Wastewater-Based Epidemiology of Stimulant Drugs: Functional Data Analysis Compared to Traditional Statistical Methods

Background

Wastewater-based epidemiology (WBE) is a new methodology for estimating the drug load in a population. Simple summary statistics and specification tests have typically been used to analyze WBE data, comparing differences between weekday and weekend loads. Such standard statistical methods may, however, overlook important nuanced information in the data. In this study, we apply functional data analysis (FDA) to WBE data and compare the results to those obtained from more traditional summary measures.

Methods

We analysed temporal WBE data from 42 European cities, using sewage samples collected daily for one week in March 2013. For each city, the main temporal features of two selected drugs were extracted using functional principal component (FPC) analysis, along with simpler measures such as the area under the curve (AUC). The individual cities’ scores on each of the temporal FPCs were then used as outcome variables in multiple linear regression analysis with various city and country characteristics as predictors. The results were compared to those of functional analysis of variance (FANOVA).

Results

The three first FPCs explained more than 99% of the temporal variation. The first component (FPC1) represented the level of the drug load, while the second and third temporal components represented the level and the timing of a weekend peak. AUC was highly correlated with FPC1, but other temporal characteristic were not captured by the simple summary measures. FANOVA was less flexible than the FPCA-based regression, and even showed concordance results. Geographical location was the main predictor for the general level of the drug load.

Conclusion

FDA of WBE data extracts more detailed information about drug load patterns during the week which are not identified by more traditional statistical methods. Results also suggest that regression based on FPC results is a valuable addition to FANOVA for estimating associations between temporal patterns and covariate information.     

Assessment of the daily ragweed pollen concentration with previous-day meteorological variables using regression and quantile regression analysis for Szeged,Hungary

Time-varying parametric linear and time-varying nonparametric regression models as well as a time-varying nonparametric median regression model are developed to predict the daily pollen concentration for Szeged in Hungary using previous-day meteorological parameters and the daily pollen concentration. The models are applied to rainy days and non-rainy days, respectively. The most important predictor is the previous-day pollen concentration level, and the only other predictor retained by a stepwise regression procedure is the daily mean global solar flux for rainy days and the daily mean temperature for non-rainy days. Although the variance percentage explained by these two predictors is higher for non-rainy (55.2%) days than for rainy (51.9%) days, the prediction rate is slightly better for rainy than for non-rainy days. Nonparametric regression yields substantially better estimates, especially for rainy days indicating a nonlinear relationship between the predictors and the pollen concentration. The explained variance percentage is 71.4 and 64.6% for rainy and non-rainy days, respectively. Concerning the mean absolute error, the nonparametric median regression provides the best estimate. The quantile regression shows that probability distribution of daily ragweed concentration is much more skewed for non-rainy days, while the more concentrated probability distribution for rainy days exhibits relatively stable ragweed pollen concentrations. The possible lowest limits of concentrations are also calculated. Under highly favorable conditions for peak concentrations, the pollen level reaches at least 350 grains m⁻³ and 450 grains m⁻³ for rainy and non-rainy days, respectively. These values again underline the excessive ragweed pollen load over the area of Szeged.     

Structural and functional studies of the first tripartite protein complex at the Trypanosoma brucei flagellar pocket collar

The flagellar pocket (FP) is the only endo- and exocytic organelle in most trypanosomes and, as such, is essential throughout the life cycle of the parasite. The neck of the FP is maintained enclosed around the flagellum via the flagellar pocket collar (FPC). The FPC is a macromolecular cytoskeletal structure and is essential for the formation of the FP and cytokinesis. FPC biogenesis and structure are poorly understood, mainly due to the lack of information on FPC composition. To date, only two FPC proteins, BILBO1 and FPC4, have been characterized. BILBO1 forms a molecular skeleton upon which other FPC proteins can, theoretically, dock onto. We previously identified FPC4 as the first BILBO1 interacting partner and demonstrated that its C-terminal domain interacts with the BILBO1 N-terminal domain (NTD). Here, we report by yeast two-hybrid, bioinformatics, functional and structural studies the characterization of a new FPC component and BILBO1 partner protein, BILBO2 (Tb927.6.3240). Further, we demonstrate that BILBO1 and BILBO2 share a homologous NTD and that both domains interact with FPC4. We have determined a 1.9 Å resolution crystal structure of the BILBO2 NTD in complex with the FPC4 BILBO1-binding domain. Together with mutational analyses, our studies reveal key residues for the function of the BILBO2 NTD and its interaction with FPC4 and evidenced a tripartite interaction between BILBO1, BILBO2, and FPC4. Our work sheds light on the first atomic structure of an FPC protein complex and represents a significant step in deciphering the FPC function in Trypanosoma brucei and other pathogenic kinetoplastids.     

