically important bacteria. Zn-based NPs were similarly effective in killing planktonic and biofilm-associated Staphylococcus aureus, Klebsiella oxytoca, and Pseudomonas aeruginosa cells. However, S. aureus was more susceptible to these investigational therapeutics. Although further studies are warranted, our findings suggest the possibility of future use of Zn-based NPs in the treatment of skin infections or preservation of food.

This work proposes a comprehensive two-dimensional gas chromatography method for the resolution and quantification of 27 amino acids, including 17 enantiomeric pairs, as stable N-trifluoroacetyl-O-methyl ester derivatives. The derivatization approach in combination with enantioselective two-dimensional gas chromatography has proven to be highly responsive with a method detection limit of 1-7 pg even for sterically hindered amino acids such as α,α-dialkylated, and N-alkylated amino acids. Accurate determination of the enantiomeric excess was achieved with errors in the range of ±0.5%-2.5% (1σ) at concentrations ≥10 -6 M. A thorough study of the mass spectra of the amino acid derivatives allowed the fragmentation pathways to be distinguished, enabling chromatographic peaks to be unambiguously assigned. The proposed method is particularly suited for applications that require the precise determination of enantiomeric excesses such as those concerning the role of d-amino acid enantiomers in humans, animals, and the environment, as well as for analyses of extraterrestrial samples aimed at understanding the selection of amino acids in stereochemical l-configuration., (© 2022 The Authors. Journal of Separation Science published by Wiley-VCH GmbH.)

Propylene oxide, the first chiral molecule recently detected in the interstellar medium, has once again raised the question whether biomolecular chirality might have cosmic origins. However, accurate chiroptical properties of propylene oxide in the ultraviolet spectral range necessary to suggest possible asymmetric synthetic routes in the gas phase are scarce. Here, we report on the first experimental measurements of the anisotropy spectra of gas-phase propylene oxide in the vacuum ultraviolet spectral range. Our experimental results provide novel insights into the handedness of interstellar circular polarization at the dawn of molecular evolution of our star- and planet-forming region. Besides the fundamental importance of this new investigation for understanding the origin and evolution of homochirality on Earth, our high-resolution experimental electronic circular dichroism data will inspire new efforts in quantum computational spectroscopy.

Transgender people commonly experience discrimination from clinicians, which directly contributes to worse mental and physical health outcomes among this population. This article describes mechanisms by which stigma perpetuates health inequity among transgender patients and highlights its unique effects on transgender patients of color. The article concludes with recommendations to cisgender clinicians on how to help prevent stigmatizing interactions with transgender patients., (Copyright 2022 American Medical Association. All Rights Reserved.)

Non-human primates (NHPs) are precious resources for cutting-edge neuroscientific research, including large-scale viral vector-based experimentation such as optogenetics. We propose to improve surgical outcomes by enhancing the surgical preparation practices of convection-enhanced delivery (CED), which is an efficient viral vector infusion technique for large brains such as NHPs'. Here, we present both real-time and next-day MRI data of CED in the brains of ten NHPs, and we present a quantitative, inexpensive, and practical bench-side model of the in vivo CED data. Our bench-side model is composed of food coloring infused into a transparent agar phantom, and the spread of infusion is optically monitored over time. Our proposed method approximates CED infusions into the cortex, thalamus, medial temporal lobe, and caudate nucleus of NHPs, confirmed by MRI data acquired with either gadolinium-based or manganese-based contrast agents co-infused with optogenetic viral vectors. These methods and data serve to guide researchers and surgical team members in key surgical preparations for intracranial viral delivery using CED in NHPs, and thus improve expression targeting and efficacy and, as a result, reduce surgical risks.

