The beginning of 2020 brought us information about the novel coronavirus emerging in China. Rapid research resulted in the characterization of the pathogen, which appeared to be a member of the SARS-like cluster, commonly seen in bats. Despite the global and local efforts, the virus escaped the healthcare measures and rapidly spread in China and later globally, officially causing a pandemic and global crisis in March 2020. At present, different scenarios are being written to contain the virus, but the development of novel anticoronavirals for all highly pathogenic coronaviruses remains the major challenge. Here, we describe the antiviral activity of previously developed by us HTCC compound (N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride), which may be used as potential inhibitor of currently circulating highly pathogenic coronaviruses – SARS-CoV-2 and MERS-CoV.INTRODUCTION
Coronaviruses mainly cause respiratory and enteric diseases in humans, other mammals, and birds. However, some species can cause more severe conditions such as hepatitis, peritonitis, or neurological disease. Seven coronaviruses infect humans, four of which (human coronavirus [HCoV]-229E, HCoV-NL63, HCoV-OC43, and HCoV-HKU1) cause relatively mild upper and lower respiratory tract disease. Other three zoonotic coronaviruses – the severe acute respiratory syndrome coronaviruses (SARS-CoV and SARS-CoV-2) and the Middle East respiratory syndrome coronavirus (MERS-CoV) are associated with severe, life-threatening respiratory infections and multiorgan failure 1–7.
SARS-CoV-2 emerged in the Hubei province of China by the end of 2019 and caused an epidemic that was partially contained in China by March 2020. However, the virus rapidly spread globally and caused the pandemic 8. SARS-CoV-2 is a betacoronavirus and belongs to a large cluster of SARS-like viruses in bats, classified in Sarbecovirus subgenus. While bats are considered to be the original reservoir, it is believed that there is an intermediate host, and pangolins were suggested as such 9. The virus is associated with a respiratory illness that, in a proportion of cases, is severe. The mortality rate varies between locations, but at present, is estimated to reach 3-4% globally. The virus infects primarily ciliated cells and type II pneumocytes in human airways, hijacking the angiotensin-converting enzyme 2 (ACE2) to enter the cell, similarly as SARS-CoV and HCoV-NL63.
MERS-CoV is related to SARS-CoV-2, but together with some bat viruses forms a separate Merbecovirus subgenus. Bats are believed to serve as an original reservoir also in this case 10, but camels were identified as the intermediate host 11. The virus never fully crossed the species border, as the human-to-human transmission is limited, and almost all the cases are associated with animal-to-human transmission. The entry receptor for MERS-CoV is the dipeptidyl peptidase 4 (DPP4) 12,13. In humans, MERS-CoV causes a respiratory illness with severity varying from asymptomatic to potentially fatal acute respiratory distress 14–16. To date, MERS-CoV infection was confirmed in 27 countries, with over 2,000 cases and a mortality rate of ~35%.
Currently, there are no vaccines or drugs with proven efficacy to treat coronavirus infection, and treatment is limited to supportive care. However, a range of therapeutics have been experimentally used in clinic to treat SARS-CoV-2 and MERS-CoV-infected patients, and their use is based on the knowledge obtained in previous years. The most promising drug candidates include broad-spectrum polymerase inhibitors (remdesivir) 17 and some re-purposed drugs (e.g., HIV-1 protease inhibitors). However, until today none of these has proven effective in randomized controlled trials. On the other hand, the antiviral potential of several small molecules have been demonstrated in cell lines in vitro, but their effectiveness in vivo have not been confirmed 18,19. For the ones that reached animal models, some promising drug candidates have been shown to exacerbate the disease (ribavirin, mycophenolic acid) 20.
Drug development for SARS-CoV-2 and MERS-CoV is of great importance, but considering the diversity of coronaviruses and the proven propensity to cross the species barrier, the SARS-CoV-2 epidemic is probably not the last one. Warnings about the possibility of another SARS-CoV epidemic have appeared in the scientific literature for a long time 21. Consequently, broad-spectrum antivirals are essential in long-term perspective.
Previously, we have demonstrated an antiviral potential of the HTCC polymer (N-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride), which efficiently hampered infection of all low pathogenic human coronaviruses in vitro and ex vivo 22 and several animal coronaviruses (unpublished data). Furthermore, using several functional and structural assays, we dissected the mechanism of the HTCC antiviral activity. We showed that the polymer interacts with the coronaviral Spike (S) protein and blocks its interaction with the cellular receptor 22–24. Here, we analyzed the HTCC activity against SARS-CoV-2 and MERS-CoV in vitro using permissive cell lines and ex vivo, using a model of human airway epithelium (HAE). The study showed that the replication of both viruses was efficiently hampered. Overall, our data show that HTCC polymers are potent broad-spectrum anticoronavirals and may be very promising drug candidates for SARS-CoV-2 and MERS-CoV.RESULTS AND DISCUSSIONHTCCs hamper MERS-CoV and SARS-CoV-2 replication in cell lines
Previously, we showed that HTCC with different degrees of substitution (DSs) is a potent inhibitor of all four low pathogenic HCoVs 22. DSs are expressed as the fraction of NH2 groups of gluocosamine units in chitosan substituted with glycidyltrimethylammonium chloride (GTMAC). The DS of the studied HTCC polymers varied between 57% and 77%; thus, the polymers were named HTCC-57, HTCC-62, HTCC-63, HTCC-65, and HTCC-77. The synthesis and characterization of polymers are described elsewhere 22,23. The analysis showed that HTCC-63 demonstrated the most significant inhibitory effect on HCoV-NL63, HCoV-OC43, and HCoV-HKU1. On the other hand, HTCC-62 and HTCC-77 proved to be effective inhibitors of HCoV-229E infection. Further, HTCC-65 effectively inhibited the replication of HCoV-NL63 and HCoV-OC43, while HTCC-62 showed a potent antiviral effect on HCoV-HKU1 infection.
The study on the MERS-CoV using the Vero cells revealed that all HTCC variants inhibit virus replication to a similar extent (~100-1000-time decrease in viral yields), at non-toxic concentration (Figure 1A, C). The inhibition of the SARS-CoV-2 infection in Vero E6 cells was even more pronounced, and all HTCC variants inhibited virus replication by ~10,000 times at non-toxic concentration (Figure 1B, C). In this case, the HTCC-63 was arbitrarily selected for further studies on MERS-CoV, while HTCC-77 was selected for SARS-CoV-2.biorxiv;2020.03.29.014183v1/FIG1F1fig1Figure 1.<italic toggle= »yes »>In vitro</italic> inhibition of MERS-CoV and SARS-CoV-2 by HTCC at non toxic concentration.
Vero cells were infected with MERS-CoV (A), and Vero E6 cells were infected with SARS-CoV-2 (B). Briefly, cultures were inoculated with a given virus in the presence of HTCC (100 μg/ml) or control PBS. Replication of viruses was evaluated at 48 h post-inoculation using RT-qPCR. The data are presented as Log Removal Value (LRV) compared to the untreated sample. The assay was performed in triplicate, and average values with standard errors are shown. (C) Cytotoxicity of HTCCs with DS ranging from 57% to 77% in vitro at 100 μg/ml. Cell viability was assessed with XTT assay. Data on the y-axis represent the percentage of values obtained for the untreated reference samples. All assays were performed in triplicate, and average values with standard errors are presented. The differences in cytotoxicity of HTCCs were not statistically significant.
Next, the dose-dependence was tested for the HTCCs. The inhibitory activity of selected polymers was verified for three different concentrations, and obtained data are shown in Figure 2.biorxiv;2020.03.29.014183v1/FIG2F2fig2Figure 2.Dose-dependent inhibition of MERS-CoV and SARS-CoV-2 replication.
Vero (MERS-CoV) or Vero E6 cells (SARS-CoV-2) were inoculated with a given virus in the presence of different concentrations of HTCC. Replication of viruses was evaluated at 48 h post-inoculation using RT-qPCR. The data are presented as Log Removal Value (LRV) compared to the untreated sample. The assay was performed in triplicate, and average values with standard errors are displayed.
Based on the data obtained, the basic parameters were calculated and are presented in Table 1.biorxiv;2020.03.29.014183v1/TBL1T1tbl1Table 1.
50% Cytotoxic concentration (CC50), 50% inhibitory concentration (IC50), and the selectivity index (SI) of two most effective HTCCs: HTCC-63 (for MERS-CoV) and HTCC-77 (for SARS-CoV-2).
The parameters observed for the SARS-CoV-2 appear to be favorable with SI above 12. It is also worth to note that HTCC was previously administered by inhalation in rats, and no adverse reactions were observed 25. In that study, HTCC was used as a carrier for the active substance, and as such, was reported to be promising for local sustained inhalation therapy of pulmonary diseases.HTCCs hamper MERS-CoV and SARS-CoV-2 replication in human airway epithelium
While the Vero cells constitute a convenient model for antiviral research, it is of utmost importance to verify whether the results obtained are not biased due to the artificial system used. This is especially important for compounds, which activity is based on electrostatic interaction. To verify whether the natural microenvironment, which is rich in sugars and charged molecules, does not abrogate the effectiveness of the inhibitors, we employed HAE cultures that mirror the fully differentiated layer linng the conductive airways, as well as the site of coronavirus replication. Briefly, fully differentiated HAE cultures were infected with a given virus 26 in the presence of previously selected HTCCs (200 μg/ml) or control PBS. Following inoculation, apical lavage samples were collected daily, and replication kinetics for each virus was investigated. The analysis revealed that the polymer efficiently hampered SARS-CoV-2 and MERS-CoV also in this model. For MERS-CoV, the inhibitory effect was the most evident at 72 h p.i., while for SARS-CoV-2 the most substantial decline of virus progeny was observed at 24 h p.i. (Figure 3). Whether such kinetics will be reflected in vivo it is to be investigated.biorxiv;2020.03.29.014183v1/FIG3F3fig3Figure 3.<italic toggle= »yes »>Ex vivo</italic> inhibition of MERS-CoV and SARS-CoV-2 by HTCC in human airway epithelium cultures.
HAE cultures were exposed to MERS-CoV or SARS-CoV-2 in the presence of HTCC-63 (for MERS-CoV) or HTCC-77 (for SARS-CoV-2) at 200 μg/ml or control PBS. To analyze virus replication kinetics, each day post infection, 100 μl of 1 × PBS was applied to the apical surface of HAE cultures and collected after 10 min of incubation at 37°C. Replication of viruses was evaluated using quantitative RT-qPCR. The data are presented as Log Removal Value (LRV) compared to the untreated sample. The assay was performed in triplicate, and average values with standard errors are shown.HTCCs inhibits MERS-CoV and SARS-CoV-2 entry into susceptible cells
Our previous research showed that the HTCC-mediated inhibition of coronaviral replication results from the electrostatic interaction between the polymer and the Spike protein of coronaviruses. We hypothesize that the selectivity of the inhibitors yields from the fitting charge distributions on the polymer and on the S proteins on the viral surface. While the interaction of a single charged moiety is relatively weak, the concatemeric nature of the virus and the polymer stabilizes the binding. Such structure-based interaction may be an interesting entry point for further fine-tuning of the polymeric inhibitors of viral replication.
To ensure that the observed effect was a result of coronavirus entry inhibition by HTCC, two experiments were performed. First, HAE cultures were inoculated with MERS-CoV in the presence of HTCC-63 (200 μg/ml) or control PBS and incubated for 2 h at 37°C. Next, cells were fixed, immunostained for MERS-CoV N protein and actin. Virus entry was analyzed with confocal microscopy. To visualize the effect, the signal attributed to intracellular MERS-CoV was quantified, and the results show that the internalization of MERS-CoV was drastically decreased (Figure 4A).biorxiv;2020.03.29.014183v1/FIG4F4fig4Figure 4.Coronavirus internalization into susceptible cells is hampered by HTCC.
(A) Pre-cooled HAE cultures were incubated with ice-cold MERS-CoV suspension in the presence or absence of HTCC-63 (200 μg/ml) for 2 h at 37°C. Next, cells were fixed in PFA and immunostained for MERS-CoV N protein and actin. Virus entry was analyzed with confocal microscopy. The data shown are representative of three independent experiments, each performed in triplicate. * P < 0.05. (B) A549 cells overexpressing ACE2 were incubated with lentiviral particles bearing GFP reporter gene, pseudotyped with SARS-CoV-2 Spike (S-SARS-CoV-2), VSV control G protein (VSV-G) or particles without an envelope protein (∆Env) in the presence of HTCC-77 or control PBS. After 2 h at 37°C cells were washed with PBS and overlaid with fresh medium. Following 72 h incubation, GFP signal was measured using fluorometer and pseudovirus entry is presented as Relative Fluorescence Units per ml. The assay was performed in triplicate, and average values with standard errors are presented. ** P < 0.005.
Due to the limited availability of tools for the SARS-CoV-2 we were not able to replicate the experiment for this virus. Here, we employed a surrogate system based on lentiviral vectors pseudotyped with full-length Spike protein of SARS-CoV-2. A549 cells overexpressing the ACE2 protein were incubated with pseudovirions harboring SARS-CoV-2 Spike or control VSV-G protein in the presence of HTCC-77 (100 μg/ml) or control PBS for 2 h at 37°C. After 72 h p.i. cells were lysed, and pseudovirus entry was quantified by measurement of the reporter GFP protein. The analysis showed a significant reduction in SARS-CoV-2 Spike pseudoviruses internalization in the presence of the polymer, while no inhibition was observed for the control VSV-G (Figure 4B).