Forecasting airborne Platanus pollen in the Madrid region

This study analyses the atmospheric concentration of Platanus pollen in four stations in the Madrid region over a period of 10 years (1994–2003). Various statistical analyses (regression analysis and decision tree) were used to prepare a forecasting model for possible application as a preventive measure in pollinosis. The data comes from the PALINOCAM network and the samplers used were Hirst type (Burkard pollen trap). Platanus pollen is present in the atmosphere during a short period of time in spring, and the maximum concentrations are detected during the last two weeks of March and the first week of April. Regression analysis shows that the pollen concentration of the two previous days is the best predictive variable. The models obtained for the four stations analysed account for between 37 and 61% of the variation in pollen levels in the air. The decision trees show how the introduction of meteorological variables improves prediction for this pollen type.     

Relationship between pollen emission and fruit production in olive (Olea europaea L.)

Aerobiological data of pollen emission concentrations is used to predict fruit production. The principal aim of this work was to study the relationship between pollen emission patterns, emission homogeneity and fruit production in olive (Olea europaea L.). Data of daily pollen concentrations in the atmosphere during the flowering period, collected over a 20‐year period in Perugia (Central Italy), and the corresponding fruit production data were analysed. Correlation and regression analyses on the partial pollen amounts (subdivisions of the whole flowering period), their statistical variability (expressed as coefficients of variation of the daily pollen concentration), and the production values in the different years demonstrate that pollen emission, during the seven to ten day period immediately preceding the maximum pollen emission day, appears to be most closely related to fruit production. Moreover, the pollen emission homogeneity (minimal variability in daily pollen concentrations) during the “critical” flowering period is very important for fertilization.     

Selection, integration, and conflict monitoring; assessing the nature and generality of prefrontal cognitive control mechanisms

Prefrontal cortex (PFC) supports flexible behavior by mediating cognitive control, though the elemental forms of control supported by PFC remain a central debate. Dorsolateral PFC (DLPFC) is thought to guide response selection under conditions of response conflict or, alternatively, may refresh recently active representations within working memory. Lateral frontopolar cortex (FPC) may also adjudicate response conflict, though others propose that FPC supports higher order control processes such as subgoaling and integration. Anterior cingulate cortex (ACC) is hypothesized to upregulate response selection by detecting response conflict; it remains unclear whether ACC functions generalize beyond monitoring response conflict. The present fMRI experiment directly tested these competing theories regarding the functional roles of DLPFC, FPC, and ACC. Results reveal dissociable control processes in PFC, with mid-DLPFC selectively mediating resolution of response conflict and FPC further mediating subgoaling/integration. ACC demonstrated a broad sensitivity to control demands, suggesting a generalized role in modulating cognitive control.     

Poaceae pollen in the atmosphere of Tetouan (NW Morocco): effect of meteorological parameters and forecast of daily pollen concentration

The Poaceae pollen season has been characterized in Tetouan during a 7-year period, and the effect of weather conditions on daily concentrations was examined. The forecast models were produced using a stepwise multiple regression analyses. Firstly, three models were constructed to predict daily Poaceae pollen concentrations during the main pollen season, as well as the pre-peak and post-peak periods with data from 2008 to 2012 and tested on data from 2013 and 2014. Secondly, the regression models using leave-one-out cross-validation were produced with data obtained during 2008–2014 taking into account meteorological parameters and mean pollen concentrations of the same day in other years. The duration of the season ranged from 70 days in 2009 to 158 days in 2012. The highest amount of Poaceae pollen was detected in spring and the first fortnight of July. The annual sum of airborne Poaceae pollen concentrations varied between 2100 and 6251. The peak of anthesis was recorded in May in six of the other years studied. The regression models accounted for 36.3–85.7% of variance in daily Poaceae pollen concentrations. The models fitted best when the mean pollen concentration of the same day in other years was added to meteorological variables, and explained 78.4–85.7% of variance of the daily pollen changes. When the year 2014 was used for validating the models, the lowest root-mean-square errors values were found between the observed and estimated data (around 13). The reasonable predictor variables were the mean pollen concentration of the same day in other years, mean temperature, precipitations, and maximum relative humidity.     

