Eight essential elements and two heavy metals were analyzed using ICP-MS after microwave digestion. For bivariate analyses of variables, we use nonparametric Spearman's correlation coefficient test. The mean concentrations of essential elements and heavy metals were as follows: sodium 330 ± 417 mg/L, magnesium 32.6 ± 15.5 mg/L, phosphorus 156 ± 46.2 mg/L, potassium 488 ± 146 mg/L, calcium 193 ± 53.2 mg/L, iron 1.65 ± 1.43 mg/L, copper 0.54 ± 0.46 mg/L, zinc 2.89 ± 3.23 mg/L, arsenic < 1 μg/L, and lead < 1 μg/L. Concentrations of heavy metals in maternal breast milk may have the important implication that it is not affected by environmental pollution in this province. This study provides reliable information about maternal breast milk concentrations of nursery mothers residing in Şanlıurfa, Turkey, and also compares the relations between essential elements and socioeconomic conditions, residing areas, and using copper equipment for food preparation of which some have not previously been reported.Maternal breast milk is a unique biological matrix that contains essential micronutrients. Potentially heavy metals may also affect infants' health and growth through maternal breast milk. The purpose of this study was to determine and compare the essential elements and heavy metals of maternal breast milk of nursery mothers residing in Şanlıurfa province, Turkey. Maternal breast milk concentrations of sodium, magnesium, phosphorus, potassium, calcium, iron, copper, zinc, arsenic, and lead were analyzed in a random sample of the first time in urban and suburban nursery Turkish mothers ( Maternal breast milk (MBM) is a unique biological matrix of nutrition for infants. Breastfeeding is vital for optimal health, growth, and development with its macro- and micronutrients, including essential elements . World HEssential elements in MBM are crucial for ideal growth and development of infants. There are a lot of benefits associated with MBM, such as decreasing the morbidity, mortality, and incidences of diseases . The comConcentration data on some essential elements and heavy metals in MBM of nursery mothers from different countries analyzed with different techniques such as flame photometry , ion chrThe aim of this study is to provide and access the information of concentrations of essential elements in MBM from nursery mothers who are residing in Şanlıurfa province of Turkey using a sensitive method and also compare the relations between essential elements and socioeconomic conditions, residing areas, and using copper equipment for food preparation.Samples were collected from 42 nursery mothers with an average age of 28.3 ± 7.08 years delivered at full term (38–40 weeks) of gestation. All samples were obtained from nursery mothers' 3–520 lactation days postpartum during the hours of 08:00–11:30 a.m. MBM samples were manually expressed from the breast using a manual pump to sterile falcon tube after 3 mL was discarded and stored at −20°C until analyses. Based on self-reporting on the socioeconomic levels, residing areas, and using copper equipment for food preparation, participants were divided into groups .Mixed calibration standards of sodium, magnesium, phosphorus, potassium, calcium, iron, copper, zinc, arsenic, and lead were prepared from dilutions of stock solutions (Agilent Japan (Lot Number: 10-160YPY2)). TraceSELECT grade of nitric acid  and hydrogen peroxide  were used for sample digestion.Ultrapure water was provided in the laboratory (18.2 MΩ/cm at 25°C) by MES MP Mini pure, (Turkey) device. Every plastic and glassware were cleaned in acid bath ) for 24 h and rinsed with ultrapure water and dried before use. The gas 99.9990% Argon was supplied by Linde Gases . Whole solutions were prepared with ultrapure water.Elements were analyzed on Agilent 7500 ce with an octopole reaction system inductively coupled plasma-mass spectrometer  with an autosampler (Cetac ASX-520) and a nebulizer .3, and 2.0 mL of 30% (v/v) H2O2 in PTFE vessels. Microwave acid digestion procedure was as follows: firstly up to 120°C for 15 min, for the second step constant for 10 min; up to 160°C in 20 min; and finally constant for 15 min; at the end a cooling stage (30 min) was done in room temperature and the samples and blanks were diluted to 50 mL with ultrapure water.MBM samples were analyzed by ICP-MS after microwave-assisted acid digestion. 1.0 mL of each sample and blank samples was digested with 4.0 mL of 65% (v/v) HNO23Na+, 24Mg+, 26Mg+, 31P+, 39 K+, 44Ca+, 56Fe+, 63Cu+, 66Zn+, 75As+, and 208Pb+ were detected. All samples were analyzed in triplicate. These isotopes were preferred to maximize the sensitivity and minimize interferences.The isotopes Limit of quantification (LOQ) and limit of detection (LOD) were calculated for ten times and the recovery of 12 elements  in raw milk samples was between 92.4 and 124.1% as shown in With a view to impose the integrity of the analyses, the same concentrations of each element as in the original sample were added to a sample and performed in the same microwave digestion procedure as the samples.Statistical analyses were carried out using SPSS 11.00 . For bivariate analyses of variables, nonparametric Spearman's correlation coefficient was used.Ethics committee approval was obtained from Medical Faculty of Harran University (74059997.050.01.04/65). Informed consents were obtained and signed by the participating nursery mothers to protect the participants' privacy, personal and clinical data, and MBM samples used only for this study purpose.The nursery mothers included in this study were on the average age of 17–44 (28.3 ± 7.08) years at the time of lactation period average of 3–520 (163 ± 139) day. r: 0.719; P < 0.001), iron , and magnesium concentrations . Magnesium concentrations in MBM were in a range of 12.7–85.9 mg/L with a mean level of 32.6 ± 15.5 mg/L. Magnesium and zinc levels were significantly correlated .Sodium levels in MBM samples ranged between 44.7 and 1703 mg/L and significantly correlated with copper , lactation period , and positively correlated with nursery mothers' age .MBM phosphorus concentrations were between the level of 84–300 mg/L, with a mean level of 156 ± 46.3 mg/L. Phosphorus concentrations were negatively correlated with iron concentrations , sodium , magnesium , and copper .Potassium concentrations of MBM varied between 161 and 903 mg/L. The content of potassium was positively correlated with zinc . Also, calcium levels were significantly correlated with magnesium  and potassium . Concentrations of calcium in MBM from suburban nursery mothers were significantly lower than concentrations of calcium in urban nursery mothers.Calcium concentrations of MBM were in the range of 90–276 mg/L with a mean level of 193 ± 53.2 mg/L. Calcium was positively correlated with the residing area , magnesium , and lactation period  and with using copper equipment in food preparation .The mean concentration of iron in MBM was 1.65 ± 1.43 mg/L. Copper concentration of MBM was positively correlated with sodium  and potassium  concentrations of MBM samples.The zinc concentrations were positively correlated with sodium  of Şanlıurfa tended to have lower intake concentrations of calcium assessed as compared with nursery mothers residing in urban areas . As a result, MBM from nursery mothers residing in suburban area (Eyyübiye) were of poorer nutritional quality that has obvious implication for infants nutrition. Arising from this result, family population was crowded and socioeconomic levels of these families were low. The results of this study were parallel with Qian et al.s'  study whCalcium is the principal element of skeleton system that improves bone mineral density in infants. Our findings show that maternal calcium intake was lower in suburban nursery mothers than other participants of this study. Qian et al.  reportedThere is evidence that sodium levels in infancy may have a long-term effect on blood pressure. The observation was that mean concentrations of sodium were higher than the previous studies , 14. MeaMBM samples seem to contain enough concentration of magnesium as there has not been a report of magnesium deficiency in breastfed infants . The levWe also found that phosphorus concentrations were in a negative correlation with lactation period. The measured phosphorus concentrations in MBM samples ranged within 84–300 mg/L with an average level of 156 ± 46.3 mg/L which was similar to Japan nursery mothers P levels with an average level of 150 ± 38 mg/L .Potassium is an intracellular cation . AverageThere have been few reports about iron content in MBM , 15, 17.The mission of essential elements in biological functions makes them especially significant for nutrition of infant. It is noted that an infant is well protected so long as certain age by maternal homeostatic processes in the event of iron, copper, and zinc . InfantsZinc and copper are essential elements which support development and represent significant levels in MBM . We founIt is presumed that the transition of copper from blood to milk regulated transfer by the mammary gland epithelium . ConsequBy detecting the concentrations of heavy metals (arsenic and lead) of MBM, we estimated that no influences from the maternal intake or environmental exposure to these elements by reason of the concentrations were below the detection limit. This result may be because the Şanlıurfa province is away from the industry. Compared with concentrations reported in the literature , 19 the We should be noted that MBM samples were obtained in one-time collection. Next research needed the composition of MBM varying on a time to time basis. Serial analyses of MBM samples in repeated times should address the concentrations of essential elements and heavy metals of Turkish mothers.In brief, our results agree on the whole with previous reports about the essential element composition of maternal breast milk but it was clearly understood that there is a large standard variation in the composition of essential elements such as Ca, Fe, and Zn. We confirmed that the composition of human milk is affected by various factors such as residing area and using copper equipment.In conclusion, we found a significant correlation in MBM concentrations between nursery mothers from urban and suburban areas of Şanlıurfa. More researches at multiple-time compositional studies are warranted to detect the concentrations for identifying appropriate interventions to ensure healthy growth and development of breastfed infants.


