Indiana University-Purdue University Indianapolis (IUPUI) / During 2020, the Indiana State Department of Health conducted a longitudinal study of novel severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) virus, the cause of COVID-19 disease, to understand the number of past and current infections as well as the prevalence of disease in the State of Indiana by conducting a survey to participants as well as administering testing for exposure to SARS-COV-2. The study consisted of 3 waves of testing, each spread months apart, consisting of a random sample and a non-random sample. The non-random sample was used to ensure the sample population was representative of the state of Indiana and was used as stratum in the logistic regression model, allowing for the adjustment for nonresponse. These finding indicate that persons of non-White race and persons of Hispanic ethnicity had highest risk of exposure to the virus. Understanding the disparity in health in various racial and ethnic populations and addressing how different communities are impacted by the pandemic, as well as working with the community is paramount when attempting to mitigate a pandemic. In addition, understanding the data from the ambient pandemic when instituting measures to mitigate the spread of viruses is also extremely important for managing health emergencies such as the COVID-19 pandemic.

After implementing a reliable mass spectrometry based kinetic study the indole conjugation with different organometallic indoles led to questions about the electronical and sterical influences on reactivity. The substitution pattern of the ferrocene functionalized indoles at the six-membered ring determines the electron density on the C3 atom, which reacts with the formed Schiff base. Since the experimental results showed the exact opposite trend, covalent docking studies were performed elucidating the importance of surface interactions. These studies were in harmony with the experimental results and determined lysine 33 as most preferable conjugation site as well as substitution in 6-position as most favourable pattern. The amine motif in compounds 6, 7 and 8 proofed to be easily fragmented by the ESI method used. The amide linker in 10 remains intact but shows a lower conversion. Those two inherent characteristics are however preferable for well-defined and site-specific bioconjugation. The synthesis and evaluation of piano stool complex derivatives with manganese and rhenium metal centre 15, 16, 18 and 22 gave additional guidance by the interpretation of applicable structural motifs. The electron-withdrawing carbonyl groups lead to the hindrance of fulvene formation and thus to no fragmentation as seen with the ferrocene group. The total conversion is low compared to 8, only 22 shows a good enough conversion to mainly monoconjugate of 45% and a possible radio-labelling application as 99mTc analogue. As consequence manganese complexes with a stable facial tricarbonyl unit and a tridentate chelator with 4-, 5- and 6-substituted aminomethylindole conjugated through an amide bond were synthesized and consecutively evaluated. The resulting organometallic indole derivatives 29, 30 and 31 all showed a total conversion around 40% similar to 16, but at the same time a rate constant in the range of 10-4 s-1 like the organic indole. Besides the similar conversion, the rate constants followed the trend of the 6-substituted derivative as fastest and then 5- and 4- substituted derivative with decreasing reactivity. For underlining the usage as technetium label for the best out of the series 31, a rhenium analogue was prepared. The resulting compound 32 was especially interesting, because the conversion was even higher than the 70% of 8 with a total of 88%. Additionally, the rate constant was a tenfold higher as well. This rendered compound 32 as best possible 99mTc analogue for further application as radio-label. After the success of 32 and realizing the sterical benefits resulting from the flexible tridentate ligand-system, substitution at the five-membered ring was explored. The complexes 33, 34 and 35 are based on indole-2-carboxylic acid and with the difference of the length of the alkyl spacer between amide and complex to probe for the influence and sterical hindrance, but all three derivatives showed no conjugation which excludes functionalization in 2-position. As the C3 is used for the actual bioconjugation, the last possible derivatization was realized on the indole-N1 by using 1-(3-bromopropyl)indole as building block during the synthesis of the ligand-system. The corresponding manganese 36 and rhenium 37 complexes both showed similar properties of a moderate conversion like 22 and a rate constant in the range of 10-5 s-1. In conclusion the rhenium complex 32 with the 6-substitution pattern at the tridentate indole-bearing ligand remains the most promising structure. The here developed liquid chromatography coupled mass spectrometry-based assay for the determination of inhibitory activity