Virus-prokaryote infection pairs associated with prokaryotic production in a freshwater lake.

Viruses infect and kill prokaryotic populations in a density- or frequency-dependent manner and affect carbon cycling. However, the effects of the stratification transition, including the stratified and de-stratified periods, on the changes in prokaryotic and viral communities and their interactions remain unclear. We conducted a monthly survey of the surface and deep layers of a large and deep freshwater lake (Lake Biwa, Japan) for a year and analyzed the prokaryotic production and prokaryotic and viral community composition. Our analysis revealed that, in the surface layer, 19 prokaryotic species, accounting for approximately 40% of the total prokaryotic abundance, could potentially contribute to the majority of prokaryotic production, which is the highest during the summer and is suppressed by viruses. This suggests that a small fraction of prokaryotes and phages were the key infection pairs during the peak period of prokaryotic activity in the freshwater lake. We also found that approximately 50% of the dominant prokaryotic and viral species in the deep layer were present throughout the study period. This suggests that the "kill the winner" model could explain the viral impact on prokaryotes in the surface layer, but other dynamics may be at play in the deep layer. Furthermore, we found that annual vertical mixing could result in a similar rate of community change between the surface and deep layers. These findings may be valuable in understanding how communities and the interaction among them change when freshwater lake stratification is affected by global warming in the future.IMPORTANCEViral infection associated with prokaryotic production occurs in a density- or frequency-dependent manner and regulates the prokaryotic community. Stratification transition and annual vertical mixing in freshwater lakes are known to affect the prokaryotic community and the interaction between prokaryotes and viruses. By pairing measurements of virome analysis and prokaryotic production of a 1-year survey of the depths of surface and deep layers, we revealed (i) the prokaryotic infection pairs associated with prokaryotic production and (ii) the reset in prokaryotic and viral communities through annual vertical mixing in a freshwater lake. Our results provide a basis for future work into changes in stratification that may impact the biogeochemical cycling in freshwater lakes.

Evaluating the necessity of post-processing techniques on d4PDF data for extreme climate assessment.

The occurrence and severity of extreme precipitation events have been increasing globally. Although numerous projections have been proposed and developed for evaluating the climate change impacts, most models suffer from significant bias error due to the coarse resolution of the climate datasets, which affects the accuracy of the climate change assessment. Therefore, in this study, post-processing techniques (interpolation and bias correction methods) were adopted on the database for Policy Decision Making for Future Climate Change (d4PDF) model for extreme climatic flood events simulation in the Chao Phraya River Basin, Thailand, under + 4-K future climate simulation. Due to the limited number of the rain gages, the gradient plus inverse distance squared interpolation method (combination of multiple linear regression and distance weighting methods) was applied in this study. In the bias correction methods, the additional setting of monthly and seasonal periods was adjusted. The proposed bias correction approach deployed gamma distribution combined with generalized Pareto distribution setting with the seasonal period for the rainy season datasets, whereas only the gamma setting was applied with the monthly period during the dry season. The outcomes revealed that the proposed method could react to extreme rainfall events, expand dry days during dry season, and intensify rainfall amount during rainy season. The post-processing d4PDF trends of six sea surface temperature (SST) patterns (consists of 90 ensemble members) of two periods (near future: 2051-2070 and far future: 2091-2110) recorded the highest and lowest amounts of annual rainfalls of 4,450 mm/year in mid-stream of Nan River and 710 mm/year in the lower CPRB, respectively. Notably, the significant variances noted in the rainfall patterns among ensembles, demanding further investigation in future climate change, impact studies. The findings of the study provided novel insights on the importance of proper post-processing techniques for improving the robustness of d4PDF in climate change impacts assessment.

Spatiotemporal impact of COVID-19 on Taiwan air quality in the absence of a lockdown: Influence of urban public transportation use and meteorological conditions.