Life on Earth employs chiral amino acids in stereochemical L-form, but the cause of molecular symmetry breaking remains unknown. Chiroptical properties of amino acids - expressed in circular dichroism (CD) - have been previously investigated in solid and solution phase. However, both environments distort the intrinsic charge distribution associated with CD transitions. Here we report on CD and anisotropy spectra of amino acids recorded in the gas phase, where any asymmetry is solely determined by the genuine electromagnetic transition moments. Using a pressure- and temperature-controlled gas cell coupled to a synchrotron radiation CD spectropolarimeter, we found CD active transitions and anisotropies in the 130-280 nm range, which are rationalized by ab initio calculation. As gas phase glycine was found in a cometary coma, our data may provide insights into gas phase asymmetric photochemical reactions in the life cycle of interstellar gas and dust, at the origin of the enantiomeric selection of life's L-amino acids., (© 2022. The Author(s).)

Homochiral proteins orchestrate biological functions throughout all domains of life, but the origin of the uniform l-stereochemistry of amino acids remains unknown. Here, we describe enantioselective adsorption experiments of racemic alanine and leucine onto homochiral d - and l -quartz as a possible mechanism for the abiotic emergence of biological homochirality. Substantial racemate resolution with enantiomeric excesses of up to 55% are demonstrated to potentially occur in interstitial pores, along grain boundaries or small fractures in local quartz-bearing environments. Our previous hypothesis on the enhanced enantioselectivity due to uranium-induced fission tracks could not be validated. Such capillary tubes in the near-surface structure of quartz have been proposed to increase the overall chromatographic separation of enantiomers, but no systematic positive correlation of accumulated radiation damage and enantioselective adsorption was observed in this study. In general, the natural l -quartz showed stronger enantioselective adsorption affinities than synthetic d -quartz without any significant trend in amino acid selectivity. Moreover, the l-enantiomer of both investigated amino acids alanine and leucine was preferably adsorbed regardless of the handedness of the enantiomorphic quartz sand. This lack of mirror symmetry breaking is probably due to the different crystal habitus of the synthetic z-bar of d -quartz and the natural mountain crystals of l -quartz used in our experiments.

Targeting the protein-protein interaction (PPI) between nuclear factor erythroid 2-related factor 2 (Nrf2) and Kelch-like ECH-associated protein 1 (Keap1) is a potential therapeutic strategy to control diseases involving oxidative stress. Here, six classes of known small-molecule Keap1-Nrf2 PPI inhibitors were dissected into 77 fragments in a fragment-based deconstruction reconstruction (FBDR) study and tested in four orthogonal assays. This gave 17 fragment hits of which six were shown by X-ray crystallography to bind in the Keap1 Kelch binding pocket. Two hits were merged into compound 8 with a 220-380-fold stronger affinity ( K i = 16 μM) relative to the parent fragments. Systematic optimization resulted in several novel analogues with K i values of 0.04-0.5 μM, binding modes determined by X-ray crystallography, and enhanced microsomal stability. This demonstrates how FBDR can be used to find new fragment hits, elucidate important ligand-protein interactions, and identify new potent inhibitors of the Keap1-Nrf2 PPI.

Purpose: Our goal was to (1) evaluate physician assistant (PA) students' knowledge, skills, and attitudes toward transgender patients, (2) develop and deliver a lecture on transgender health care, and (3) determine changes in knowledge, skills, and attitudes postlecture., Methods: A 32-item survey was developed and emailed to PA students. Students were then invited to a 2-hour lecture on transgender-specific psychosocial topics and clinical skills plus medical treatment for gender dysphoria. Lecture attendees retook the survey after 2 weeks., Results: Initial mean self-assessment scores regarding clinical knowledge and skills were below 3 on a 7-point Likert scale. All measures improved following the lecture (p < .001 for all). Overall, 86.7% of students were interested in the addition of transgender health into their didactic curriculum., Conclusions: Our results illustrate insufficient student competence in transgender medicine. Our proposed transgender curriculum may be a useful guide for educators wanting to integrate this topic into their instruction., Competing Interests: The authors declare no conflict of interest., (Copyright © 2021 Physician Assistant Education Association.)