of drug candidates against the 3CLpro of the sever acute respiratory syndrome coronavirus type 2 was successfully implemented and especially designed to give, due to the available absorption spectra and corresponding mass traces, further insight in the otherwise through fluorescence resonance energy transfer-based assays neglected influences on the inhibition results. Starting with a literature-known quinolone containing covalent inhibitor 42 an N1-methylated derivative 43 and their analogues 44 and 45 in which the benzoic acid was exchanged for ferrocene carboxylic acid were synthesized. The inhibition of 3CLpro was evaluated by the concentration of initial 15mer peptide left after incubation and for that purpose the for 280 nm defined molar attenuation coefficient of (26.41±0.59) L*mol-1*cm-1 determined and used. The results showed a reaction of DL dithiothreitol with the less stable benzoic acid esters leading to a moderate inhibitory effect. The methylation in N1-position showed an increase in stability. The methylated and with ferrocene carboxylic acid functionalized derivative showed a complete inhibition during the timeframe of the assay. In search of a fluorescent and therefore traceable inhibitor, 4 hydroxycoumarin was used to synthesize the analogue with benzoic acid 49 and ferrocene carboxylic acid 50. Both derivatives were less stable than their analogues but exhibited the same trend of a more stable ferrocene-derived compound, which exerted a higher inhibition as well. After preparing and testing the model thioester 53 and showing an inactivation of the established inhibitor ebselen, it was concluded that the reaction with DL dithiothreitol reduces the concentration of active intact inhibitor and therefore decreases the inhibition rate during the assay. The next step was proofing the reducing agent as non-essential for the fast assay conducted in a timeframe of 5 min to circumvent the negative influence of DL dithiothreitol. By excluding every inhibition-altering part, the resulting method is the perfect tool for precise statements in relation of inhibitory activity. Then the inhibition assay was repeated for ebselen and the best out of the here introduced organometallic inhibitors 45. Both give equivalent results of a complete inhibition during the measurement. The implemented liquid chromatography coupled mass spectrometry-based assay has many advantages over the fluorescence resonance energy transfer-based assays in which all the information and insight accumulated by the evaluation of uv/vis traces and mass spectra are not available leading to wrong or deviating results regarding the inhibitory capacity of inhibitor candidates. / Nach der erfolgreichen Implementierung einer auf Massenspektrometrie basierenden Methode, um kinetische Studien über die Indolkonjugation durchzuführen, kamen Fragen über die elektronischen und sterischen Einflüsse auf die Reaktivität auf. Das Substitutionsmuster der Ferrocen-funktionalisierten Indole am sechsgliedrigen Ring bestimmt die Elektronendichte am C3-Atom, welches wiederum mit der zuvor entstandenen Schiffbase reagiert. Da die experimentellen Daten einen der Elektronendichte entgegengesetzten Trend aufwiesen, wurden Docking-Studien durchgeführt, um die Einflüsse der Wechselwirkungen mit der Proteinoberfläche näher zu beleuchten. Diese entsprachen den experimentellen Werten und zeigten, dass Lysin 33 die bevorzugte Konjugationsstelle ist und dass die Substitution in 6-Position die energetisch günstigste darstellt. Das sekundäre Amin der Verbindungen 6, 7 und 8 wurde unter den ESI-Bedingungen leicht fragmentiert. Die Amid-Bindung von 10 dagegen bleibt intakt und ein langsamerer Umsatz, welcher für einen wohl definierten Umsatz sorgt, wurde beobachtet. Die Synthese und Evaluation der Halbsandwich-Komplexe 15, 16, 18 und 22 gab zusätzlichen Aufschluss über die strukturellen Einflüsse. Die elektronenziehenden Carbonyle behindern die Bildung der Fulvene, welche für die Ferrocen-Derivate beobachtet worden war. Der Gesamtumsatz ist vergleichsweise gering und nur Substanz 22 zeigt eine annehmbare Konversion von 45% an Monokonjugat auf, welches sie wiederum für den Einsatz als 99mTc-Analog qualifiziert. Als direkte Konsequenz wurden Mangankomplexe mit einer stabilen facialen Tricarbonyl-Einheit und einem Indol beinhaltenden tridentaten Liganden synthetisiert. Dies wurde für die 4-, 5- und 6-Substitution durchgeführt um die Komplexe 29, 30 und 31 zu erhalten. Alle drei zeigten eine Konversion von ungefähr 40% und lagen mit einer Ratenkonstante von 10-4 s-1 in dem Bereich des organischen Indols. Die Ratenkonstanten folgten hingegen wiederum dem Trend der begünstigten 6-Position, gefolgt von der 5- und dann 4-Position. Um einen Verglich zu einer analogen Technetium-Verbindung zu ziehen wurde der beste Komplex 31 als Rhenium-Analog 32 hergestellt. Dies wies den bisher höchsten Gesamtumsatz von 88% auf, welches die Konversion von 70% der Verbindung 8 überschritt. Dadurch qualifizierte sich Verbindung 32 als