Next, to verify whether the mechanism of action for the highly pathogenic betacoronaviruses is similar to that observed for alphacoronaviruses 22, and is based on locking the interaction between the virus and the entry receptor, we analyzed MERS-CoV co-localization with its entry receptor, DPP4 in the presence or absence of the HTCC. For this, human cell line Huh7 was inoculated with the virus or mock in the presence of HTCC-63 (100 μg/ml) or control PBS and incubated for 2 h at 4°C. Subsequently, cells were fixed and immunostained for the DPP4 and MERS-CoV N using specific antibodies. Virus co-localization with its receptor was examined using confocal microscopy. Obtained results demonstrated that in the control samples, virions co-localize with the DPP4 protein, while in the presence of the polymer, this interaction is blocked (Figure 5).biorxiv;2020.03.29.014183v1/FIG5F5fig5Figure 5.HTCC blocks interaction between the virus and its entry receptor.
Pre-cooled Huh7 cells were incubated for 3 h at 4°C with ice-cold MERS-CoV or mock in the presence or absence of HTCC-63 (100 μg/ml). Next, cells were fixed with PFA and immunostained for MERS-CoV-N (green), DPP4 (red) and nuclear DNA (blue). MERS-CoV interaction with the DPP4 protein was analyzed with confocal microscopy. Co-localisation of DPP4 with MERS-CoV-N was determined by confocal microscopy and is presented as Manders’ M2 coefficient. The decrease in colocalization was statistically significant (P < 0.0005). Each image represents maximum projection of axial planes. Representative images are shown.
Taking together, we show here that the previously developed and described polymeric HTCC anticoronaviral compounds based on chitosan are able to efficiently inhibit infection with emerging coronaviruses. We believe that the HTCC can be fine-tuned to target any coronavirus, and this interaction is specific to viruses that belong to the Coronaviridae family. One may speculate that the inhibition results from the concatemeric nature of the virus surface and the fact that the polymer with appropriate charge distribution can interact with multiple sites on this surface. While the interaction of the monomer is relatively weak, and no inhibition is observable for monomers, the sum of interactions stabilizes the binding and specific inhibition is observed. Considering that the extended chain length for the HTCC used is ~700 nm this scenario seems realistic 27. If that would be true, HTCC would constitute a first structure-specific inhibitor of viral replication. The major disadvantage of the HTCC is that, at present, it is not registered for use in humans. However, previous experience with HTCC in different laboratories shows that it may be delivered topically to the lungs, it is not associated with toxicity, and it does not worsen the lung function 25. We believe that HTCC is a promising drug candidate that should be further studied, as it provides a ready-to-use solution for SARS-CoV-2 and future emerging coronaviruses.MATERIALS AND METHODSThe active compound
The HTCC was prepared in the same manner as previously described 22,23,28.Plasmid constructs
The codon-optimised full-length SARS-CoV-2 S gene was designed and purchased from GeneArt (Thermo Fisher Scientific, Poland). The gene was cloned into pCAGGS vector sequence verified that was a gift from Xingchuan Huang. psPAX (Addgene plasmid # 12260) and pMD2G (Addgene plasmid # 12259) was a gift from Didier Trono. Lego-G2 vector (Addgene plasmid #25917) was a gift from Boris Fehs.Cell culture
Vero and Vero E6 (Cercopithecus aethiops; kidney epithelial; ATCC: CCL-81 and CRL-1586), Huh7 (Homo sapiens; hepatocellular carcinoma; ECACC: 01042712) and A549 cells with ACE2 overexpression (A549/ACE2)29 were cultured in Dulbecco’s MEM (Thermo Fisher Scientific, Poland) supplemented with 3% fetal bovine serum (heat-inactivated; Thermo Fisher Scientific, Poland) and antibiotics: penicillin (100 U/ml), streptomycin (100 μg/ml), and ciprofloxacin (5 μg/ml). Cells were maintained at 37°C under 5% CO2.Human airway epithelium (HAE) cultures
Human airway epithelial cells were isolated from conductive airways resected from transplant patients. The study was approved by the Bioethical Committee of the Medical University of Silesia in Katowice, Poland (approval no: KNW/0022/KB1/17/10 dated 16.02.2010). Written consent was obtained from all patients. Cells were dislodged by protease treatment, and later mechanically detached from the connective tissue. Further, cells were trypsinized and transferred onto permeable Transwell insert supports (ϕ = 6.5 mm). Cell differentiation was stimulated by the media additives and removal of media from the apical side after the cells reached confluence. Cells were cultured for 4-6 weeks to form well-differentiated, pseudostratified mucociliary epithelium. All experiments were performed in accordance with relevant guidelines and regulations.Cell viability assay
HAE cultures were prepared as described above. Cell viability assay was performed by using the XTT Cell Viability Assay (Biological Industries, Israel) according to the manufacturer’s instructions. On the day of the assay, 100 μl of the 1 × PBS with the 50 μl of the activated XTT solution was added to each well/culture insert. Following 2 h incubation at 37°C, the solution was transferred onto a 96-well plate, and the signal was measured at λ = 490 nm using the colorimeter (Spectra MAX 250, Molecular Devices). The obtained results were further normalized to the control sample, where cell viability was set to 100%.Virus preparation and titration
MERS-CoV stock (isolate England 1, 1409231v, National Collection of Pathogenic Viruses, Public Health England, United Kingdom) was generated by infecting monolayers of Vero cells. SARS-CoV-2 stock (isolate 026V-03883; kindly granted by Christian Drosten, Charité – Universitätsmedizin Berlin, Germany by the European Virus Archive – Global (EVAg); https://www.european-virus-archive.com/) was generated by infecting monolayers of Vero E6 cells. The virus-containing liquid was collected at day 3 post-infection (p.i.), aliquoted and stored at −80°C. Control Vero or Vero E6 cell lysate from mock-infected cells was prepared in the same manner. Virus yield was assessed by titration on fully confluent Vero or Vero E6 cells in 96-well plates, according to the method of Reed and Muench. Plates were incubated at 37°C for 3 days and the cytopathic effect (CPE) was scored by observation under an inverted microscope.Virus infection
In in vitro experiments, fully confluent Vero, Vero E6, or Huh7 cells in 96-well plates (TPP) were exposed to MERS-CoV, SARS-CoV-2 or mock at a TCID50 of 400 per ml in the presence of tested polymer or control medium. Following a 2 h incubation at 37°C, unbound virions were removed by washing with 100 μl of 1 × PBS and fresh medium containing dissolved respective polymer was added to each well. Samples of cell culture supernatant were collected at day 3 p.i. and analyzed using RT-qPCR.
For the ex vivo study, fully differentiated human airway epithelium (HAE) cultures were exposed to the tested polymer or control PBS for 30 min at 37°C, following inoculation with MERS-CoV or SARS-CoV-2 at a TCID50 of 400 per ml in the presence of the polymer or control PBS. Following 2 h incubation at 37°C, unbound virions were removed by washing with 200 μl of 1 × PBS and HAE cultures were maintained at an air—liquid interphase for the rest of the experiment. To analyze virus replication kinetics, each day p.i., 100 μl of 1 × PBS was applied at the apical surface of HAE and collected following the 10 min incubation at 32°C. All samples were stored at −80°C and analyzed using RT-qPCR.Isolation of nucleic acids and reverse transcription (RT)
Viral DNA/RNA Kit (A&A Biotechnology, Poland) was used for nucleic acid isolation from cell culture supernatants, according to the manufacturer’s instructions. cDNA samples were prepared with a High Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific, Poland), according to the manufacturer’s instructions.Quantitative PCR (qPCR)
Viral RNA yield was assessed using real-time PCR (7500 Fast Real-Time PCR; Life Technologies, Poland). cDNA was amplified in a reaction mixture containing 1 × qPCR Master Mix (A&A Biotechnology, Poland), in the presence of probe (100 nM) and primers (450 nM each).biorxiv;2020.03.29.014183v1/TBL1AT2tbl1aTable 1.Primers and probes.
The reaction was carried out according to the scheme: 2 min at 50°C and 10 min at 92°C, followed by 40 cycles of 15 s at 92°C and 1 min at 60°C. In order to assess the copy number for N gene, DNA standards were prepared, as described before 26.Immunostaining and confocal imaging
Fixed cells were permeabilized with 0.1% Triton X-100 in 1 × PBS and incubated overnight at 4°C in 1× PBS supplemented with 5% bovine serum albumin (BSA) and 0.5% Tween 20. To visualize MERS-CoV particles, cells were incubated for 2 h at room temperature with mouse anti-MERS-CoV N IgGs (1:000 dilution, Sino Biological, China), followed by 1 h of incubation with Alexa Fluor 488-labeled goat anti-mouse IgG (2.5 µg/ml; Thermo Fisher Scientific, Poland). Actin filaments was stained using phalloidin coupled with Alexa Fluor 633 (0.2 U/ml; Thermo Fisher Scientific, Poland). Nuclear DNA was stained with DAPI (4’,6’-diamidino-2-phenylindole) (0.1 µg/ml; Sigma-Aldrich, Poland). Immunostained cultures were mounted on glass slides in ProLong Gold antifade medium (Thermo Fisher Scientific, Poland). Fluorescent images were acquired under a Leica TCS SP5 II confocal microscope (Leica Microsystems GmbH, Mannheim, Germany) and a Zeiss LSM 710 confocal microscope (Carl Zeiss Microscopy GmbH). Images were acquired using Leica Application Suite Advanced Fluorescence LAS AF v. 2.2.1 (Leica Microsystems CMS GmbH) or ZEN 2012 SP1 software (Carl Zeiss Microscopy GmbH) deconvolved with Huygens Essential package version 4.4 (Scientific Volume Imaging B.V., The Netherlands) and processed using ImageJ 1.47v (National Institutes of Health, Bethesda, MD, USA). At the time of the study, no antibodies specific to SARS-CoV-2 were available to us.Pseudovirus production and transduction
293T cells were seeded on 10 cm2 dishes, cultured for 24 h at 37°C with 5% CO2 and transfected using polyethyleneimine (Sigma-Aldrich, Poland) with the lentiviral packaging plasmid (psPAX), the VSV-G envelope plasmid (pMD2G) or SARS-CoV-2 S glycoprotein (pCAGGS-SARS-CoV-2-S) and third plasmid encoding GFP protein (Lego-G2). Cells were further cultured for 72 h at 37°C with 5% CO2 and pseudoviruses were collected every 24 h and stored at 4°C.
A549/ACE2 cells were seeded in 48-wells plates, cultured for 24 h at 37°C with 5% CO2 and transduced with pseudoviruses harboring VSV-G or S-SARS-CoV-2 proteins or lacking the fusion protein (∆Env) in the presence of polybrene (4 µg/ml; Sigma-Aldrich, Poland) and HTCC-77 (100 μg/ml) or control PBS. After 4 h incubation at 37°C unbound virions were removed by washing thrice with 1 × PBS and cells were further cultured for 72 h at 37°C with 5% CO2. Cells were lysed in RIPA buffer (50 mM Tris, 150 mM NaCl, 1% Nonidet P-40, 0.5% sodium deoxycholate, 0.1% SDS, pH 7.5) and transferred onto black 96-wells plates. Fluorescence levels were measured on a microplate reader Gemini EM (Molecular Devices, UK).Statistical analysis
All the experiments were performed in triplicate, and the results are presented as mean ± standard deviation (SD). To determine the significance of the obtained results Student t test was carried out. P values of < 0.05 were considered significant.ACKNOWLEDGEMENTS
This work was supported by the subsidy from the Polish Ministry of Science and Higher Education for the research on the SARS-CoV-2 and a grant from the National Science Center UMO-2017/27/B/NZ6/02488 to KP.
The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript.
The technology is owned by the Jagiellonian University (Krakow, Poland) and protected by a patent no WO2013172725A1 and associated documents.AUTHOR CONTRIBUTIONS STATEMENT
A.M., Y.C., A.S., E. B. D., X.G., Y.G., J.L., L.C. conducted the experiments. M.O., M.U., and S.R.M provided materials and methods for the study. A.M., K.P. designed the study and experiments, analysed the data and wrote the manuscript. K.L., D.L., F.Z., M.N., K.S. analysed the data. K.P. supervised the study. All authors reviewed the manuscript and approved the submitted version. All authors agreed to be personally accountable for their own contributions and to ensure that questions related the accuracy or integrity of any part of the work are appropriately investigated, resolved, and the resolution documented in the literature.ADDITIONAL INFORMATIONCompeting interests
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A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant. MBio 5, e00047–00014, doi:10.1128/mBio.00047-14 (2014).19Ratia, K. et al. A noncovalent class of papain-like protease/deubiquitinase inhibitors blocks SARS virus replication. Proc Natl Acad Sci U S A 105, 16119–16124, doi:10.1073/pnas.0805240105 (2008).20Barnard, D. L. et al. Enhancement of the infectivity of SARS-CoV in BALB/c mice by IMP dehydrogenase inhibitors, including ribavirin. Antiviral Res 71, 53–63, doi:10.1016/j.antiviral.2006.03.001 (2006).21Cheng, V. C., Lau, S. K., Woo, P. C. & Yuen, K. Y. Severe acute respiratory syndrome coronavirus as an agent of emerging and reemerging infection. Clin Microbiol Rev 20, 660–¨C202C, doi:10.1128/CMR.00023-07 (¨C203C).22Milewska, A. et al. HTCC: Broad Range Inhibitor of Coronavirus Entry. PLoS One 11, e0156552, doi:10.1371/journal.pone.0156552 (2016).23Milewska, A. et al. Novel polymeric inhibitors of HCoV-NL63. Antiviral Res 97, 112–121, doi:10.1016/j.antiviral.2012.11.006 (2013).24Kamil, K., Aleksandra, M., Maria, N., Krzysztof, P. & Krzysztof, S. THE USE OF CHITOSAN POLYMER IN THE TREATMENT AND PREVENTION OF INFECTIONS CAUSED BY CORONAVIRUSES.25Yang, T. T. et al. Cyclosporine A/porous quaternized chitosan microspheres as a novel pulmonary drug delivery system. Artif Cells Nanomed Biotechnol 46, 552–564, doi:10.1080/21691401.2018.1463231 (2018).26A, M. et al.27Kühtreiber, W. M., P., L. R., Chick, W. L. & View ORCID Profil Anna Kula-Pacurar, J. W., Agnieszka Suder, Artur Szczepanski, ¨C251C ¨C252C, ¨C253C ¨C254C, ¨C255C ¨C256C, Tomasz Stacel View ORCID Profile. Cell Encapsulation Technology and Therapeutics. (¨C257C).28Ciejka, J., Wolski, K., Nowakowska, M., Pyrc, K. & Szczubiałka, K. Biopolymeric nano/microspheres for selective and reversible adsorption of coronaviruses. Mater Sci Eng C Mater Biol Appl ¨C269C, ¨C270C–¨C271C, doi:10.1016/j.msec.2017.03.047 (¨C272C).29Milewska, A. et al. Human coronavirus NL63 utilizes heparan sulfate proteoglycans for attachment to target cells. J Virol 88, 13221–13230, doi:10.1128/JVI.02078-14 (2014).