The unprecedented outbreak of COVID-19 significantly improved the atmospheric environment for lockdown-imposed regions; however, scant evidence exists on its impacts on regions without lockdown. A novel research framework is proposed to evaluate the long-term monthly spatiotemporal impact of COVID-19 on Taiwan air quality through different statistical analyses, including geostatistical analysis, change detection analysis and identification of nonattainment pollutant occurrence between the average mean air pollutant concentrations from 2018-2019 and 2020, considering both meteorological and public transportation impacts. Contrary to lockdown-imposed regions, insignificant or worsened air quality conditions were observed at the beginning of COVID-19, but a delayed improvement occurred after April in Taiwan. The annual mean concentrations of PM10, PM2.5, SO2, NO2, CO and O3 in 2020 were reduced by 24%, 18%, 15%, 9.6%, 7.4% and 1.3%, respectively (relative to 2018-2019), and the overall occurrence frequency of nonattainment air pollutants declined by over 30%. Backward stepwise regression models for each air pollutant were successfully constructed utilizing 12 meteorological parameters (R2 > 0.8 except for SO2) to simulate the meteorological normalized business-as-usual concentration. The hybrid single-particle Lagrangian integrated trajectory (HYSPLIT) model simulated the fate of air pollutants (e.g., local emissions or transboundary pollution) for anomalous months. The changes in different public transportation usage volumes (e.g., roadway, railway, air, and waterway) moderately reduced air pollution, particularly CO and NO2. Reduced public transportation use had a more significant impact than meteorology on air quality improvement in Taiwan, highlighting the importance of proper public transportation management for air pollution control and paving a new path for sustainable air quality management even in the absence of a lockdown.

Complete Genome Sequence of a Phage Infecting Sphingomonadaceae.

We isolated a phage infecting a member of the Sphingomonadaceae family from a freshwater lake. The phage has a DNA genome of 41,771 bp, with a GC content of 61.7%. The genome harbors 50 predicted protein-coding genes and an auxiliary metabolic gene, which encodes a protein belonging to the radical S-adenosylmethionine superfamily.

Draft Genome Sequences of Sphingomonadaceae Strains Isolated from a Freshwater Lake.

We isolated two bacterial strains (Sphingomonadaceae family) from Lake Biwa, Japan. Based on whole-genome sequencing results, one strain (BSN-002) was assigned to the Sphingopyxis genus and the other (BSN-004) to Sphingomonas aquatilis.

Application of artificial intelligence methods for monsoonal river classification in Selangor river basin, Malaysia.

Rivers in Malaysia are classified based on water quality index (WQI) that comprises of six parameters, namely, ammoniacal nitrogen (AN), biochemical oxygen demand (BOD), chemical oxygen demand (COD), dissolved oxygen (DO), pH, and suspended solids (SS). Due to its tropical climate, the impact of seasonal monsoons on river quality is significant, with the increased occurrence of extreme precipitation events; however, there has been little discussion on the application of artificial intelligence models for monsoonal river classification. In light of these, this study had applied artificial neural network (ANN) and support vector machine (SVM) models for monsoonal (dry and wet seasons) river classification using three of the water quality parameters to minimise the cost of river monitoring and associated errors in WQI computation. A structured trial-and-error approach was applied on input parameter selection and hyperparameter optimisation for both models. Accuracy, sensitivity, and precision were selected as the performance criteria. For dry season, BOD-DO-pH was selected as the optimum input combination by both ANN and SVM models, with testing accuracy of 88.7% and 82.1%, respectively. As for wet season, the optimum input combinations of ANN and SVM models were BOD-pH-SS and BOD-DO-pH with testing accuracy of 89.5% and 88.0%, respectively. As a result, both optimised ANN and SVM models have proven their prediction capacities for river classification, which may be deployed as effective and reliable tools in tropical regions. Notably, better learning and higher capacity of the ANN model for dataset characteristics extraction generated better predictability and generalisability than SVM model under imbalanced dataset.

Seasonal Variation in Viral Infection Rates and Cell Sizes of Infected Prokaryotes in a Large and Deep Freshwater Lake (Lake Biwa, Japan).