Drosophila suzukii. The close phylogenetic relationship of D. suzukii with Drosophila melanogaster allows, through genomic and functional studies, an insight into the mechanisms directing stress tolerance in D. suzukii.Neuropeptides are central to the regulation of physiological and behavioural processes in insects, directly impacting cold and desiccation survival. However, little is known about the control mechanisms governing these responses in 44) and DH31 neuropeptides demonstrated a high level of conservation between D. suzukii and D. melanogaster with respect to peptide sequences, neuronal expression, receptor localisation, and diuretic function in the Malpighian tubules. Despite D. suzukii's ability to populate cold environments, it proved sensitive to both cold and desiccation. Furthermore, in D. suzukii, Capa acts as a desiccation‐ and cold stress‐responsive gene, while DH44 gene expression is increased only after desiccation exposure, and the LK gene after nonlethal cold stress recovery.Capability (Capa), leucokinin (LK), diuretic hormone 44 (DHDrosophila species, providing evidence that similar signalling pathways control fluid secretion in the Malpighian tubules. Identifying processes governing specific environmental stresses affecting D. suzukii could lead to the development of targeted integrated management strategies to control insect pest populations. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.This study provides a comparative investigation into stress tolerance mediation by neuroendocrine signalling in two  Both cold and desiccation are closely linked at the molecular level to adjustments in the expression of genes involved in key physiological responses, for example in ion transport, carbohydrate metabolism, antioxidant production, immunity, signalling and gene expression pathways.D. melanogaster, several families of neuropeptides have been identified that regulate diuresis in the MTs, including the diuretic hormones DH3144,D. suzukii have allowed identification of orthologous peptides and receptors, which in turn facilitates functional studies to identify suitable targets for the development of novel control strategies against this invasive pest.The ability to rapidly respond to, and tolerate over time, abiotic stresses such as temperature fluctuations , in the short term and/or as a seasonal variation, requires insect osmoregulatory systems, including the gut and Malpighian  tubule (MT), to act in a dynamic and appropriate manner to effect the physiological processes necessary to ensure the animals' survival. Insect osmoregulation, that is, the regulation of cellular ion and water homeostasis via transporters and water channels, is subject to highly sophisticated endocrine control.D. melanogaster can readily be used to identify fundamental similarities that may exist in D. suzukii. Here, we describe the peptide sequences, morphology and organisation of the Capa, LK, and DH44‐producing neurons in the larval and adult central nervous system (CNS) of D. suzukii to gain better insight into the structure–function relationship of these neuropeptides. We also determined the MT‐specific cell types that receive Capa, LK, DH44 and DH31 neuropeptide signals and their physiological role in modulating tubule fluid homeostasis. Furthermore, we investigated tolerance to desiccation and cold in both D. melanogaster and D. suzukii, and we provide data suggesting that neuroendocrine responses to these stresses are potentially altered by the action of neuropeptides acting on the MT to regulate water and ion homeostasis critical for the animal's survival.The depth of knowledge garnered from the genetic model organism 22.1Drosophila melanogaster wild‐type Canton‐S (CS) flies were obtained from Bloomington Stock Center  and maintained on a standard Drosophila medium at 22 °C (unless otherwise stated) and 45‐55% relative humidity with a 12:12 h light:dark photoperiod. Drosophila suzukii flies, originally collected in Italy, were obtained from the laboratory of R. Elwyn Isaac  and reared on standard cornmeal agar medium supplemented with a blueberry fruit, at 26 °C and 60% humidity with a 12:12 h light:dark photoperiod. Adult Drosophila (1‐2 weeks post‐eclosion) of both sexes were used in this study.2.244 . Samples were mounted on poly‐l‐lysine (Sigma‐Aldrich)‐covered 35‐mm glass‐bottomed dishes  in Vectashield . Confocal images were taken using an LSM 880 inverted microscope (Zeiss) and processed with Zen black/blue software  and Adobe Photoshop/Illustrator CS 5.1.Immunostaining procedures were performed as described previously.44 1:2000, rhodami44 1:2000, and rab44 1:2000 were use2.3Drosophila Capa‐1, LK, DH44 and DH31  were synthesised by Cambridge Peptides . The amino acid sequences used to synthesise the neuropeptides were based on the D. melanogaster sequences as most of the peptides predicted and identified from D. suzukii appear to be identical to those previously characterised from D. melanogaster.5‐maleimide‐Capa‐1, Alexa‐488‐C5‐maleimide‐LK, TMR‐C5‐maleimide‐DH44 and TMR‐C5‐maleimide‐DH31. Ligand receptor binding assays were performed on live MTs dissected from 6 − 7‐day‐old adult females. Tubules were mounted on poly‐L‐lysine coated glass‐bottomed dishes in phosphate‐buffered saline (PBS) and then incubated for 2 minutes in a 1:1 mixture of Schneider's medium:Drosophila saline containing 500 ng/ml DAPI and 10‐7 m fluorescent‐tagged neuropeptide and subsequently with 10−5 m unlabelled neuropeptides. As the staining is transitory, the samples were imaged immediately after treatment. All images were taken using a Zeiss LSM 810 inverted confocal microscope.2.4D. suzukii flies were dissected under Schneider's medium and isolated into 10‐µl drops of a 1:1 mixture of Schneider's medium:Drosophila saline. Intact tubules were left to secrete for approximately 30 min, with non‐secreting tubules being replaced if necessary, to produce a set of 10 − 15 working tubules. Secretion rates were measured every 10 min, then the diuretic Drosophila Capa‐1, LK, DH44 and DH31 neuropeptides were added to 10−7 m, and secretion rates were measured for a further 30 min. Data are plotted as mean ± standard error of the mean (SEM).Secretion assays were performed as described previously.2.5D. melanogaster and D. suzuki 6‐day‐old normally fed female flies were anaesthetised and weighed on an AND GR‐202  precision balance . Another group of flies of each Drosophila species were weighed after 18 h of desiccation with no food and no water. Water loss over 18 h was calculated for each group by subtracting the water content at 18 h from that at 0 h. Experiments were run in triplicate with at least 10 flies of each Drosophila species.To measure wet body weight, 2.62.6.1Drosophila suzukii 6‐day‐old female flies were untreated (control), chilled at 4 °C for 6 h (cold), or chilled at 4 °C for 6 h and subsequently allowed to recover for 24 h (recovery). Before treatment, both D. suzukii and D. melanogaster were maintained at a culture temperature of 26 °C to enable a direct species comparison and avoid any potential effects of rearing temperature on inherent thermal tolerance. No mortality was observed for the different treatment groups.2.6.2Drosophila suzukii 6‐day‐old female flies were either untreated (control), placed in empty vials (no food or water) for 18 h at 21 °C and 45‐55% relative humidity (desiccation), or exposed to desiccation for 18 h and subsequently allowed to recover for 24 h (recovery). Before treatment, both D. suzukii and D. melanogaster were maintained at a culture temperature of 26 °C to enable a direct species comparison and avoid any potential effects of rearing temperature and relative humidity on inherent thermal and desiccation resistance. Following treatment, groups of 10 flies were transferred immediately to microcentrifuge tubes containing TRIzol Reagent (Thermo Fisher Scientific) for RNA extraction. For D. suzukii whole‐fly mRNA expression, quantitative reverse transcriptase − polymerase chain reaction (qRT‐PCR) amplifications were performed in an ABI StepOnePlus Detection System  with Brilliant III Ultra‐Fast SYBR Green QPCR master mix  using the following primer pairs: Capa, 5'‐CCGAGTCTGGCAAACAGTCTG‐3' and 5'‐ GACACCAACAAGGGACAAAAGG‐3'; LK, 5'‐CAGTTTCTGGACAGGATTCGGAG‐3' and 5'‐ CAGGTCTCGCATTGTAGGCATC‐3; DH31, 5'‐ATGCCCAGCCAATCCAATGGTG‐3' and 5'‐ TGCGTCCAAAGCGAGTCATC‐3'; DH44, 5'‐CTTCTGCTGGAAATCGCACG‐3' and 5'‐ GTTGCTCCAAACCCTCCA‐3'; Ribosomal protein L18 (rpl18), 5'‐GTTGCTCCAAACCCTCCA‐3' and 5'‐ GATCCGTCTAACACCTCCC ‐3'. Data were normalised against the D. suzukii rpl18 standard and expressed as fold change compared with controls ± SEM (n = 3).2.72.7.150), i.e. the low temperature that results in 50% mortality of a test population. Adults of D. suzukii and D. melanogaster, both reared at 26 °C, were selected at 6 days post‐eclosion, respectively, and exposed to a range of low temperatures  using a direct plunge method.2 and placed in Eppendorf tubes at densities of ten adults per tube, which, in turn, were placed within a glass boiling tube held within an alcohol bath  pre‐set to the desired temperature. Adults were held at the desired exposure temperature for 1 h. Following exposure, adults were allowed to recover in culture vials containing a food source and survival was assessed after 48 h. The procedure was repeated for each exposure temperature.Cold tolerance was determined via calculation of the lower lethal temperature , and the open end of the tube was sealed with parafilm . Vials were checked hourly until no living flies remained. All experiments were run in triplicate with at least 30 flies in each run for each Drosophila species.2.8http://www.neurostresspep.eu/diner) and/or from the National Center for Biotechnology Information (NCBI).Neuropeptide precursor sequences were obtained from DINeR , was used. For mRNA level quantification, one‐way analysis of variance (ANOVA) followed by Tukey's multiple comparisons of means with a significance level of P < 0.05 (for three independent groups: nonstressed versus stressed versus recovered) was used to compare neuropeptide mRNA levels in cold or desiccated flies. For survival curves obtained in desiccation assays, significance was assessed by the log‐rank (Mantel–Cox) test. Log‐rank tests were conducted for each pairwise comparison. The statistical tests were performed using GraphPad Prism 7.0 software . For cold shock experiments, the temperature resulting in 50% mortality of experimental populations (LLT50) was determined for low exposure temperatures using Probit analysis in MINITAB, version 17 . Handling controls resulted in approximately 99% survival across all treatments. The natural response rate was therefore assumed to