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By an unknown mechanism, alpha-synuclein (α-syn) inhibits autophagy in yeast and human cells. Herein, using the yeast Saccharomyces cerevisiae, we tested the hypothesis that α-syn disrupts autophagy by inhibiting the required association of sorting nexin 4 (Snx4) with phagophores. Snx4 contains a phox (PX) homology domain that selectively binds membranes enriched in phosphatidylinositol 3-phosphate (PI3P). Using fluorescence microscopy, we show that upon nitrogen starvation, 70% of the cells exhibited green puncta (phagophores); whereas identically treated cells expressing α-syn exhibited a significantly lower percentage of cells (30%) with such puncta. Our interpretation is that α-syn outcompetes Snx4 for binding to membranes enriched in PI3P, resulting in fewer phagophores and consequently inefficient induction of autophagy. As a control, we tested whether α-syn disrupts the binding of Vps27-GFP to late endosomes/multivesicular bodies (MVBs). Vps27 contains a PI3P-binding domain called FYVE. α-Syn did not disrupt the binding of Vps27-GFP to late endosomes. α-Syn likely inhibits the binding of PX- but not FYVE-containing proteins to PI3P because FYVE domains bind more than two-orders of magnitude tighter than PX domains. We propose that in all cells, whether yeast or human, α-syn has the potential to inhibit protein trafficking pathways that are dependent on PX-domain proteins such as sorting nexins.

22 pags., 8 figs., 1 tab., Massive stars disrupt their natal molecular cloud material through radiative and mechanical feedback processes. These processes have profound effects on the evolution of interstellar matter in our Galaxy and throughout the universe, from the era of vigorous star formation at redshifts of 1-3 to the present day. The dominant feedback processes can be probed by observations of the Photo-Dissociation Regions (PDRs) where the far-ultraviolet photons of massive stars create warm regions of gas and dust in the neutral atomic and molecular gas. PDR emission provides a unique tool to study in detail the physical and chemical processes that are relevant for most of the mass in inter-and circumstellar media including diffuse clouds, proto-planetary disks, and molecular cloud surfaces, globules, planetary nebulae, and star-forming regions. PDR emission dominates the infrared (IR) spectra of star-forming galaxies. Most of the Galactic and extragalactic observations obtained with the James Webb Space Telescope (JWST) will therefore arise in PDR emission. In this paper we present an Early Release Science program using the MIRI, NIRSpec, and NIRCam instruments dedicated to the observations of an emblematic and nearby PDR: the Orion Bar. These early JWST observations will provide template data sets designed to identify key PDR characteristics in JWST observations. These data will serve to benchmark PDR models and extend them into the JWST era. We also present the Science-Enabling products that we will provide to the community. These template data sets and Science-Enabling products will guide the preparation of future proposals on star-forming regions in our Galaxy and beyond and will facilitate data analysis and interpretation of forthcoming JWST observations., Support for JWST-ERS program ID 1288 was provided through grants from the STScI under NASA contract NAS5-03127 to STScI (K.G., D.V.D.P., M.R.), Univ. of Maryland (M.W., M.P.), Univ. of Michigan (E.B., F.A.), and Univ. of Toledo (T.S.-Y.L.). O.B. and E.H. are supported by the Programme National “Physique et Chimie du Milieu Interstellaire” (PCMI) of CNRS/INSU with INC/INP co-funded by CEA and CNES, and through APR grants 6315 and 6410 provided by CNES. E. P. and J.C. acknowledge support from the National Science and Engineering Council of Canada (NSERC) Discovery Grant program (RGPIN-2020-06434 and RGPIN-2021-04197 respectively). E.P. acknowledges support from a Western Strategic Support Accelerator Grant (ROLA ID 0000050636). J.R.G. and S.C. thank the Spanish MCINN for funding support under grant PID2019-106110GB-I00. Work by M.R. and Y.O. is carried out within the Collaborative Research Centre 956, subproject C1, funded by the Deutsche Forschungsgemeinschaft (DFG)—project ID 184018867. T.O. acknowledges support from JSPS Bilateral Program, grant No. 120219939. M.P. and M.W. acknowledge support from NASA Astrophysics Data Analysis Program award #80NSSC19K0573. C.B. is grateful for an appointment at NASA Ames Research Center through the San José State University Research Foundation (NNX17AJ88A) and acknowledges support from the Internal Scientist Funding Model (ISFM) Directed Work Package at NASA Ames titled: “Laboratory Astrophysics—The NASA Ames PAH IR Spectroscopic Database.”

Animal models are at the forefront of biomedical research for studies of viral transmission, vaccines, and pathogenesis, yet the need for an ideal large animal model for COVID-19 remains. We used a meta-analysis to evaluate published data relevant to this need. Our literature survey contained 22 studies with data relevant to the incidence of common COVID-19 symptoms in rhesus macaques (Macaca mulatta), cynomolgus macaques (Macaca fascicularis), African green monkeys (Chlorocebus aethiops), and ferrets (Mustela putorius furo). Rhesus macaques had leukocytosis on Day 1 after inoculation and pneumonia on Days 7 and 14 after inoculation, in frequencies that were similar enough to humans to reject the null hypothesis of a Fisher exact test. However, the differences in overall presentation of disease were too different from that of humans to successfully identify any of these 4 species as an ideal large animal of COVID-19. The greatest limitation to the current study is a lack of standardization in experimentation and reporting. To expand our understanding of the pathology of COVID-19 and evalu- ate vaccine immunogenicity, we must extend the unprecedented collaboration that has arisen in the study of COVID-19 to include standardization of animal-based research in an effort to find the optimal animal model.