Keto Advanced come with all necessary information about Keto Advanced BHB Weight Loss Pills. Nowadays most of us are busy with our works all day long. It is hard for us to find our time to do physical exercises to lose weight.
Besides, many of us love foods such as Burger, French fries, chicken fries and want to eat them without gaining weight. For these people, there is no other best solution like Keto Advanced. This weight loss formula contains BHB salts that burn body fat via a ketogenic diet pill. You can try this revolutionary supplement and get a fit body.
What is Keto Advanced Pill?
It is hard for us to keep our bodies fit without doing any exercise. Besides, we love to eat fast food such as cake, pizza, cold drinks, etc. But these foods have serious side effects on our body. It is also hard for us to find out necessary times to take care of our weight.
As a result, we have fat and get overweighted. But at this time, it is not easy to lose weight without having a fantastic formula such as Keto advanced weight loss pill.
To solve this significant problem, you can try this supplement. To keep our bodies fit and energetic, we need to burn fat. We bring Keto Advanced reviews from our real-life experience. This supplement burns fat from the body. The pills provide beta-hydroxybutyrate to the body naturally.
This dietary solution is perfect for searching for the right and easy way to burn fat. We know that a keto diet may contain moderate proteins and no carbs. When we take fewer carbohydrates, we can quickly burn our fat. Advanced Keto plus is such a type of medication.
How Does Keto Advanced Weight Loss Work?
When we want to take a supplement, we search for the correct information because some do not work. But this weight loss formula works well. It is a guaranteed product that works 100%. For healthy weight loss, it can be an ideal supplement. It works in a healthy manner. It works effectively and makes your body healthy.
If you want to follow diet plans, it is pretty tricky. With a bit of effort, you can lose weight if you take Keto Advanced pills. However, this supplement follows the ketogenic process and works as a ketogenic diet. The keto pills contain BHB Ketones that drive your body into ketosis. Your body will under ketosis and effectively lose fat like a ketogenic diet.
So, when you start to take this supplement, you should take a low-carb diet. It is essential because it helps the process to obtain proteins, vitamins. Thus, the fat of your body can burn quickly, and you get energy. Then this energy is used in your body functions. Therefore, this supplement works and provides us excellent benefits. It not only makes our body slim but also keeps us energetic the whole day long.
Keto Advanced Weight Loss Pills Ingredients
From this keto advanced weight loss review, we also get familiar with the ingredients of this supplement. The elements do not have any harmful enzymes. It is essential to know the ingredients when we purchase any supplement. However, let’s see them.
- Garcinia Cambogia:
It is a natural ingredient that comes from a plant that contains HCA. This thing controls hunger cravings. If you take this appetite suppressant, it helps you to reduce extra calories from your body. In this way, you can lose weight and get a fit figure without any side effects.
- Forskolin Extract
You can call it mint extract. This powerful ingredient is a responsible one that delivers healthy nutrients to the human body. It is an antioxidant and removes toxic things from your body, and improves your metabolic power. Besides, it is also helpful to improve the immune system of your body.
- BHB Ketones
BHB ketones are very effective in improving the conjugative function of your body. It accelerates metabolic rate and increases the energy level. Thus, your body can function well and can work better throughout the day.
These are the main ingredients of Keto Advanced diet pills. All of them make this dietary formula an amazing one.
Keto Advanced Weight Loss Benefits
Our body is made of some tissues. The types of them are different. For this reason, the tissues need extra nutrients and strength, and other things. We need all of the nutrients to keep our bodies healthy and fit. When we go through a weight loss process, some processes also go through our bodies. The impact of them is positive for our body. However, the good effects of advanced keto pills are below:
- This dietary pill controls fat storage and also eliminates the stored fat from the body.
- As it releases a lot of energy, it helps the body to perform well.
- It also improves our digestive and immune systems.
- It suppresses hunger and reduces hunger cravings.
- As this supplement contains a lot of necessary nutrients, antioxidants, our body is nourished by it.
- To ensure a healthy body, it improves metabolism.
- It also prevents harmful cholesterol levels.
- To have a perfect body shape, there is no alternative to this product.
- This supplement produces more energy and ensures your body lean.
- As this product is made of natural ingredients, it has no harmful side effects on our body.
Keto Advanced Fat Burner Side Effects
Keto Advanced pills are the best way to lose fat and weight naturally. We know that most of the supplements have some side effects. Like others, this formula also has some side effects. All of our body functions are not the same. Our body needs some requirements and maintenance.
For this reason, some people face a low amount of side effects, and some do not. With a sensitive body, you can face some side effects. However, the side effects are minor such as fatigue, diarrhea, and constipation, headache, etc. If you see that it does not support suit with you, you should stop taking the pills.
Is Keto Advanced Safe?
You already know from these advanced dietary keto reviews that the ingredients of this supplement contain BHB ketones. This fluid produces natural ketosis and does not contain any chemicals or artificial ingredients. Besides, the elements are already familiar to our bodies. So, there is no significant risk of pills like fake pills. Not only that, these keto pills are clinically proven that increases metabolism and produce more energy.
So, it is clear that the supplement works in a natural way to burn fat and keep you fit for the day long. When you take the pills, you will stay in a ketosis stage to burn fat and lose weight. The only workout cannot give you such a quick transform of the body.
Pros and Cons of Keto Advanced Diet Pills
Everything has pros and cons. Let’s see the pros and cons of this product.
- It is made of natural ingredients.
- The ketosis activator really works well.
- It helps you to lose weight faster.
- This product also boosts energy level.
- You will get a long-term result if you take it.
- It is hard to find out the disadvantages of this product.
- If your body condition is not so good, you can face some minor side effects.
Is it an Advanced Keto Diet Pill scam?
Keto advanced reviews show that this product is not a scam. It is a famous dietary formula for weight loss. You can lose weight faster with this ketogenic diet. The ingredients of this pill contain BHB and increase energy levels. BHB is a natural source to work for cellular respiration.
Besides, it produces energy as an alternative to carbohydrates. As it increases the ketosis metabolic rate, it is a great supplement to lose weight quickly. But some risks are found online about Keto advanced supplements. The progressive keto shark tank TV shows are fraudulent because they are not from the company.
So, Keto Advanced Shark Tank is fake. Keto advanced weight loss 800 mg, Keto Advanced 1500 are two forms of the supplement. You can get anyone of them.
To know more about this product, you can search for keto advanced weight loss pills reviews
at different websites.
How to take Keto Advanced Weight Loss pills
It would help if you took the Keto Advanced pills incorrectly way to get perfect results. You will get the pills in capsule form. You should consume one capsule two times a day. It is recommended to take 30 days course. Take one pill in the morning and another one in the evening.
The FAQs About Keto Advanced
Let’s see some frequently asked questions from advanced dietary keto reviews of different consumers.
Is Advanced Keto Plus Safe?
Yes, Keto Plus is safe. As a natural formula, without some minor side effects in some cases, it is secure.
What Does Keto Advanced Weight Loss Do?
Keto Advanced Weight loss pills work based on ketosis. The ingredients of it bring the body under a ketosis process. It increases metabolism level and produces more energy by burning fat from our body. It also prevents us from taking more carbs. For this reason, we can lose weight fast.
Does Advanced Keto Work?
Yes, of course, Advanced Keto works.
What is The Best Keto Product?
There are many keto products in the market. But among them, I have tried the Keto Advanced diet pills. I think it can be the best choice for you as the best keto product. To be more sure, you can see other Keto advanced weight loss reviews.
How Fast Can You Lose Weight on Keto?
I think Keto works faster than workouts. When you start to take the Keto Advanced pill, you will see the result within a week. It is not possible to do a regular diet plan. The ingredients take your body into ketosis, and your body starts to burn fat and produce more energy. Thus, your weight loss target fulfills soon.
From Keto Advanced reviews, we now know about this fantastic supplement properly. We can say that it is a great product to lose weight naturally. This product is available online at the lowest price. So, if you have overweight with a good health condition, you can start taking this product. And get a noticeable result within a week.