Applying modern analogs to understand the pollen content of coprolites

Most scientists working with coprolites from archaeological contexts assume that human fecal specimens reflect the mixing of the pollen ingested during the period in which the contribution to the coprolite, 19 to 37.5 h, was ingested, that the amount of pollen in a fecal sample directly reflects the amount of pollen originally ingested during that interval, and that differences between the amounts of pollen in different fecal specimens reflect differences in the quantities of pollen ingested at different times. These assumptions were tested and found wanting in an experiment in which two persons sequentially ate separate quantities of 15 pollen types in meals over a four-day interval. The pollen was retrieved and analyzed from feces produced during those four days and five days of subsequent fecal production. Pollen ingested first appeared in relatively small amounts, usually the day after it was ingested. Its concentration per gram of sample then increased rapidly and remained high over a one to three day interval relative to the amounts in previous and subsequent fecal specimens deposited. When pollen concentrations declined some pollen was retained in the gastrointestinal system and much lower concentrations per gram of sample of each type continued to appear in fecal samples for several days. These relatively low pollen concentrations appeared in fecal samples approximately twice as often as did higher concentrations. Our results indicate that comparatively high pollen concentrations can be used to determine that a given pollen type was ingested, but comparisons between pollen concentrations of the same pollen type in different fecal specimens or between different pollen types in the same fecal specimen, cannot be used to determine whether different amounts of pollen were ingested, or what was the relative amount of each ingested. Because pollen concentrations per gram of sample varied widely with time since ingestion, percentages of given pollen types did not occur in predictable patterns and could actually increase as the concentration of the pollen type decreases. Hence, percentages should not be used in coprolite pollen analysis. The experimental results also suggest that variations in the pollen content of different portions of a coprolite are meaningful only in terms of the overall pattern of a sequential group of coprolites.     

Late Holocene palaeoenvironment change in central Tianshan of Xinjiang,northwest China

The northern slopes of central Tianshan Mountains in Xinjiang, northwestern China can provide an ideal database to research palaeoclimate as disturbance by human impact is relatively low and the vegetation zones reflect climatic gradients. In order to establish the correlation between modern climatic factors and surface pollen assemblages and to reconstruct palaeoclimate on the northern slope of central Tianshan Mountains, three Holocene sections in Daxigou, Huashuwozi and Sichanghu located at different elevations and vegetation zones were chosen for study. A total of 80 surface pollen samples in 86 vegetation quadrats were collected for pollen‐vegetation relationship analysis. The Warmth Index (WI) and Moisture Index (MI) were calculated based on averaged modern climate data during 1951 – 2000 at eight weather stations in the study area. Pollen percentages of Picea, Artemisia, Chenopodiaceae, Ephedra, and Tamarix, as well as A/C (Artemisia/Chenopodiaceae) and AP/NAP (arboreal/nonarboreal pollen) ratio were selected as pollen variables and WI and MI were chosen as climatic variables. The relationship between pollen percentages (Picea, Artemisia, Chenopodiaceae and Tamarix), A/C, AP/NAP ratio, WI and MI values were estimated (95% confidence interval) using stepwise multiple linear regression analysis. WI and MI values for the three sections were calculated using these regression equations, and palaeoclimate for the study area could be reconstructed. The results showed periods of both cool‐humid and warm‐dry conditions on the northern slopes of Tianshan Mountain during the late Holocene.     