We report the first spectroscopic observations of a background star seen through the region between the ionization front and the dissociation front of the nebula IC 63. This photodissociation region (PDR) exhibits intense extended red emission (ERE) attributed to fluorescence by large molecules/ions. We detected strong diffuse interstellar bands (DIB) in the stellar spectrum, including an exceptionally strong and broad DIB at $\lambda$4428. The detection of strong DIBs in association with ERE could be consistent with the suggestion that the carriers of DIBs and ERE are identical. The likely ERE process is recurrent fluorescence, enabled by inverse internal conversions from highly excited vibrational levels of the ground state to low-lying electronic states with subsequent transitions to ground. This provides a path to rapid radiative cooling for molecules/molecular ions, greatly enhancing their ability to survive in a strongly irradiated environment. The ratio of the equivalent widths (EW) of DIBs $\lambda$5797 and $\lambda$5780 in IC 63 is the same as that observed in the low-density interstellar medium with UV interstellar radiation fields (ISRF) weaker by at least two orders of magnitude. This falsifies suggestions that the ratio of these two DIBs can serve as a measure of the UV strength of the ISRF. Observations of the nebular spectrum of the PDR of IC 63 at locations immediately adjacent to where DIBs were detected failed to reveal any presence of sharp emission features seen in the spectrum of the Red Rectangle nebula. This casts doubts upon proposals that the carriers of these features are the same as those of DIBs seen at slightly shorter wavelengths., Comment: 13 pages, 9 figures

The human chemokines CCL19 and CCL21 bind to the G-protein coupled receptor (GPCR) CCR7 and play an important role in the trafficking of immune cells, as well as cancer metastasis. Conserved binding sites for sulfotyrosine residues on the receptor contribute significantly to the chemokine/GPCR interaction and have been shown to provide promising targets for new drug-discovery efforts to disrupt the chemokine/GPCR interaction, and consequently tumor metastasis. Here, we report the first X-ray crystal structure of a truncated CCL19 (residues 7–70) at 2.50 Å resolution, revealing molecular details crucial for protein-protein interactions. Although the overall structure is similar to the previously determined NMR model, there are important variations, particularly near the N-terminus and the so-called 30’s and 40’s loops. Computational analysis using the FTMap server indicates the potential importance of these areas in ligand binding, and the differences in binding hot spots compared to CCL21. NMR titration experiments using a CCR7-derived peptide (residues 5–11, TDDYIGD) further demonstrates potential receptor recognition sites, such as those near the C-terminus and 40’s loop, that consist of both positively charged and hydrophobic residues which may be important for receptor binding. Taken together, the X-ray, NMR, and computational analysis herein provides insights into the overall structure and molecular features of CCL19 and enables investigation into this chemokine’s function and inhibitor development.

Having an early family history of coronary artery disease (CAD) is strongly associated with incident cardio-vascular disease. In younger patients with acute coronary syndrome, an association between family history and the severity of CAD, determined by the number of affected coronary arteries diagnosed through coronary angiography, has been reported. However, it is unknown if these findings apply to unselected patients with a first-time myocardial infarction (MI) and whether an early family history of other atherosclerotic cardiovascular diseases (ASCVD) affect CAD severity. Therefore, our objective was to investigate the association between a family history of ASCVD and the extent of CAD determined by coronary angiography findings in a large nationwide sample of patients with a first-time MI. Material and methods This study included a consecutive sample of 22,686 one-year survivors of first-ever MI aged, 18 to 76 years, identified in the national register SWEDEHEART. Attending the standardized 1-year revisit after MI, from 2006 through 2013. First-degree relatives, defined as parents and full siblings, were identified in the Swedish Multi-Generation Register and data on historical hospitalisations and deaths were extracted. The exposure was defined as having a first-degree relative with records of early ASCVD, defined as a register-verified hospitalisation due to myocardial infarction, any angina with coronary revascularization, stroke or cardiovascular death before the age of 55 in male or 65 years in female relatives. The primary outcome was severity of CAD categorized to four groups: zero-, single-, two- or three-vessel disease as registered in SWEDE-HEART. Left main stem disease was included in the three-vessel category. The distribution of the CAD severity categories was compared between those with and without an early family history of ASCVD using multinomial logistic regression, adjusted for age and sex and using single-vessel disease as the reference group. Results We included 22 686 participants (median age 63 [IQR 57–68] years; 27% women), of which 1450 had a family history of early ASCVD. A total of 2134 patients did not have any significant obstructive findings at the first coronary angiography, 10 951 patients had single-vessel disease, 5516 patients had two-vessel disease and 3970 patients had three-vessel or main stem disease. Both two-vessel disease (RRR 1.31, 95% CI 1.14–1.50) and three vessel and/or left main stem disease (RRR 1.29, 95% CI 1.1–1.51) were more common in patients with early family history of ASCVD when adjusting for age and sex, whereas no significant angiographic disease was less common (RRR 0.76, 95% 0.61–0.95). Conclusion Patients with a first-time MI and a family history of early ASCVD have an approximately 30% higher risk of advanced coronary artery disease, with more than two affected coronary vessels or left main stem disease. Funding Acknowledgement Type of funding sources: Foundation. Main funding source(s): The Swedish Heart and Lung Association (Riksförbundet HjärtLung) Graph 1

We tested the ability of alpha-synuclein (α-syn) to inhibit Snx3-retromer-mediated retrograde trafficking of Kex2 and Ste13 between late endosomes and the trans-Golgi network (TGN) using a Saccharomyces cerevisiae model of Parkinson’s disease. Kex2 and Ste13 are a conserved, membrane-bound proprotein convertase and dipeptidyl aminopeptidase, respectively, that process pro-α-factor and pro-killer toxin. Each of these proteins contains a cytosolic tail that binds to sorting nexin Snx3. Using a combination of techniques, including fluorescence microscopy, western blotting and a yeast mating assay, we found that α-syn disrupts Snx3-retromer trafficking of Kex2-GFP and GFP-Ste13 from the late endosome to the TGN, resulting in these two proteins transiting to the vacuole by default. Using three α-syn variants (A53T, A30P, and α-synΔC, which lacks residues 101–140), we further found that A53T and α-synΔC, but not A30P, reduce Snx3-retromer trafficking of Kex2-GFP, which is likely to be due to weaker binding of A30P to membranes. Degradation of Kex2 and Ste13 in the vacuole should result in the secretion of unprocessed, inactive forms of α-factor, which will reduce mating efficiency between MATa and MATα cells. We found that wild-type α-syn but not A30P significantly inhibited the secretion of α-factor. Collectively, our results support a model in which the membrane-binding ability of α-syn is necessary to disrupt Snx3-retromer retrograde recycling of these two conserved endopeptidases.