beste Option für eine Umsetzung als Radiomarker. Nach den Erkenntnissen über die begünstigte Sterik der flexiblen tridendaten Liganden wurde versucht den Erfolg auf den Fünfring zu übertragen. Eine auf Indol-2-säure basierende Reihe an Komplexen (33,34,35) mit unterschiedlich langen Alkyl-Ketten zwischen Indol und Metallkomplex sollte Aufschluss über den sterischen Anspruch geben, allerdings war keine Konjugation für alle drei Derivate zu beobachten. Als Konsequenz wurde die 2-Position für weitere Funktionalisierung nicht weiter berücksichtigt. Die letzte verbliebene Stelle war das Indol-amin selbst, welche durch den Einsatz von 1-(3-bromopropyl)indol bei der Ligandensynthese erfolgreich alkyliert wurde. Der resultierende Mangan- 36 und Rhenium-Komplex 37 zeigten eine mit 22 vergleichbare moderate Konversion und um eine Größenordnung kleinere Ratenkonstante. Durch diese Erkenntnisse verblieb der Rhenium-Komplex 32 weiterhin der beste Kandidat für eine Umsetzung als radioaktiver Marker. Der hier beschriebene liquid chromatography coupled mass spectrometry basierte Assay für die Bestimmung der Inhibitions-Wirkung gegenüber der 3CLpro des sever acute respiratory syndrome coronavirus type 2 wurde erfolgreich etabliert. Durch die zusätzlichen Absorptions , sowie Massenspektren wurden weitere Informationen gewonnen, welche in den oft angewendeten Fluoreszenz-Resonanzenergietransfer Assays vernachlässigt werden und zu Verfälschungen der Inhibitionsergebnisse führen. Ausgehend von dem literaturbekannten kovalenten Inhibitor 42 wurde ein N1-methyliertes Derivat 43 hergestellt. Durch den Austausch von Benzoesäure durch Ferrocensäure wurden die Analoge 44 und 45 erhalten. Die Restkonzentration des 15mer Substrats nach der Inkubation wurde als Maß der Inhibition herangezogen. Hierfür wurde der molare Extinktionskoeffizient bei 280 nm mit (26.41±0.59) L*mol-1*cm-1 bestimmt. Die Methylierung führte zu einer erhöhten Stabilität. Der Wirkstoffkandidat 45 zeigte außerdem eine vollständige Inhibition im Rahmen des durchgeführten Assays. 4-Hydroxycoumarin wurde verwendet, um die fluoreszenten Analoga 49 und 50 herzustellen. Beide waren instabiler, wobei das Ferrocen-Derivat wie zuvor eine erhöhte Inhibition aufzeigte. Nach der Synthese und Auswertung des Assays für eine Thioester Modellverbindung 53 und einer nachgewiesenen Inaktivierung des etablierten Inhibitors Ebselen wurden die Nebenreaktionen mit DL-Dithiothreitol als Ursache für einen Konzentrationsabfall der aktiven und intakten Inhibitoren und infolgedessen verringerte Inhibition angenommen. Als nächstes wurde bewiesen, dass der Zusatz eines Reduktionsmittels wie keinen Einfluss auf die Enzymaktivität während des kurzen Assays hat. So wurden alle Einflüsse auf die Inhibitionswirkung ausgeschlossen und der nun makellose Assay für Ebselen und den hier vorgestellten neuartigen organometallischen Inhibitor 45 wiederholt, welche beide eine vollständige Inhibition aufwiesen.

Introducción: La pandemia por SARS-Cov-2 ha tenido un impacto negativo en múltiples aspectos de la vida humana, tanto en lo físico, psicológico, económico, social y cultural. Las afecciones de la calidad de vida están asociadas al confinamiento y la libertad de salir, de pasar tiempo con sus amigos o familiares, o de realizar actividades; por lo que se ven privados de la mayor parte de su interacción social. El objetivo de este presente artículo es brindar información sobre cómo afecta el confinamiento y aislamiento social por la pandemia a la calidad de vida y salud mental.

Im sechsten Semester des Medizinstudiums an der Julius-Maximilians-Universität Würzburg findet das verpflichtende Praktikum „Impfkurs“ statt. Im Rahmen dieses Kurses wurde vom Sommersemester 2020 bis zum Sommersemester 2021 ein standardisierter online Fragebogen erhoben, der unter anderem demographische Daten sowie Expositionsmöglichkeiten gegenüber SARS-CoV-2 im privaten, beruflichen und universitären Umfeld erfragte. Zusätzlich wurde im gleichen Zeitraum der SARS-CoV-2 Serostatus der Medizinstudierenden erhoben und ausgewertet und dieser mit den Daten des Fragebogens zusammengeführt. Dafür wurden Blutproben entnommen, welche im Labor des Instituts für Virologie der Universität Würzburg mittels Western Blot auf IgG/IgM/IgA Antikörper gegen SARS-CoV-2 untersucht wurden. / In the sixth semester of medical studies at the Julius-Maximilians-Universität Würzburg, the compulsory internship “vaccination course” takes place. As part of this course, a standardized online questionnaire was collected from the summer semester 2020 to the summer semester 2021, which, among other things, collected demographic data and exposure to SARS-CoV-2 in the private, professional and university environment. In addition, the SARS-CoV-2 serostatus of the medical students was collected and evaluated during the same period and merged with the data from the questionnaire. For this purpose, blood samples were taken, which were tested for IgG/IgM/IgA antibodies against SARS-CoV-2 by Western blot.

