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Comparison of approaches to quantify SARS-CoV-2 in wastewater using RT-qPCR: Results and implications from a collaborative inter-laboratory study in Canada

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Journal of Environmental Sciences 2021年第9期33卷 218-229页

作者： Alex H.S.Chik melissa B.Glier mark Servos Chand S.mangat Xiao-Li Pang Yuanyuan Qiu Patrick m.D’Aoust Jean-Baptiste Burnet Robert Delatolla Sarah Dorner Qiudi Geng John P.Giesy Jr Robert mike mcKay michael R.mulvey Natalie Prystajecky Nivetha Srikanthan Yuwei Xie Bernadette Conant Steve E.Hrudey Canadian SARS-CoV-Inter-Laboratory Consortium Consultant to Canadian Water Network Inc. KitchenerCanada Presently at Ontario Clean Water Agency MississaugaCanada Environmental microbiology BC Centre for Disease ControlVancouverCanada Department of Biology University of WaterlooWaterlooCanada National microbiology Laboratory Public Health Agency of CanadaWinnipegCanada Public Health Laboratory Alberta Precision LaboratoryEdmontonCanada Department of Laboratory medicine&Pathology Faculty of Medicine and DentistryUniversity of AlbertaEdmontonAB T6G 2G3Canada Faculty of Engineering University of OttawaOttawaCanada Département des génies civil géologique et des minesPolytechnique MontréalMontréalCanada Veterinary Biomedical Sciences University of SaskatchewanSaskatoonCanada Toxicology Centre University of SaskatchewanSaskatoonCanada Great Lakes Institute for Environmental Research University of WindsorWindsorCanada Department of medical microbiology University of ManitobaWinnipegCanada Department of Pathology and Laboratory medicine University of British ColumbiaVancouverCanada Canadian Water Network Inc. WaterlooCanada Group author details below

Detection of SARS-CoV-2 RNA in wastewater is a promising tool for informing public health decisions during the COVID-19 ***,approaches for its analysis by use of reverse transcription quantitative polymerase chain reaction(RT-q PCR)are still far from standardized *** characterize inter-and intra-laboratory variability among results when using various methods deployed across Canada,aliquots from a real wastewater sample were spiked with surrogates of SARS-CoV-2(gamma-radiation inactivated SARS-CoV-2 and human coronavirus strain 229E[HCoV-229E])at low and high levels then provided"blind"to eight *** estimates reported by individual laboratories were consistently within a 1.0-log_(10) range for aliquots of the same spiked *** laboratories distinguished between low-and high-spikes for both *** expected,greater variability was observed in the results amongst laboratories than within individual laboratories,but SARS-CoV-2 RNA concentration estimates for each spiked condition remained mostly within 1.0-log_(10) *** no-spike wastewater aliquots provided yielded non-detects or trace levels(mL)of SARS-CoV-2 *** appear linked to methods that included or focused on the solids fraction of the wastewater matrix and might represent in-situ SARS-CoV-2 to the wastewater ***-229E RNA was not detected in the no-spike ***,all methods yielded comparable results at the conditions *** behavior of SARS-CoV-2 and spiked surrogates in wastewater should be considered to evaluate method effectiveness.A consistent method and laboratory to explore wastewater SARS-CoV-2 temporal trends for a given system,with appropriate quality control protocols and documented in adequate detail should succeed.

关键词： COVID-19 Wastewater surveillance Public health Quality assurance Quality control

Evaluating Potential Vaccine Antigens in both the Chlamydia trachomatisand Chlamydia muridarumIntravaginal mouse Challenge models

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World Journal of Vaccines 2019年第2期9卷 49-69页

作者： Robin m. Kaufhold melissa A. Boddicker Jodie A. Field Bob J. Lucas Joseph m. Antonello Amy S. Espeseth Julie m. Skinner Jon H. Heinrichs Jeffrey G. Smith Department of Infectious Diseases/Vaccines MRL (West Point PA) Merck & Co. Inc. Kenilworth NJ USA Present Address Teva Pharmaceuticals West Chester Pennsylvania USA Department of Biometrics Research MRL (West Point PA) Merck & Co. Inc. Kenilworth NJ USA Present Address Sanofi Pasteur Swiftwater Pennsylvania USA Present Address Moderna Therapeutics Cambridge Massachusetts USA

