is a foodborne protozoan parasite that causes outbreaks of diarrheal illness (cyclosporiasis) with clear seasonality worldwide. In the environment,
oocysts are very robust, and contact with ...contaminated soil may serve as an important vehicle in the transmission of this organism, and it is considered a risk factor for this infection. The present study evaluated a flotation concentration method, previously shown to provide the best detection results when compared with DNA isolation directly from soil samples, in two main types of farm soil, silt loam soil and sandy clay loam, as well as in commercial potting mix samples inoculated with different numbers of
oocysts. The flotation method was able to detect as few as 10 oocysts in 10 g of either type of farm soil without modifications, but needed an extra wash and samples of reduced size for the processing of the commercial potting mix to be able to detect 20 oocysts/5 g. A recently modified real-time PCR method for the detection of
based on a mitochondrial gene target was also evaluated using selected samples of each type of soil. This comparative study confirmed that the concentration of oocysts in soil samples by flotation in high-density sucrose solutions is a sensitive method that can detect low numbers of oocysts in different types of soil.
Cyclospora cayetanensis is a protozoan parasite that causes foodborne outbreaks of diarrheal illness (cyclosporiasis) worldwide. Contact with soil may be an important mode of transmission for C. ...cayetanensis and could play a role in the contamination of foods. However, there is a scarcity of detection methods and studies for C. cayetanensis in soil. Traditional parasitology concentration methods can be useful for the detection of C. cayetanensis, as found for other protozoa parasites of similar size. The present study evaluated a concentration method using flotation in saturated sucrose solution, subsequent DNA template preparation and qPCR following the Bacteriological Analytical Manual (BAM) Chapter 19b method. The proposed flotation method was compared to three commercial DNA isolation kits (Fast DNATM 50 mL SPIN kit for soil (MP Biomedicals, Irvine, CA, USA), Quick-DNATM Fecal/Soil Microbe Midiprep kit (Zymo Research, Irvine, CA, USA) and DNeasy® PowerMax® Soil Kit (Qiagen, Hilden, Germany)) for the isolation and detection of DNA from experimentally seeded C. cayetanensis soil samples (5−10 g with 100 oocysts). Control unseeded samples were all negative in all methods. Significantly lower cycle threshold values (CT) were observed in the 100 oocyst C. cayetanensis samples processed via the flotation method than those processed with each of the commercial DNA isolation kits evaluated (p < 0.05), indicating higher recovery of the target DNA with flotation. All samples seeded with 100 oocysts (n = 5) were positive to the presence of the parasite by the flotation method, and no inhibition was observed in any of the processed samples. Linearity of detection of the flotation method was observed in samples seeded with different levels of oocysts, and the method was able to detect as few as 10 oocysts in 10 g of soil samples (limit of detection 1 oocyst/g). This comparative study showed that the concentration of oocysts in soil samples by flotation in high-density sucrose solutions is an easy, low-cost, and sensitive method that could be implemented for the detection of C. cayetanensis in environmental soil samples. The flotation method would be useful to identify environmental sources of C. cayetanensis contamination, persistence of the parasite in the soil and the role of soil in the transmission of C. cayetanensis.
Cyclosporiasis, caused by the coccidian parasite Cyclospora cayetanensis, has emerged as an increasing global public health concern, with the incidence of laboratory-confirmed domestically acquired ...cases in the US exceeding 10,000 since 2018. A recently published qPCR assay (Mit1C) based on a mitochondrial target gene showed high specificity and good sensitivity for the detection of C. cayetanensis in fresh produce. The present study shows the integration and verification of the same mitochondrial target into a fully automated and streamlined platform that performs DNA isolation, PCR, hybridization, results visualization, and reporting of results to simplify and reduce hands-on time for the detection of this parasite. By using the same primer sets for both the target of interest (i.e., Mit1C) and the internal assay control (IAC), we were able to rapidly migrate the previously developed Mit1C qPCR assay into the more streamlined and automated format Rheonix C. cayetanensisTM Assay. Once the best conditions for detection were optimized and the migration to the fully automated format was completed, we compared the performance of the automated platform against the original “bench top” Mit1C qPCR assay. The automated Rheonix C. cayetanensis Assay achieved equivalent performance characteristics as the original assay, including the same performance for both inclusion and exclusion panels, and it was able to detect as low as 5 C. cayetanensis oocysts in fresh produce while significantly reducing hands-on time. We expect that the streamlined assay can be used as a tool for outbreak and/or surveillance activities to detect the presence of C. cayetanensis in produce samples.