As viruses regulate prokaryotic abundance and the carbon cycle by infecting and lysing their prokaryotic hosts, the volume of infected prokaryotes is an important parameter for understanding the impact of viruses on aquatic environments. However, literature regarding the seasonal and spatial variations in the cell volume of infected prokaryotes is limited, despite the volume of the prokaryotic community varying dynamically with season and water column depth. Here, we conducted a field survey for two annual cycles in a large and deep freshwater lake (Lake Biwa, Japan), where large prokaryotes inhabit the deeper layer during the stratified period. We used transmission electron microscopy to reveal the seasonal and spatial variation in the frequency of viral infection and cell volume of infected prokaryotes. We found that the viral infection rate in the surface layer increased when estimated contact rates increased during the middle of the stratified period, whereas the infection rate in the deeper layer increased despite low estimated contact rates during the end of the stratified period. In addition, in the deeper layer, the fraction of large prokaryotes in the total and infected prokaryotic communities increased progressively while the number of intracellular viral particles increased. We suggest different ways in which the viral abundance is maintained in the two water layers. In the surface layer, it is speculated that viral abundance is supported by the high viral infection rate because of the high activity of prokaryotes, whereas in the deeper layer, it might be supported by the larger number of intracellular viral particles released from large prokaryotes. Moreover, large prokaryotes could contribute as important sources of organic substrates via viral lysis in the deeper layer, where labile dissolved organic matter is depleted.

Biological sources of variations of tartrate-resistant acid phosphatase 5b in a healthy Japanese population.

BACKGROUND: Tartrate-resistant acid phosphatase 5b (TRACP5b) is a bone resorption marker that is mainly used in clinical management of osteoporosis. For proper interpretations of test results for serum TRACP5b, we explored their biological sources of variation, esp. age-related changes, and associations with other bone-related markers in healthy Japanese adults. METHODS: During the 2009 East-Southeast Asian multicentre study for determination of reference intervals, 72 major laboratory tests were measured by centralized assays in 3541 well-defined healthy volunteers. The current study included 1980 test results in Japanese subjects for five bone-related markers: TRACP5b, bone alkaline phosphatase, intact parathyroid hormone, calcium and inorganic phosphate. Information on sources of variation, including body mass index, smoking habits and ABO-blood group, were obtained from a health status questionnaire. RESULTS: Gender-specific profiles of age-related changes were observed for each parameter. Increased values starting from 40 years of age in females were most prominent for TRACP5b, followed by bone alkaline phosphatase and inorganic phosphate. TRACP5b in males decreased with body mass index, bone alkaline phosphatase and TRACP5b were higher in blood type-O subjects, especially in males. TRACPT5b was closely correlated with bone alkaline phosphatase, and moderately correlated with adjusted calcium and inorganic phosphate, especially in females aged ≥45 years. Reference intervals for each analyte were determined parametrically based on gender and age. CONCLUSIONS: This study elucidated sources of variation of TRACP5b and related bone markers in healthy Japanese subjects and demonstrated a specific age profile for each marker. These results are of relevance for better clinical usage and interpretations of serum levels of bone markers.

Verifying the applicability of SWAT to simulate fecal contamination for watershed management of Selangor River, Malaysia.

The Soil and Water Assessment Tool (SWAT) ecohydrological model was utilized to simulate fecal contamination in the 1937 km2 Selangor River Watershed in Malaysia. The watershed conditions posed considerable challenges owing to data scarcity and tropical climate conditions, which are very different from the original conditions that SWAT was developed and tested for. Insufficient data were compensated by publicly available data (e.g., land cover, soil, and weather) to run SWAT. In addition, field monitoring and interviews clarified representative situations of pollution sources and loads, which were used as input for the model. Model parameters determined by empirical analyses in the USA (e.g., surface runoff, evapotranspiration, and temperature adjustment for bacteria die-off) are thoroughly discussed. In particular, due consideration was given to tropical climate characteristics such as intense rainfall, high potential evapotranspiration, and high temperatures throughout the year. As a result, the developed SWAT successfully simulated fecal contamination ranging several orders of magnitude along with its spatial distribution (i.e., Nash-Sutcliffe Efficiency (NSE) = 0.64, Root Mean Square Error-Observations Standard Deviation Ratio (RSR) = 0.64 at six mainstem sites, and NSE = 0.67 and RSR = 0.57 at 12 major tributaries). Moreover, mitigation countermeasures for future worsening of fecal contamination (i.e., E.coli concentration > 20,000 CFU/100 mL for 690 days during nine years at a raw water intake point for Kuala Lumpur [KL] residents) were analyzed through scenario simulations, thereby contributing to discussing effective watershed management. The results propose improving decentralized sewage treatment systems and treating chicken manure with effective microorganisms in order to guarantee water safety for KL residents (i.e., E.coli concentrations <20,000 CFU/100 mL throughout the period, considering Malaysian standards). Accordingly, this study verified the applicability of SWAT to simulate fecal contamination in areas that are difficult to model and suggests solutions for watershed management based on quantitative evidence.