be close to zero and was not included in the model. Significant differences in mortality were identified by non‐overlapping 95% fiducial limits.For fluid secretion analysis and for the water retention assay, a two‐tailed Student's 33.1D. suzukii protein databases revealed the precursor sequences of Capa,44 and DH31 neuropeptide families. A comparison of D. suzukii and D. melanogaster Capa, LK, DH44 and DH31 peptide precursors is shown in Fig. D. suzukii predicted peptides are identical to the sequences found in D. melanogaster. Furthermore, the presence of these peptides and/or peptides precursors in D. suzukii has previously been confirmed by mass spectrometric analysis. Amino acid BLAST searches of the 3.244 neuropeptides in D. suzukii, we determined the localisation of the neurons producing these neuropeptides and their projections. As the peptide precursor sequences are very similar between D. suzukii and D. melanogaster, we used available antisera against D. melanogaster Capa, LK, and DH44 on the D. suzukii larval and adult CNS.To gain insight into the physiological function of Capa, LK and DHThe antiserum to the Capa precursor recognised several neurons in the larval and adult CNS Fig. a and b, 26,D. suzukii CNS  driven by DH44‐GAL4.44 neurons of the PI also express the LK receptor.Using a specific anti‐DHCNS Fig. e and f, 3.3D. melanogaster that Capa, LK, DH44 and DH31 neuropeptides are released into the circulation from neurohaemal release sites, whereupon they bind their cognate G protein‐coupled receptors localised at the MTs.D. suzukii MTs, we were able to demonstrate specificity of binding of Capa‐1 (one of the peptides encoded by the Capa gene), LK, DH44 and DH31 neuropeptides to tubule PCs and SCs. As in D. melanogaster, application of Capa‐1‐F, DH44‐F and DH31‐F maps their receptors to the basolateral membrane of PCs . In desiccating environments, a key mechanism used by insects to maintain water balance is to reduce the rate of water loss.Drosophila species . During desiccation stress, water loss in D. suzukii was significantly higher than in D. melanogaster . During cold exposure, D. melanogaster proved significantly more cold tolerant than D. suzukii, as determined by non‐overlapping 95% fiducial limits . For D. melanogaster, 5 − 95% mortality occurred over a temperature range of 1.1 °C for females and 1.9 °C for males. For D. suzukii, the corresponding mortality level occurred over a temperature range of 3.7 and 4.8 °C, respectively.As Capa and kinin have been previously shown to modulate stress tolerance in 3.644 and DH31 signalling in desiccation and cold stress were explored , while LK and DH31 expression remained unchanged. Following cold exposure, Capa mRNA levels were up‐regulated , while cold exposure had no significant effect on LK, DH44 or DH31 expression. Surprisingly, LK mRNA levels were up‐regulated during recovery after cold challenge  and with substantial mortality occurring above the supercooling point.D. suzukii and D. melanogaster differ when in their respective winter phenotypes is presently unknown. However, at least in their summer morphs, our lab‐reared Canton‐S D. melanogaster is significantly more cold tolerant than D. suzukii.It should be noted that experiments were performed on individuals of both species maintained at temperatures >20 °C, corresponding to summer conditions. Recent research has shown that D. melanogaster, with some studies observing that males demonstrate enhanced cold toleranceD. melanogaster and D. suzukii and observed no significant difference in cold tolerance between the sexes for both species. This apparent discrepancy may be a consequence of variation in the indices applied to the individual cold tolerance assays, i.e. chill coma, supercooling point, lethal time, lethal temperature etc.,Previous studies have presented contradictory descriptions comparing levels of cold tolerance between the sexes in D. melanogaster proved hardier in response to cold stress, we also observed that D. melanogaster was significantly more tolerant to desiccation than D. suzukii. Indeed, D. suzukii proved highly sensitive to desiccation, with complete lethality occurring in both sexes after approximately 24 h of desiccation (as opposed to D. melanogaster which survived approximately twice as long). Our data suggest that this increased sensitivity to desiccation is a consequence of the observed higher level of fluid loss in D. suzukii as compared with D. melanogaster. It has been shown that, under desiccation conditions, diuretic neuropeptide receptor expression in D. melanogaster tubules is decreased, with a concomitant reduction of the basal tubule fluid secretion rate, therefore potentially limiting water loss.D. suzukii is surprising, and, although the reduced rate of water loss in desiccated Drosophila may be caused by combined reduction in respiratory, cuticular, and excretory water loss, it is possible that, given the unchanged D. suzukii basal secretion during desiccation stress, the MTs may play an important role in water balance during desiccation resistance.In keeping with our observation that D. suzukii during desiccation, with females exhibiting enhanced desiccation resistance, surviving for 8 h longer than males. This is supported by previous work which found female D. suzukii to be significantly more desiccation‐resistant. D. melanogaster males survived starvation longer than females, despite the smaller size of males.Interestingly, whilst no difference was apparent in the cold tolerance responses of males and females, a significant sexually dimorphic effect was observed in D. melanogaster MTs to modulate fluid homeostasis have also been found to modulate stress tolerance. Roles for Capa, LK and DH44 neuropeptide signalling in cold and desiccation has been recently demonstrated in D. melanogaster. Our data for D. suzukii indicate the involvement of Capa and DH44 signalling in desiccation, while Capa signalling is also involved in response to cold stress, as the expression levels for both neuropeptides were increased in response to these stresses and returned to basal levels within the recovery period. Surprisingly, LK mRNA levels were up‐regulated during the recovery period following a cold challenge, suggesting that LK neurons may release LK peptide during recovery from cold stress. As LK does have demonstrated roles in feeding behaviourD. suzukii would be a useful subject for future investigations. Diuretic neuropeptides that act on Drosophila, several peptide hormones have been identified as pivotal to regulating ion transport and fluid secretion in the MTs and gut.D. suzukii the neuropeptide sequences for Capa, LK, DH44 and DH31, the neuroendocrine cells that produce these neuropeptides, their processes and the potential neural network that affects water homeostasis , showing it to be similar to that of D. melanogaster. To fully determine the importance of these neuropeptides and receptor genes in D. suzukii biology, it will be necessary to develop functional genomic tools that are currently employed in D. melanogaster. Germline transformation techniques and Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR‐associated proteins (CRISPR/Cas9)‐mediated mutagenesis have already been successfully deployed in D. suzukii,In


Moreover, we provide global distributions of VOC formation potentials, which can be used as simple tools for field studies to estimate photochemical VOC emissions depending on location and season.The surface of the oceans acts as a global sink and source for trace gases and aerosol particles. Recent studies suggest that photochemical reactions at this air/water interface produce organic vapors, enhancing particle formation in the atmosphere. However, current model calculations neglect this abiotic source of reactive compounds and account only for biological emissions. Here we show that interfacial photochemistry serves as a major abiotic source of volatile organic compounds (VOCs) on a global scale, capable to compete with emissions from marine biology. Our results indicate global emissions of 23.2–91.9 TgC yr Volatile organic compounds are photochemically produced in the ocean surface microlayer, but estimates are missing. Here the authors combine experiments and observations to quantify photochemical emissions of volatile organic compounds and show that they are comparable to biological production. Nonetheless, the most prominent example of a surfactant-enriched surface is probably the ocean, covering more than 70% of Earth. Studies have shown that this air/water interface is almost ubiquitously covered by a thin film of amphiphilic compounds, which are enriched there with respect to the bulk water7. Such sea surface microlayers (SMLs) were found to have significant effects on marine biogeochemical as well as climate related mechanisms by directly affecting processes such as exchange of trace gases , heat, and aerosol particles3. In addition, recent field studies9 confirmed for the first time previous laboratory observations15 suggesting that irradiation of this air/water interface by sunlight produces organic vapors, known to enhance particle formation in the atmosphere. These emissions were attributed to purely photochemical reactions occurring in the SML. However, current model calculations neglect this abiotic source of reactive compounds and account only for organic vapors that are produced directly by biological processes20.Life on Earth is a constant and versatile source of surface-active compounds  in the ambient environment. Biogenic surfactants are, therefore, omnipresent on surfaces exposed to the open atmosphere, ranging from lakes and rivers to cloud droplets, and aerosol particles15, to identify locations and time periods in which such photochemistry is of major importance for marine VOC levels. Moreover, we explore the global extent of such yet underestimated VOC emissions as well as their implications for organic aerosol (OA) mass in marine environments. Our estimations are based on a collection of photochemical VOC production parameters, surface UV irradiation, biological activity, and surface wind speeds. Using these variables, we estimate SML coverage of the oceans and VOC formation potentials from interfacial photochemistry on a 1° × 1° global grid. We infer potential OA mass contributions from photochemically produced VOCs using aerosol partitioning theory and a one-dimensional volatility basis set approach23. As we demonstrate, accounting for abiotic photochemical VOC production closes current gaps in our understanding of atmospheric chemistry in remote regions and underlies the importance of the interplay between biology and chemistry for climatic processes. We infer global emissions of 