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| 2. |
- Beal, Jacob, et al.
(författare)
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Robust estimation of bacterial cell count from optical density
- 2020
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
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Tidskriftsartikel (refereegranskat)abstract
- Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.
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| 3. |
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- 2019
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Tidskriftsartikel (refereegranskat)
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| 4. |
- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)abstract
- In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
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| 5. |
- Zhang, Huai, et al.
(författare)
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A global survey on the use of the international classification of diseases codes for metabolic dysfunction-associated fatty liver disease.
- 2024
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Ingår i: Hepatology international. - 1936-0541.
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Tidskriftsartikel (refereegranskat)abstract
- With the implementation of the 11th edition of the International Classification of Diseases (ICD-11) and the publication of the metabolic dysfunction-associated fatty liver disease (MAFLD) nomenclature in 2020, it is important to establish consensus for the coding of MAFLD in ICD-11. This will inform subsequent revisions of ICD-11.Using the Qualtrics XM and WJX platforms, questionnaires were sent online to MAFLD-ICD-11 coding collaborators, authors of papers, and relevant association members.A total of 890 international experts in various fields from 61 countries responded to the survey. We also achieved full coverage of provincial-level administrative regions in China. 77.1% of respondents agreed that MAFLD should be represented in ICD-11 by updating NAFLD, with no significant regional differences (77.3% in Asia and 76.6% in non-Asia, p=0.819). Over 80% of respondents agreed or somewhat agreed with the need to assign specific codes for progressive stages of MAFLD (i.e. steatohepatitis) (92.2%), MAFLD combined with comorbidities (84.1%), or MAFLD subtypes (i.e., lean, overweight/obese, and diabetic) (86.1%).This global survey by a collaborative panel of clinical, coding, health management and policy experts, indicates agreement that MAFLD should be coded in ICD-11. The data serves as a foundation for corresponding adjustments in the ICD-11 revision.
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| 6. |
- Huang, Shan, et al.
(författare)
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Dipyridamole enhances the anti-cancer ability of aspirin against colorectal cancer by inducing apoptosis in an unfolded protein response-dependent manner
- 2023
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Ingår i: Cellular Oncology. - : Springer Science and Business Media LLC. - 2211-3428 .- 2211-3436. ; 46:4, s. 953-967
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Tidskriftsartikel (refereegranskat)abstract
- PURPOSE: Available evidence indicates that dipyridamole enhances the anti-thrombotic effects of aspirin for the prevention of secondary strokes. Aspirin is a well-known non-steroid anti-inflammatory drug. This anti-inflammatory property has turned aspirin into a potential drug for inflammation-related cancers such as colorectal cancer (CRC). Here, we aimed to explore whether the anti-cancer effect of aspirin against CRC could be improved by combined administration with dipyridamole.METHODS: Population-based clinical data analysis was conducted to assess a possible therapeutic effect of combined dipyridamole and aspirin treatment in inhibiting CRC compared with either monotherapy. This therapeutic effect was further verified in different CRC mouse models, i.e. an orthotopic xenograft mouse model, an AOM/DSS mouse model, an Apc min/+ mouse model and a patient derived xenograft (PDX) mouse model. The in vitro effects of the drugs on CRC cells were tested using CCK8 and flow cytometry assays. RNA-Seq, Western blotting, qRT-PCR and flow cytometry were used to identify the underlying molecular mechanisms. RESULTS: We found that dipyridamole combined with aspirin had a better inhibitory effect on CRC than either monotherapy alone. The enhanced anti-cancer effect of the combined use of dipyridamole with aspirin was found to rely on the induction of an overwhelmed endoplasmic reticulum (ER) stress and subsequent pro-apoptotic unfolded protein response (UPR), which was different from the anti-platelet effect.CONCLUSIONS: Our data indicate that the anti-cancer effect of aspirin against CRC may be enhanced by combined administration with dipyridamole. In case further clinical studies confirm our findings, these may be repurposed as adjuvant agents.