Extracellular vesicles from mesenchymal stem cells of dental pulp and adipose tissue display distinct transcriptomic characteristics suggestive of potential therapeutic targets.

Objective: Mesenchymal stem cells (MSCs) are isolated from various human tissues and used for therapy, in which beneficial effects are attributed mainly to mesenchymal stem cell-derived extracellular vesicles (MSC-EVs). Whereas MSCs of diverse tissue types share cardinal stem cell features, it is becoming evident that MSCs of each tissue type possess unique properties as well. For designing efficient stem cellbased therapies, it is crucial to understand the unique properties associated with MSCs and MSC-EVs of each tissue type. Such unique properties can be analyzed through transcriptomic approaches using comprehensive gene expression databases and sophisticated analytical tools. Here, we comparatively studied the transcriptomes in MSC-EVs of dental pulp and adipose tissue. Additionally, the transcriptomes of MSC-EVs were compared with the cellular transcriptomes of MSCs for the same tissue types. Methods: MSCs were cultured from human dental pulp and adipose tissue specimens. Conditioned culture media were collected to prepare MSC-EVs, from which RNAs were isolated and subjected to next-generation sequencing for transcriptomic analysis. Gene expression signatures in MSC-EVs of each tissue type were investigated using gene set analysis. Results: MSC-EVs obtained from dental pulp-derived MSCs showed distinct transcriptomic signatures of neurogenesis and neural retina development while MSC-EVs of adipose tissue-derived MSCs showed signatures of mitochondrial activity and skeletal system development. The transcriptomes of MSC-EVs resembled the cellular transcriptomes of MSCs, and the genes associated with neurogenesis were highly expressed in both MSCs and MSC-EVs of dental pulp. Adipose tissue-derived MSCs and MSC-EVs highly expressed genes associated with angiogenesis, hair growth, and dermal matrices. Conclusion: The clear and distinct signatures of neurogenesis and neural retina development in dental pulp-derived MSC-EVs imply neurodegenerative disorders and retinal diseases as putative therapeutic targets. In contrast, the transcripts in adipose tissue-derived MSC-EVs could be useful in rejuvenating the skin and musculoskeletal system. Further insights into MSC-EVs of divergent tissue types may expand the list of potential therapeutic targets.

Comparison among different ASEAN water quality indices for the assessment of the spatial variation of surface water quality in the Selangor river basin, Malaysia.

The assessment of surface water quality is often laborious, expensive and tedious, as well as impractical, especially for the developing and middle-income countries in the ASEAN region. The application of the water quality index (WQI), which depends on several independent key parameters, has great potential and is a useful tool in this region. Therefore, this study aims to find out the spatial variability of various water quality parameters in geographical information system (GIS) environment and perform a comparative study among the ASEAN WQI systems. At present, there are four ASEAN countries which have implemented the WQI system to evaluate their surface water quality, which are (i) Own WQI system-Malaysia, Thailand and Vietnam-and (ii) Adopted WQI system: Indonesia. A spatial distribution of 12 water quality parameters in the Selangor river basin, Malaysia, was plotted and then applied into the different ASEAN WQI systems. The WQI values obtained from the different WQI systems have an appreciable difference, even for the same water samples due to the disparity in the parameter selection and the standards among them. WQI systems which consider all biophysicochemical parameters provide a consistent evaluation (Very Poor), but the system which either considers physicochemical or biochemical parameters gives a relatively lenient evaluation (Fair-Poor). The Selangor river basin is stressed and impacted by all physical, biological and chemical parameters caused by both the aridity of the climate and anthropogenic activities. Therefore, it is crucial to include all these aspects into the evaluation and corresponding actions should be taken.