32.5–129 Tg C yr–1 (23.2–91.9 TgC yr–1) of organic vapors from the oceans into the marine atmosphere. Moreover, our results suggest a potential contribution to organic aerosol mass of more than 60% over the remote ocean. In addition, to support planning and design of future field studies with a focus on VOC emissions from interfacial photochemistry, we provide global distributions of VOC emission potentials as well as a small calculation tool based on Matlab, to estimate photochemical VOC emissions depending on location and season.Here, we combine for the first time results on the formation and presence of SMLs with observations of photochemical production of organic vapors from irradiation of surfactant-enriched air/water interfaces24 estimated to wind speeds of 8–10 m s–1, however, recent field observations25 suggest SML coverage even up to wind speeds of 13 m s–1.Since VOC formation from interfacial photochemistry will only occur in the presence of surfactants, our starting point is the global SML coverage of the oceans. The exact spatial and temporal dynamics of these surface films are largely dependent on local biological and meteorological conditions. In particular, high wind speeds at the ocean’s surface were shown to disturb the formation of such SMLs by breaking waves. The upper limit for the formation of SMLs was previously2, we use biological net productivity and mean surface wind speeds to derive global SML distributions. As shown in Fig. –1 , solar radiation UVa,b at the Earth’s surface , and a correction factor Fsurfactant to account for SML presence and surfactant concentration. In addition, we add a factor kg to account for the impact of wind on the gas transfer velocities across the air–sea interface, which are largely determined by sea-surface roughness7. We keep µphoto in units of mW s cm–2 day–1 to allow for convenient conversion to daily VOC fluxes, as explained later. Here, we use averaged surfactant concentrations from field observations2 and normalize µphoto to the highest surfactant concentrations, commonly observed in eutrophic regions. Fsurfactant can therefore be expressed ascsurfactant , giving values between 0 and 1. For the gas transfer coefficient kg, we apply the wind speed dependent parameterization of McGillis and co-workers26 and normalize it to laboratory conditions of Ciuraru et al.10 and Brüggemann et al.14:U10 at 10 m height, and the sample flow speed under laboratory conditions Ulab = 5.31 × 10–2 m s–1 . Moreover, we perform all calculations at wind speed limits of 8, 10, and 13 m s–1 to account for the range of global SML abundances.Assuming a linear relationship between photochemical VOC production and solar radiation as well as a logarithmic decay with decreasing surfactant concentrationsµphoto and, thus, photochemical VOC formation is expected to occur especially in coastal waters in the tropics throughout the entire year. However, in these regions elevated direct biological VOC emissions are also anticipated due to high microbiological activity in surface waters. Thus, total annual photochemical VOC emissions are of minor importance in such locations, despite constantly high values for µphoto. Nonetheless, on shorter time scales interfacial photochemistry might play a pivotal role here, e.g., during the decay of microalgae blooms. In contrast, for remote regions of the oceans with high solar radiation but lower biological activity, interfacial photochemistry can serve as a major source of VOCs over longer periods. The extent of this abiotic source for organic vapors typically varies with the season. For example, the southern Indian Ocean exhibits an elevated µphoto for the seasons Jan–Mar and Oct–Dec, whereas the remote North Pacific shows similar photochemical potentials for the seasons Apr–Jun and Jul–Sep. We note that the extent and variability of µphoto eventually depends on the SML formation dynamics, and thus also on the wind speed limit for SML formation. For example, the high values obtained for the Southern Ocean for the season of Oct–Dec rapidly decline to zero, if we set the wind speed limit for SML formation to 10 m s–1  in the Arctic during July–August8. In general, the observed OVOCs exhibited a higher degree of oxidation compared to laboratory studies. However, this difference might simply be due to some initial oxidation reactions of the originally emitted VOCs in the atmosphere, as sampling occurred ~15 m above the sea surface. Moreover, the applied acetate chemical ionization mass spectrometer is in particular sensitive towards acidic compounds, such as the detected organic acids. In addition to such OVOC emissions, Burkart et al. found enhanced new particle formation in the Arctic summer at low wind speeds and sunny conditions9. However, since new particle formation is a complex process, we note that besides interfacial photochemistry several other factors might have affected these observations significantly. Nevertheless, these field observations are in good agreement with our calculations, which show a maximum for µphoto for the measurement location and time . However, it is important to note that Kim et al.27 observed high wind speeds of up to 25 m s–1 during their study period, with an average wind speed of 11 m s–1. Formation and presence of prevalent SMLs, and hence, photochemical VOC formation at the air/sea interface, are unlikely under such conditions. Thus, our calculations for an ocean surface that is largely free of SMLs  in the Supplementary Information  directly yields estimates on VOC emissions (in molecules cm–2 day–1). Thus, allowing us to approximate photochemical VOC emission fluxes for a certain region and season, even for single compounds. As a simple example, we estimate global annual isoprene emissions of 1.11 (0.70–1.52) Tg yr–1 from interfacial photochemistry , hence, contributing significantly to marine VOC emissions.A further benefit of try Fig. , using SWe note that the approach presented here can solely give an estimate on average monthly VOC fluxes from interfacial photochemistry. Thus, more sophisticated models with higher time and space resolution will be necessary, to predict photochemical VOC fluxes on a daily or even hourly basis. Nonetheless, as shown in Table 30 observed emission fluxes of acetone and acetaldehyde of 7.11 × 1014 and 3.9 × 1014 molecules cm–2 day–1, respectively, during a cruise in the tropical Atlantic (3–39° N). However, our calculations merely predict emission fluxes in the range of 0.58–1.1 × 1014 and 0.20–0.60 × 1014 molecules cm–2 day–1 from interfacial photochemistry for acetone and acetaldehyde, respectively.In particular, photochemical production and emission of larger (unsaturated) VOCs, such as isoprene, seems important, since few other abiotic formation pathways are known for such compounds in the marine environment. In contrast, the contribution to fluxes of smaller VOCs, such as acetone, or acetaldehyde, is probably less significant, since a large variety of strong sources exists in both the atmosphere and the ocean. As an example, Yang et al.31  from unsaturated VOC oxidation products. To approximate background aerosol concentrations, primary OA emissions are estimated based on biological activity and surface wind speedsDespite remaining uncertainties, our study demonstrates that abiotic VOC production from interfacial photochemistry might serve as a major contributor to marine VOC levels and OA mass loadings. Isoprene emissions from photochemistry are expected to be in the same range as direct biological emissions and might even dominate depending on location and season. Moreover, SOA formation from oxidation of photochemically produced VOCs might explain field observations of new particle formation events which are occurring decoupled from biological activity. Therefore, we suggest that VOC and SOA production from interfacial photochemistry should be taken into account to accurately model and predict atmospheric chemistry and related processes, such as ozone cycling and cloud formation.https://www.eoas.ubc.ca/~rich/map.html) for the production of global maps. A general overview on all data sources and calculations is given in Supplementary Fig. All data processing was conducted using Matlab , including the m_map tools package  for the air/water interface of each 1-by-1 degree grid cell (Eq. (tsun (in s), solar radiation UVa,b at the Earth’s surface (in mW cm–2 day–1), and a unitless correction factor Fsurfactant to account for SML coverage and surfactant concentration . As exptsun and UVa,b we used climatological data on solar irradiation at the surface of the Earth in the UV-a (315–400 nm) and UV-b (280–315 nm) range from the NCAR TUV model (https://www2.acom.ucar.edu/modeling/tropospheric-ultraviolet-and-visible-tuv-radiation-model), giving monthly averaged UV irradiance values  for the period of 1979–2000. The data was refined from 1° × 1.25° resolution to 1° × 1° resolution by linear interpolation for all further calculations.As depicted in Supplementary Fig. csurfactant) and SML coverage was conducted according to the method described by Wurl et al.2. First, oligo-, meso- and eutrophic regions of the ocean were identified from global maps of net primary production (NPP), based on the Vertically Generalized Production Model38. Monthly data on NPP were obtained from the ocean productivity webpage of the Oregon State University (http://www.science.oregonstate.edu/ocean.productivity), ranging from July 2002 to November 2016  from field observations2. The assigned surfactant concentrations are as follows: for oligotrophic waters csurfactant = 320 µg Teq L–1 , for mesotrophic waters csurfactant = 502 µg Teq L–1 , and for eutrophic waters csurfactant = 663 µg Teq L–1 .Global mapping of the ocean’s surfactant concentrations  were obtained from NCEP/NCAR reanalysis project (http://www.esrl.noaa.gov) and averaged for each month . Furthermore, we assumed photochemical VOC production to exhibit a logarithmic decay with decreasing surfactant concentrations11. Fsurfactant was therefore expressed as shown in Eq. , measured photochemical VOC emissions from laboratory studies VOClab (in molecules mW–1 s–1), and the photochemical potential µphoto. Values for VOClab can readily be obtained from photochemical experiments under controlled conditions, using common VOC measurement techniques such as proton transfer reaction mass spectrometry (PTR-MS). In this study we used such data from previous work of George and co-workers14. For example, to estimate global emissions of isoprene from interfacial photochemistry, we used previously reported values for VOClab from marine SML and biofilm samples14, which are in the range of 3.71–6.19 × 107 molecules mW–1 s–1.To infer estimates on