The purpose of this study is to understand the role of α-synuclein in the regulation of the trafficking and release of modulators of metastasis in melanoma. Metastasis causes melanoma-induced death and 5-year survival rates, subsequent to metastasis to regional lymph nodes or distal organs, of 9% and 4%, respectively, hence novel therapeutic targets are required to control metastasis. Emerging evidence suggests that α-synuclein, a 14.4 KDa protein, which is a key player in Parkinson’s disease, is significantly correlated with melanoma progression. Our hypothesis is that α-synuclein positively regulates the release of extracellular vesicles. Extracellular vesicles (EVs) have been shown to be important in tumor metastasis, immune modulation and chemoresistance. The rationale for this hypothesis is that α-synuclein is a unique membrane-associated catalyst of endocytosis and exocytosis, and, specifically, synuclein regulates neurotransmitter release through SNARE complex assembly and the dilation of the fusion pore. The objective of the study is to measure the concentration of extracellular vesicles in the conditioned media from wild type SK-MEL-28 melanoma cells, SNCA knock out cells, and rescued SNCA-KO cells (where α-synuclein is re-expressed in KO cells via lentivirus) using Nanoparticle Tracking Analysis. The expected result was that knocking out SNCA would decrease the number of EVs released per cell relative to wild type control cells. To this end, we grew wild type, SNCA-KO and rescued SNCA-KO cells for 48 hours at 37°C and collected conditioned media. EVs were extracted from each conditioned medium using the ExoQuick-TC kit (Systems Bioscience) and stored at -80°C. EVs were characterized by PAGE followed by western blotting for common EV-markers and EV samples were shipped to the University of Colorado for Nanoparticle Tracking. Here we report a significant reduction in the concentration of EVs in SNCA-KO cell lines compared to control cells (P=0.0123), an effect that was rescued upon re-expression of α-synuclein in knock-out cells (P Citation Format: Nirjhar M. Aloy, Michael W. Graner, Christina Coughlan, Sergey V. Slepenkov, Sahar Shekoohi, Stephan N. Witt. Knocking out alpha synuclein causes decreased release of extracellular vesicles in melanoma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1575.

Malignant melanoma is one of the most aggressive and fifth most prevalent cancer among men and women. Clinical and epidemiological studies shows that Parkinson’s disease (PD) patients have higher risk of developing invasive melanoma, and, reciprocally, patients with invasive melanoma have a higher risk of developing PD. Alpha-synuclein (α-syn; SNCA), an intrinsically disordered protein expressed in neurons, is also highly expressed in malignant melanoma and is a common molecular signature between Parkinson’s disease and melanoma. We have previously reported that SK-Mel-28 SNCA-knockout (KO) cells exhibit suppressed tumor growth and proliferation in vivo and in vitro, and that putting back SNCA via lenti virus restored the phenotype to that of the control cells. The present study is designed to investigate the role of α-syn in melanoma progression in SK-MEL-28 cells in the context of metastasis and the epithelial-to-mesenchymal transition (EMT). The SK-Mel-28 SNCA-KO clones showed a significant decrease in the cell-surface protein L1CAM, a major cell adhesion molecule highly expressed in various melanoma and correlates with melanoma progression, whereas the knock in clones resembled the control. Migration and invasion assay were performed using transwell chambers showed a profound migratory and invasive defect in SNCA-KO clones. To explore further whether this is a proliferative or migratory defect, a single cell phagokinetic motility assay was performed with colloidal gold which showed a marked decrease in cell motility in knockout clones compared to control and knock in clones, which was consistent with our migration and invasion experiments. To determine the proteins involved in the inhibition of invasion mediated by α-syn, we immunoblotted for the mesenchymal markers N-cadherin and vimentin, and these were significantly downregulated in the SNCA-KO clones. These data reinforce that knocking out SNCA impedes melanoma cell migration and invasion in SK-Mel- 28 cell line, implying that α-syn might be a novel therapeutic target for malignant melanoma. Citation Format: Nithya Gajendran, Santhanasabapathy Rajasekaran, Nirjhar M. Aloy, Sahar Shekoohi, Stephan N. Witt. Knocking out SNCA in SKMEL28 melanoma cells suppresses EMT and prevents invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2443.

The protein alpha-synuclein (α-syn) is unusual because, depending on its conformation and the type of cell in which it is expressed, it is pro-death or pro-survival, triggering neurodegeneration in Parkinson’s disease and enhancing cell survival of some melanomas. To probe the function of α-syn in melanoma, we used CRISPR/Cas9 to knockout SNCA, the gene that codes for α-syn, in SK-Mel-28 melanoma cells. The SNCA-knockout clones in culture exhibited a decrease in the transferrin receptor 1 (TfR1), an increase in ferritin, an increase of reactive oxygen species and proliferated slower than control cells. These SNCA-knockout clones grafted into SCID mice grew significantly slower than the SK-Mel-28 control cells that expressed α-syn. In the excised SNCA-knockout xenografts, TfR1 decreased 3.3-fold, ferritin increased 6.2-fold, the divalent metal ion transporter 1 (DMT1) increased threefold, and the iron exporter ferroportin (FPN1) decreased twofold relative to control xenografts. The excised SNCA-KO tumors exhibited significantly more ferric iron and TUNEL staining relative to the control melanoma xenografts. Collectively, depletion of α-syn in SK-Mel-28 cells dysregulates cellular iron metabolism, especially in xenografts, yielding melanoma cells that are deficient in TfR1 and FPN1, that accumulate ferric iron and ferritin, and that undergo apoptosis relative to control cells expressing α-syn.