Cyclospora cayetanensis is a foodborne protozoan parasite that causes outbreaks of diarrheal illness (cyclosporiasis) with clear seasonality worldwide. In the environment, C. cayetanensis oocysts are ...very robust, and contact with contaminated soil may serve as an important vehicle in the transmission of this organism, and it is considered a risk factor for this infection. The present study evaluated a flotation concentration method, previously shown to provide the best detection results when compared with DNA isolation directly from soil samples, in two main types of farm soil, silt loam soil and sandy clay loam, as well as in commercial potting mix samples inoculated with different numbers of C. cayetanensis oocysts. The flotation method was able to detect as few as 10 oocysts in 10 g of either type of farm soil without modifications, but needed an extra wash and samples of reduced size for the processing of the commercial potting mix to be able to detect 20 oocysts/5 g. A recently modified real-time PCR method for the detection of C. cayetanensis based on a mitochondrial gene target was also evaluated using selected samples of each type of soil. This comparative study confirmed that the concentration of oocysts in soil samples by flotation in high-density sucrose solutions is a sensitive method that can detect low numbers of oocysts in different types of soil.
Cyclospora cayetanensis is a protozoan parasite that causes foodborne outbreaks of diarrheal illness (cyclosporiasis) worldwide. Contact with soil may be an important mode of transmission for C. ...cayetanensis and could play a role in the contamination of foods. However, there is a scarcity of detection methods and studies for C. cayetanensis in soil. Traditional parasitology concentration methods can be useful for the detection of C. cayetanensis, as found for other protozoa parasites of similar size. The present study evaluated a concentration method using flotation in saturated sucrose solution, subsequent DNA template preparation and qPCR following the Bacteriological Analytical Manual (BAM) Chapter 19b method. The proposed flotation method was compared to three commercial DNA isolation kits (Fast DNAsup.TM 50 mL SPIN kit for soil (MP Biomedicals, Irvine, CA, USA), Quick-DNAsup.TM Fecal/Soil Microbe Midiprep kit (Zymo Research, Irvine, CA, USA) and DNeasysup.® PowerMaxsup.® Soil Kit (Qiagen, Hilden, Germany)) for the isolation and detection of DNA from experimentally seeded C. cayetanensis soil samples (5–10 g with 100 oocysts). Control unseeded samples were all negative in all methods. Significantly lower cycle threshold values (Csub.T) were observed in the 100 oocyst C. cayetanensis samples processed via the flotation method than those processed with each of the commercial DNA isolation kits evaluated (p < 0.05), indicating higher recovery of the target DNA with flotation. All samples seeded with 100 oocysts (n = 5) were positive to the presence of the parasite by the flotation method, and no inhibition was observed in any of the processed samples. Linearity of detection of the flotation method was observed in samples seeded with different levels of oocysts, and the method was able to detect as few as 10 oocysts in 10 g of soil samples (limit of detection 1 oocyst/g). This comparative study showed that the concentration of oocysts in soil samples by flotation in high-density sucrose solutions is an easy, low-cost, and sensitive method that could be implemented for the detection of C. cayetanensis in environmental soil samples. The flotation method would be useful to identify environmental sources of C. cayetanensis contamination, persistence of the parasite in the soil and the role of soil in the transmission of C. cayetanensis.
Cyclosporiasis, caused by the coccidian parasite Cyclospora cayetanensis, has emerged as an increasing global public health concern, with the incidence of laboratory-confirmed domestically acquired ...cases in the US exceeding 10,000 since 2018. A recently published qPCR assay (Mit1C) based on a mitochondrial target gene showed high specificity and good sensitivity for the detection of C. cayetanensis in fresh produce. The present study shows the integration and verification of the same mitochondrial target into a fully automated and streamlined platform that performs DNA isolation, PCR, hybridization, results visualization, and reporting of results to simplify and reduce hands-on time for the detection of this parasite. By using the same primer sets for both the target of interest (i.e., Mit1C) and the internal assay control (IAC), we were able to rapidly migrate the previously developed Mit1C qPCR assay into the more streamlined and automated format Rheonix C. cayetanensissup.TM Assay. Once the best conditions for detection were optimized and the migration to the fully automated format was completed, we compared the performance of the automated platform against the original “bench top” Mit1C qPCR assay. The automated Rheonix C. cayetanensis Assay achieved equivalent performance characteristics as the original assay, including the same performance for both inclusion and exclusion panels, and it was able to detect as low as 5 C. cayetanensis oocysts in fresh produce while significantly reducing hands-on time. We expect that the streamlined assay can be used as a tool for outbreak and/or surveillance activities to detect the presence of C. cayetanensis in produce samples.