photochemical VOC production and emission of single compounds from interfacial photochemistry at the surface of the ocean, 21 and a one-dimensional volatility basis set22, lumping VOC oxidation products in 10 volatility bins with saturation vapor pressures of log C*(300 K) = {−3, −2, −1, 0, 1, 2, 3, 4, 5, 6} µg m–3. Since so far no parameterization of SOA yields for oxidation of VOCs from interfacial photochemistry is available, here we accounted solely for oxidation of unsaturated VOCs by OH radicals, using SOA yields for alkenes previously given by Tsimpidi et al.23 . Moreover, we assumed pseudo-first order kinetics for the reaction of VOCs with OH radicals. We attempted to approximate the reaction rate constant kOH for the mixture of unsaturated VOCs by averaging known kOH constants of similar compounds39. An overview of the compounds and rate constants used for this approximation is given in Supplementary Table kOH = 109.76 × 1012 cm3 molecule–1 s–1, which was then applied to estimate formation of VOC oxidation products. For all calculations, we used an OH concentration of cOH = 1.5 × 106 molecules cm–3. Unless stated otherwise, the aging time for SOA formation and growth was set to 12 h per day at a temperature of 15 °C.As depicted in Supplementary Fig. 39. Monthly mean data on sea surface temperature and chlorophyll-a concentrations were obtained from MODIS ocean products (https://modis.gsfc.nasa.gov/). POA emissions were instantaneously diluted into the marine boundary layer (MBL). Monthly mean MBL heights were obtained from the NOAA-CIRES 20th Century Reanalysis project (https://www.esrl.noaa.gov/psd/data/20thC_Rean/)  mass fluxes and concentrations above the ocean, in order to model accurately the partitioning of VOC oxidation products. POA was, however, solely regarded as background aerosol, which was not taking part in partitioning, i.e., POA was defined as non-volatile. Here we applied established POA emission schemes, which infer POA fluxes from sea surface temperature, chlorophyll-a concentration, and surface wind speed–3 , an ambient temperature of 20 °C, an MBL height of 600 m, and an emission rate of unsaturated VOCs of VOCambient = 4.1×109 molecules cm–2 s–1 . Both VOC formation from interfacial photochemistry and oxidation of the gas-phase products start at time 0. The resulting mixing ratio of, yet unreacted, unsaturated VOCs is depicted in the left panel of the figure, reaching a steady state of ca. 4 × 108 molecules cm–3  after 8 h. This equilibrium mixing ratio might seem quite low, however, we note that here we assume instantaneous mixing of the emitted VOCs into the entire volume of the MBL. Therefore, ambient mixing ratios in direct proximity of the air/water interface  are likely to be much larger, whereas our calculations solely represent a lower limit.As a simplified example, Supplementary Fig. –3 is observed after a total aging time of 12 h, i.e., a relative increase of 22% in OA particle mass. Again, this value represents solely a lower limit of enhanced SOA formation, since we excluded ozone chemistry, which would generate additional low volatility oxidation products, probably enhancing OA particle mass concentrations.Nonetheless, even for this dilute scenario we observe significant SOA formation from VOC oxidation products, as depicted in the right panel of Supplementary Fig. The Matlab code of the Photochemistry At Liquid/Air interfaces containing Surfactants (PhotochemAtLAS) model, including instructions, is publicly available on the online community repository Bitbucket (https://bitbucket.org/bruggemann_m/photochematlas).Further data sets generated during this study are available from the corresponding author upon reasonable request.Supplementary Information


The aim of this study is to evaluate the effect of growth hormone therapy (rGH) on mitochondrial function on peripheral muscle and to correlate with exercise capacity in subjects with severe adult growth hormone deficiency (GHD).Six months, double-blind, randomized, crossover, placebo-controlled trial of subcutaneous rGH in 17 patients with GHD.Quadriceps muscle biopsies were obtained at baseline, 3 months, and 6 months to measure succinate dehydrogenase (SDH) to assess mitochondrial activity. Exercise capacity was measured with cardiopulmonary exercise testing. Lipids, glycemic parameters, and body fat levels were also measured.p < 0.05) and normalized with rGH in the active phase (p < 0.005). Patients showed an increase in SDH (p < 0.01) from base line that differed between placebo and rGH therapy treatment groups (p < 0.05): those treated by rGH followed by placebo showed a significant increase in SDH (p < 0.001) followed by a decrease, with a significant between group difference at the end of 6 months (p < 0.05). No significant improvements or correlation with exercise capacity was found.Serum insulin-like growth factor 1 (IGF1) levels reduced fat mass by 3.2% (Short-term rGH for 3 months normalized IGF1 levels, reduced fat mass, and had a significant effect on mitochondrial function, but exercise capacity was unchanged.Number ISRCTN94165486. Patients with hypopituitarism have reduced life expectancy when compared to the general population, mainly attributed to cerebrovascular and cardiovascular disease , 2. The 2 max) is widely accepted as the single best measure of cardiovascular fitness and maximal aerobic power and represents the maximum capacity of an individual’s body to use oxygen during incremental exercise. Studies assessing VO2max in GHD have reported a reduction in aerobic capacity associated with a decreased VO2max up to 28% ; however, available evidence also suggests a GH-independent IGF1 effect as the growth response is better when GH-deficient patients are treated with GH in comparison to IGF1 treatment in patients with GH insensitivity .GH binding to its receptors modulate its effects in the mitochondria , 12 throTo date, it is unknown if GHD is associated with muscle mitochondrial dysfunction. Therefore, the aim of this study is to evaluate the effects of short term administration of GH on peripheral muscle oxidative capacity using SDH activity as a surrogate marker and whether this correlated with exercise capacity and cardiovascular risk markers.Seventeen patients  with hypopituitarism resulting from pituitary tumors who were treated with surgery, radiotherapy, or both were recruited. Severe GHD was confirmed by a peak GH response to insulin-induced hypoglycemia of less than 9 mU/L (3 ng/mL). The average time from diagnosis of the pituitary lesion to inclusion in the trial was 33 months with confirmation of severe GHD 12 months prior to inclusion in the trial. Subjects were on stable hormone replacement doses for thyroid, adrenal, and testosterone deficiencies for the previous 6 months prior to the study and throughout the study, and no adjustment of the doses were needed during the study. None of them had been previously treated with rGH  versus placebo . The patients were randomized with a random generator table and, using a crossover design, allocated to study group A or B. Daily rGH or placebo injections were prepared by a pharmacist, which was separate from the trial. Nine patients in one arm and eight in the second arm were randomized to either placebo or rGH for 3 months before being crossed over to the second arm of the study following a 2-week washout period for a further 3-month period , and total cholesterol, triglycerides, and high-density lipoprotein cholesterol levels were measured enzymatically using a Synchron LX20 analyzer . IGF1 was measured using a solid-phase, enzyme-labeled chemiluminescent immunocentric assay on an Immulite 2000 analytical platform . IGF1 SDS measurements were calculated from the online calculator for the Immulite assay platform (http://ticemed_sa.upmc.fr/sd_score/).At the beginning and end of each phase, following an overnight fast, weight and blood pressure were measured, and blood samples were collected. Blood pressure was measured after the patients had been seated quietly for at least 5 min with the right arm supported at heart level. Blood pressure measurements were performed using an automated device  during each study visit. Two readings were obtained at the beginning of each visit at least 1 min apart, and the mean value was taken. Total body fat was measured with the bioelectrical impedance analysis technique using Tanita scales . Fasting venous blood samples were collected, separated by centrifugation at 2,000 Quadriceps muscle biopsies were taken to assess changes in SDH activity using immunohistochemistry. The biopsies were frozen immediately after excision, mounted onto cork disks, frozen in 2-methylbutane at −196°C, and finally 12-µm sections were cut using a cryostat and mounted onto polysine (VWR) microscope slides, which were stored at −25°C until use. All samples were analyzed in a single batch. Thawed sections were incubated for 1 h at 37°C in 0.5 M Sorenson’s buffer containing 0.5 M sodium succinate and 1 mg/mL nitroblue tetrazolium. After rinsing in water, the sections were fixed in formal saline and mounted in glycergel for optical density analysis using a light microscope equipped with a Cool-Snap digital camera and Image Pro plus an image analysis software . Four random fields of view at 100× magnification were used from each section. Black and white images were captured, background corrected, and calibrated for incident light. An average optical density value for each was measured.2max) following cardiopulmonary exercise test, using a modified Bruce protocol. During the test, patients wore a tightly fitting facemask to which was connected a capnograph and a sample tube enabling online measurement of ventilation and metabolic gas exchange. A respiratory exchange ratio (RER) >1 was taken to suggest a maximal effort together with an attainment of at least 85% of maximal heart rate.Exercise capacity was assessed calculating the maximal oxygen consumption  for the combined arms comparing baseline just prior to rGH therapy to 3 months rGH therapy . There was a difference between placebo and rGH-treated groups  , slope of the relation between ventilation and carbon dioxide production (VE/VCO2), anaerobic threshold, RER, exercise time, or pulse at maximal exercise were noticeable (Table p > 0.05).No significant effects in peak oxygen consumption peak VO, slope op < 0.05). IGF1 levels did not differ with 3 months of placebo therapy and normalized with 3 months rGH therapy; these patients were then continued on growth hormone at the end of this period. For those on rGH treatment that was started on rGH (IGF1 120 ± 51 μg/L) and crossed over to placebo, IGF1 levels fell to those of pretreatment after rGH was withdrawn  before