Extended Red Emission (ERE) is a widely observed optical emission process, present in a wide range of circumstellar and interstellar environments in the Milky Way galaxy as well as other galaxies. Definitive identifications of the ERE carriers and the ERE process are still a matter of debate. Numerous models have been proposed in recent decades, often developed without consideration of the growing body of observational constraints, which by now invalidate many of these models. This review focuses on the most well-established observational constraints which should help to delineate the way toward a generally accepted explanation of the ERE and an understanding of its place in the radiation physics of the interstellar medium., Comment: 10 pages, 2 figures, preprint to be published in the Topical Collection on "Unexplained Spectral Phenomena in the Interstellar Medium" of Astrophysics and Space Science

We probed the role of alpha-synuclein (α-syn) in modulating sorting nexin 3 (Snx3)-retromer-mediated recycling of iron transporters in Saccharomyces cerevisiae and Caenorhabditis elegans. In yeast, the membrane-bound heterodimer Fet3/Ftr1 is the high affinity iron importer. Fet3 is a membrane-bound multicopper ferroxidase, whose ferroxidase domain is orthologous to human ceruloplasmin (Cp), that oxidizes external Fe+2 to Fe+3; the Fe+3 ions then channel through the Ftr1 permease into the cell. When the concentration of external iron is low (1 µM), Fet3/Ftr1 is maintained on the plasma membrane by retrograde endocytic-recycling; whereas, when the concentration of external iron is high (10 µM), Fet3/Ftr1 is endocytosed and shunted to the vacuole for degradation. We discovered that α-syn expression phenocopies the high iron condition: under the low iron condition (1 µM), α-syn inhibits Snx3-retromer-mediated recycling of Fet3/Ftr1 and instead shunts Fet3/Ftr1 into the multivesicular body pathway to the vacuole. α-Syn inhibits recycling by blocking the association of Snx3-mCherry molecules with endocytic vesicles, possibly by interfering with the binding of Snx3 to phosphatidylinositol-3-monophosphate. In C. elegans, transgenic worms expressing α-syn exhibit an age-dependent degeneration of dopaminergic neurons that is partially rescued by the iron chelator desferoxamine. This implies that α-syn-expressing dopaminergic neurons are susceptible to changes in iron neurotoxicity with age, whereby excess iron enhances α-syn-induced neurodegeneration. In vivo genetic analysis indicates that α-syn dysregulates iron homeostasis in worm dopaminergic neurons, possibly by inhibiting SNX-3-mediated recycling of a membrane-bound ortholog of Cp (F21D5.3), the iron exporter ferroportin (FPN1.1), or both.

Background Clostridioides difficile (C difficile) infection (CDI) is a major health problem in the United States and despite updated guidelines, the laboratory diagnosis remains vexed. A multistep algorithm is recommended to diagnose CDI that includes antigen, toxin and toxin gene Nucleic Acid Amplification (NAAT) assays. This study was done to assess severity of CDI based on toxin B and NAAT statuses. The other objective was to analyze if antibiotics and PPI/H2B (Proton Pump Inhibitors and H2 blockers) affected severity of CDI. Methods Retrospective analysis of all adult patients admitted to a tertiary medical center with diarrhea and a positive C difficile antigen test from 01/2017- 12/2017. From more than 2000 stool samples submitted to the lab, C diff antigen was positive in 265 patients. 191 were diagnosed with CDI based on the 2-step algorithm. Clinical data was available for 168 patients. Severity of CDI was determined based on published guidelines. Fischer’s exact test was used for statistical analysis. Results The mean age at diagnosis was 55.96. Toxin B was detected in 34% (57/168) patients and Toxin NAAT positive in 66% (111/168) patients. 57% of CDI was health care onset compared to 43% with community onset. 42% (72/168) were classified as severe out of which 40.2% (29) were toxin B positive, and 59.8% (43) were NAAT positive. There were no significant differences in severity of CDI based on toxin B and NAAT status (50.9% vs 38.4%, p=0.14). 46% of cases from community vs 39.6% from hospitals were classified as severe CDI (p=0.415). 72% of cases had antibiotic use in the last 30 days. Use of antibiotics was significantly associated with severe CDI (82% vs 64%, p=0.015). 62.5% (105) patients had history of PPI/H2B use and severity was not significantly associated with its use (p=0.872). Conclusion Our study shows that the presence of toxin did not significantly impact the clinical severity of CDI. The use of antibiotics did not affect the presence of toxin although the total number of CDI cases with previous antibiotic exposure was high. Patients who had recent antibiotic exposure were more likely to have severe clinical presentation. More toxin positive cases were health care onset but the effect was not pronounced. Severity of CDI did not significantly depend on health care onset or on exposure to PPI/H2B. Disclosures Atul Kothari, MD, Ansun Biopharma (Consultant)

Many institutions overwhelm the first year seminar with “one-shot” library instruction sessions, which are not necessarily linked to any form of assignment or assessment. So how can librarians maintain information literacy instruction throughout a student's academic career? Data collected by the Rivier University librarians showcases the ability to implement information literacy more effectively by streamlining and leveling it out over a four-year period.

Sirtuin 5 (SIRT5) is a mitochondrial NAD+-dependent protein deacylase which regulates the enzymatic activity of numerous mitochondrial proteins due to increased succinylation and malonylation, including enzymes of energy substrate oxidation and mitochondrial antioxidant enzymes. Since energy depletion and mitochondrial oxidative stress contribute to myocardial IR injury, it was our objective to evaluate the potential role of SIRT5 in IR injury. In regular Langendorff heart perfusions, 8 week-old cardiomyocyte-selective SIRT5−/− mice showed no difference in LV developed pressure or dp/dt max compared to wildtype mice. However, recovery of LV developed pressure and dp/dt max following 25 min of ischemia was lower by 34% and 20% in SIRT5−/− mice compared to WT mice, respectively. In contrast, postischemic recovery of cardiac function was not impaired and even improved in mice with cardiomyocyte-selective overexpression of SIRT5 compared to WT mice undergoing IR. Mitochondrial H2O2 generation was significantly increased in SIRT5−/− mice compared to WT mice following IR, and mitochondria-targeted antioxidant treatments (MnTBAP or SS-31) during heart perfusion completely normalized recovery of contractile parameters in SIRT5−/− mice following IR. In conclusion, SIRT5 is not required to maintain cardiac function under physiological conditions. However, lack of SIRT5 aggravates myocardial IR injury, likely by increasing mitochondrial oxidative stress. SIRT5 agonism may thus represent a potential therapeutic strategy to attenuate myocardial IR injury.