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| 7. |
- Jin, Ying-Hui, et al.
(författare)
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Chemoprophylaxis, diagnosis, treatments, and discharge management of COVID-19 : An evidence-based clinical practice guideline (updated version)
- 2020
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Ingår i: Military Medical Research. - : Springer Science and Business Media LLC. - 2054-9369. ; 7:1
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Tidskriftsartikel (refereegranskat)abstract
- The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of a rapidly spreading illness, coronavirus disease 2019 (COVID-19), affecting more than seventeen million people around the world. Diagnosis and treatment guidelines for clinicians caring for patients are needed. In the early stage, we have issued "A rapid advice guideline for the diagnosis and treatment of 2019 novel coronavirus (2019-nCoV) infected pneumonia (standard version)"; now there are many direct evidences emerged and may change some of previous recommendations and it is ripe for develop an evidence-based guideline. We formed a working group of clinical experts and methodologists. The steering group members proposed 29 questions that are relevant to the management of COVID-19 covering the following areas: chemoprophylaxis, diagnosis, treatments, and discharge management. We searched the literature for direct evidence on the management of COVID-19, and assessed its certainty generated recommendations using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach. Recommendations were either strong or weak, or in the form of ungraded consensus-based statement. Finally, we issued 34 statements. Among them, 6 were strong recommendations for, 14 were weak recommendations for, 3 were weak recommendations against and 11 were ungraded consensus-based statement. They covered topics of chemoprophylaxis (including agents and Traditional Chinese Medicine (TCM) agents), diagnosis (including clinical manifestations, reverse transcription-polymerase chain reaction (RT-PCR), respiratory tract specimens, IgM and IgG antibody tests, chest computed tomography, chest x-ray, and CT features of asymptomatic infections), treatments (including lopinavir-ritonavir, umifenovir, favipiravir, interferon, remdesivir, combination of antiviral drugs, hydroxychloroquine/chloroquine, interleukin-6 inhibitors, interleukin-1 inhibitors, glucocorticoid, qingfei paidu decoction, lianhua qingwen granules/capsules, convalescent plasma, lung transplantation, invasive or noninvasive ventilation, and extracorporeal membrane oxygenation (ECMO)), and discharge management (including discharge criteria and management plan in patients whose RT-PCR retesting shows SARS-CoV-2 positive after discharge). We also created two figures of these recommendations for the implementation purpose. We hope these recommendations can help support healthcare workers caring for COVID-19 patients.
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| 8. |
- Zhang, Kang-Ping, et al.
(författare)
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Global Leadership Initiative on Malnutrition criteria as a nutrition assessment tool for patients with cancer
- 2021
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Ingår i: Nutrition (Burbank, Los Angeles County, Calif.). - : Elsevier. - 0899-9007 .- 1873-1244. ; 91-92
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Tidskriftsartikel (refereegranskat)abstract
- Objectives: Since the launch of Global Leadership Initiative on Malnutrition (GLIM), there has been an urgent need to validate the new criteria, especially in patients with cancer. The aim of this study was to evaluate and validate the use of the GLIM criteria in patients with cancer.Method: This multicenter cohort study compared the GLIM with the scored Patient-Generated Subjective Global Assessment (sPG-SGA). The 1-y survival rate, multivariate Cox regression analysis, k-value, sensitivity, specificity, receiver operating characteristic (ROC) curve, and time-dependent ROC analysis were applied to identify the performance of the GLIM.Results: Among the 3777 patients in the study, 50.9% versus 49.1% or 36.3% versus 63.7% of the patients were defined as well-nourished and malnourished by GLIM or sPG-SGA, respectively. GLIM presented moderate consistency (k = 0.54, P < 0.001), fair sensitivity and specificity (70.5 and 88.3%) compared with sPG-SGA. There was no difference in the 1-y survival rate in malnourished patients (76.9 versus 76.4%, P = 0.711), but it was significantly different in well-nourished patients (85.8 versus 90.3%, P < 0.001) between GLIM and sPG-SGA. The above difference was eliminated after omitted nutritional risk screening (NRS)-2002 screening before GLIM (88.1 versus 90.3%, P = 0.078). Omitting NRS-2002 screening before GLIM did not change the 1-y survival rate in well-nourished or malnourished patients by GLIM with NRS-2002 screening (76.9 versus 78.9%, P = 0.099; 85.8% versus 88.1%, P = 0.092) although it significantly raised the rate of malnutrition to 72.5%. The combination of "weight loss and cancer" showed better performance than other combinations.Conclusions: GLIM could be a convenient alternative to sPG-SGA in nutrition assessment for patients with cancer. The combination of "weight loss and cancer" was better than other combinations. Considering the higher risk for malnutrition in patients with cancer, NRS-2002 screening may not be needed before GLIM.