they too were recommenced on rGH therapy at the end of the study period. No significant differences were observed in body mass index (BMI), waist–hip ratio, blood pressure, glucose, HbA1c, total cholesterol, or triglycerides (Table p > 0.05). IGF1 SDS scores at baseline, 3 months active, and then 3 months placebo were −1.21 ± 0.9, 1.29 ± 0.9, and −1.07 ± 1.0, respectively, for the group that had active treatment first followed by placebo. IGF1 SDS scores at baseline, 3 months placebo, and then 3 months active were −2.2 ± 1.1, −2.1 ± 1.2, and 0.1 ± 0.8, respectively, for the group who had placebo followed by active treatment. While the baseline IFG-1 SDS scores differed at baseline, this difference was not significant (p < 0.06).Recombinant GH replacement normalized IGF1 levels within the reference range. The incremental difference in total body fat mass determined by Tanita measurement between active and placebo arms was calculated for each patient (placebo 0.2 ± 0.2% versus 3.2 ± 0.3% active) and was significantly reduced  after 6 months therapy (Our study suggests that at the end of the 3-month rGH treatment period, SDH reflective of mitochondrial oxidative activity in the peripheral muscle was higher in the rGH group compared to the placebo group that would be in accord with the reduced oxidation that was seen for rGH replacement over a 6-week period . However therapy , 22. In y weight –26; howe therapy , due to  therapy . SDH is  therapy  intermit therapy , 31. Fur therapy . It shouThe adequacy of rGH replacement in this study was shown by the normalized IGF1 levels. However, this dose and/or the length of treatment was inadequate to improve weight, central obesity, and hypercholesterolemia. It has been suggested that patients with the most adverse lipid profile benefit the most following rGH replacement  with chaOur study has a number of limitations. This was a small study although offset by the crossover design. The duration of GH therapy was short that may have been insufficient for the full spectrum of cardiovascular benefit to have accrued. However, the effect of GH on oxidation may be seen after 6 weeks of GH replacement therapy and therefore 3 months treatment would have been sufficient for muscle mitochondrial function changes to be evident , and indAll the patients normalized IGF1 within reference range, but due to the study design, it was not possible to further titrate the rGH; however, normalization of IGF1 with a reduction of the body fat suggests that the patients were replaced adequately. The length of GH deficiency in the study may have been too short to have allowed more of a GH deficiency spectrum to develop after the active treatment phase.In summary, short-term, fixed rGH for 3 months had a significant effect on mitochondrial function and favorably improved total fat mass and IGF1 levels although cardiovascular risk factors and exercise capacity did not differ over this time period.The study followed the declaration of Helsinki guidelines and was approved by the Hull and East Riding ethics committee. The trial registration number is ISRCTN94165486.SG did the data analysis and wrote the manuscript. TS, ZJ, and SA revised and edited the manuscript. All authors approved the manuscript for submission.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Human behavior is frequently described both in abstract, general terms and in concrete, specific terms. We asked whether these two ways of framing equivalent behaviors shift the inferences people make about the biological and psychological bases of those behaviors. In five experiments, we manipulated whether behaviors are presented concretely  or abstractly . People judged concretely framed behaviors to be less biologically based and, on some dimensions, more psychologically based than the same behaviors framed in the abstract. These findings held true for both mental disorders (Experiments 1 and 2) and everyday behaviors (Experiments 4 and 5), and yielded downstream consequences for the perceived efficacy of disorder treatments (Experiment 3). Implications for science educators, students of science, and members of the lay public are discussed. In everyday life, we tend to frame behaviors in different ways. Sometimes we talk about behavior in general terms . At other times, we talk about those same behaviors with reference to particular people in the context of their lives . The question is whether these different kinds of descriptions matter; that is, does framing affect the inferences we make about those behaviors? Although these abstract and concrete descriptions seem to essentially depict the same behaviors, we found that the two levels of description lead to different judgments about how to explain the behavior. Across five studies, participants favored biological explanations  more for abstract descriptions than for concrete cases and they favored some psychological explanations  more for concrete cases than for abstract descriptions. These shifts in people’s preferences occurred both for ordinary behaviors  and mental disorder symptoms (e.g. delusions). As neuroscience and genetics research have increasingly been capturing the public’s attention, we argue that these results have important implications for science education and for public health communication.A Beautiful Mind; Howard, I Know This Much Is True; Lamb, In the real world, unusual human behaviors (e.g. the symptoms of schizophrenia) are often described at one of two distinct levels of abstraction. At one level, behaviors are described in the abstract, as generalized across individuals. For example, when we google the word “schizophrenia,” the websites that immediately come up—from the National Institute of Mental Health, Mental Health America, National Alliance for the Mentally Ill, Wikipedia, schizophrenia.com, and so on—provide abstract descriptions of schizophrenia and its symptoms (e.g. delusions). Abstract descriptions are also found when we search through an encyclopedia, dictionary, or medical handbook. At another level, we also talk about specific instances of the same behaviors (e.g. a woman who strongly believes that the next-door neighbor is her husband when in fact they have not met). One might learn about the concrete symptoms of schizophrenia via the depiction of a particular person in a film  are not tangible in nature, laypeople may actually see them as being more abstract than biological states, which refer to tangible things such as the physical brain. Furthermore, from a reductionist viewpoint, biological explanations would be considered lower level explanations for behaviors than psychological explanations for the same behaviors. Within the hierarchy of levels of explanation, psychological explanations are more abstract than biological ones, being relatively lacking in concrete, physically grounded detail  and non-generic statements  judgments are affected by the abstract versus concrete framing of behaviors and, if so, in what direction judgments are affected. We tested these hypotheses by measuring people’s endorsements of various biological and psychological explanations for behavior, across a range of equivalent abstract and concrete cases.Diagnostic and Statistical Manual of Mental Disorders , as in nosologies such as the Our prediction is that biological explanations are more strongly endorsed in the abstract than in the concrete, and that psychological explanations of behavior are more strongly endorsed in concrete cases than in the abstract. That is, we would expect laypeople to think that brain chemistry, neural structure, and so on are better explanations of calm performance in general than of Allen’s calm performance in particular. Conversely, we predict that explanations attributing calm performance to intentions or emotions would be endorsed more for Allen’s calm performance than for calm performance in general.We tested these predictions across five experiments. Experiments 1 and 2 compared laypeople’s judgments of the biological  bases of various mental disorders. Each disorder was described in a concretely or abstractly framed vignette, judged by pretest participants to be essentially equivalent. Experiment 3 tested whether these inferences have downstream consequences for how people would choose to intervene on disordered behavior—by using medication or by using psychotherapy. Finally, Experiments 4 and 5 extended these results beyond the domain of mental disorders, examining lay judgments for behaviors that are uncommon (and hence in need of explanation) but not the result of mental disorders.Experiment 1 tested whether laypeople’s causal attributions for disordered behavior are shifted by abstract versus concrete framing. Although clinicians tend to view behaviors as more biologically based in the abstract than in the concrete, and more psychologically based in the concrete than in the abstract .Fifty-one participants were recruited via Amazon Mechanical Turk ; the mean rating for the “good example” question was 8.21 (SD = 0.29). Mean ratings by item were all at least 7.60. Thus, these pretest results verified that each pair of abstract and concrete versions is fairly equivalent.Because we are testing the effect of abstract versus concrete framing of the same behavior, we recruited a separate group of 40 participants from Amazon Mechanical Turk to complete a pretest, measuring whether the abstract and concrete version of each behavior correspond to each other. Each behavior was shown on a separate page and the two versions of each behavior, abstract and concrete, were presented side by side on the page. As an attention check, two filler items not designed to be equivalent were also included. Four participants failed this check. Of the remaining 36 pretest participants, 15 judged whether the abstract version was “a good abstract description” of the concrete version on a scale of 1–9 , while 21 judged whether the concrete version was “a good example” of the abstract version on a scale of 1–9 . The mean rating for the “good abstract description” question was 7.97  to allow for attention and comprehension checks. If participants paid attention to the task, these filler items should receive very high ratings on biological questions and very low ratings on psychological questions. Five participants who did not show this pattern for the two filler items  were excluded from the final data analyses.Q1. Do you think [their/her/his] brain chemistry is different from that of people who [are not like this/do not do this]?Q2. Do you think [their/her/his] brain structures are different from those of people who [are not like this/do not do this]?Q3. Do you think there is a genetic basis for this?For the main experiment, nine questions