I cannot recommend Transform Studio and its staff highly enough. I started attending personal trainings when, following spinal surgery, I had to rebuild the muscles around the base of the spine and improve my general fitness. Adnan showed a deep understanding of the biomechanics involved and designed a programme that started moderately and then increased in intensity. There was no undue pressure and I felt very comfortable returning to the gym. One of the great strengths of the studio is that there is a very sociable atmosphere and the only competition is with yourself! All the staff have a great sense of humour while remaining totally professional and focused. For me the sessions were as much psychological as physical as I worked my way back to full movement and capacity. All the staff really know what they are doing and their advanced level qualifications are evident in the way they work. After 6 months of slow but steady work, friends and colleagues were commenting on the evident change in my shape and posture. Last summer, the trainings took place outdoors at the athletics track, which was an added bonus. Frankly I should have taken this step and started regular trainings long ago and before surgery as it has done me a lot of good. Whatever your aims for physical fitness I have no hesitation in recommending Transform Studio and its excellent staff.


With a strong emphasis on quality, innovation, and patient satisfaction, Prevest DenPro’s new oral care line promises to deliver exceptional results and promote optimal oral health.
Good oral hygiene is essential for maintaining healthy teeth and gums, preventing dental issues such as cavities, gum disease, and bad breath. Prevest DenPro’s oral care products are designed to empower individuals to take charge of their oral health, providing them with effective and easy-to-use solutions.
One of the standout features of Prevest DenPro’s oral care line is the incorporation of advanced formulations that target specific oral health concerns. Whether it’s tooth sensitivity, gum inflammation, or enamel protection, their products are carefully crafted to address these issues effectively. By focusing on targeted solutions, individuals can choose the products that best suit their needs, resulting in a personalized and optimized oral care routine.
Prevest DenPro’s oral care range includes mouthwash, dental floss, and other complementary products, all of which are formulated with a strong emphasis on quality ingredients. The company ensures that their products are free from harmful chemicals and additives, prioritizing the well-being of their consumers. With Prevest DenPro’s oral care products, individuals can trust that they are using safe and reliable solutions that contribute to their overall oral health.
Furthermore, Prevest DenPro’s oral care products are designed to deliver exceptional results. Their toothpaste effectively cleans and protects teeth, removing plaque and surface stains while strengthening the enamel. The mouthwash provides a refreshing and germ-fighting rinse, promoting fresh breath and a healthy oral environment. The dental floss is designed to glide smoothly between teeth, removing plaque and debris from hard-to-reach areas. By incorporating these products into a daily oral care routine, individuals can enjoy the benefits of improved oral health and a confident smile.
Prevest DenPro’s commitment to innovation is also evident in their packaging and design. The oral care products are packaged in user-friendly containers with precise dispensers, making them easy to use and ensuring minimal waste. The sleek and modern design adds a touch of elegance to the overall oral care experience, further enhancing the satisfaction of the users.
With the launch of their new oral care line, Prevest DenPro Limited reinforces its position as a trusted leader in the dental industry. Their focus on quality, innovation, and patient-centric solutions sets them apart. Individuals can rely on Prevest DenPro’s oral care products to deliver exceptional results and contribute to their overall oral health and well-being.
As the importance of oral hygiene continues to be emphasized, Prevest DenPro’s new range of oral care products comes at the perfect time. By providing individuals with effective and reliable solutions, the company empowers them to take proactive steps towards maintaining optimal oral health. With Prevest DenPro’s oral care products, individuals can enjoy a healthy and confident smile that lasts a lifetime.