Detection of DNA damage formation by natural organic matter using EGFP-fused MDC1-expressing cells.

Studies have been conducted on the genotoxicity and carcinogenicity of disinfection by-products formed from natural organic matter (NOM) and mitigation effect for mutagens and clastogens by NOM. Whereas, reportedly, synthetic humic acid in high concentration has induced genotoxicity in human cells, and NOM samples have provoked mild oxidative and other physiological responses in aquatic organisms. Our group developed a novel detection method for DNA damage formation, namely enhanced green fluorescent protein (EGFP)-fused mediator of DNA damage checkpoint 1 (MDC1)-expressing human cells as simple and high-sensitive system. By using this method, a significant increase in the foci area was observed after 3 h, but not 24 h for 130 mgC L-1 Suwannee River fulvic acid (SRFA), 38 mgC L-1 humic acid (SRHA), and 19 mgC L-1 NOM (SRNOM). The SRNOM concentration is the original environmental one; therefore, it was suggested that the formation and repair of DNA damage associated with γ-H2AX, a biomarker for DNA double-strand breaks by mild oxidative stress, in Suwannee River (SR) were detected for the first time. The increase in the foci area was not observed for 18 mgC L-1 Lake Biwa fulvic acid (LBFA) and 50 mg L-1 catechin after both 3 h and 24 h. The difference between the SR and Lake Biwa (LB) samples may result from the differences in their electron-accepting capacity. The application of this methodology is expected to elucidate oxidative stress and toxicological effects shortly and in detail for many water samples.

Elucidation of stability profiles of common chemistry analytes in serum stored at six graded temperatures.

Background Many reports address the stability of biochemical analytes in serum. However, studies covering a wide range of storage temperatures are unavailable. Using equipment enabling precise temperature control, we investigated the effect of six different storage temperatures on serum analytes. Methods Serum specimens from seven healthy volunteers were obtained and divided into multiple aliquots for storage at -30, -20, -10, 0, 4, and 25 °C. On days 1, 3, 7, 14, 28 and 56, the aliquots stored at each temperature were relocated to a deep freezer maintained at -80 °C. On day 60, all aliquots were measured collectively for 13 major chemistry analytes. Results (1) At 25 °C, alanine aminotransferase (ALT), creatine kinase (CK), aspartate aminotransferase (AST) and total bilirubin (TBil) were very unstable especially on day 7 and later. (2) At ≤4 °C, alkaline phosphatase (ALP), γ-glutamyltransferase (GGT), amylase (AMY), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), triglyceride (TG), TBil and complement component-4 (C4) were generally stable and were very stable at 25 °C until day 14. (3) Between -20 and 4 °C, especially at -10 °C, test results of ALT, AST and lactate dehydrogenase (LDH) showed prominent decreases, but their stability was greatly improved at -30 °C. (4) In contrast, the value of complement component-3 (C3) increased at ≥- 20 °C. (5) At -30 °C, test results of all analytes were generally very stable except for ALT and CK, which showed noticeable reductions in activity after 14 days. Conclusions This is the first study to assess the stability of serum analytes at six graded temperatures simultaneously. Each analyte has a unique stability pattern for a range of temperatures.

Evaluation of algal photosynthesis inhibition activity for dissolved organic matter with the consideration of inorganic and coloring constituents.

The effect of waterborne ingredient on ecosystem has been of great interest. In the present study, the evaluation method using algal photosynthesis inhibition assay with dual-channel pulse amplitude modulation (PAM) system was established for a series of water samples to elucidate the potential effect of the total body of organic compounds including natural organic matter (NOM) on aquatic ecosystems. The more sensitive and less time-consuming monitoring method compared with algal growth inhibition assay was suggested, especially considering inorganic and coloring constituents. Algal photosynthesis inhibition activity was detected with high sensitivity for photosystem II (PSII) inhibitors, whereas the IC10 of the other chemicals was over the environmental standard concentration for Chlamydomonas moewusii (Chlorophyceae) and Pheodactylum tricornutum (Diatomea). The photosynthesis inhibition activity of Lake Biwa dissolved organic matter (LBDOM) and fulvic acid (LBFA) was significantly detected at ≥10 times the concentration and >10 mgC L-1, respectively, whereas prominent activity was confirmed for Suwannee River NOM (SRNOM) on the river original concentration (>30 mgC L-1) for both algae. Significant inhibition activity was detected in both algae at least in twice-concentration for water samples from a wastewater treatment pilot plant. There was no great difference in the activity between sewage secondary effluent and its filtrate with ultrafiltration (UF), and physically washing water for the UF membrane.