The entorhinal cortex (EC) is a critical element of the hippocampal formation located within the medial temporal lobe (MTL) in primates. The EC has historically received attention for being the primary mediator of cortical information going into and coming from the hippocampus proper. In this review, we highlight the significance of the EC as a major player in memory processing, along with other associated structures in the primate MTL. The complex, convergent topographies of cortical and subcortical input to the EC, combined with short-range intrinsic connectivity and the selective targeting of EC efferents to the hippocampus, provide evidence for subregional specialization and integration of information beyond what would be expected if this structure were a simple conduit of information for the hippocampus. Lesion studies of the EC provide evidence implicating this region as critical for memory and the flexible use of complex relational associations between experienced events. The physiology of this structure's constituent principal cells mirrors the complexity of its anatomy. EC neurons respond preferentially to aspects of memory-dependent paradigms including object, place, and time. EC neurons also show striking spatial representations as primates explore visual space, similar to those identified in rodents navigating physical space. In this review, we highlight the great strides that have been made toward furthering our understanding of the primate EC, and we identify paths forward for future experiments to provide additional insight into the role of this structure in learning and memory.

A new electrochemical methodology has been developed for the preparation of a wide variety of functionalized orthoesters under mild and green conditions from easily accessible dithiane derivatives. The new methodology also offers an unprecedented way to access tri(fluorinated) orthoesters, a class of compound that has never been studied before. This provides the community with a rapid and general method to prepare libraries of functionalized orthoesters from simple and readily available starting materials.

Most chronic wounds are related to comorbidities, for which no clinical trials are performed. This retrospective propensity matched-cohort study examined data from 2 074 000 lower extremity wounds across 644 institutions to determine the effectiveness of TheraSkin plus standard of care (SOC; n = 1997) versus SOC alone (n = 1997). Multivariate modelling comparing outcomes such as healing rates, percent area reductions (PARs), amputations, recidivism, treatment completion, and medical transfers were evaluated. A higher proportion of wounds in the treatment group compared with the controls were more likely to close (68.3% versus 60.3%), particularly wounds with exposed structures (64% versus 50.4%) and with lower recidivism at 6 months (24.9% versus 28.3%). The control group was 2.75x more likely to require amputation than the treatment group. The combination of propensity matching and logistic regression analysis on a particularly large database demonstrated that wounds treated with TheraSkin had higher healing rates, higher PARs (78.7% versus 68.9%), fewer amputations, lower recidivism, higher treatment completion (61.0% versus 50.6%), and lower medical transfers (16.1% versus 23.5%) than SOC alone. This study considered data from complex wounds typically excluded from controlled trials and supports the idea that real-world evidence studies can be valid and reliable., (© 2019 The Authors. International Wound Journal published by Medicalhelplines.com Inc and John Wiley & Sons Ltd.)

A new electrochemical methodology has been developed for the generation of oxycarbonyl radicals under mild and green conditions from readily available hemioxalate salts. Mono- and multi-functionalised γ-butyrolactones were synthesised through exo-cyclisation of these oxycarbonyl radicals with an alkene, followed by the sp 3 -sp 3 capture of the newly formed carbon-centred radical. The synthesis of functionalised valerolactone derivatives was also achieved, demonstrating the versatility of the newly developed methodology. This represents a viable synthetic route towards pharmaceutically important fragments and further demonstrates the practicality of electrosynthesis as a green and economical method to activate small organic molecules., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Inhibiting the protein-protein interaction (PPI) between the transcription factor Nrf2 and its repressor protein Keap1 has emerged as a promising strategy to target oxidative stress in diseases, including central nervous system (CNS) disorders. Numerous non-covalent small-molecule Keap1-Nrf2 PPI inhibitors have been reported to date, but many feature suboptimal physicochemical properties for permeating the blood-brain barrier, while others contain problematic structural moieties. Here, we present the first side-by-side assessment of all reported Keap1-Nrf2 PPI inhibitor classes using fluorescence polarization, thermal shift assay, and surface plasmon resonance-and further evaluate the compounds in an NQO1 induction cell assay and in counter tests for nonspecific activities. Surprisingly, half of the compounds were inactive or deviated substantially from reported activities, while we confirm the cross-assay activities for others. Through this study, we have identified the most promising Keap1-Nrf2 inhibitors that can serve as pharmacological probes or starting points for developing CNS-active Keap1 inhibitors.