1. Excellent Reputation: Bastyr University is recognized as a leader in natural health education, known for its rigorous curriculum and high standards of excellence.
2. Accredited Programs: All programs offered at Bastyr University are accredited by the appropriate accrediting bodies, ensuring that you will receive a quality education.
3. Integrative Approach: Bastyr University takes an integrative approach to health and wellness, combining the best practices from both Western and natural medicine.
4. Focus on Holistic Health: The university’s programs focus on holistic health, emphasizing the well-being of the mind, body, and spirit.
5. Faculty Expertise: The faculty at Bastyr University are experts in their fields, bringing a wealth of knowledge and experience to the classroom.
6. Research Opportunities: Bastyr University offers a range of research opportunities, allowing students to contribute to the advancement of natural health sciences.
7. Clinical Training: Through Bastyr University’s teaching clinic, students have the opportunity to gain hands-on experience working with patients under the guidance of experienced practitioners.
8. Natural Medicine Dispensary: The university operates a natural medicine dispensary where students can learn about and dispense natural remedies.
9. Strong Alumni Network: Bastyr University has a strong and active alumni network, providing opportunities for networking and mentorship.
10. Beautiful Campus: Located in the Pacific Northwest, Bastyr University’s campus offers stunning views and a peaceful environment for learning.


I am a UKCP Registered Psychotherapist (registration number 04156573). I trained at Metanoia Institute and qualified in 2004 with an MA in Integrative Psychotherapy (prior to which, in 1998 and 2000, I completed a diploma and a BA Hons in Integrative Counselling at Chichester Univerisity). I am EMDR (Eye Movement Desensitisation and Reprocessing) trained to Advanced Level and I have also trained in Sensorimotor Psychotherapy for the Treatment of Trauma.
I have expertise in sexualised trauma, domestic abuse, victim behaviour, sex offending and the interviewing of victims of sexual crimes. Additionally I have considerable knowledge regarding the therapeutic needs of victims/survivors of rape, sexual assault, child sexual exploitation/abuse and domestic abuse.
I have over 22 years experience of working therapeutically with women and men who have experienced sexualised trauma(s). I have provided training and consultation to all police forces in England and Wales; and many other professionals including Judges, Magistrates, Lawyers, Social Workers, Teachers, Forensic Medical Examiners (FMEs), Independent Sexual Violence Advisers (ISVAs), Counsellors and Crisis Workers. I have been commissioned by a number of police forces and CPS areas to provide expert testimony, explaining victim behaviour in rape, kidnap and domestic abuse cases.


The finest currants produced anywhere in the world: full of flavor, beautiful vine fruit aroma, juicy, nutritious and healthy. 100% natural, sulfite free, without preservatives or additives, dried naturally under the sun.
DIOTERRA Vostizza Currants PDO original and historical harvest of Greece, cultivated exclusively in the area of Aeghialeia in the Peloponnese-southern Greece, is rendered unique gastronomic treasure thanks to the ideal combination of rich soil, sea breeze and sunshine. Vostizza currants coming from the grapes of the Greek vine variety Vitis Vinifera L, are cultivated since the 4th century B.C.
They have been integral part of the Mediterranean diet, of high nutritional value, awarded Protected Designation of Origin (P.D.O.) by the EU and ranked as the best thanks to the excellent aroma and rich flavor.
Today Vostizza Currants follow exactly the same pattern as has been happening for centuries; upon harvest, they are naturally dried under the sun, selected one by one by hand.
Vostizza currants have been part of the Mediterranean eating habits since antiquity thanks to their excellent properties; they are rich in potassium - higher even than that of the banana - rich in fibre, fat-free and natural source of iron, phosphorus and vitamin B6, contributing to normal energy-yielding metabolism, to normal function of the muscles, to normal oxygen transport in the body, to normal formation of red blood cells and hemoglobin and to the maintenance of normal blood pressure. Vostizza currants constitute natural source of antioxidants, high in polyphenols, flavonoids and anthocyanines, neutralizing free radicals in the body and contributing to the general well being.
Enjoy Vostizza currants:
• as a snack all day long
• with yogurt, honey and walnuts
• in confectionery, in cakes, muffins and scones
• in oat or cereals
• with cheese and dry-cured ham
• in green salads
• in main courses as stuffing or side dish
• in desserts or in fruit salads
Paper box 200g (7oz)
Gluten free, GMO free, Kosher, Halal
Store in a cool and dry place. Refrigerate after opening.
Finest selected, Greek farm grown Sultana Raisins: extra juicy, packed with natural sweetness and flavor. Nutritious and healthy, 100% sulfite free, without added sugar and preservatives, handpicked and dried naturally under the sun.


Dr. Paula Moraga (https://paula-moraga.github.io/) is an Assistant Professor of Statistics at King Abdullah University of Science and Technology (KAUST). Previously, she held academic positions at Lancaster University, Harvard School of Public Health, London School of Hygiene and Tropical Medicine, Queensland University of Technology, and University of Bath.
Dr. Moraga has worked in statistical research for over a decade, with a strong focus on spatial epidemiology and modeling. She has developed innovative statistical methods and open-source software for disease surveillance, and her work has directly informed strategic policy in reducing disease burden in several countries. Past projects include the development of modeling architectures to understand the spatio-temporal patterns and identify targets for intervention of malaria in Africa, leptospirosis in Brazil, and cancer in Australia. Dr. Moraga has worked on the development of a number of R packages for disease modeling, detection of clusters, and travel-related spread of disease, and is the author of SpatialEpiApp, a Shiny web application for the analysis of spatial and spatio-temporal disease data.
Dr. Moraga has taught statistics and spatial epidemiology courses at both undergraduate and graduate levels at universities in the United Kingdom, Australia, Ethiopia and Saudi Arabia, and has been invited to deliver training courses on disease mapping and R at international workshops. She has also created online educational materials that impact learning on a large scale, and has published extensively on statistical methodology, software, and health and environmental applications.
Dr. Moraga received her Ph.D. in Mathematics from the University of Valencia, Spain, and her Master’s in Biostatistics from Harvard University, USA.


To some people, a family pet is simply as precious as a kid. Taking care of your family pet is very essential, therefore is providing the best food. Good food and the ideal animal items can help your pet stay healthy and fit.
Animal nutrition in our industry plays a big function in our society although not all people recognize this. The majority of customers think that purchasing animal items such as canned foods are simply equivalent to the meat that you can feed them every day. They also believe that the canned foods just include toxins and preservatives which can hurt the pet's health one day.
If you want to offer your dog natural, healthy food, however you do not have adequate time to prepare raw meals for your animal, you can buy a few of the food such as those ready-to-eat freezes dried and frozen products or raw meals. Your animals will enjoy this kind of raw food. This can be a replacement if you can't feed him with a bone. These food items are extremely special that they have food supplements.
One of the options for feeding pet's raw food is Life's Abundance, which is a natural kibble animal food for your pet dogs and felines . It consists of only natural, healthy components and is nutrition thick with the essential vitamins, fats, minerals and fatty acids which are best dog puppies important for the best health of your canine or cat.
Dr. Jane Bicks, have actually challenged the requirement to increase illness and bad health in our family pets. If that is so, we also need to take good care of our family pets.
If you are looking for all natural healthy family pet items, they can be discovered in the regional pet stores. Often, some foods are not proper to the pets that you have.
If you are not satisfied with your search, you can look for the animal items online. It is really easy to look for animal products online.
Great food and the best pet products can help your pet remain healthy and fit.
The majority of customers think that buying family pet products such as canned foods dog buying sites are just comparable to the meat dogs favorite food that you can feed them every day. If you desire to offer your pet natural, healthy food, however you do not have sufficient time to prepare raw meals for your animal, you can purchase some of the food products such as those ready-to-eat freezes dried and frozen materials or raw meals. One of the alternatives for feeding pet's raw food is Life's Abundance, which is a natural kibble animal food for your canines and cats . If you are looking for all natural healthy animal products, they can be discovered in the local pet stores.


Safe Hands, Hand Sanitiser 70% Alcohol Gel, Antibacterial & Antiviral, Kills 99.99% Of Bacteria & Viruses, Meets Official European Standards
PROVEN TO KILL BACTERIA & VIRUSES – Our antibacterial and antiviral formula is effective at killing greater than 99.99% of bacteria and viruses and meets official European Standard BS EN 14476. Safe Hands is a truly unique hand sanitiser that is specially formulated to give peace of mind.
70% ALCOHOL & RAPID ACTION FORMULA – Safe Hands rapidly cleanses your hands of all germs. Our sanitiser contains an optimal alcohol percentage of 70% ensuring a quick-acting, fast-drying and non-sticky user experience. No water is needed, just pop open the bottle, squeeze into palms, rub hands together thoroughly, and you’re good to go!
MOISTURISES & SOFTENS SKIN – Safe Hands is tough on germs but kind to skin. Our formula has been specially formulated with moisturiser to protect your skin from drying. Gone are the days of abrasive sanitisers that leave hands feeling dry and dull, using our sanitiser will leave your skin feeling nourished, smooth and safe from all harmful viruses and bacteria.
OFFICIALLY CERTIFIED TO EUROPEAN STANDARDS – Safe Hands has received three official certifications from European Standards. Our sanitiser passes European standard BS EN 14476 (effective antiviral protection), BS EN1276 (effective antibacterial protection) and BS EN 1500 (effective at killing 99.9% of germs on the skin).
PRODUCED IN THE UK – We pride ourselves on our British heritage, from design to formulation to packaging, Safe Hands is proudly produced to the highest standards in the United Kingdom.”


Una Nueva Solución para el SCI: Las bacterias-el eslabón perdido en el tratamiento del síndrome del colon irritable de Mark Pimentel M.D. está disponible para descargar en formato PDF y EPUB. Aquí puedes acceder a millones de libros. Todos los libros disponibles para leer en línea y descargar sin necesidad de pagar más.
Mark Pimentel M.D. con Una Nueva Solución para el SCI: Las bacterias-el eslabón perdido en el tratamiento del síndrome del colon irritable
A New IBS Solution offers a revolutionary look at the way Irritable Bowel Syndrome (IBS) is currently being treated. Today, over 60 million people of all ages across the country are suffering from IBS, which is characterized by abdominal pain, bloating, and altered bowel habits. But because patients may find it difficult to discuss their bowel problems with their physicians, they often suffer in silence or even worse, “learn to live with it.”
According to Dr. Mark Pimentel, Director of the Gastrointestinal Motility Program at Cedars-Sinai Medical Center, the majority of IBS cases can be treated successfully. Dr. Pimentel believes that the root cause of most IBS symtoms can be attributed to an overgrowth of bacteria in the small intestine. A New IBS Solution takes you through the historical evolution of conventional medicine's view on IBS in a way that can be easily understood. In addition, Dr. Pimentel presents a simple treatment protocol that will not only help you resolve your IBS symptoms, but will also prevent their recurrence.