Epidermal growth factor-nanoparticle conjugates change the activity from anti-apoptotic to pro-apoptotic at membrane rafts.

The proliferation epidermal growth factor (EGF) is known to acquire contradictory apoptotic activities upon conjugation with gold nanoparticles (GNPs) through hitherto unknown mechanisms. Here, we identified an essential role of membrane rafts in the drastic activity switching of EGF-GNPs through the following intracellular signaling. (1) In contrast to the rapid diffusion of activated EGF receptor after the soluble EGF stimulation, the receptor is confined within membrane rafts upon binding to the EGF-GNPs. (2) This initial receptor confinements switch its endocytosis process from normal clathrin-mediated endocytosis to caveolin-mediated one, changing the phosphorylation dynamics of essential downstream kinases, i.e., extracellular signal-regulated kinase and AKT. Importantly, the destruction of membrane rafts by ß-cyclodextrin reversed this trafficking and signaling, restoring EGF-GNPs to lost anti-apoptotic property. These results reveal the importance of GNP-mediated signal condensation at membrane rafts in conferring the unique apoptotic activity on EGF-nanoparticle conjugates. STATEMENT OF SIGNIFICANCE: Epidermal growth factor (EGF) is a small secretory protein that induces cell proliferation upon binding to its receptor existed on cellular plasma membranes. One interesting feature of the protein in the nanobiology field is, its acquisition of apoptosis-inducing (cellular suicide) activity rather than proliferative one upon conjugation to gold nanoparticles through hitherto unknown mechanisms. Here, we identified the involvement of membrane rafts, plasma membrane nanodomains enriched with cholesterol, in the apoptosis processes by changing the receptor trafficking and downstream signal transduction pathways. Moreover, the destruction of lipid rafts restored the EGF-nanoparticle conjugates with lost anti-apoptotic activity. These finding highlight potential applications of EGF-nanoparticle conjugates to cancer therapy, as the EGF receptor are highly expressed in cancer cells.

Biological sources of variation of serum adiponectin among healthy individuals in comparison with related nutritional and inflammatory markers.

BACKGROUND: Serum adiponectin (AN) is a nutritional and inflammatory marker of various diseases. Its biological sources of variation (SVs) have been widely evaluated, but its relationships with other related markers remain to be studied comprehensibly by use of multivariate analyses including factor analysis. METHODS: Serum specimens from 752 well-defined healthy subjects (295 males; 457 females) obtained from Japan, Hong Kong, Taiwan, and Vietnam were tested for total AN (tAN) and high-molecular-weight AN (hmAN) together with 71 major analytes. We chose test results of 11 analytes known as nutritional and inflammatory markers: insulin, HDL-C, LDL-C, TG, ALT, GGT, UA, TTR, CRP, C3, and C4. Factor analysis (FA) was performed to elucidate commonality among the analytes. Multiple regression analysis (MRA) was performed analyte by analyte to evaluate its biological SVs including age, BMI, and levels of alcohol consumption, smoking, and exercise. RESULTS: Both serum ANs showed female predominance, reduction with increased BMI, and regional difference (lower in Southeast Asia). The ratio of hmAN to tAN was higher in females. MRA showed no clear associations of tAN or hmAN with the levels of drinking alcohol or cigarette smoking. FA revealed that both tAN and hmAN have a commonality in biological variations with HDL-C, TG, CRP, and insulin but not with ALT, GGT, and TTR. CONCLUSIONS: No apparent differences were observed between tAN and hmAN with regard to commonality or association with related analytes by FA or MRA. It appears not necessary to distinguish hmAN from tAN when interpreting test results among healthy individuals.

Time required for resetting postural effects on serum constituents in healthy individuals.