A finales de 2020 se aprobaron las vacunas desarrolladas en el mundo occidental contra el virus SARS-CoV-2, que ya están siendo inoculadas, conjuntamente con vacunas chinas y rusas. Mientras tanto, estamos en una segunda oleada de la enfermedad y el nuevo coronavirus se ha ido transformando para permitirse una mejor propagación, alojamiento y replicación en el ser humano. La enfermedad se manifiesta ahora con nueva sintomatología, mayor contagio, inclemencia y variación en el número de fallecimientos. La infección de la gestante por coronavirus se está presentando con severidad y consecuencias materno perinatales. Ya se inició la vacunación en gestantes y madres lactantes, previa conversación con su ginecólogo sobre los riesgos y beneficios. Este artículo ofrece un breve rel los acontecimientos que tuvieron lugar durante la transición de 2020 a 2021. / In late 2020, vaccines developed in the Western world against the SARS-CoV-2 virus were approved and are currently being inoculated, together with Chinese and Russian vaccines. In the meantime, we are in a second wave of the disease and the new coronavirus has been transforming to allow for better propagation, harboring and replication in humans. The disease now manifests itself with new symptoms, greater contagiousness, severity and variation in the number of deaths. Coronavirus infection of pregnant women is occurring with harshness and maternal and perinatal consequences. Vaccination has been initiated in pregnant women and nursing mothers, after discussion with their gynecologists about risks and benefits. This article provides a b ok place during the transition from 2020 to 2021.

Boletín que incluye información científica sobre el COVID-19, incluye artículos científicos y artículos preprint actualizados al 27 de Mayo de 2020.

The development of molecular tools that enable precise manipulation and control of biological systems would allow for a broader understanding of cellular functions and applications in biotechnology, synthetic biology, and therapeutic research. The discovery of CRISPR/Cas systems and the understanding and repurposing of their mechanisms have revolutionized the field of molecular biology. Here, I identified and characterized novel CRISPR/Cas systems and applied them for different in vivo and in vitro applications. In this work, I interrogated various Cas13 effector proteins and identified the most efficient Cas13 effector (CasRx) for in planta applications. I adapted CasRx to engineer plant immunity against different plant RNA viruses. CasRx showed robust activity and specificity against RNA viruses, demonstrating its suitability for studying key questions relating to virus biology. To expand the Cas13 toolbox and enable new applications, I performed a homology search of Cas13 enzymes in prokaryotic genomes and metagenomes, and identified previously uncharacterized, novel CRISPR/Cas13 effector proteins. I first identified and functionally characterized a small size, miniature Cas13 effector (named here as mCas13) and combined it with isothermal amplification to develop a simple and sensitive CRISPR-based SARS-CoV-2 diagnostic platform. In addition, I discovered and biochemically characterized the first known thermostable Cas13 proteins and showed that these thermostable proteins are phylogenetically related. I harnessed the unique features of these thermostable enzymes to develop the first one-pot, RT-LAMP coupled Cas13-based nucleic acid detection assay, which was utilized for highly sensitive, specific, and easily programmable detection of SARS-CoV-2 and other viruses. Lastly, I utilized CRISPR/Cas12a to develop a detection assay of plant ssDNA geminiviruses with easy-to-interpret visual readouts, making it suitable for point-of-use applications. In addition, I leveraged the self vs. non-self-discrimination and pre-crRNA processing capabilities of CRISPR/Cas12a, with the allosteric transcription factors (aTFs)- regulated expression of CRISPR array to engineer a field-deployable small molecule detection platform. I demonstrated the ability of the developed platform to detect different tetracycline antibiotics with high sensitivity and specificity. In conclusion, my work demonstrates that the discovery and characterization of programmable nucleic acid targeting systems could enable their utility for biotechnological innovations, including technologies for inhibition of viral replication and diagnostics.