We study the anomalous microwave emission (AME) in the Lynds Dark Nebula (LDN) 1780 on two angular scales. Using available ancillary data at an angular resolution of 1 degree, we construct an SED between 0.408 GHz to 2997 GHz. We show that there is a significant amount of AME at these angular scales and the excess is compatible with a physical spinning dust model. We find that LDN 1780 is one of the clearest examples of AME on 1 degree scales. We detected AME with a significance > 20$\sigma$. We also find at these angular scales that the location of the peak of the emission at frequencies between 23-70 GHz differs from the one on the 90-3000 GHz map. In order to investigate the origin of the AME in this cloud, we use data obtained with the Combined Array for Research in Millimeter-wave Astronomy (CARMA) that provides 2 arcmin resolution at 30 GHz. We study the connection between the radio and IR emissions using morphological correlations. The best correlation is found to be with MIPS 70$\mu$m, which traces warm dust (T$\sim$50K). Finally, we study the difference in radio emissivity between two locations within the cloud. We measured a factor $\approx 6$ of difference in 30 GHz emissivity. We show that this variation can be explained, using the spinning dust model, by a variation on the dust grain size distribution across the cloud, particularly changing the carbon fraction and hence the amount of PAHs., Comment: 14 pages, 11 figures, submitted to MNRAS

Background An antimicrobial stewardship program (ASP) strategy to minimize the use of overly broad antimicrobials is to suppress specific antimicrobial susceptibility results when isolates are sensitive to narrow antibiotics. There is limited data on possible adverse outcomes of this method. Patients with febrile neutropenia (FN) and gram-negative bacteremia (GNB) whose culture is sensitive to non-pseudomonal antibiotics still require broader pseudomonal coverage to treat the syndrome of FN. We evaluated if ASP suppression of anti-pseudomonal antibiotics adversely affects patients with FN and GNB. Methods In February 2018, our institution’s ASP began suppressing cefepime and meropenem susceptibility results from E. coli, Klebsiella spp, and Proteus spp when sensitive to cefepime (MIC ≤ 2), ceftriaxone and ceftazidime. We performed a retrospective analysis of patients with FN and GNB from 2016 – 2020 to evaluate the appropriateness of antibiotic regimens before and after the ASP intervention. Antibiotic regimens were deemed inappropriate if the patient was de-escalated to a narrow-spectrum, non-pseudomonal agent while neutropenic. Of 338 inpatient encounters identified with any bacteremia and FN, 49 were due to non-Pseudomonas, non-ESBL GNB, 20 before and 29 after the intervention. Sixteen of the 29 post-intervention patients were excluded, as their isolates did not meet suppression criteria. This resulted in a total of 13 patients in the post-intervention group. Results After culture susceptibility reports were released, 3 out of 20 patients in the pre-intervention group (15%) and 4 out of 13 patients in the post-intervention group (30.8%) were inappropriately tailored to narrow-spectrum antibiotics (p=0.39). There was no significant difference in 30-day mortality, 10.0% pre- and 0% post-intervention (p=0.50), or amount of meropenem prescribed, 45% pre- and 38.5% post-intervention (p=0.74). Conclusion These data show no significant difference in inappropriate antibiotic regimens prescribed for patients with FN and GNB after ASP antibiotic suppression was implemented. 30-day mortality was also not affected. The ASP intervention did not decrease meropenem prescriptions in this patient group, which may be appropriate. Larger studies are needed to verify these findings. Disclosures Ryan K. Dare, MD, MS, Accelerate Diagnostics, Inc (Research Grant or Support) Mary Burgess, MD, Pfizer Inc (Grant/Research Support)

Retromer is a phylogenetically conserved, multisubunit coat complex that controls endosomal protein trafficking and sorting. Mutations in the retromer gene VPS35 cause late-onset Parkinson disease, suggesting that trafficking defects cause neurodegeneration. Sorting nexins assist retromer to guide cell surface proteins to their assigned destinations, and our interest here is sorting nexin 3 (Snx3). Snx3 binds to membranes via a phox homolog (PX) domain that binds phosphatidylinositol 3-phosphate (PI3P), and in human cells its cargo proteins are the transferrin and Wnt receptors and the divalent metal ion transporter, whereas in yeast the best characterized cargo is the iron permease Ftr1. We recently discovered that α-synuclein inhibits Snx3-retromer recycling of Ftr1 in an unexpected way: α-synuclein, which avidly binds to negatively charged lipids, blocks the association of Snx3 to early endosomes. Here, we discuss mechanisms by which α-synuclein can disrupt Snx3-retromer-mediated recycling.

Background: The discovery of the mutation causing Huntington’s disease (HD) in 1993 allowed direct mutation analysis and predictive testing to identify currently unaffected carriers with a sensitivity and specificity of virtually 100%. Objective: The present study was designed to comprehensively profile the participants who sought predictive testing for HD between 1993 and 2009 in our Huntington centre. Methods: Using a retrospective design, we analysed the written documentation of the counselling sessions for all referrals for predictive mutation testing in this time span. Six hundred sixty-three individuals at risk for HD requested predictive testing. Roughly half (n = 333) completed the protocol and received their test result. Results: In general our findings are in accordance with other reports: most participants share an a priori risk of 50% (91.1%); more females request testing (58.5%); and those who ask for the result are mostly in their 30 s (mean = 35.1 years). Of those at 50% or 25% prior risks, 47.4% and 22.7%, respectively, tested positive in accordance with the respective risk of inheriting HD. Generally, more participants with an affected mother than father sought genetic testing (52.5% versus 47.5%). Interestingly, this difference was especially evident in the group of females who finally withdrew from testing (59.1%, p = 0.040). Men, in particular those who decided in favour of the test, were more often accompanied by their partner in the pre-test counselling session than vice versa (67.9% versus 44.7%, p = 0.003). On the other hand, significantly more men who were being tested did not have a companion in the pre-test session as compared with men who decided against the test (40.0% versus 25.7%, p = 0.012). During the first four years of predictive testing (1993–1996) more participants completed the protocol and received their test result as in later years. Yet, in this early time span significantly fewer females finally decided in favour of the test (48.4%, p = 0.005). These findings are discussed longitudinally and in the context of the experience in other centres. Conclusions: We present new gender-specific aspects of decision-making for predictive HD tests.