Influence of meteorological parameters on Olea flowering date and airborne pollen concentration in four regions of Portugal

For calculating the total annual Olea pollen concentration, the onset of the main pollen season and the peak pollen concentration dates, using data from 1998 to 2004, predictive models were developed using multiple regression analysis. Four Portuguese regions were studied: Reguengos de Monsaraz, Valença do Douro, Braga and Elvas. The effect of some meteorological parameters such as temperature and precipitation on Olea spatial and temporal airborne pollen distribution was studied. The best correlations were found when only the pre‐peak period was used, with thermal parameters (maximum temperature) showing the highest correlation with airborne pollen distribution. Independent variables, selected by regression analysis for the predictive models, with the greatest influence on the Olea main pollen season features were accumulated number of days with rain and rainfall in the previous autumn, and temperatures (average and minimum) from January through March. The models predict 59 to 99% of the total airborne pollen concentration recorded and the initial and peak concentration dates of the main Olea pollen season.     

A new weather factor predicting airborne pollen concentration: Péczely’s macrosynoptic weather types

In the present study determinative factors for ragweed (Ambrosia elatior) pollination were studied in Budapest between 1991 and 1996. The aim was to create a model to predict the day-to-day pollen count variation. The pollen concentration is determined mostly by the potential concentration and the mean concentration of the three previous days. These two variables can explain 56% of the total variance. Daily mean and maximum temperature, daily temperature fluctuation and the number of hours of sunshine in the previous day have a significant positive effect on the pollen count. The amount of precipitation on the previous day, relative humidity on the actual and on the previous day influence it negatively. When a cyclone prevails, pollen concentration is usually lower than the seasonal average. Some anticyclonal situations have a similar effect, while other anticyclonal types promote pollination. A predictive model was then created by multiple regression using the potential concentration, the mean concentration of the three previous days, the daily temperature fluctuation and the synoptic type as independent variables. This model can explain more than 68% of the total variance, and its accuracy is >71%. The model seems to predict accurately the trends during the pollen seasons, thus it will probably be a good tool in the practical prediction in Budapest, and the methodology will hopefully be applicable to other sites of the Carpathian Basin too.     

Holocene Palynological Records in Lake Selincuo,Northern Xizang

Selincuo (31°34′–31°57′N, 88°31′–89°21′E)is one of the great lakes in northern Xizang surrounded by alpine grassland, composed mainly of Stipa purpurea, S, subssessifolia var. bassipulomosa. A 3.08 m long core has been taken from a water depth of 27 m, 3 km to the north from the lake. Palynological records of the core combined with surface sample results have revealed from the regional as well as local vegetational and environmental history since the last 12 000 a BP, a very low pollen concentration (19–209 grains/g), mainly of airborne pollen, which indicated a treeless alpine sparse vegetation. A great vegetation change took place at Ca. 9 600 a BP when alpine sparse vegetation was replaced by alpine grassland lasting until Ca. 6 000 a BP. Quite a number of tree pollen grains found in this time interval might be explained as a result of expansion of forests somewhere around the Xizang Plateau. These changes must have caused by climatic warming during the Early Holocene. During the last 6 000 years the vegetation had remained as the alpine grassland, although probably more luxurouste than before, but their was an obvious increase of arboreal pollen, such as the increase of Abies during Ca. 6 000–4 000 a BP, Picea 4 000–2 200 a BP, Pinus 2 200–1 000 a BP which indicated continuous expansion of forests growing around the plateaus. During the last 1 000 years both the concentration and proportion of the arboreal pollen decreased and those of Cypraceae pollen increased sharply. This might mean a reduction of the forests around the plateau and an expansion of swamps around the lake.     

2012—2016年海淀区气传花粉物候特征及其与气象要素的关系

利用重力沉淀法对2012—2016年海淀区气传花粉种类及含量进行监测,分析海淀区的花粉种类、峰值分布特征和含量变化、以及花粉浓度的不连续变化特征,在此基础上采用集合经验模态分解法对花粉浓度进行多时间尺度分析,并分析了花粉浓度与气象要素之间的关系.结果表明: 研究期间,海淀区的主要气传花粉种类已经发生改变,柏科、杨柳科等木本植物代替草本植物成为含量最多的气传花粉类别;花粉浓度的年内峰值分布并无明显改变;近5年花粉浓度整体呈下降趋势,但橘科等草本植物花粉浓度呈上升趋势;统计时段内,花粉浓度在4月上旬、5月下旬及8月上旬共发生3次不连续变化;花粉浓度存在准2 d、准51 d、准128 d的变化周期;温度对于花粉浓度的影响在各气象要素中占有主导地位,16～18 ℃范围内花粉浓度明显升高;气温的变化对逐日花粉浓度的影响具有一定的滞后性和持续性,通常与滞后2～7 d的花粉浓度相关性较高,日照时数和风速对当天的花粉浓度影响最显著.     