Identifying relevant animal challenge models adds to the complexity of human vaccine development. murine challenge models have been the most utilized animal model for Chlamydia trachomatis vaccine development. The question arises as to whether the C. trachomatis or C. muridarum pre-clinical model is optimal. We compared C. muridarum and C. trachomatis intravaginal challenge models in a combined total of seventy-five studies evaluating potential vaccine candidates. In 100% (42/42) of C. muridarum studies, mice immunized with Chlamydia elementary bodies (EB) demonstrated a significant reduction in urogenital bacterial shedding as measured by qPCR (p C. trachomatis studies. We have evaluated proposed vaccine antigens in both models and observed immunization with Chlamydia major outer membrane protein (mOmP) vaccine formulations to be protective (p C. trachomatis model, and immunization with PmpD p82 translocator domain was not protective in either model. We also observed in both models that depletion of CD4+ T-cells in mOmP-immunized mice resulted in diminished protective immunity but animals were still able to reduce the infection level. In contrast, mice immunized with live EBs by intraperitoneal route did not require CD4+ T-cells to resolve urogenital infection from intravaginal challenge in either model. Overall, we have found the C. muridarum model to be a more robust, reliable, and reproducible model for vaccine antigen discovery.

关键词： Chlamydia mouse Challenge model Intravaginal Challenge Sexually Transmitted Diseases

Social-epigeneticmediators for racial disparities in pulmonary impairment among childhood cancer survivors

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Cancer Communications 2023年第2期43卷 290-293页

作者： Nan Song Qian Dong Cheng Chen Qian Li Heather mulder Emily Plyler John Easton Emily Walker Scott Olson Geoffrey Neale Kevin RKrull Deo Kumar Srivastava Kirsten K.Ness Jinghui Zhang melissa m.Hudson Leslie L.Robison I-Chan Huang Zhaoming Wang Department of Epidemiology and Cancer Control St.Jude Children’s Research HospitalMemphisTN 38105USA College of Pharmacy Chungbuk National UniversityCheongjuChungbuk 28160South Korea School of Public Health Shanghai Jiaotong UniversityShanghai 200025P.R.China Department of Biostatistics St.Jude Children’s Research HospitalMemphisTN 38105USA Department of Computational Biology St.Jude Children’s Research HospitalMemphisTN 38105USA Hartwell Center St.Jude Children’s Research HospitalMemphisTN 38105USA Department of Oncology St.Jude Children’s Research HospitalMemphisTN 38105USA

Dear Editor Previous research found that childhood cancer survivors of African ancestry have significantly higher morbidity and mortality than those of European ancestry[1].However,after adjusting for socio-economic factors,the magnitudes of racial health disparities are either substantially decreased or become statistically non-significant[1],suggesting that social and economic determinants may contribute to racial health ***,we conducted epigenome-wide association studies(EWAS)for three key social determinants of health(SDOHs),namely,personal educational attainment,personal income,and neighborhood deprivation among survivors of childhood cancer,where 130 epigenome-wide significant SDOH-CpG associationswere identified among European ancestry survivors,and 25 of which were also validated in African ancestry survivors[2].Notably,many SDOH-associated CpG sites are also associated with tobacco use.

关键词： vivo cancer mortality

Effectiveness of Ledipasvir/Sofosbuvir with / without Ribavarin in Liver Transplant Recipients with Hepatitis C

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Journal of Clinical and Translational Hepatology 2017年第2期5卷 101-108页

作者： Sammy Saab Justin Rheem melissa A.Jimenez Tiffany m.Fong michelle H.mai Caterina A.Kachadoorian Negin L.Esmailzadeh Sherona N.Bau Susan Kang Samantha D.Ramirez Jonathan Grotts Gina Choi Francisco A.Durazo mohammed m.El-Kabany Steven-Huy B.Han Ronald W.Busuttil Departments of medicine at the University of California at Los Angeles Los Angeles California USA Departments of Surgery at the University of California at Los Angeles Los Angeles California USA Department of medicine at Harbor-University of California at Los Angeles medical Center Torrance California USA Departments of Surgery at the University of California at Los Angeles Los Angeles California USA Department of Biostatistics at the University of California at Los Angeles Los Angeles California USA