Dear candidates of grade 2020
Recently, the new type of coronavirus infection of pneumonia affects the hearts of the people of the whole country. The Party Central Committee, the State Council, the Ministry of education, the provincial Party committee, the provincial government and the provincial education department have made special arrangements for the prevention and control of the epidemic situation. The Party committee of our University attaches great importance to the life, health and safety of teachers and students, puts forward clear requirements for the implementation of the work deployment of higher authorities, and formulates the work plan for epidemic prevention and control. According to the notice of Shaanxi Provincial Education Examination Institute, we will postpone the publication of the preliminary examination results of postgraduate enrollment examination in 2020. The school will, according to the situation of epidemic prevention and control and the arrangement of superior competent departments, organize and arrange the follow-up work such as the re examination of postgraduate enrollment on the premise of ensuring the health of candidates and openness, fairness and justice. Please rest assured to prepare for the re examination, wait patiently and pay attention to the official notice of the school's official website and wechat public number. In the face of the epidemic, please strengthen confidence, scientific prevention and control, consciously at home, reduce going out, and effectively strengthen personal and family protection. At the same time, we should treat the epidemic situation correctly and keep a sunny and healthy attitude. Let's work together to tide over the difficulties. After the epidemic is overcome, we will wait for you in Xi'an University of Posts and telecommunications!
Graduate School of Xi'an University of Posts and Telecommunications
February 3, 2020


Rivier University ranked #1 in the state and top tier in the nation for nursing
The University has received top rankings for its nursing programs, including #1 Nursing School in New Hampshire.
Following a comprehensive evaluation of U.S. nursing schools for 2022, Nursing Schools Almanac ranked Rivier University as the #1 Nursing School in New Hampshire and #10 in New England. Rivier also ranks #47 nationwide among private nursing schools.
“Rivier is gratified to learn of the University’s exceptional national, regional, and state rankings for nursing,” says Sister Paula Marie Buley, IHM, Rivier’s President. “This further demonstrates the transformational nature of a Rivier education and the outstanding nursing faculty and curriculum that students experience.”
Nursing Schools Almanac assessed 3,000 schools on three critical dimensions: academic prestige and perceived value, breadth and depth of nursing programs offered, and student success, particularly on the NCLEX licensure examination. Rivier Bachelor of Science in Nursing students achieved an NCLEX first-time pass rate of 92% in 2021, far exceeding the national average of 82%.
The University’s nursing labs were recently transformed with a $5 million total renovation. The Nursing Simulation and Clinical Education Center is open and fully operational for hands-on learning and high-tech simulation for nurses and nurse practitioners. State-of-the-art skills and simulation labs offer the newest technology for clinical assessment, community nursing, and telehealth simulation.
“Our new Nursing Simulation and Clinical Education Center has elevated the quality of nursing education and patient care,” shares Dr. Paula Williams, Dean of the Division of Nursing and Health Professions. “The Center is equipped with high-fidelity simulators and state-of-the-art technology that allow students to gain real-world experience in controlled clinical settings.”
Rivier offers on-campus and online nursing degrees from the associate to the doctoral level, and all programs are accredited by the Accreditation Commission for Education in Nursing (ACEN). For complete information, view the University’s expansive nursing program offerings.
Prospective students are invited to contact the Office of Admissions at email@example.com or (603) 897-8507 to learn about the application process and the University’s competitive financial aid assistance. High school students are invited to schedule an in-person campus tour and interview.


One compelling argument as to why feet can smell worse than armpits is that the feet contain more sweat glands than the armpits, which can lead to more bacteria growth and a stronger odor. While both the feet and armpits contain apocrine glands, which produce sweat that can mix with bacteria and create an odor, the feet contain more eccrine glands, which produce a higher volume of sweat. This excess sweat can create a moist environment in shoes and socks, which can further promote bacterial growth and contribute to foot odor.
In addition, the feet are often covered in socks and shoes, which can trap moisture and sweat, making it difficult for the feet to breathe and further contributing to the growth of odor-causing bacteria. This can be exacerbated by certain footwear choices, such as closed-toe shoes or boots, which can trap heat and sweat and create an ideal environment for bacterial growth.
Another factor that can contribute to foot odor is the presence of fungi, such as athlete’s foot. Fungal infections can cause a foul smell, as well as itching and peeling of the skin.
While both feet and armpits can develop unpleasant odors, the higher number of sweat glands in the feet, combined with the warm and moist environment created by socks and shoes, can make them more prone to developing a stronger and more persistent odor than the armpits.
In conclusion, while the exact causes of foot and armpit odor may vary from person to person, the higher number of sweat glands and the warm, moist environment of the feet make them more likely to develop a stronger and more persistent odor than the armpits.


At the Rafael Nadal Foundation Centers, we strive to instill positive nutritional habits among the children and adolescents we serve. We recognize that proper nutrition is not only crucial for physical development but also for emotional and mental well-being. In this regard, we focus on two key aspects: nutritional education and the prevention of potential eating disorders.
Nutritional Education: Mindful Eating
In our centers, we dedicate time to impart fundamental knowledge about nutrition. Our approach goes beyond simply listing food groups or counting calories. We delve into the essence of nutrition, emphasizing the importance of a balanced and varied diet.
We teach children and adolescents to identify foods rich in essential nutrients such as vitamins, minerals, and antioxidants. Additionally, we educate them on the importance of maintaining proper hydration and how to choose fresh and natural foods over processed and ultra-processed ones. Through practical examples and interactive activities, we capture the attention of our young participants, making nutrition accessible and relevant to their daily lives.
Prevention of Eating Disorders: Healthy Relationship with Food
Alongside nutritional education, we care about the mental and emotional health of our young people and focus on preventing potential eating disorders.
We recognize that in today’s society, there is considerable pressure related to body image and food, especially among adolescents.
We work with participants to foster a positive and balanced relationship with food. This helps them understand the importance of listening to their bodies and recognizing hunger and satiety cues. We also address delicate issues such as the influence of media and social networks on body image perception and unrealistic beauty standards.
Through reflection exercises and open discussions, we create a safe space where young people can express their concerns and receive support. Our holistic approach promotes comprehensive well-being, recognizing the connection between mind, body, and spirit.
Engaging Families: Extending Impact to Homes and Communities
We acknowledge that family influence is crucial in shaping healthy eating habits. Therefore, we extend our educational sessions to the families of our participants. We believe in the power of community learning and mutual support among family members, children, and educators.
By involving families, we strengthen the impact of our work, creating a supportive home environment that reinforces positive messages about food and well-being. We provide resources and practical tools for parents to promote a healthy relationship with food at home, from meal planning to food preparation and the encouragement of physical activity.
At the Rafael Nadal Foundation, we believe that healthy eating is more than just a list of foods we should or should not eat. It is a journey towards physical, emotional, and mental well-being.
In our centers, we provide the children and adolescents we serve with the necessary support to develop a positive and balanced relationship with food and their bodies.


Reduce Violence Against Emergency Nurses
Violence in emergency departments has reached epidemic levels and emergency nurses are particularly vulnerable. The 24‐hour accessibility of the emergency department, the lack of adequately trained, armed or visible security guards, and an overall stressful environment are among the chief reasons why emergency nurses are victims of violence at such a high rate.
The Emergency Nurses Association (ENA) will continue to advocate for laws that strengthen the criminal penalties for assaulting or battering emergency nurses and other health care workers in emergency departments.
Reduce Workplace Injury
Nurses are at risk of harm from the environment in which they work. Factors influencing this situation include ergonomic injuries, needle stick injuries, increasing morbid obesity levels, and an aging workforce.
ENA will support federal legislation and programs aimed at reducing workplace injuries and illnesses for health care personnel and patients, while improving the safety of patient care delivery.
Establish Safe Emergency Department Staffing Guidelines
Maintaining the appropriate level of emergency nurse staff is critical to the ability of emergency departments to provide quality healthcare for patients.
As such, ENA supports policies that empower nurses to create appropriate staffing plans specific to each emergency department. This approach recognizes the many factors that determine the appropriate level of staffing, as well as the flexibility needed to account for changing circumstances in emergency departments.
Increase Funding for Nursing Education and Training
ENA urges Congress to increase its investment to alleviate the growing nursing shortage that is impacting every aspect of the U.S. healthcare system and negatively affecting patient care.
ENA will support legislation that reauthorizes and funds the Nursing Workforce Development programs under Title VIII of the Public Health Service Act. The Title VIII programs provide the main federal funding for nursing education, practice, and retention.
Protect and Recognize Emergency Nursing Scope of Practice
ENA will support public policies that remove restriction on the role and scope of practice of RNs and advance practice registered nurses (APRNs) in appropriate healthcare settings.
ENA will support legislation that allows APRNs to practice autonomously, and independently prescribe medication, and oppose restrictions on administering procedural sedation medications by qualified emergency department nurses. In addition, ENA will support legislation that recognizes APRNs as full practice providers in the Veterans Administration’s
health care system.
Support Programs to Enhance Access and Improve Care for Mental Health Patients
ENA will support federal government initiatives aimed at enhancing access and improving care for mental health patients. Further, ENA supports policies that alleviate the holding or boarding of such patients in the emergency department.
ENA will support comprehensive mental health reform legislation, including bills that improve Medicare and Medicaid reimbursement for mental health services and enhance community-based resources for the mentally ill.
Increase support for trauma systems and trauma centers
Trauma is the leading cause of death for people ages 1‐44, and 35 million people are treated for traumatic injuries every year. Nevertheless, more than 45 million people lack access within one hour to a trauma center able to treat severe traumatic injuries.
ENA will continue to fight for legislation and funding to improve the trauma care in the U.S. This includes both the development of optimally functioning trauma systems and quick access to state‐of‐the‐art trauma centers.
Maintain funding for poison control centers
According to the Centers for Disease Control and Prevention, more than 40,000 people die annually from poisoning and approximately 2,000 are treated each day in emergency departments.
ENA will support programs and funding that provide resources for the nation’s 56 poison control centers.
Improve treatment of patients who abuse prescription opioids and heroin
The U.S. is experiencing a historic epidemic in opioids and heroin abuse. In 2014, drug overdoses were the leading cause of accidental death in the U.S.
ENA will support comprehensive legislation to combat the ongoing national crisis of addiction to opioid drugs, including provisions to encourage states and local communities to pursue a full array of proven strategies to combat addiction and support individuals in recovery.
Fund Research to Improve the Delivery of Emergency Care
ENA will continue to strive for improved patient safety and enhanced quality of care by collaborating with appropriate stakeholders to promote and conduct research to enhance emergency care for all patients.
ENA was a strong advocate of the establishment in 2012 of the Office of Emergency Care Research (OECR) at the National Institutes of Health. The Office is a focal point for basic and clinical research on emergency care across NIH. ENA will continue to support the research initiatives and innovations fostered by OECR.
In addition, ENA will support the work of the National Institute of Nursing Research (NINR) at the National Institutes of Health and urges adequate federal funding for NINR within the overall NIH budget.
Continue and expand federal research and support for pediatric emergency care
ENA will strongly support policies that will enhance emergency medical services for children. As such, ENA urges Congress to continue to support the Emergency Medical Services for Children (EMSC) program at the Health Resources and Service Administration. EMSC, which has provided funds to every state, is the only federal program dedicated to improving emergency care for children.
ENA will support legislation that reauthorizes and provides adequate federal funding for the EMSC program.