OBJECTIVE: We investigated postural effects on concentrations of commonly measured serum analytes and sought for optimal duration of sitting before venipuncture in an outpatient setting. METHODS: Seven healthy volunteers changed posture serially at 30-minute (min) intervals from lying, sitting, slowly walking, sitting, and to lying again. At the end of the first lying posture, wing-needle was inserted into cubital vein and 3mL of blood was drawn every 5min. All the serum was stored at -80°C until the time of collective measurements for 16 major chemistry analytes two weeks later. RESULTS: Concentrations of macromolecular analytes (albumin, total cholesterol, alkaline phosphatase, etc.) in supine position were increased (on average 5-7%) after 30min of sitting and further increased (10-14%) during 30min of walking. After slow walking, time required for return of values to the levels of sitting posture took approximately 20min. Concentrations of small molecules (sodium, urea, etc.) did not show any variation by the postural changes. Potassium and inorganic phosphate showed walking-related increase in concentrations. Creatinine was lowered in sitting posture, which implicates postural changes in renal flow. CONCLUSION: It is generally recommended to sit for 20min or more before venipuncture, although the effect of prior physical movement remains a little longer for potassium and inorganic phosphate.

A global multicenter study on reference values: 1. Assessment of methods for derivation and comparison of reference intervals.

OBJECTIVES: The IFCC Committee on Reference Intervals and Decision Limits coordinated a global multicenter study on reference values (RVs) to explore rational and harmonizable procedures for derivation of reference intervals (RIs) and investigate the feasibility of sharing RIs through evaluation of sources of variation of RVs on a global scale. METHODS: For the common protocol, rather lenient criteria for reference individuals were adopted to facilitate harmonized recruitment with planned use of the latent abnormal values exclusion (LAVE) method. As of July 2015, 12 countries had completed their study with total recruitment of 13,386 healthy adults. 25 analytes were measured chemically and 25 immunologically. A serum panel with assigned values was measured by all laboratories. RIs were derived by parametric and nonparametric methods. RESULTS: The effect of LAVE methods is prominent in analytes which reflect nutritional status, inflammation and muscular exertion, indicating that inappropriate results are frequent in any country. The validity of the parametric method was confirmed by the presence of analyte-specific distribution patterns and successful Gaussian transformation using the modified Box-Cox formula in all countries. After successful alignment of RVs based on the panel test results, nearly half the analytes showed variable degrees of between-country differences. This finding, however, requires confirmation after adjusting for BMI and other sources of variation. The results are reported in the second part of this paper. CONCLUSION: The collaborative study enabled us to evaluate rational methods for deriving RIs and comparing the RVs based on real-world datasets obtained in a harmonized manner.

A global multicenter study on reference values: 2. Exploration of sources of variation across the countries.

OBJECTIVES: The intent of this study, based on a global multicenter study of reference values (RVs) for serum analytes was to explore biological sources of variation (SVs) of the RVs among 12 countries around the world. METHODS: As described in the first part of this paper, RVs of 50 major serum analytes from 13,396 healthy individuals living in 12 countries were obtained. Analyzed in this study were 23 clinical chemistry analytes and 8 analytes measured by immunoturbidimetry. Multiple regression analysis was performed for each gender, country by country, analyte by analyte, by setting four major SVs (age, BMI, and levels of drinking and smoking) as a fixed set of explanatory variables. For analytes with skewed distributions, log-transformation was applied. The association of each source of variation with RVs was expressed as the partial correlation coefficient (rp). RESULTS: Obvious gender and age-related changes in the RVs were observed in many analytes, almost consistently between countries. Compilation of age-related variations of RVs after adjusting for between-country differences revealed peculiar patterns specific to each analyte. Judged fromthe rp, BMI related changes were observed for many nutritional and inflammatory markers in almost all countries. However, the slope of linear regression of BMI vs. RV differed greatly among countries for some analytes. Alcohol and smoking-related changes were observed less conspicuously in a limited number of analytes. CONCLUSION: The features of sex, age, alcohol, and smoking-related changes in RVs of the analytes were largely comparable worldwide. The finding of differences in BMI-related changes among countries in some analytes is quite relevant to understanding ethnic differences in susceptibility to nutritionally related diseases.