Glucokinase phosphorylated a series of C-1 fluorinated α-d-gluco-heptuloses. These phosphorylated products were discovered to be inhibitors of α-phosphomannomutase/phosphoglucomutase (αPMM/PGM) and β-phosphoglucomutase (βPGM). Inhibition potency with both mutases inversely correlated to the degree of fluorination. Structural analysis with αPMM demonstrated the inhibitor binding to the active site, with the phosphate in the phosphate binding site and the anomeric hydroxyl directed to the catalytic site., Competing Interests: The authors declare no competing financial interest.

The biomolecular homochirality in living organisms has been investigated for decades, but its origin remains poorly understood. It has been shown that circular polarized light (CPL) and other energy sources are capable of inducing small enantiomeric excesses ( ee s) in some primary biomolecules, such as amino acids or sugars. Since the first findings of amino acids in carbonaceous meteorites, a scenario in which essential chiral biomolecules originate in space and are delivered by celestial bodies has arisen. Numerous studies have thus focused on their detection, identification, and enantiomeric excess calculations in extraterrestrial matrices. In this review we summarize the discoveries in amino acids, sugars, and organophosphorus compounds in meteorites, comets, and laboratory-simulated interstellar ices. Based on available analytical data, we also discuss their interactions with CPL in the ultraviolet (UV) and vacuum ultraviolet (VUV) regions, their abiotic chiral or achiral synthesis, and their enantiomeric distribution. Without doubt, further laboratory investigations and upcoming space missions are required to shed more light on our potential extraterrestrial molecular origins.

Background: Airports may represent significant sources of secondhand smoke (SHS) exposure for both travelers and employees. While previously common smoking rooms have largely disappeared from US airports, smoking continues to occur outdoors at terminal entrances. SHS may be especially high at arrival areas, since they oftentimes are partially enclosed by overhead departures, creating stagnant microenvironments. This study assessed particulate matter <2.5 microns in diameter (PM2.5), a common surrogate for SHS, at airport terminal locations to evaluate both outdoor exposure risk and possible indoor drift of SHS from outdoor sources., Methods: A convenience sample of nine airport terminal arrival areas in the US state of Florida was surveyed between February and July 2018. PM2.5 levels were assessed outdoors and indoors at terminal entrances and at control areas far into terminal interiors. We also examined the impact of smoking location on SHS exposure by correlating cigarette and passing vehicle counts with PM2.5 levels at terminals with contrasting proximity of designated smoking locations to terminal entrances., Results: Although outdoor PM2.5 levels (mean 17.9, SD 6.1  µ g/m 3 ) were significantly higher than indoors ( p < 0.001), there was no difference between indoor areas directly inside terminal entrances and areas much further interior (mean 8.8, SD 2.6 vs mean 8.5, SD 3.0  µ g/m 3 , p =0.49). However, when smoking areas were in close proximity to terminal entrances, the number of lit cigarettes and vehicular traffic per minute predicted 70% of the variance of PM2.5 levels ( p < 0.001), which was attributable mostly to the cigarette number ( β  = 0.83; 95% CI (0.55 to 1.11); p < 0.001). This effect was not observed at smoking areas further away., Conclusion: PM2.5 data did not suggest indoor drift from outside smoking. Nevertheless, absolute exposure outdoors was high and correlated with the location of designated smoking areas. Further studies are needed to examine the effect of microclimate formation on exposure risk.