Inhibition of Pkhd1 impairs tubulomorphogenesis of cultured IMCD cells

Fibrocystin/polyductin (FPC), the gene product of PKHD1, is responsible for autosomal recessive polycystic kidney disease (ARPKD). This disease is characterized by symmetrically large kidneys with ectasia of collecting ducts. In the kidney, FPC predominantly localizes to the apical domain of tubule cells, where it associates with the basal bodies/primary cilia; however, the functional role of this protein is still unknown. In this study, we established stable IMCD (mouse inner medullary collecting duct) cell lines, in which FPC was silenced by short hairpin RNA inhibition (shRNA). We showed that inhibition of FPC disrupted tubulomorphogenesis of IMCD cells grown in three-dimensional cultures. Pkhd1-silenced cells developed abnormalities in cell-cell contact, actin cytoskeleton organization, cell-ECM interactions, cell proliferation, and apoptosis, which may be mediated by dysregulation of extracellular-regulated kinase (ERK) and focal adhesion kinase (FAK) signaling. These alterations in cell function in vitro may explain the characteristics of ARPKD phenotypes in vivo.     

The influence of temperature,relative humidity and rainfall on the occurrence of pollen allergens (Betula,Poaceae, Ambrosia artemisiifolia) in the atmosphere of Bratislava (Slovakia)

The occurrence of pollen grains in the atmosphere markedly relates to meteorological factors. In the study we have evaluated a correlation between the concentration of pollen grains in the atmosphere of Bratislava and temperature, relative humidity and rainfall during the vegetation period of 1995 and 1997. For our analysis we have selected one representative of each phytoallergen group (trees, grasses, weeds). We have chosen the Betula genus of trees, the whole Poaceae family of grasses and ragweed Ambrosia artemisiifolia L. to represent weeds. The taxons mentioned represent the most significant allergens in Slovakia. The concentration of pollen grains has been monitored by a Lanzoni volumetric pollen trap. The data obtained, the average daily concentration in 1 m(3), have been included in the statistical analysis together with values for the average daily temperature, relative humidity and total rainfall in 24 h. The correlation between the concentration of pollen grains in the atmosphere and selected meteorological variables from daily monitoring has been studied with the help of linear regression and correlation coefficients. We have found the average daily temperature and relative humidity (less than temperature) to be significant factors influencing the occurrence of pollen grains in the atmosphere of Bratislava. The total daily rainfall does not seem to be significant from the statistical point of view.     

Modelling the pollen season start in <Emphasis Type="Italic">Corylus avellana</Emphasis> and <Emphasis Type="Italic">Alnus glutinosa</Emphasis>

Hazel (Corylus avellana L.) and black alder (Alnus glutinosa (L.) Gaertn.) are important sources of airborne pollen and represent an allergen threat during the flowering period. Researches on airborne pollen concentrations in both species are useful in allergology, as well as for fruit production for hazel. The aims of the present study were: (1) to investigate the relationships between environmental conditions and the airborne pollen concentration of hazel and black alder during the flowering period by correlation and multiple regression analysis and (2) to predict the pollen season start (PSS) by using a sequential model, in order to obtain a helpful tool in allergology and hazel cultivation. In this study, the applied method defines the pollen season as the period in which 90 % of the total season’s catch occurred, using a data set of 18 years (1996–2014). The relationships between daily meteorological parameters (temperature, humidity, rainfall and wind speed) during the 14-day period that precedes the PSS and the PSS of hazel and black alder (day of the year) were investigated. The results showed that mean temperature and the number of rainy days before the PSS are the main factors influencing PSS for both taxa. Moreover, the chilling and heat needed to break dormancy were estimated in order to predict the PSS of both species. Different years and different thresholds of temperature and chill days were used to calibrate and validate the model.     