Barnard 207 (B207, LDN 1489, LBN 777), also known as the Vulture Head nebula, is a cometary globule in the Taurus-Auriga-Perseus molecular cloud region. B207 is known to host a Class I protostar, IRAS 04016+2610, located at a projected distance of ~8,400 au from the dense core centre. Using imaging and photometry over a wide wavelength range, from UV to sub-mm, we study the physical properties of B207 and the dust grains contained within. The core density, temperature, and mass are typical of other globules found in the Milky Way interstellar medium (ISM). The increase in the dust albedo with increasing optical wavelengths, along with the detection of coreshine in the near infrared, indicates the presence of larger dust grains in B207. The measured optical, near-, mid- and far-infrared intensities are in agreement with the CMM+AMM and CMM+AMMI dust grain type of The Heterogeneous dust Evolution Model for Interstellar Solids (THEMIS), suggesting mantle formation on the dust grains throughout the globule. We investigate the possibility of turbulence being responsible for diffusing dust grains from the central core to external outer layers of B207. However, in situ formation of large dust grains cannot be excluded., 13 pages, 11 figures. Accepted for publication in Astronomy & Astrophysics

Phosphatidylethanolamine (PE) is the second most abundant phospholipid in mammalian cells. PE comprises about 15–25% of the total lipid in mammalian cells; it is enriched in the inner leaflet of membranes, and it is especially abundant in the inner mitochondrial membrane. PE has quite remarkable activities: it is a lipid chaperone that assists in the folding of certain membrane proteins, it is required for the activity of several of the respiratory complexes, and it plays a key role in the initiation of autophagy. In this review, we focus on PE’s roles in lipid-induced stress in the endoplasmic reticulum (ER), Parkinson’s disease (PD), ferroptosis, and cancer.

We analysed new wide-field, wide- and narrow-band optical images of IC 59 and IC 63, two nebulae which are externally illuminated by the early B-star {\gamma} Cas, with the objective of mapping the extended red emission (ERE), a dust-related photoluminescence process that is still poorly understood, in these two clouds. The spatial distribution of the ERE relative to the direction of the incident radiation and relative to other emission processes, whose carriers and excitation requirements are known, provides important constraints on the excitation of the ERE. In both nebulae, we find the ERE intensity to peak spatially well before the more extended distribution of mid-infrared emission in the unidentified infrared bands, supporting earlier findings that point toward far-ultraviolet (11 eV < E$_\mathrm{{photon}}$ < 13.6 eV) photons as the source of ERE excitation. The band-integrated absolute intensities of the ERE in IC 59 and IC 63 measured relative to the number density of photons available for ERE excitation are lower by about two orders of magnitude compared to ERE intensities observed in the high-latitude diffuse interstellar medium (ISM). This suggests that the lifetime of the ERE carriers is significantly reduced in the more intense radiation field prevailing in IC 59 and IC 63, pointing toward potential carriers that are only marginally stable against photo-processing under interstellar conditions. A model involving isolated molecules or molecular ions, capable of inverse internal conversion and recurrent fluorescence, appears to provide the most likely explanation for our observational results., Comment: 19 pages, 11 figures, accepted for publication in MNRAS

The Parkinson’s disease-associated protein alpha-synuclein (α-syn) is often highly expressed in melanoma. α-Syn appears to exert a pro-survival function in melanoma cells, possibly by maintaining autophagy homeostasis. The long-term goal of our work is to determine the mechanism by which α-syn confers survival in melanoma cells. Herein, we used CRISPR (clustered regularly interspaced short palindromic repeats) to disrupt SNCA, the gene that codes for α-syn, in the melanoma cell line SK-Mel-28. Western blotting and real-time-qPCR verified that we generated four α-syn knockout (KO) clones. Three clones showed no expression of α-syn, whereas one had about 50% less α-syn than the control cells. DNA sequencing is consistent with three of the clones having both alleles of SNCA knocked out, whereas one clone has only one allele knocked out. α-Syn KO clones were tested in a variety of assays to ascertain the function of α-syn. First, the percentage of cells with exofacial phosphatidylserine (PS) was determined by annexin V-FITC/PI staining in conjunction with fluorescence-activated cell sorting. On average, there was a statistically significant 50% increase in exofacial PS (annexin V+/PI-) in three of the four α-syn KO clones compared to control cells. Exofacial PS was increased in the α-syn KO clones without a concomitant increase in apoptotic cells (annexin V+/PI+). Second, the clones were tested for growth in vivo using SCID mice. All four clones showed a ~65% reduction in the rate of tumor growth, followed over 12 weeks. Third, immunohistochemical staining of excised tumors from the SCID mice showed that the proliferation marker Ki-67 was significantly (p< 0.0001) lower in the tumors generated from the KO clones compared to tumors from control cells. In conclusion, given that α-syn avidly binds to PS, one possibility is that α-syn is like molecular Velcro that binds to PS molecules embedded in the inner leaflet, and this binding helps ATP-driven flippases keep PS in the inner leaflet. Loss α-syn then increases exofacial PS, which is a marker of senescence. Our working model is that α-syn is an anti-senescence protein, and that the α-syn KO clones grow slower than control cells in vivo because the clones have entered a senescent state. Citation Format: Sahar Shekoohi, Santhanasabapathy Rajasekaran, Shu Yang, Sureshbabu Nagarajan, Dhaval Patel, Xiuping Yu, Stephan N. Witt. Knocking out the gene (SNCA) that codes for the Parkinson’s disease-related protein alpha-synuclein in SK-Mel-28 melanoma cells significantly retards tumor growth in SCID mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 896.

Commission 21 was one of the oldest and smallest in the IAU yet one which underwent the most evolution since its inception. It began in 1955 as Light of the Night Sky (Lumière du Ciel Nocturne) under the Presidentship of Jean Dufay (University Observatory at Lyon). As the name suggested, in the years before space observations, its focus was on observations of atmospheric light which, at the time, was the domain of astronomers. Thus the early proceedings of the Commission were dominated by reports of the daytime and nighttime emission as observed from different locations and different conditions. Our knowledge of the Earth's atmosphere is still shaped by these early observations (Meier R.R., 1991, Space Sci. Rev. 58, 1). Members of Commission 21 were key to organizing interational collaborations to set up atmospheric stations at different latitudes, including in the Souther hemisphere and in ensuring a consistent calibration between the instruments. The Commission also bought in results from Soviet scientists in an era where communication was limited by both politics and language.