Hydrogels have been studied as promising materials in different biomedical applications such as cell culture in tissue engineering or in wound healing. In this work, we synthesized different nanocellulose-alginate hydrogels containing cellulose nanocrystals, TEMPO-oxidized cellulose nanocrystals (CNCTs), cellulose nanofibers or TEMPO-oxidized cellulose nanofibers (CNFTs). The hydrogels were freeze-dried and named as gels. The nanocelluloses and the gels were characterized by different techniques such as Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), and dynamic mechanical thermal analysis (DMTA), while the biological features were characterized by cytotoxicity and cell growth assays. The addition of CNCTs or CNFTs in alginate gels contributed to the formation of porous structure (diameter of pores in the range between 40 and 150 μm). TEMPO-oxidized cellulose nanofibers have proven to play a crucial role in improving the dimensional stability of the samples when compared to the pure alginate gels, mainly after a thermal post-treatment of these gels containing 50 wt % of CNFT, which significantly increased the Ca 2+ crosslinking density in the gel structure. The morphological characteristics, the mechanical properties, and the non-cytotoxic behavior of the CNFT-alginate gels improved bioadhesion, growth, and proliferation of the cells onto the gels. Thus, the alginate-nanocellulose gels might find applications in tissue engineering field, as for instance, in tissue repair or wound healing applications.

Antimicrobial resistance (AMR) in bacteria and associated human morbidity and mortality is increasing. The use of antimicrobials in livestock selects for AMR that can subsequently be transferred to humans. This flow of AMR between reservoirs demands surveillance in livestock and in humans. We quantified and characterized the acquired resistance gene pools (resistomes) of 181 pig and 178 poultry farms from nine European countries, sequencing more than 5,000 Gb of DNA using shotgun metagenomics. We quantified acquired AMR using the ResFinder database and a second database constructed for this study, consisting of AMR genes identified through screening environmental DNA. The pig and poultry resistomes were very different in abundance and composition. There was a significant country effect on the resistomes, more so in pigs than in poultry. We found higher AMR loads in pigs, whereas poultry resistomes were more diverse. We detected several recently described, critical AMR genes, including mcr-1 and optrA, the abundance of which differed both between host species and between countries. We found that the total acquired AMR level was associated with the overall country-specific antimicrobial usage in livestock and that countries with comparable usage patterns had similar resistomes. However, functionally determined AMR genes were not associated with total drug use.

Recent studies have shown that hippocampal "time cells" code for sequential moments in temporally organized experiences. However, it is currently unknown whether these temporal firing patterns critically rely on upstream cortical input. Here we employ an optogenetic approach to explore the effect of large-scale inactivation of the medial entorhinal cortex on temporal, as well as spatial and object, coding by hippocampal CA1 neurons. Medial entorhinal inactivation produced a specific deficit in temporal coding in CA1 and resulted in significant impairment in memory across a temporal delay. In striking contrast, spatial and object coding remained intact. Further, we extended the scope of hippocampal phase precession to include object information relevant to memory and behavior. Overall, our work demonstrates that medial entorhinal activity plays an especially important role for CA1 in temporal coding and memory across time., (Copyright © 2017 Elsevier Inc. All rights reserved.)

We present a consistent linear response formulation of the density functional based tight-binding method for long-range corrected exchange-correlation functionals (LC-DFTB). Besides a detailed account of derivation and implementation of the method, we also test the new scheme on a variety of systems considered to be problematic for conventional local/semilocal time-dependent density functional theory (TD-DFT). To this class belong the optical properties of polyacenes and nucleobases, as well as charge transfer excited states in molecular dimers. We find that the approximate LC-DFTB method exhibits the same general trends and similar accuracy as range-separated DFT methods at significantly reduced computational cost. The scheme should be especially useful in the determination of the electronic excited states of very large molecules, for which conventional TD-DFT is supposed to fail due to a multitude of artificial low energy states.