were developed to measure people’s judgments of the biological and psychological bases of behaviors. Three biological questions were designed to probe beliefs about biological causes of behaviors:Q4. Do you think this is caused by cognitive factors ?Q5. Do you think this is caused by [their/her/his] emotions and desires?Q6. Do you think this is caused by [their/her/his] [personalities/personality]?Q7. Do you think [they are/she is/he is] intentionally [like this/doing this]?Q8. Do you think [they/she/he] should be [held responsible for/given credit for] [being like this/doing this]?Q9. Do you think the causes of this are under [their/her/his] control?Because naïve biology is likely to be limited, only three questions could be developed . In contrast, because the existing literature suggests that naïve psychology encompasses a number of aspects of behavior  were derived from tables of contents of Introductory Psychology textbooks as factors that are frequently addressed in the study of individual differences. Q7, Q8, and Q9  were derived from questions measuring beliefs about agency . For each version of each behavior, the nine questions were presented in randomized order across participants and across items. For each item, participants completed the nine explanation judgments on the same screen, with each item presented on a separate screen.Qualtrics  × 2  within-subjects design.We first computed a biological score for each item by averaging each participant’s responses to the three biological measures , and a psychological score for each item by averaging each participant’s responses to the six psychological measures (α = 0.97).F = 95.68, p < 0.001, ηp2 = 0.70, as shown in Fig. M = 5.37, SD = 1.23) than for the concrete versions , t(42) = −6.32, p < 0.001, d = −0.96, while psychological attributions were higher for the concrete versions  than for the abstract versions , t(38) = 10.85, p < 0.001, d = 1.65.We predicted that biological attributions would be greater for the abstract version than for the concrete version and that psychological attributions would be greater for the concrete version than for the abstract version. To test this, we conducted a 2 (concrete or abstract) × 2  repeated measures ANOVA on each participant’s mean across items. This analysis revealed the predicted interaction, F,42 = 95.Figure F = 17.32, p = 0.009, ηp2 = 0.78. Biological attributions were higher for the abstract versions  than for the concrete versions , t(5) = −2.58, p = 0.049, d = −1.05, while psychological attributions were higher for the concrete versions  than for the abstract versions , t(5) = 5.04, p = 0.004, d = 2.06.The interaction effect also held up in a by-item analysis, using each item’s mean score across participants. A 2 (abstract or concrete) × 2  repeated measures ANOVA on these scores revealed a significant interaction, Experiment 1 found that biological attributions were higher for abstract than concrete descriptions and psychological attributions were higher for concrete than abstract descriptions for the same behaviors. Remarkably, although neither the abstract nor the concrete version explicitly mentioned anything about the causes of the behaviors, attributions were strongly affected by the framing manipulation. Thus, not only expert clinicians .A total of 124 participants were recruited via Amazon Mechanical Turk, of whom nine were excluded . The six psychological questions were split into two groups to equate the total number of questions received across all participants. Sample sizes were determined by power analyses on the data from Experiment 1, with 95% power subject to a minimum of 40 participants per condition (prior to excluding random responders and repeat participants).The stimulus materials were the same as in Experiment 1. Unlike in Experiment 1, the nine questions were grouped into three sets: Biological  = 54.83, p < 0.001, ηp2 = 0.50, as shown in Fig. M = 5.31, SD = 1.20) than for the concrete versions , t(40) = −7.47, p < 0.001, d = −1.67. Conversely, psychological attributions were higher for the concrete than for the abstract versions, both for Psychological Set 1 , t(37) = 3.44, p = 0.001, d = 0.56, and for Psychological Set 2 , t(35) = 8.36, p < 0.001, d = 1.38. As shown in Fig. p < 0.05, two-tailed; cognitive factors reached marginal significance in the predicted direction, p < 0.10).We conducted a 2 × 3 mixed-model ANOVA on each participant’s mean across items, with framing (concrete or abstract) as a within-subjects factor and attribution type  as a between-subjects factor. This analysis revealed the predicted interaction, F = 22.51, p = 0.005, ηp2 = 0.15. Biological attributions were higher for the abstract versions  than for the concrete versions , t(5) = −3.04, p = 0.029, d = −1.24, while psychological attributions were significantly higher for the concrete versions  than for the abstract versions , t(5) = 3.90, p = 0.011, d = 1.59.The interaction effect also held up in a by-item analysis. A 2 (abstract or concrete) × 2  repeated measures ANOVA on the item means revealed a significant interaction, These results show that the strong shifts in attribution shown in Experiment 1 cannot have occurred due to demand to inversely rate biological and psychological causes. Rather, these shifts occur independently, reflecting both a stronger belief in biological causation in the abstract and a stronger belief in psychological causation in the concrete.In Experiment 3, we tested whether the effect of abstract versus concrete framing on biological versus psychological attributions might have a downstream effect on the perceived efficacy of treatments for mental disorders. Such a finding would have implications both for psychiatric intervention and for public health, since perceived treatment efficacy can influence actual treatment efficacy  to 9 (“completely”). Participants were told that psychotherapy refers to “treatment by psychological means, involving repeated verbal interactions between a clinician and a client,” and that medication refers to “treatment by psychiatric, psychoactive, or psychotropic drugs.” These judgments were always made on the same page and their order was counterbalanced so that some participants always made medication judgments first and other participants always made psychotherapy judgments first. The abstract versus concrete framing was a within-subject factor with the order of the items counterbalanced as in Experiment 1, so that the abstract and concrete versions of the same item would appear in separate halves of the experiment.F = 9.61, p = 0.004, ηp2 = 0.20, as shown in Fig. M = 5.71; SD = 1.64) rather than concretely , t(39) = 3.70; p = 0.001; d = 0.58. However, judgments of the effectiveness of psychotherapy did not reliably differ between the abstract  and concrete versions , t(39) = 0.79, p = 0.43, d = 0.13.We conducted a 2 (concrete or abstract) × 2 (medication or psychotherapy) repeated-measures ANOVA on individual participants’ means across items. This analysis revealed the predicted interaction, F1,9 = 9.61,When behaviors are described more abstractly, and biological explanations thereby seem more plausible (as shown in Experiments 1 and 2), the current results suggest that people come to believe that biological interventions on that causal system are more likely to influence those behaviors. These results generalize the effect of abstract and concrete framing on biological attributions to a new measure. That said, it is difficult to say whether or not the effect on treatment decisions is statistically mediated by attributions, since the effect was found for medication but not for psychotherapy. A test for mediation would require a design that measured both attributions and treatments simultaneously.Why did the effect not extend to psychotherapy efficacy judgments? Although it is possible that this occurred because the effect of abstract/concrete framing on psychological explanations is less stable than the effect on biological explanations, we think this is not the most likely explanation. The abstractness manipulation was sufficient to find robust differences for both psychological and biological explanations in Experiments 1 and 2 and this same manipulation was used here in Experiment 3. Instead, the null effect on psychotherapy judgments is likely the result of a ceiling effect: Participants’ judgments for the psychotherapy items were between 6.5 and 7 on a nine-point scale, which may be at ceiling given people’s moderate perceptions of the degree to which psychotherapy has the potential to be effective Jorm, . In contExperiments 1–3 showed that biological and psychological attributions shift depending on abstract versus concrete framing not only for clinicians .Forty-nine lay participants were recruited via Amazon Mechanical Turk, of whom ten were excluded . A separate group of 18 participants judged whether the concrete version was “a good example” of the abstract version on a scale of 1–9 , yielding a mean rating of 7.99 (SD = 0.23). Mean ratings by behavior were all at least 7.33.As for Experiment 1, we conducted a pretest of these items to determine whether the abstract and concrete versions of each behavior were perceived to correspond to each other as intended. We recruited a separate group of 41 participants for this pretest, of whom five were excluded for failing the attention check. Of the remaining 36 pretest participants, 18 judged whether the abstract version was “a good abstract description” of the concrete version on a scale of 1–9 , yielding a mean rating of 7.61  = 33.95, p < 0.001, ηp2 = 0.47, as shown in Fig. M = 4.81, SD = 1.22) than for the concrete versions , t(38) = −4.36, p < 0.001, d = −0.70, while psychological attributions were higher for the concrete versions  than for the abstract versions , t(38) = 4.84, p < 0.001, d = 0.78.Each participant’s biological  and psychological (α = 0.85) attributions were averaged separately. We conducted a 2 (concrete or abstract) × 2  repeated measures ANOVA on each participant’s mean across items. This analysis revealed the predicted interaction, p < 0.05 level (two-tailed t-test against 0), one biological factor reached marginal significance , and two psychological factors did not significantly differ from 0 .As shown in Fig. F = 16.62, p = 0.005, ηp2 = 0.70. Biological attributions were higher for the abstract versions  than for the concrete versions , t(7) = −4.27, p = 0.004, d = −1.51, while psychological attributions were higher for the concrete versions  than for the abstract versions , t(7) = 2.65, p = 0.033, d = 0.94.The interaction effect also held up in a by-item analysis, using each item’s mean score across participants. A 2 (abstract or concrete) × 2  repeated