While much has been done to educate food handlers on safe food-handling practices, continued training in food allergies is needed by workers to recognize the initial signs of allergic reactions – and the best practices for safely preparing food for and serving food allergic guests.
What Are Major Food Allergens?
Although most food allergies cause relatively mild and minor symptoms, some food allergies can cause severe reactions, and may even be life-threatening.
There is no cure for food allergies. Strict avoidance of food allergens — and early recognition and management of allergic reactions to food — are important measures to prevent serious health consequence.
While more than 160 foods can cause allergic reactions in people with food allergies, the law identifies the eight most common allergenic foods. These foods account for 90 percent of food allergic reactions, and are the food sources from which many other ingredients are derived.The eight foods identified by the law are:
- Fish (e.g., bass, flounder, cod)
- Crustacean shellfish (e.g., crab, lobster, shrimp)
- Tree nuts (e.g., almonds, walnuts, pecans)
These eight foods, and any ingredient that contains protein derived from one or more of them, are designated as “major food allergens” by FALCPA.
FDA’s Role: Labeling
To help Americans avoid the health risks posed by food allergens, FDA enforces the Food Allergen Labeling and Consumer Protection Act of 2004 (the Act). The Act applies to the labeling of foods regulated by FDA which includes all foods except poultry, most meats, certain egg products, and most alcoholic beverages which are regulated by other Federal agencies. The Act requires that food labels must clearly identify the food source names of any ingredients that are one of the major food allergens or contain any protein derived from a major food allergen.As a result, food labels help allergic consumers identify offending foods or ingredients so they can more easily avoid them.
Video: Food Allergies – Reducing the Risks
Severe Food Allergies Can Be Life-Threatening
Following ingestion of a food allergen(s), a person with food allergies can experience a severe, life-threatening allergic reaction called anaphylaxis.
This can lead to:
- constricted airways in the lungs
- severe lowering of blood pressure and shock (“anaphylactic shock”)
- suffocation by swelling of the throat
Each year in the U.S., it is estimated that anaphylaxis to food results in:
- 30,000 emergency room visits
- 2,000 hospitalizations
- 150 deaths
Prompt administration of epinephrine by autoinjector (e.g., Epi-pen) during early symptoms of anaphylaxis may help prevent these serious consequences.
Know the Symptoms
Symptoms of food allergies typically appear from within a few minutes to 2 hours after a person has eaten the food to which he or she is allergic. Allergic reactions can include:
- Flushed skin or rash
- Tingling or itchy sensation in the mouth
- Face, tongue, or lip swelling
- Vomiting and/or diarrhea
- Abdominal cramps
- Coughing or wheezing
- Dizziness and/or lightheadedness
- Swelling of the throat and vocal cords
- Difficulty breathing
- Loss of consciousness
Mild Symptoms Can Become More Severe
Initially mild symptoms that occur after ingesting a food allergen are not always a measure of mild severity. In fact, if not treated promptly, these symptoms can become more serious in a very short amount of time, and could lead to anaphylaxis.
Food Allergy Training – Federal & General Resources
- AAAAI/ACAAI Practice Parameters
- Centers for Disease Control and Prevention
- Department of Agriculture
- Department of Health and Human Services
- Food Allergies – Healthfinder
- European Academy of Allergy and Clinical Immunology: International Consensus (ICON) Papers
- FDA – Food Allergens
- FDA – Food Allergen Labeling & Consumer Protection Act – FAQs
- National Institutes of Health (NIH)
- Food Allergy – National Institute of Allergy and Infectious Diseases (NIAID)
- Addendum Guidelines for the Prevention of Peanut Allergy (2017)
- National Institute for Allergy & Infectious Diseases: Guidelines for the Diagnosis and Management of Food Allergy in the United States: Summary for Patients, Families and Caregivers (May 2011)
- National Institute of Allergy and Infectious Diseases, National Institutes of Health: Food Allergy
- National Library of Medicine


What Happens After the Lasik Procedure?
After you are totally prepared, the Lasik treatment takes less compared to fifteen mins to complete for both eyes. This will certainly seem a remarkably short time to have a permanent modification to your vision, as well as hereof Lasik seems nearly as well excellent to be real. Furthermore, most Lasik individuals observe enhanced vision promptly or within a few hours after the Lasik treatment has been done.
This does not mean that you ought to anticipate to walk out of the Lasik clinic with perfect vision and with no should treat your eyes meticulously for the following several days. It also suggests that some Lasik clients will certainly require even more time to see the complete outcomes for the Lasik treatment, in some cases as high as 6 months for their vision to stabilize permanently. Prepare for excellent vision, as well as put in the time and take care of the operation to create its ideal outcome.
Normally the Lasik doctor will offer the post-procedure patient a safety shield for their eyes. He could additionally recommend sunglasses throughout the day if you experience sensitivity to light after the Lasik treatment is done.
Many clients at the Lasik centers often get eye drops to keep their eyes moist for time after the Lasik procedure is done. Once again, this varies by patient and by physician, so inquire about your certain situation, specifically if you are vulnerable to eye dryness on a periodic basis even prior to the Lasik procedure. Also, it might be useful to keep any ceiling followers or other air flow gadgets off in the home for the initial couple of days.
Many clients can return to work and typical everyday activities the day after the Lasik treatment is done, and do not call for any additional help from other pals or member of link the family. There is normally little to no post-operative pain after Lasik has been performed. It is advised that clients go to sleep as rapidly as feasible after the Lasik surgery in order to decrease any type of post-operative pain. Upon waking, boosted vision from the Lasik improvements ought to currently begin to show up.
This boosted vision may not be the final product of the Lasik procedure. The enhancement to nearsightedness after Lasik is usually quick as well as significant, though there dig this could be some issues in checking out quickly for the very first few days after the Lasik operation. This is completely typical, as well as ought to clean up prior to the week is out.
Individuals that make use of Lasik to enhance their farsightedness typically find a dramatic renovation the day after the Lasik surgery. It might be that there is a short-term blurring of objects distant, yet this will certainly settle itself. If this condition continues to be for more than a couple of days, the Lasik physician can advise and suggest temporary glasses until vision is stabilized.
These are all common post-operative suggests for a Lasik client, in order to feel comfy with what to expect after the Lasik procedure. As with any medical therapy, get all of your concerns responded to by the team of your Lasik facility for your individual instance.
In enhancement, most Lasik individuals discover boosted vision immediately or within a couple of hours after the Lasik click to read treatment has actually been performed.
It likewise suggests that some Lasik people will certainly need even more time to see the complete outcomes for the Lasik treatment, sometimes as much as six months for their vision to stabilize completely. Lots of people at the Lasik centers commonly obtain eye goes down to keep their eyes moist for some time after the Lasik treatment is done. People that utilize Lasik to enhance their farsightedness usually discover a significant improvement the day after the Lasik surgical procedure.


This morning, more than 20,000 volunteers and fundraisers gathered to run, walk and ride the main arteries in and out of downtown Toronto in an effort to raise awareness and funds for heart health. Now in its’ 29th year, the annual Becel Heart & Stroke Ride, supporting the Heart and Stroke Foundation, raised nearly $7 million for life saving research, health promotion, and advocacy efforts.
Personally, this was my sixth time participating in the ride, and having witnessed both an uncle and grandfather suffer from heart health challenges, it certainly will not be my last. This year I was joined for the first time by my brother-in-law and father-in-law, who were both riding in honour of a family member and avid cyclist who recently suffered a heart attack. Fortunately, this family member is now on the road to recovery and we’ll hopefully see him at the Ride for Heart next year.
Here’s a quick snapshot of the Ride for Heart by the numbers:
- 5 and 10 kilometers participants ran or walked on the Gardiner Expressway
- 29 years the annual Ride for Heart has been held in Toronto
- 25, 50 and 75 kilometers cyclists rode on the Don Valley Parkway and Gardiner Expressway
- 80 percentage of premature heart diseases and strokes that are preventable by adopting healthy behaviours
- 5,000 runners and walkers that participated in the event
- 15,000 cyclists that participated in the event
- 66,000 Canadians that die each year from heart disease and stroke
- 1.3 million Canadians living with heart disease (estimate)
- 7 million dollars raised at this year’s Becel Heart & Stroke Ride
- 2.5 billion heart beats during an average human lifetime.
- 20.9 billion annual cost of heart disease and strokes on the Canadian economy