The objective of this study was to investigate the effect of substituting soybean meal products with yeast-derived microbial protein (YMP) on lactation performance in diets containing 2 forage-to-concentrate ratios. Sixteen Holstein cows (4 primiparous and 12 multiparous) were randomly assigned to multiple 4 × 4 Latin squares with a 2 × 2 factorial arrangement of treatments. Diets contained low (LF; 45% of diet DM) or high forage (HF; 65% of diet DM) and YMP at 0 (NYMP) or 2.25% (WYMP) of the diet. The forage mix consisted of 67% corn silage and 33% alfalfa hay on a DM basis. No interactions of forage and YMP were noted for any of the production parameters measured. Feed efficiency (energy-corrected milk/dry matter intake) was greater for cows fed NYMP compared with WYMP. Regardless of the addition of YMP, cows fed LF had greater dry matter intake and produced more milk than cows fed HF. In addition, cows fed LF produced more energy-corrected milk than those fed HF. Milk fat percentage was lower in cows fed LF compared with HF, whereas fat yield was similar between forage concentrations. Fat yield tended to decrease with feeding YMP. Interactions of forage and YMP were observed for propionate concentration, acetate and propionate proportion, and acetate-to-propionate ratio. A tendency for an interaction of forage and YMP was also noted for ruminal pH. Cows fed HF diets had greater ruminal ammonia and butyrate concentrations, as well as proportion of butyrate. Arterial concentrations of Ile, Leu, Met, Thr, and Val were greater in cows fed LF. Cows fed NYMP had greater arterial concentrations of Ile, Lys, Trp, and Val than cows fed WYMP. Substitution of soybean proteins with YMP did not improve performance or feed efficiency of high-producing dairy cows regardless of the forage-to-concentrate ratio of the diet., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to determine if increased dietary fat from dried distillers grains with solubles (DDGS) in diets of growing heifers affected dry matter intake, average daily gain (ADG), growth performance, and nutrient digestibility. Thirty-three Holstein heifers (133±18 d old) were used in a 24-wk randomized complete block design. Treatments were (1) control (CON) containing ground corn and soybean products, (2) low-fat (LFDG) containing low-fat, high-protein DDGS and ground corn, and (3) high-fat (HFDG) with traditional DDGS. All diets contained 39.8% grass hay, 24.8% corn silage, and 1.5% vitamins and minerals. The HFDG diet was formulated to contain 4.8% fat compared with 2.8% in the CON and LFDG diets, which were greater in nonfibrous carbohydrate. Diets had a net energy gain of 1.0Mcal/kg of dry matter and were limit-fed at 2.45% of body weight. Heifers were weighed every 2wk and rations were adjusted accordingly. Heart girth, hip and wither heights, body length, and body condition score were recorded every 2wk. Total-tract digestion of nutrients was evaluated during wk16 using fecal grab sampling and an external marker. No treatments by time interactions were found. Dry matter intakes, body weights, ADG, and gain-to-feed ratio were similar among treatments; however, ADG averaged 0.96kg/d among treatments, which is greater than recommended. All body frame measurements and body condition scores were similar among treatments. Total-tract digestibilities of dry matter and organic matter were not different among treatments. However, crude protein and neutral detergent fiber digestibility were increased in the HFDG diet compared with the CON and LFDG diets. These results demonstrate that using DDGS or low-fat DDGS with corn in growing heifer rations can maintain performance. Utilizing the fat in DDGS as a dietary energy source in replacement of starch from corn did not influence growth performance or negatively affect nutrient digestion., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to determine if increased dietary fat from dried distillers grains with solubles (DDGS) in diets of growing heifers affected metabolic profile, plasma fatty acid profile, and reproductive maturation. Thirty-three Holstein heifers (133±18 d of age) were used in a 24-wk randomized complete block design with 3 treatment diets. Treatment diets were (1) control (CON) containing ground corn (15.9% of DM) and soybean products (17.9%), (2) low-fat (LFDG) containing low-fat DDGS (21.9%) and ground corn (11.9%), or (3) high-fat (HFDG) with traditional DDGS (33.8%). Diets were isonitrogenous and isocaloric, but the HFDG diet was formulated to contain 4.8% fat compared with 2.8% in the CON and LFDG diets. All 3 diets were limit-fed to 2.45% of body weight on a dry matter basis, and resulted in a mean average daily gain of 0.96kg/d across treatments. Every 4wk, jugular blood was collected for analysis of metabolites and metabolic hormones. During wk20 of the feeding period, blood samples were collected for analysis of plasma fatty acid profiles. When heifers weighed between 200 and 300kg of body weight, coccygeal blood samples were taken twice weekly for analysis of progesterone to determine if puberty