measures ANOVA on these scores revealed the predicted interaction, Experiment 4 found that shifts in attribution occur not only for mental disorders, but for a much broader range of human behaviors. These shifts were consistent across the three biological measures , but somewhat more variable across the psychological measures. Although four of our psychological measures shifted significantly in the predicted direction, two others—cognitive factors and personality—did not.Since all psychological measures shifted significantly in Experiments 1 and 2 depending on framing, it is worth considering why shifts were not seen for cognitive factors and personality in Experiment 4. We speculate that these somewhat less consistent effects of psychological attributions may be due in part to a weaker manipulation of abstractness that we used in Experiment 4, compared to Experiments 1–3. Whereas those previous experiments described the behaviors at the level of a category  that did not invoke any individuals, Experiment 4 described the behaviors in terms of a group of individuals engaging in the behavior. Because even the abstract versions referred to human agents, they might have somewhat triggered psychological explanations. Furthermore, people may consider cognitive factors (e.g. beliefs and intelligence) and personality to be more immutable than the other, more transient psychological factors we tested, such as emotions and intentions. Nonetheless, significant shifts were still obtained for a majority of our measures of psychological attribution—and all measures of biological attribution —testifying to the robustness of the attributional shifts in the face of this weaker manipulation.Experiment 5 sought to replicate the framing effects on attributions for ordinary behaviors, using a between-subjects design as in Experiment 2.N = 9 due to taking similar studies in the past and N = 12 due to random responses on filler items). Thus, data from 219 participants were used for the analyses.Two hundred and forty participants were recruited via Amazon Mechanical Turk, of whom 21 were excluded (N = 36), the Psychological Set 1 (N = 145), or the Psychological Set 2 (N = 38) questions. Sample sizes were determined by power analyses on the data from Experiment 4, with 95% power subject to a minimum of 40 participants per condition (prior to excluding random responders and repeat participants).The stimulus materials were the same as in Experiment 4. The design and the procedure were the same as in Experiment 2 in that participants received either the Biological  = 51.15, p < 0.001, ηp2 = 0.31, as shown in Fig. M = 5.29, SD = 1.11) than for the concrete versions , t(35) = −6.81, p < 0.001, d = −1.13, whereas the responses to the Psychological Set 2 questions were higher for the concrete  than for the abstract versions , t(37) = 5.16, p < 0.001, d = 0.84. The responses to the Psychological Set 1 questions did not differ between the concrete and abstract versions , t(144) = 1.18, p = 0.24, d = 0.10, because cognitive abilities and personality—the two psychological measures that did not reach significance in Experiment 1—were unaffected by the manipulation.  as a within-subjects factor and attribution  as a between-subjects factor. This analysis revealed the predicted interaction, F = 38.80, p < 0.001, ηp2 = 0.85. Biological attributions were higher for the abstract versions  than for the concrete versions , t(7) = −5.33, p = 0.001, d = −1.88, while psychological attributions were marginally higher for the concrete versions  than for the abstract versions , t(7) = 2.15, p = 0.069, d = 0.76. Follow-up analyses conducted separately on the two sets of psychological measures showed that this marginally significant effect on psychological attributions occurred because concrete items were rated significantly higher than abstract items on the Psychological Set 2 measures , t(7) = 2.49, p = 0.041, d = 0.88, while the concrete and abstract items were rated similarly on the Psychological Set 1 measures , t(7) = 0.59, p = 0.57, d = 0.21.The interaction effect also held up in a by-item analysis. A 2 (abstract or concrete) × 2  repeated measures ANOVA on the item means revealed a significant interaction, In sum, the results of Experiment 5 fully replicate the findings of Experiment 4, where biological attributions were consistently stronger in the abstract and psychological attributions were typically stronger in the concrete (with two of six measures failing to reach significance). Finding these same effects in a between-subjects design shows that the framing shifts cannot be due to a perceived demand to rate the psychological and biological explanations inversely.In daily life, people often describe behaviors at differing levels of abstraction—as abstract generalizations across individuals or as concrete behaviors of individuals. We hypothesized that this distinction between abstract and concrete framing would lead to different explanatory preferences; namely, a stronger preference for biological explanations in the abstract and more reluctance to accept biological explanations for concrete cases.The results across Experiments 1, 2, 4, and 5 corroborated this hypothesis. Both in contemplating disordered (Experiments 1 and 2) and everyday behaviors (Experiments 4 and 5), participants generally judged explanations in terms of genetics, neural chemistry, and brain structure to be more appropriate when faced with abstract descriptions of behavior than when faced with concrete cases. These differing explanatory stances also had downstream consequences such that people preferred a more “biological” intervention (medication) for treating disorders when described abstractly than when described in terms of a concrete case (Experiment 3).It should also be noted that our claims are only about whether endorsement of biological and psychological explanations was influenced by abstract descriptions relative to concrete descriptions. Thus, we are not claiming that abstract framing would increase endorsement of biological explanations to the extent that they would be preferred to psychological explanations. In fact, this was not the case in Experiments 3–5. Similarly, we are not claiming that concrete framing would make psychological explanations be endorsed more than biological explanations; again, the current results failed to show that consistently (Experiments 1 and 2). Preferences for biological versus psychological explanations can vary greatly simply due to the nature of the events. For instance, “Don is full of himself” would be difficult to explain in terms of biological factors and thus although an abstract framing like “Some people are full of themselves” may make biological accounts more plausible, psychological accounts may still be more dominant than biological accounts even in the abstract framing.In addition, we acknowledge that other factors may influence the availability of biological versus psychological explanations, including individual differences in theory of mind  = −0.38, p = 0.013, and failed to reach significance in Experiment 3, r(37) = 0.34, p = 0.16. Taken together, these findings speak against the possibility that those who are more likely to endorse mind–body dualism are more likely to be subject to the abstract/concrete framing effect. Nonetheless, these null results should be taken with caution, in part because the dualism scale may have become a less valid measure of dualist beliefs in recent years. That is, the pervasiveness of biological accounts of human behaviors may have made laypeople deny mind–body dualism when confronted explicitly, as is the case in the dualism scale. Future research, using more implicit measures of dualism, can help us better understand the shape and the scope of dualist beliefs that laypeople hold.That said, our results do not present any direct demonstrations of Cartesian dualism, the claim that mind and body are distinct substances. We collected participants’ dualists beliefs at the end of Experiments 1 and 4, presenting them with the dualism scale from Stanovich , and fouPeople hold lay theories across many domains that differ dramatically from more formal scientific theories, including theories in biology (Shtulman, Very little is known, however, about what circumstances lead individuals to apply their formal versus intuitive theories to a problem when these theories disagree. We speculate that people may be more likely to rely on their formal theories in the abstract and more likely to default to their earlier, intuitive theories in the concrete. This idea can provide a further mechanism for the current findings. Whereas folk psychology is a natural and early-emerging mode of explanation (e.g. Gergely & Csibra, One way to test the formal education hypothesis would be to ask whether an analogous effect arises in other domains. Would people apply different lay economic theories in contemplating one individual country versus countries in general? Would people apply different lay theories of evolution in contemplating one particular species versus species in general? Would people give different advice about how to maximize happiness if the advice is applied to a particular person versus people in general? To the extent that formal and intuitive theories may give different verdicts, these questions may be of considerable practical importance.A second way to test the hypothesis would be to conduct developmental studies. Presumably, young children do not have a formal education in biology or neuroscience, so if the effect is indeed driven by formal education, it should not arise among young children. By contrast, if the effect is driven by an intuition that biological explanations are tied to immutability and hence essentialism, it might arise much earlier in development. For instance, Cimpian and Markman  found thWe found that, like clinicians (Kim et al., Our finding also has implications for science education more broadly. Science educators have long debated the relative value of abstract and concrete teaching materials (see Fyfe, McNeil, Son, & Goldstone, We explain human behaviors in multiple ways. We can emphasize the importance of responsibility, controllability, intentions, beliefs, and desires. We can also explain human behavior in terms of biological forces, such as genes, neural chemistry, and brain structure. Our results showed that biological theories of behavior are more privileged when contemplating abstract descriptions rather than concrete cases. Thus, even though abstract and concrete descriptions of behavior are both ubiquitous in the world, and often seemingly equivalent, they can nonetheless lead to very different inferences about the causes underlying the behavior.