MNJ Technologies offers a full range of healthcare IT services, including cloud, security, network, connectivity, and technology sourcing support.
IT adoption in healthcare has risen dramatically as hospitals, health systems, and patients increasingly rely on digital health technologies. Health systems are swiftly moving to virtual care, remote work, and more coordinated communication and data management. Cybersecurity is also a high priority as ransomware attacks increase in the sector.
CIO & IT expertise is also in high demand as healthcare organizations take on a new age of cybersecurity, meet telehealth and electronic health record demand, and take on a new era of big data, AI, and ML initiatives. MNJ is well-positioned to assist with the healthcare industry’s needs in these areas.
Healthcare providers are now leveraging the cloud to gain efficiencies, optimize workflows, lower the costs associated with healthcare delivery, and offer personalization in care plans to improve outcomes. Healthcare organizations are also generating, consuming, storing, and sharing more data in the cloud.
Cloud computing is lowering costs, improving interoperability, and powering a new age of analytics and telemedicine. At the same time, it is increasing concerns over privacy, security, and the impact of downtime.
MNJ provides cloud professionals, tools, and services to meet the growing needs of healthcare.
Our teams can help you plan, manage, and execute cloud initiatives including assisting in design, integration, and support of cloud solutions.
MNJ offers customized managed connectivity solutions to meet each client’s specific needs.
Telehealth and real-time patient monitoring are putting increasing connectivity demand on healthcare organizations. Doctors and staff are increasingly dealing with patients remotely, and the need for communications and connectivity is at an all-time high.
MNJ has a team of experienced healthcare IT services professionals and relationships with over sixty carriers. We’re well-positioned to meet the ever-increasing connectivity demands of healthcare.
Advanced surgical instruments and implants, diagnostic tools, pharmaceutical delivery solutions, and digital health platforms put increasing pressure on network bandwidth. Investment in innovation and next-gen capabilities is ramping as well.
MNJ’s healthcare IT services deliver maximum uptime while delivering higher bandwidth, lower costs, simplified management, and an improved user experience.
MNJ’s skilled people, automated processes, and management tools deliver consistent network performance.
Our solutions offer high performance and availability to meet today’s needs and the bandwidth to enable transformation in healthcare.
MNJ offers expertise and advanced threat intelligence to assist healthcare providers with cybersecurity
Since the global pandemic hit, the healthcare sector has seen a surge in cybersecurity attacks. The pandemic pushed healthcare to enable remote delivery of healthcare services rapidly, creating unforeseen vulnerabilities.
There has been a significant increase in phishing attacks, ransomware, web application attacks, and other threats targeting healthcare providers. The trend has put an enormous strain on healthcare IT & security organizations.
MNJ offers expertise and advanced threat intelligence to assist healthcare providers with cybersecurity by performing assessments, establishing governance programs, prioritizing actions based on risk profiles, and building holistic security programs.
Healthcare transformation requires modernizing IT infrastructure. MNJ offers technology sourcing to ease this burden on healthcare organizations. Our experienced professionals work with IT teams, helping them understand the latest vendor offerings and options to meet transformation needs.
MNJ’s technology sourcing services include procuring hardware and software that best meets a project’s needs—assisting with software licensing, distribution, management, and renewal—configuring, testing, and deploying solutions.
MNJ also provides SaaS subscription management overseeing usage and compliance.
Large projects like remote patient monitoring or telehealth can be achieved painlessly with the help of MNJ’s professional services.
Designing solutions, implementing, managing, and supporting them is also a considerable burden on already understaffed healthcare IT organizations.
MNJ’s professional IT services help healthcare overcome these challenges by supplementing in-house IT with the skills and expertise to take maximum advantage of today’s technology.
Plan · Implement · Navigate with MNJ Technologies


Announcing a Leadership Transition at Georgetown University School of Nursing and Health Studies
May 9, 2019
Dear Members of the Georgetown University Community:
I write to share important news about our School for Nursing and Health Studies (SNHS). Patricia (Pat) Cloonan, Ph.D., RN, Dean of the SNHS, has informed me that she will conclude her term of service as Dean and return to the faculty at the end of this summer. Pat has provided leadership for the SNHS since 2014, first in an interim capacity and for the last three years as Dean of SNHS. I am deeply grateful to Pat for her steadfast commitment to our Georgetown community, and I wish to express my most sincere appreciation for all she has done for the SNHS community over the past five years.
During her time as Dean, Pat strengthened both our undergraduate and graduate programs, advanced our commitment to academic excellence, and oversaw the launch of several new academic programs, including the Executive Master of Science in Health Systems Administration, the Clinical Nurse Leader Master of Science, the Doctor of Nursing Practice, and the Doctor of Nurse Anesthesia Practice. During her tenure, applications to the SNHS have risen to record- breaking numbers, and she has also provided tremendous leadership in growing our online nursing education program, which has enabled more than 1,000 advanced nursing leaders to obtain degrees over the last eight years, many of whom have gone on to provide critically needed care in medically underserved areas.
Pat has been deeply engaged in strengthening our work to promote health equity, and better understand the social determinants that have perpetuated health inequality for vulnerable communities on a local and national level. She has continued to pursue her scholarly work on improving health literacy and developing equitable systems of care. She directs a funded project focused on developing new models for engaging community health workers in the District of Columbia. Under her leadership, graduates of our SNHS have pursued careers oriented towards service, guided by an enduring commitment to health access, equity, and quality.
Prior to her tenure as Dean, Pat had already been a distinguished member of our Georgetown community for more than 20 years. As chair and associate professor of Health Systems Administration, she helped launch the Georgetown University Open School Chapter of the Institute for Healthcare Improvement, leading students through projects related to quality improvement and patient safety. For many years, she also co-led the Health Care Leadership Institute with colleagues from the McDonough School of Business. She has served on a wide range of community boards, including MedStar Georgetown University Hospital.
Over the course of her decades with our University, Pat has left an indelible mark on our students and faculty who are dedicated to pursuing careers in healthcare. She has grounded our work in the conviction that health is inextricably linked to the ability to flourish, and has inspired many of our students to not only provide care to individual patients, but to strive to enact system-wide change on a policy level. In 2014, Pat was recognized with the Georgetown University Medical Center’s Outstanding Service Award. In 2018, she received the Distinguished Healthcare Leader Award from the National Association of Health Services Executives.
In the time ahead, Dr. Edward B. Healton, Executive Vice President for Health Sciences and Executive Dean of our School of Medicine, and I will share a plan for interim leadership as we begin a search for Pat’s successor. I look forward to the opportunity to come together as a community and recognize the many ways that Pat has enriched and strengthened our School of Nursing and Health Studies, and our University.
Please join me in expressing our appreciation to Pat, for her tireless efforts and her continuing dedication to our Georgetown community, as she prepares to conclude her service as Dean.
John J. DeGioia


Preventing your dogs from getting ticks this summer can keep both your pets and your family healthy. Ticks can carry many diseases like Lyme disease and tapeworms into your house, which can make both you and your pets very ill. There are many simple things you can do to protect your dogs and cats from ticks. Read on to learn more.
Medications, such as ones that you give your dog orally are helpful in preventing ticks. There is also spot on treatment, which is a liquid that you apply to the back of your pets and provide month-long protection. However, oral treatments carry fewer risks than spot treatments because spot treatments can be dangerous for small children who come into contact with them.
Shampoo is another method of protecting your dog from ticks. A tick shampoo usually kills ticks upon contact, but you will need to wash your dog often because the active ingredients in tick shampoos are not as long lasting as the oral medicine.
Cats vs. Dogs
Cats are a bit different than dogs, so before using any medications of these types on your cats, you should consult your veterinarian. Cats are much more sensitive to the chemicals in these medications than dogs may be. Many pet owners opt for a tick collar for their pets in the summer months. Safe to use, these collars are often only effective to prevent ticks on your pet’s head and neck.
It is also important to protect your house as well. Start by vacuuming the carpets in your house and by washing your pet’s bedding as well as any fabric where they may spend time. If you have a serious tick infestation, you can consider having your lawn and home sprayed. There are several classes of pesticides that can help control the remaining number of ticks in your yard and help keep any new ticks away.
Protecting your pets from ticks will help protect your family from the diseases that ticks carry, and there are many options for the treatment of tick infestations. To find out more about protecting your pets and home from ticks, contact Clegg’s online or via phone at 888-672-5344.


This Keto peanut butter smoothie is a light and refreshing keto smoothie with a delicious peanut butter flavor. Plus, this is a dairy-free low-carb smoothie with only 4.7 grams of net carbs per serving.
IS PEANUT BUTTER KETO-FRIENDLY?
Yes, you can have peanut butter on a keto diet. Obviously, it has to be pure peanut butter made of 100% peanuts with no added sugar and no added oil.
In fact, vegetable oils are not keto-friendly, and many brands add oils or sugar to their products.
So make sure to check the ingredient list and look for peanuts only or peanuts and salt.
So is this chocolate peanut butter smoothie keto? Yes, it very much is! Use it to top up your macros, and it can help you with your weight loss.
To learn how to calculate your macros, check out my macro calculator!
CAN I TAKE A PEANUT BUTTER SMOOTHIE AS A PROTEIN SHAKE?
Absolutely! Many Keto-friendly smoothies can be used as protein shakes. Just make sure to include enough good protein sources!
One way to add some healthy, keto-friendly proteins is to add a scoop of low-carb protein powder. Most of them would not alter the total carbs but would boost your proteins!
HOW TO MAKE KETO PEANUT BUTTER SMOOTHIE
This is an easy keto breakfast smoothie ready in 5 minutes using only 6 wholesome ingredients.
Bonus, it’s also a vegan smoothie if you are using canned coconut cream instead of heavy cream!
Let’s have a look at the ingredients you need to make this magical refreshing keto drink.
- Peanut butter – as mentioned above, make sure you use natural peanut butter, no added sugar, no added oil.
- Sugar-free crystal sweetener – I love to use Erythritol or Monk fruit blend, but xylitol will work as well. Learn more about keto sweeteners.
- Unsweetened cocoa powder – this adds a delicious chocolate flavor for the best chocolate smoothie.
- Cream – heavy cream or canned coconut cream for a dairy-free alternative. Learn why cream is a good keto-friendly dairy product.
- Unsweetened almond milk
- Ice cubes – play with the amount of ice and change the texture! For an icier, frothier low-carb smoothie recipe, add a few more ice cubes!