had been reached. Plasma concentrations of nonesterified fatty acids were similar among treatments and consistent over the duration of the study. Plasma concentrations of β-hydroxybutyrate, insulin, insulin-like growth factor-1, and leptin were similar among heifers fed each treatment diet, but increased over the duration of the feeding period. Serum concentrations of glucose tended to be less in heifers fed HFDG compared with heifers fed the CON diet. Glucose concentrations fluctuated throughout the feeding period, but no treatment by time interactions were noted. Plasma urea N concentrations were less in heifers fed LFDG compared with heifers fed HFDG and CON diets. The concentrations of plasma urea N increased over the duration of the feeding period, with no treatment by week interaction. Total plasma cholesterol was greater in heifers fed HFDG compared with the CON and LFDG diets, and a significant week effect and a week by treatment interaction were observed. Fatty acid profiles also differed among treatments based on the supply of fatty acids from the diet. Progesterone analysis indicated that heifers fed HFDG tended to be pubertal at a younger age than heifers on CON. These results demonstrate that dietary fat from DDGS can be used in high-plane of nutrition rations for growing heifers and maintain metabolic energy status compared with starch from corn, but alters the concentrations of different blood lipids., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this research was to determine the subsequent effects on lactation and reproductive performance from feeding fat from distillers dried grains with solubles (DDGS) to growing dairy heifers. During the prepubertal growth phase, 33 Holstein heifers (133±18 d old) were used in a 24-wk randomized complete block design. Treatments included (1) a control diet (CON) containing ground corn and soybean products, 2) a diet with low-fat DDGS (LFDG), and (3) a higher-fat diet with traditional DDGS (HFDG). All diets contained 39.8% grass hay, 24.8% corn silage, and 1.5% vitamins and minerals. Previous results demonstrated that growth performance was maintained across treatments, but plasma cholesterol and fatty acids were greater and puberty may occur earlier in heifers fed HFDG. It was hypothesized that differences among treatments in metabolic profile and puberty may influence reproductive and first-lactation performance. Posttrial data on reproductive performance and milk production for the first 4 mo of lactation were collected for each heifer from dairy herd records. At 3wk prepartum and at calving, body weights, body condition scores, and body measurements were taken. No differences were observed among treatments for age at conception or age at calving. At calving, heifers fed the HFDG were shorter in withers height compared with heifers fed the other diets. Milk yields and components were similar or improved in heifers fed the distillers grains diets compared with heifers fed CON. Heifers fed LFDG had greater milk production and a tendency for greater milk protein yields compared with the heifers fed CON. Energy-corrected milk yields were similar among treatments. Feeding increased dietary fat from DDGS during the prepubertal growth phase did not negatively affect milk production, despite earlier attainment of puberty compared with other treatments. The overall ADG for all 3 treatments was 0.96kg/d during the prepubertal period, which is greater than recommended and may have compromised milk yield across treatments. Based on these findings, producers can feed either DDGS or LFDG with corn in replacement of soybean products and corn to prepubertal heifers and maintain or enhance subsequent reproductive and lactation performance. Dietary fat from DDGS can replace starch from corn as an energy source for prepubertal heifers without detriment to later performance., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Following injury to the CNS, astrocytes undergo a broad range of biochemical, morphological, and molecular changes collectively referred to as reactive astrogliosis. Reactive astrocytes exert both inflammatory and protective effects that inhibit and promote, respectively, neural repair. The mechanisms underlying the diverse functional properties of reactive astrogliosis are not well understood. Achieving a greater understanding of these mechanisms is critical to developing therapeutic strategies to treat the injured CNS. Here we demonstrate a method to trigger reactive astrogliosis in the adult mouse forebrain using a forebrain stab lesion. This lesion model is simple, reliable, and requires only a stereotaxic device and a scalpel blade to produce the injury. The use of stab lesions as an injury model in the forebrain is well established and amenable to studies addressing a broad range of neuropathological outcomes, such as neuronal degeneration, neuroinflammation, and disruptions in the blood brain barrier (BBB). Thus, the forebrain stab injury model serves as a powerful tool that can be applied for a broad range of studies on the CNS response to trauma.