Blue luminescence (BL) and extended red emission (ERE) are observed as diffuse, optical-wavelength emissions in interstellar space, resulting from photoluminescence by ultraviolet(UV)-illuminated interstellar grains. Faintness and the challenge of separating the BL and ERE from the frequently much brighter dust-scattered continuum present major observational hurdles, which have permitted only slow progress in testing the numerous models that have been advanced to explain these two phenomena. Both the ERE, peaking near 680 nm (FWHM ~ 60 - 120 nm) and the BL, asymmetrically peaking at ~ 378 nm (FWHM ~ 45 nm), were first discovered in the Red Rectangle nebula. Subsequently, ERE and BL have been observed in other reflection nebulae, and in the case of the ERE, in carbon-rich planetary nebulae, H II regions, high-latitude cirrus clouds, the galactic diffuse ISM, and in external galaxies. BL exhibits a close spatial and intensity correlation with emission in the aromatic emission feature at 3.3 micron, most likely arising from small, neutral polycyclic aromatic hydrocarbon (PAH) molecules. The spectral characteristics of the BL also agree with those of fluorescence by PAH molecules with 13 to 19 carbon atoms. The BL phenomenon is thus most readily understood as the optical fluorescence of small, UV-excited aromatic molecules. The ERE, by contrast, though co-existent with mid-IR PAH emissions, does not correlate with emissions from either neutral or ionized PAHs. Instead, the spatial ERE morphology appears to be strictly governed by the density of far-UV (E ≥ 10.5 eV) photons, which are required for the ERE excitation. The most restrictive observational constraint for the ERE process is its exceptionally high quantum efficiency. If the ERE results from photo-excitation of a nano-particle carrier by photons with E ≥ 10.5 eV in a single-step process, the quantum efficiency exceeds 100%. Such a process, in which one to three low-energy optical photons may be emitted following a single far-UV excitation, is possible in highly isolated small clusters, e.g. small, dehydrogenated carbon clusters with about 20 to 28 carbon atoms. A possible connection between the ERE carriers and the carriers of DIBs may exist in that both are ubiquitous throughout the diffuse interstellar medium and both have an abundance of low-lying electronic levels with E ≤ 2.3 eV above the ground state.

Sirtuins (SIRTs) are NAD(+)-dependent enzymes that catalyze deacylation of protein lysine residues. In mammals, seven sirtuins have been identified, SIRT1-7. SIRT3-5 are mainly or exclusively localized within mitochondria and mainly participate in the regulation of energy metabolic pathways. Since mitochondrial ATP regeneration is inevitably linked to the maintenance of cardiac pump function, it is not surprising that recent studies revealed a role for mitochondrial sirtuins in the regulation of myocardial energetics and function. In addition, mitochondrial sirtuins modulate the extent of myocardial ischemia reperfusion injury and the development of cardiac hypertrophy and failure. Thus, targeting mitochondrial sirtuins has been proposed as a novel approach to improve myocardial mitochondrial energetics, which is frequently impaired in cardiac disease and considered an important underlying cause contributing to several cardiac pathologies, including myocardial ischemia reperfusion injury and heart failure. In the current review, we present and discuss the available literature on mitochondrial sirtuins and their potential roles in cardiac physiology and disease.

Protein aggregation is a common problem in protein biochemistry and is linked to many cellular pathologies and human diseases. The molecular chaperone ClpB can resolubilize and reactivate aggregated proteins. This unit describes the procedure for following reactivation of an aggregated enzyme glucose-6-phosphate dehydrogenase mediated by ClpB from Escherichia coli in cooperation with another molecular chaperone, DnaK. The procedures for purification of these chaperones are also described.

We have been investigating the role that phosphatidylethanolamine (PE) and phosphatidylcholine (PC) content plays in modulating the solubility of the Parkinson's disease protein alpha-synuclein (α-syn) using Saccharomyces cerevisiae and Caenorhabditis elegans. One enzyme that synthesizes PE is the conserved enzyme phosphatidylserine decarboxylase (Psd1/yeast; PSD-1/worms), which is lodged in the inner mitochondrial membrane. We previously found that decreasing the level of PE due to knockdown of Psd1/psd-1 affects the homeostasis of α-syn in vivo. In S. cerevisiae, the co-occurrence of low PE and α-syn in psd1Δ cells triggers mitochondrial defects, stress in the endoplasmic reticulum, misprocessing of glycosylphosphatidylinositol-anchored proteins, and a 3-fold increase in the level of α-syn. The goal of this study was to identify drugs that rescue this phenotype. We screened the Prestwick library of 1121 Food and Drug Administration-approved drugs using psd1Δ + α-syn cells and identified cyclosporin A, meclofenoxate hydrochloride, and sulfaphenazole as putative protective compounds. The protective activity of these drugs was corroborated using C. elegans in which α-syn is expressed specifically in the dopaminergic neurons, with psd-1 depleted by RNAi. Worm populations were examined for dopaminergic neuron survival following psd-1 knockdown. Exposure to cyclosporine, meclofenoxate, and sulfaphenazole significantly enhanced survival at day 7 in α-syn-expressing worm populations whereby 50-55% of the populations displayed normal neurons, compared to only 10-15% of untreated animals. We also found that all three drugs rescued worms expressing α-syn in dopaminergic neurons that were deficient in the phospholipid cardiolipin following cardiolipin synthase (crls-1) depletion by RNAi. We discuss how these drugs might block α-syn pathology in dopaminergic neurons.

Parkinson disease (PD) results from the slow, progressive loss of dopaminergic neurons in the substantia nigra. Alterations in α-synuclein (aSyn), such as mutations or multiplications of the gene, are thought to trigger this degeneration. Here, we show that aSyn disrupts mitogen-activated protein kinase (MAPK)-controlled stress signaling in yeast and human cells, which results in inefficient cell protective responses and cell death. aSyn is a substrate of the yeast (and human) polo-like kinase Cdc5 (Plk2), and elevated levels of aSyn prevent Cdc5 from maintaining a normal level of GTP-bound Rho1, which is an essential GTPase that regulates stress signaling. The nine N-terminal amino acids of aSyn are essential for the interaction with polo-like kinases. The results support a unique mechanism of PD pathology.