Background and Aims:Recurrent infection of hepatitis C virus (HCV) in liver transplant (LT) recipients is universal and associated with significant morbidity and ***:We retrospectively evaluated the safety and efficacy of ledipasvir/sofosbuvir with and without ribavirin in LT recipients with recurrent genotype 1 hepatitis ***:Eighty-five LT recipients were treated for recurrent HCV with ledipasvir/sofosbuvirwith and without ribavirin for 12 or 24 *** mean (± standard deviation [SD]) time from LT to treatment initiation was 68 (±71) *** mean (± SD) age of the cohort was 63 (±8.6) years *** recipients were male (70%).Baseline alanine transaminase,total bilirubin,and HCV ribonucleic acid (RNA) values (± SD) were 76.8 (±126)mg/dL,0.8 (±1.3) U/L,and 8,010,421.9 (±12,420,985)IU/mL,*** of 43 recipients who were treated with ribavirin required drug cessation due to side effects,with 4 of those being anemia *** recipient discontinued the ledipasvir/***-one percent of recipients had undetectable viral levels at 4 weeks after starting therapy,and all recipients had complete viral suppression at the end of *** sustained viral response at 12 weeks after completion of therapy was 94%.Conclusion:Ledipasvir and sofosbuvir with and without ribavirin therapy is an effective and well-tolerated interferon-free treatment for recurrent HCV infection after *** is not uncommon in LT recipients receiving ribavirin.

关键词： Liver transplantation Immunosuppressant Hepatitis C Direct-acting agents Sustained viral response

A Novel LNP-Based ChlamydiaSubunit Vaccine Formulation That Induces Th1 Responses without Upregulating IL-17 Provides Equivalent Protection in mice as Formulations That Induced IL-17 and Th1 Cytokines

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World Journal of Vaccines 2020年第4期10卷 55-75页

作者： melissa A. Boddicker Robin m. Kaufhold Kara S. Cox Bob J. Lucas Jinfu Xie Deborah D. Nahas Sinoeun Touch Amy S. Espeseth Kalpit A. Vora Julie m. Skinner Department of Infectious Diseases/Vaccines Merck & Co. Inc. West Point PA USA

We evaluated novel Chlamydial vaccines, consisting of major outer membrane protein (mOmP) alone or in combination with polymorphic membrane proteins D (PmpD) and G (PmpG) using a C57BL/6 mouse model. Native mOmP (nmOmP) isolated from m>Cm>. m>muridarumm> elementary bodies (EBs) and recombinant PmpD and PmpG proteins were adjuvanted with monophosphoryl lipid A (mPLA), with either lipid nanoparticles (LNPs) or the cationic lipid dimethyldioctadecylammonium bromide (DDA). Antibody titers to m>Cm>. m>muridarumm> nmOmP, and EBs were evaluated by ELISA, and T-cell responses were analyzed by intracellular cytokine staining (ICS). Protection from challenge was determined by qPCR. Vaccine immunized mice showed significantly higher antibody titers to nmOmP (P m>Cm>. m>muridarumm> EBs (P mpared to the adjuvant alone group. Antibody titers in vaccine groups with monophosphoryl lipid A (mPLA) + LNP were higher as compared to the mPLA + DDA group (P m nmOmP + PmpG + PmpD p73 + PmpD p82 + mPLA + DDA) vs (Cm nmOmP + PmpG + PmpD p73 + PmpD p82 + mPLA + LNP) for both m>Cm>. m>muridarumm> EBs and PmpG. ICS analysis showed more robust CD4 + T-cell responses (IFN-m>γm>/IL-2/TNF-a) in the DDA and LNP groups compared to the adjuvant alone group. The DDA + mPLA gave robust Th17 responses in comparison to mPLA and LNP group. mice immunized with m>Chlamydiam> antigens also showed protection from m>Cm>. m>muridarumm> challenge, by reduction in bacterial shedding for all groups (P mpared to shedding from the adjuvant control. Both vaccine formulations generated robust immunological responses, and both were protective by reducing bacterial shedding after challenge. This data indicates equal protection can be achieved without the induction of Th17 responses.