Over one-fourth of Googlers use internal, data-center-hosted virtual desktops. This on-premises offering sits in the corporate network and allows users to develop code, access internal resources, and ...use GUI tools remotely from anywhere in the world. Among its most notable features, a virtual desktop instance can be sized according to the task at hand, has persistent user storage, and can be moved between corporate data centers to follow traveling Googlers. Until recently, our virtual desktops were hosted on commercially available hardware on Google’s corporate network using a homegrown open-source virtual cluster-management system called Ganeti. Today, this substantial and Google-critical workload runs on GCP (Google Compute Platform). This article discusses the reasons for the move to GCP, and how the migration was accomplished.
Abstract Background & Aims The association between prevalence of celiac disease and geographic region is incompletely understood, but the occurrence of several autoimmune disorders has been found to ...vary along a North–South gradient. We examined geographic, demographic, and clinical factors associated with prevalence of celiac disease and gluten-free diet (GFD) in the United States (US). Methods In a population-based study, we analyzed data on gluten-related conditions from the US National Health and Nutrition Examination Survey, from 2009 through 2014, on 22,277 participants 6 years and older. We identified persons with celiac disease, based on results of serum tests for immunoglobulin A against tissue transglutaminase and endomysium or on both a health care provider diagnosis and adherence to a GFD. Gluten-avoidance without celiac disease was defined as adherence to a GFD without a diagnosis of celiac disease. We compared mean serum levels of biochemical and nutritional markers based on status of gluten-related conditions. Results We found 0.7% of participants to have celiac disease and 1.1% of participants to avoid gluten without celiac disease. Celiac disease was more common among individuals who lived at latitudes of 35–39º North (odds ratio, 3.2; 95% CI, 1.4–7.1) or at latitudes of 40º North or more (odds ratio, 5.4; 95% CI, 2.6–11.3) than individuals who lived at latitudes below 35º North independent of race or ethnicity, socioeconomic status, and body mass index. Gluten-avoidance without celiac disease was more common among individuals who lived at latitudes of 40º North or more, independent of demographic factors and body mass index. Participants with undiagnosed celiac disease (identified by positive results from serologic tests) had lower mean levels of B12 and folate (data collected from 2009 through 2012) than persons without celiac disease. Participants with a health-care provider diagnosis of celiac disease had a lower mean level of hemoglobin than persons without celiac disease. Mean levels of albumin, calcium, iron, ferritin, cholesterol, vitamin B6, and vitamin D (data collected from 2009 through 2010) did not differ between participants with gluten-related conditions and those without. Conclusions In the US population, a higher proportion of persons living at latitudes of 35º North or greater have celiac disease and/or avoid gluten than persons living south of this latitude, independent of race or ethnicity, socioeconomic status, or body mass index. Mean levels of B12 and folate are lower in individuals with undiagnosed celiac disease, and levels of hemoglobin are lower in participants with a diagnosis of celiac disease, compared to individuals without celiac disease.
Recently published randomized controlled trials (RCTs) showed improved overall survival (OS) and progression-free survival (PFS) with the combination of immunotherapy and chemotherapy as compared to ...chemotherapy alone in advanced non-small cell lung cancer (NSCLC). We aimed to provide a systematic review and meta-analysis of RCTs regarding the efficacy and safety of immunotherapy and chemotherapy combinations for advanced NSCLC.
On December 23
, 2021, we searched databases for RCTs that reported PFS and OS as primary outcomes.
We included 11 RCTs with 6,386 patients (3,850 in the combination therapy group and 2,536 in the chemotherapy group). Combination therapy was associated with an improvement in PFS (HR: 0.60; 95% CI: 0.54, 0.66; P < 0.00001) andOS (HR: 0.77; 95% CI: 0.68, 0.87; P ≤ 0.0001), compared to chemotherapy. There were no significant differences between both groups in terms of treatment-related adverse events (TRAEs) (RR: 1.07; 95% CI: 0.99, 1.16; P = 0.09).
The combination of immunotherapy and chemotherapy as first-line treatment for advanced NSCLC significantly improved PFS and OS compared to chemotherapy alone without a significant increase in the overall TRAEs.
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