The Prestwick and NIH chemical libraries were screened for drugs that protect baker’s yeast from sugar-induced cell death (SICD). SICD is triggered when stationary-phase yeast cells are transferred from spent rich medium into water with 2% glucose and no other nutrients. The rapid, apoptotic cell death occurs because reactive oxygen species (ROS) accumulate. We found that triclabendazole, which is used to treat liver flukes in cattle and man, partially protects against SICD. Characterization of triclabendazole revealed that it also protects yeast cells from death induced by the Parkinson’s disease-related protein alpha-synuclein (α-syn), which is known to induce the accumulation of ROS.

Stationary-phase Saccharomyces cerevisiae cells transferred from spent rich media into water live for weeks, whereas the same cells die within hours if transferred into water with 2% glucose in a process called sugar-induced cell death (SICD). Our hypothesis is that SICD is due to a dysregulated Crabtree effect, which is the phenomenon whereby glucose transiently inhibits respiration and ATP synthesis. We found that stationary-phase cells in glucose/water consume 21 times more O2 per cell than exponential-phase cells in rich media, and such excessive O2 consumption causes reactive oxygen species to accumulate. We also found that inorganic phosphate and succinate protect against SICD but by different mechanisms. Phosphate protects by triggering the synthesis of Fru-1,6-P2, which inhibits respiration in isolated mitochondria. Succinate protects in wild-type cells but fails to protect in dic1Δ cells. DIC1 codes for a mitochondrial inner membrane protein that exchanges cytosolic succinate for matrix phosphate. We propose that succinate depletes matrix phosphate, which in turn inhibits respiration and ATP synthesis. In sum, restoring the Crabtree effect, whether with phosphate or succinate, protects cells from SICD.

The diffuse cm-wave IR-correlated signal, the "anomalous" CMB foreground, is thought to arise in the dust in cirrus clouds. We present Cosmic Background Imager (CBI) cm-wave data of two translucent clouds, {\zeta} Oph and LDN 1780 with the aim of characterising the anomalous emission in the translucent cloud environment. In {\zeta} Oph, the measured brightness at 31 GHz is 2.4{\sigma} higher than an extrapolation from 5 GHz measurements assuming a free-free spectrum on 8 arcmin scales. The SED of this cloud on angular scales of 1{\odot} is dominated by free-free emission in the cm-range. In LDN 1780 we detected a 3 {\sigma} excess in the SED on angular scales of 1{\odot} that can be fitted using a spinning dust model. In this cloud, there is a spatial correlation between the CBI data and IR images, which trace dust. The correlation is better with near-IR templates (IRAS 12 and 25 {\mu}m) than with IRAS 100 {\mu}m, which suggests a very small grain origin for the emission at 31 GHz. We calculated the 31 GHz emissivities in both clouds. They are similar and have intermediate values between that of cirrus clouds and dark clouds. Nevertheless, we found an indication of an inverse relationship between emissivity and column density, which further supports the VSGs origin for the cm-emission since the proportion of big relative to small grains is smaller in diffuse clouds.

The diffuse high-latitude H-alpha background is widely believed to be predominantly the result of in-situ recombination of ionized hydrogen in the warm interstellar medium of the Galaxy. Instead, we show that both a substantial fraction of the diffuse high-latitude H-alpha intensity in regions dominated by Galactic cirrus dust and much of the variance in the high-latitude H-alpha background are the result of scattering by interstellar dust of H-alpha photons originating elsewhere in the Galaxy. We provide an empirical relation, which relates the expected scattered H-alpha intensity to the IRAS 100um diffuse background intensity, applicable to about 81% of the entire sky. The assumption commonly made in reductions of CMB observations, namely that the observed all-sky map of diffuse H-alpha light is a suitable template for Galactic free-free foreground emission, is found to be in need of reexamination., Comment: 26 pages, 5 figures, Accepted for publication in ApJ

Because over expression of Hsp70 molecular chaperones suppresses the toxicity of aberrantly folded proteins that occur in Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis, and various polyQ-diseases (Huntington's disease and ataxias), Hsp70 is garnering attention as a possible therapeutic agent for these various diseases. Here, I review progress in this fascinating field of molecular chaperones and neurodegeneration and describe our current understanding of the mechanisms by which Hsp70 protects cells from the PD-related protein called alpha-synuclein (alpha-syn).

Angiographic Moyamoya is a rare cerebrovascular disease most frequent in asia. Its characateristics are recurrent ischemic attacks due to progressive occlusion of ICA branches. Angiography reveals fine arterial collateralisation reminding of ascending smoke ("moyamoya" in japanese). Neurosurgical treatment strategies include direct and indirect reanastomosation procedures. Randomised trials for comparison of clinical outcome and long term survival remain missing. A 23 years old female with glycogenosis type IA was first diagnosed bilateral angiographic moyamoya with bilateral proximal stenosis of ICA after transient ischemic attack (TIA). Coincidence of both rare diseases moyamoya and glycogenosis has previously been reported in three cases, so that this metabolic dysfunction presumably is a true risk factor for moyamoya. In our case, excellent angiographic and functional results were achieved by bilateral, consecutive Enzephalo-Duro-Arterio-Myo-Synangiosis (EDAMS).

Biofilms in surface waters primarily consist of allochthonous microorganisms. Under conditions of pollution faecally derived bacteria may interact with these biofilms. Total coliform bacteria, Escherichia coli and intestinal enterococci are used to monitor source water quality, indicating faecal pollution and the possible presence of enteric pathogens. In the present study the occurrence of faecal indicators was investigated in biofilms (epilithic biofilms, sediments) of German rivers. All of the biofilms contained significant concentrations of these bacteria, which were several orders of magnitude lower compared with the total cell number and the number of culturable heterotrophic plate count bacteria indicating that faecal indicator bacteria represented a minor fraction of the whole biofilm communities. The biofilms displayed approximately two orders of magnitude higher concentrations of total coliforms, E. coli and enterococci compared with the overlying water. Identification of coliform and enterococcal isolates from the biofilms revealed the presence of species which are known to be opportunistic pathogens. Overall, the results of the present study show that faecal indicator bacteria can survive in the presence of high cell densities of the authochthonous microflora in epilithic biofilms and sediments, suggesting that these biofilms may act as a reservoir for bacterial pathogens in polluted rivers.