Previous studies have shown that female rats exhibit different patterns of drug seeking during multiple phases of cocaine addiction when compared with males. However, the underlying mechanisms for these sex differences remain largely unknown. Here, we used a cocaine self-administration/reinstatement model to examine neuronal activation, as determined by Fos expression, following cue-induced reinstatement of cocaine seeking in male and female rats. Fos expression revealed both similarities between sexes in some brain regions, as well as selective sexually dimorphic patterns. As compared to no cue control subjects, conditioned cues induced higher Fos expression in the Cg1 region of the anterior cingulate cortex, but lower expression in the nucleus accumbens in both males and females. Females exhibited higher Fos expression than males in multiple brain regions, including the agranular insular cortex, dorsal medial caudate-putamen, nucleus accumbens shell, ventral tegmental area, dorsal subiculum, and ventral CA1 and CA3 regions of the hippocampus. Notably, only Fos expression in the prelimbic cortex, nucleus accumbens shell, basolateral amygdala, and ventral subiculum correlated positively with lever responding in response to conditioned cues across males and females. These findings indicate that while sexually dimorphic Fos activation does occur, the relationship between cue-induced cocaine seeking and neuronal activation may be similar for males and females in key brain regions of the relapse circuit.

Gangliosides are a class of sphingolipids characterized by a ceramide lipid chain attached to an anionic oligosaccharide moiety that varies in complexity based on the level of sialylation. Heterogeneity in the oligosaccharide chain of gangliosides is a direct result of the monosaccharide structure, content, sequence, and connections. Gangliosides are highly concentrated in the central nervous system, and are cell type-specific as well as development-dependent and their quantities and species can undergo drastic changes during cell differentiation. Specific localization of gangliosides also allows for interaction with a variety of bioeffectors, including glycoproteins, antibodies, peptide hormones, and growth factors. There are currently no rapid analytical assays capable of identifying and quantifying gangliosides. The aim of this study is to establish a reliable chromatographic mass spectrometry based assay capable of profiling ganglioside levels in complex biological samples at high sensitivity. We describe here a chromatographic method using an amino column on which the separation is based on hydrophilic interaction with the sugar moiety of gangliosides. Several gangliosides, including GM1-3, GD1a,b, GD2-3, and GT1a,b, were efficiently separated in less than 10 min at a limit of detection ranging between 10-50 pg on column with a concentration dynamic range extending over 4 orders of magnitude. The developed method allowed the sensitive quantitation of gangliosides derived from the blood serum of patients with different esophagus diseases, including, adenocarcinoma, high-grade dysplasia, and Barrett's., (Copyright © 2013 Elsevier B.V. All rights reserved.)