La pandemia del nuevo coronavirus continúa con nosotros y lo hará por largo tiempo. Ha causado un nuevo modo de vivir, con aislamiento, protección personal, distanciamiento, empleo de la virtualidad y otros. Se ha mejorado el diagnóstico del infectado y su manejo. No existe cura aún, aunque se cuenta con vacunas aprobadas con premura. La crisis de salud ha desnudado la falta de preparación de nuestros sistemas de salud, y ha devenido en crisis políticas y económicas, empobrecimiento, muerte e inquietud emocional y psicológica. En estas páginas continuamos escribiendo de manera resumida los nuevos conocimientos sobre el SARS-CoV-2 y la enfermedad COVID-19, su diagnóstico, fisiopatología, manejo sintomático y de la enfermedad severa, la reinfección, sus secuelas y letalidad. Pero, principalmente, el compromiso de la mujer infectada durante el embarazo y el parto y puerperio, así como aspectos del alojamiento conjunto y lactancia; y qué ocurre cuando la infección afecta al neonato. Queda por saber el futuro de las madres y niños que sufrieron la infección. / The new coronavirus pandemic continues with us and will do so for a long time. It has brought a new way of life, with isolation, confinement, personal protection, distancing, use of virtuality and others. The diagnosis of the infected and its management has been improved; there is no cure yet, although there are vaccines approved in haste. The health crisis has exposed the lack of preparation of our health systems, resulting in political and economic crises, with impoverishment, death, and emotional and psychological complications. In these pages we continue writing in a summarized way the new knowledge about SARS-CoV-2 and the COVID-19 disease, its diagnosis, pathophysiology, symptomatic management and the severe disease, re-infection, its sequelae and lethality. But mainly how it affects the infected woman during pregnancy, childbirth and the puerperium, as well as aspects of rooming in and breastfeeding. And what happens when the infection affects the newbo ture of the mothers and children who suffered the infection remains to be known.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the novel coronavirus that causes coronavirus disease 2019 (COVID-19) and has escalated, becoming a pandemic in early 2020. Multiple testing modalities have been developed to detect this virus including RT-PCR, antigen, and serology testing. RT-PCR testing is the clinical gold standard and is used for diagnostic purposes of current infections. Antigen testing is rapid and requires significantly less equipment, but lacks the sensitivity of RT-PCR testing. Serology assays detect antibodies raised against SARS-CoV-2, so only detect prior exposure. It is important to note that use of antibody tests may also detect prior asymptomatic infections. For these reasons, it is imperative that all testing modalities be continuously developed and improved to better our understanding of disease transmission, helping to inform and change infection control policies and protecting both employees in the workplace and patients. We aim to quantify the seroprevalence of anti-SARS-CoV-2 IgG in the healthcare workers at Boston Medical Center, including those with asymptomatic infections. Our results show an overall seroprevalence of 5.5% with an asymptomatic seroprevalence of 1.8%. High risk groups include those who are obese, smokers, and Hispanic/LatinX. Experiencing some symptoms was associated with a higher risk of seropositivity, as was lack of social distancing amongst coworkers. In a separate study, we aim to assess the direct antigen rapid tests (DART) created by E25Bio in patients seeking care at Boston Medical Center. This study has been significantly limited by number of participants, as recruitment has been paused during both COVID-19 surges in Boston, MA. The current data shows poor positive agreement between DART and RT-PCR, but acceptable negative agreement. Each testing modality works to fill in the gaps of knowledge that still persist around SARS-CoV-2. Each of these testing types provides a unique piece of information and when used together, will help to inform strategies to overcome the SARS-CoV-2 pandemic.

La terbinafina es un fármaco que puede inducir daño hepático agudo. Presentamos el caso de un paciente varón de 40 años que desarrolló disfunción hepática después de 35 días de tratamiento con terbinafina por onicomicosis. El estudio anátomo patológico demostró: hepatitis aguda en resolución, además de ductopenia y colestasis. Estos hallazgos, sin el antecedente de hepatitis viral o autoinmune, son consistentes con el diagnóstico de daño hepático inducido por drogas (DILI). En este reporte presentamos el primer caso en nuestro país de un paciente que es afectado por una enfermedad hepática aguda: injuria hep osteriormente infección por SARS-CoV-2 en el contexto de una pandemia. / Terbinafine is a drug that can induce acute liver damage. We present the case of a 40-year-old male patient who developed liver dysfunction after 35 days of terbinafine treatment for onychomycosis. The anatomopathological study showed: acute hepatitis in resolution, in addition to ductopenia and cholestasis. These findings, without a history of viral or autoimmune hepatitis, are consistent with the diagnosis of drug-induced liver damage (DILI). In this report we present the first case in our country of a patient who is affected by an acute liv ver injury, to which SARS-CoV-2 infection was later associated in the context of a pandemic.