关键词： LNP Chlamydia IL-17 mouse model Th1 Cytokines

Nutrient Evaluation of Dining Center Food Waste and Comparison to monogastric and Ruminant Feedstuffs

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Open Journal of Animal Sciences 2023年第3期13卷 323-335页

作者： Nelsa m. Beckman Phillip A. Lancaster Haley K. Otott Rachel N. Kort Katherine m. Krauss melissa A. Schrader Jordan T. Gebhardt Charles R. Stark Chad B. Paulk Department of Grain Science and Industry Kansas State University Manhattan USA Department of Clinical Sciences Kansas State University Manhattan USA Department of Housing and Dining Kansas State University Manhattan USA Department of Diagnostic medicine Pathobiology Kansas State University Manhattan USA

The objective of this study was to analyze the nutrient composition and variability of university dining hall food waste and compare it with common feedstuffs used in ruminant and monogastric diets. Food waste was categorized into two initial streams: mainstream (mS) from the serving line and vegetable preparation (VP) from the kitchen. Waste was collected from the Kramer Dining Center, Kansas State University, resulting in 30 daily samples. Waste was weighed and ground to homogenous particle size. Daily samples of mS and VP were analyzed for nutrient composition, where results were combined to calculate the nutrient profile of a hypothetical mixed food waste stream (mX) composited by total weight. Data were analyzed using R statistical software (v 4.2.2). moisture and neutral detergent fiber (NDF) were greater in VP (P , while ether extract (EE) was less compared to mS and mX. Crude protein (CP) was greater (P mS and mX streams compared to VP. The total digestible nutrients (TDN) and energy were greater in mS food waste than in mX, which was also greater than VP (P content, measured by standard deviation, was similar (P > 0.05) among streams for NDF, nitrogen-corrected neutral detergent fiber, acid detergent insoluble crude protein, CP, ash, lignin, and digestible and metabolizable energy. Dry matter and EE variation were greater (P mS, whereas VP was less (P mpared to mX. Standard deviation increased (P mS and mX for neutral detergent insoluble crude protein, TDN, and gross energy when compared to VP. Despite having 70% - 80% moisture, dining hall food waste does have nutritive value and the potential to be included in ruminant and monogastric diets. Further research needs to be done to understand the value of including it in animal diets.

关键词： Food Waste Feedstuffs monogastric Ruminant Nutrient Variability

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少女CEO创业记

高中生（作文） 2014年第9期 40-41页

作者： melissa Locker Lina

梅根·格拉塞17岁时创立了一家名叫Yellow berry的少女内衣公司。她现在是一名高中生,课程繁忙,课外活动丰富,同时还要运营一家快速成长的企业,忙得不亦乐乎。格拉塞萌生创办公司的想法,是在一次与妹妹玛丽·玛格丽特逛商场的时候激发... 详细信息

梅根·格拉塞17岁时创立了一家名叫Yellow berry的少女内衣公司。她现在是一名高中生,课程繁忙,课外活动丰富,同时还要运营一家快速成长的企业,忙得不亦乐乎。格拉塞萌生创办公司的想法,是在一次与妹妹玛丽·玛格丽特逛商场的时候激发的。她说：“我跟妈妈和妹妹在商场买内衣,妹妹正在发育阶段,我们想为她购买她人生中的第一件内衣。

关键词： CEO 创业少女 berry 女内衣课外活动发育阶段高中生

machine Learning Highlights Downtrending of COVID-19 Patients with a Distinct Laboratory Profile

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Health Data Science 2021年第1期2021卷 11-19页

作者： He S.Yang Yu Hou Hao Zhang Amy Chadburn Lars F.Westblade Richard Fedeli Peter A.DSteel Sabrina E.Racine-Brzostek Priya Velu Jorge L.Sepulveda michael J.Satlin melissa m.Cushing Rainu Kaushal Zhen Zhao Fei Wang Department of Pathology and Laboratory medicine Weill Cornell MedicineNew YorkNYUSA New York-Presbyterian Hospital/Weill Cornell medical Campus New YorkNYUSA Department of Population Health Sciences Weill Cornell MedicineNew YorkNYUSA Department of medicine Weill Cornell MedicineNew YorkNYUSA Department of Emergency medicine Weill Cornell MedicineNew YorkNYUSA Department of Pathology School of Medicine and Health SciencesGeorge Washington UniversityWashington DCUSA Division of Infectious Disease Department of MedicineWeill Cornell MedicineNew YorkNYUSA