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| 9. |
- Zhang, Qi, et al.
(författare)
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Scored-GLIM as an effective tool to assess nutrition status and predict survival in patients with cancer
- 2021
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Ingår i: Clinical Nutrition. - : Elsevier. - 0261-5614 .- 1532-1983. ; 40:6, s. 4225-4233
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Tidskriftsartikel (refereegranskat)abstract
- Background & aims: The Global Leadership Initiative on Malnutrition (GLIM) released new universal criteria for diagnosing and grading malnutrition, and calls for further investigations not only in different clinical setting but also in GLIM itself including reference value, combination and weight of different GLIM criteria. This study aimed to weigh the GLIM criteria and develop a scored-GLIM system, and then validate as well as evaluate its application in nutritional assessment and survival prediction for patients with cancer. Design: A total of 3547 patients in the primary cohort and 415 patients in the validation cohort were included in the study. Patients' nutritional status were retrospectively assessed using the GLIM criteria. Kaplan-Meier survival curves and multivariate Cox regression analyses were performed to analyze the association between nutritional status and overall survival (OS). A nomogram was produced to quantify the GLIM criteria and develop the scored-GLIM system. C-index, receiver operating characteristic (ROC) curve and calibration curve analyses were performed to validate the predictive accuracy and discriminatory capacity of the scored-GLIM. Finally, a decision curve was applied to assess the clinical utility of the scored-GLIM system. Results: In the primary cohort, 70.3% of patients were diagnosed as malnutrition. The malnutrition severity grading according to the GLIM criteria were associated with the prognosis of patients with cancer (HR 1.42,1.23 to 1.65 for moderate malnutrition; HR 1.80,1.84 to 2.09 for severe malnutrition). The weight of each GLIM criteria was calculated, and unintentional weight loss was the most determining factor acting upon mortality (HR 1.82, 1.64 to 2.10 for stage II and HR 1.50, 1.31 to 1.73 for stage I). A nomogram was constructed by four factors of GLIM to weigh the GLIM criteria. The areas under the ROC curve were 65.3 (1-year ROC) and 65.5 (3-year ROC), and the C-index was 0.62, and the calibration curves fitted well. Decision curve analysis demonstrated the clinical usefulness of the scored-GLIM system. Conclusion: The accuracy and net clinical benefit of scored-GLIM system were similar to scored-PG-SGA but higher than GLIM both in nutrition assessment and in survival prediction for patients with cancer. These findings, along with its time-savings advantages over scored-PG-SGA, suggest scored-GLIM be a better nutritional assessment tool. (c) 2021 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.
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| 10. |
- Zheng, Yi Wu, et al.
(författare)
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Indoor Allergen Levels and Household Distributions in Nine Cities Across China
- 2015
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Ingår i: Biomedical and environmental sciences. - 0895-3988 .- 2214-0190. ; 28:10, s. 709-717
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE: Chinese allergic subjects have high levels of sensitization to house dust mite (HDM) and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households.METHODS: Dust samples were collected from nine cities. Major allergens Der p 1 and Der f 1 from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.RESULTS: HDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from <0.006 to 9.2 µg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.CONCLUSION: HDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.
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