Meteorological variables connected with airborne ragweed pollen in Southern Hungary

About 30% of the Hungarian population has some type of allergy, 65% of them have pollen sensitivity, and at least 60% of this pollen sensitivity is caused by ragweed. The short (or common) ragweed (Ambrosia artemisiifolia = Ambrosia elatior) has the most aggressive pollen of all. Clinical investigations prove that its allergenic pollen is the main reason for the most massive, most serious and most long-lasting pollinosis. The air in the Carpathian Basin is the most polluted with ragweed pollen in Europe. The aim of the study is to analyse how ragweed pollen concentration is influenced by meteorological elements in a medium-sized city, Szeged, Southern Hungary. The data basis consists of daily ragweed pollen counts and averages of 11 meteorological parameters for the 5-year daily data set, between 1997 and 2001. The study considers some of the ragweed pollen characteristics for Szeged. Application of the Makra test indicates the same period for the highest pollen concentration as that established by the main pollination period. After performing factor analysis for the daily ragweed pollen counts and the 11 meteorological variables examined, four factors were retained that explain 84.4% of the total variance of the original 12 variables. Assessment of the daily pollen number was performed by multiple regression analysis and results based on deseasonalised and original data were compared.     

软质与硬质地表对树木花粉日飘散变化的影响

以油松(Pinus tabulaeformis)、白玉兰(Magnolia denudata)、白皮松(Pinus bungeana)、臭椿(Ailanthus altissima)为被试树种,对比研究了春季静风晴朗天气中软质与硬质两种地表条件下不同树种在距树10m高度1.5m处花粉浓度的日变化特征。研究结果表明:(1)4个树种在相同地表环境的花粉浓度日变化趋势基本一致,但同一树种花粉浓度的日变化特征在软、硬两种地表条件下的差异明显。软质地面一天内空气中花粉浓度最大值出现14:00时前后,04:00时花粉浓度最低,这与全天内空气温度的变化正好一致,而与空气相对湿度的变化恰好相反。硬质地表近地空间空气中的花粉浓度则呈现"双峰型"日变化特征,两次峰值分别出现在14:00时和20:00时,硬质地面花粉浓度20:00时晚高峰的出现时间与硬质地面温度日峰值一致。(2)分析硬质地表20:00时花粉浓度高峰出现的原因可能与硬质地面的散热特性有关,硬质地面夜间释放积蓄热量的过程会在一定程度上增强近地面空气的对流运动,并辅助空气中的花粉粒子不断飘散,形成花粉浓度晚高峰。(3)相对于软质地面来讲,硬质地表对空气中花粉飘散的影响作用持续时间更长,这也在一定程度上延长了致敏花粉的危害时间,加剧了致敏花粉的污染程度。研究还进一步在花粉致敏树种栽植、地表覆盖方式等方面进行了讨论;同时建议花粉症患者根据花粉污染发生规律合理规避花粉浓度聚集高峰期出行,从而有效缓解致敏花粉对易感人群的健康威胁。     

Airborne birch and oak pollen grains and birch pollen allergens at a common sampling station in Stockholm

In the present study, the airborne concentrations of birch and oak pollen grains and birch pollen allergens have been recorded in samples from a common sampling station in Stockholm. The sampling period was between April 22^nd and May 31^st 2003. The objectives were to evaluate if analysis of allergen particles in parallel with pollen grains would be relevant to aid subjects suffering from pollinosis. Days with low birch pollen counts had relatively high nominal allergen concentrations in the beginning of the sampling period. The birch pollen grain concentration peaks, during the dry pollination culmination interval in the middle of the period, were associated with correspondingly lower nominal concentrations of allergens than grains. At the end of the sampling period very high nominal amounts of allergen appeared, as reflected by high concentrations of oak pollen grains. The high allergen concentrations were obtained as a result of inherent cross‐reactivity of anti‐ Bet v 1 antibodies with Que a antigens, which are immunologically analogous with Bet v 1. Allergen concentrations increased and decreased after light and heavy rain, respectively. Results obtained indicate that adding a pollen count forecast with allergen concentration data should aid pollen allergic subjects to avoid particulate allergens which might be expected to penetrate more easily than pollen grains into indoor environments.