*** York City(NYC)experienced an initial surge and gradual decline in the number of SARS-CoV-2-confirmed cases in 2020.A change in the pattern of laboratory test results in COVID-19 patients over this time has not been reported or correlated with patient *** performed a retrospective study of routine laboratory and SARS-CoV-2 RT-PCR test results from 5,785 patients evaluated in a NYC hospital emergency department from march to June employing machine learning ***.A COVID-19 high-risk laboratory test result profile(COVID19-HRP),consisting of 21 routine blood tests,was identified to characterize the SARS-CoV-2 *** half of the SARS-CoV-2 positive patients had the distinct COVID19-HRP that separated them from SARS-CoV-2 negative ***-CoV-2 patients with the COVID19-HRP had higher SARS-CoV-2 viral loads,determined by cycle threshold values from the RT-PCR,and poorer clinical outcome compared to other positive patients without the ***,the percentage of SARS-CoV-2 patients with the COVID19-HRP has significantly decreased from march/April to may/***,viral load in the SARS-CoV-2 patients declined,and their laboratory profile became less distinguishable from SARS-CoV-2 negative patients in the later *** longitudinal analysis illustrates the temporal change of laboratory test result profile in SARS-CoV-2 patients and the COVID-19 evolvement in a US *** analysis could become an important tool in COVID-19 population disease severity tracking and *** addition,this analysis may play an important role in prioritizing high-risk patients,assisting in patient triaging and optimizing the usage of resources.

关键词： routine evolve analysis

Limited Knowledge of Acetaminophen in Patients with Liver Disease

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Journal of Clinical and Translational Hepatology 2016年第4期4卷 281-287页

作者： Sarnrny Saab Peter G.Konyn matthew R.Viramontes melissa A.Jimenez Jonathan F.Grotts Wally Hamidzadah Veronica P.Dang Negin L.Esmailzadeh Gina Choi Francisco A.Durazo mohamed m.El-Kabany Steven-Huy B.Han myron J.Tong Department of medicine the University of California at Los Angeles Los Angeles CA USA Department of Surgery the University of California at Los Angeles Los Angeles CA USA Department of Surgery the University of California at Los Angeles Los Angeles CA USA Department of Biostatistics the University of California at Los Angeles Los Angeles CA USA Department of Surgery the University of California at Los Angeles Los Angeles CA USA California and Huntington medical Research Institutes Pasadena CA USA

Background andAims:Unintentional acetaminophen overdose remains the leading cause of acute liver failure in the United *** with underlying liver disease are at higher risk of poor outcomes from acetaminophen *** knowledge of acetaminophen may be a preventable contributor to elevated rates of overdose and thus acute liver *** purpose of this study is to assess knowledge of acetaminophen dosing and presence of acetaminophen in common combination products in patients with liver ***:We performed a cross-sectional study of patients with liver disease at the Pfleger Liver Institute at the University of California,Los Angeles between June 2015 and August *** completed a demographic questionnaire and an acetaminophen knowledge *** information was obtained from the medical ***:Of 401 patients with liver disease,30 (15.7%) were able to correctly identify that people without liver disease can safely take up to 4 g/day of *** majority of patients (79.9%-86.8%) did not know that Norco(R) (hydrocone/acetaminophen),Vicodin(R) (hydrocone/acetaminophen) and Percocet(R) (oxycodone/acetaminophen) contained *** 45.3% of the patients knew that Tylenol(R) #3 contained ***:We conclude that patients with liver disease have critically low levels of knowledge of acetaminophen,putting them at risk both of acetaminophen overdose,as well as undermedication,and inadequate management of chronic *** recommend an increase in education efforts regarding acetaminophen dosage and its safety in the setting of liver *** education for those at risk of low acetaminophen knowledge is essential to minimizing acetaminophen overdose rates and optimizing pain management.

关键词： Acetaminophen Pain management Cirrhosis