Low Birth Weight (LBW) is one of the major risk factor for death in early life. However, little is known about predictors of LBW in sub-Saharan Africa. Therefore, the aim of this study was to measure the incidence and determinants of LBW in a rural population of Ethiopia.An observational cohort study on pregnant women was conducted from December 2009 to November 2010. During the study period 1295 live birth were registered and the weights of 956 children were measured within 24 hours after birth. Socio-demographic, economic, maternal and organizational factors were considered as a predicators of LBW, defined as birth weight below 2500g. Logistic regression was used to analyze the data, odds ratio (OR) and confidence intervals (CI) are reported.The incidence of LBW was 28.3%. It is significantly associated with poverty , maternal Mid Upper Arm Circumference (MUAC) less than 23 cm , not attending ANC , mother’s experience of physical violence during pregnancy , and longer time to walk to health facility .The incidence of LBW was high in Kersa. Babies born to women who were poor, undernourished, experienced physical violence during pregnancy and who had poor access to health services were more likely to be LBW in this part of the country.In this largely poor community where ANC coverage is low, to reduce the incidence of LBW, it is essential to improve access for maternal health care. The involvement of husbands and the community at large to seek collective action on LBW is essential. Low Birth Weight (LBW) is one of the risk factors for child morbidity and mortality Though its extent varies from place to place, LBW is a common phenomenon in many parts of the world. For example, in North America, its prevalence is less than 6% In Harare teaching hospital, Zimbabwe, per 1000 live births the annual incidence of LBW was 199 LBW is routinely associated with several maternal factors such as younger and older age, rural residence, low family economic status and illiteracy Women, who have three or more children, who are not living with the father of the new born, who have multiple gestations, who have unintended pregnancy and who have a history of delivering LBW have an increased risk of delivering LBW baby Similarly pregnancy co-morbidities such as existence of hypertension, anemia, and poor maternal weight gain during the index pregnancy increases the risk of having LBW baby th day after delivery. The measurement obtained on the 7th day is different from the measurement that would be obtained on date of birth. Therefore, they are not true reflection of birth weight In Ethiopia literature on LBW is scarce. The study conducted in Addis Ababa used retrospective data from a tertiary teaching hospital. Clients who visited such hospitals are usually referred by other hospitals. Even then it is only those who can afford to stay in Addis Ababa accesses the service. Therefore, they are different from those who give birth in the usual clinical setting. The study conducted around Jimma used weight measured up to the 7On the other hand, the DHS results were based on the verbal report of the baby’s weight, and active baby weighting was not done. In addition, DHS looked for incidents of LBW for babies born in the last five years preceding the survey Thus, it was found useful to conduct a study in rural settings where 84% of the Ethiopian population lives. The aim of this study was to identify the incidence and determinants of LBW in the rural community.Identifying determinates of LBW and preventing them helps in reducing early childhood morbidity and mortality resulting from LBW. Therefore, addressing factors affecting the birth weight of babies will contribute to the achievement of the Millennium Development Goal (MDG) number 4. The knowledge gained from this study can be applied to the other parts of the country and beyond with similar characteristics.The study was conducted in Kersa Demographic Surveillance and Health Research Center (KDS-HRC) field site, Ethiopia from December 2009 to November 2010. The study site has two urban (Kersa and Weter towns) and 10 rural Kebeles . According to the first census in 2007, the study site has 48, 192 inhabitants in 10, 256 households.The health service in the district is provided by six health centers, 28 health posts and health extension workers posted in all the Kebeles. According to the health office in the district, the health coverage in 2011 is 80% The study was approved by the Ethiopian National Ethical Clearance Board. The mothers of the babies enrolled in the study were informed about the study, the nature of the study, its objectives, the expected outcomes, and the benefits and risks associated with the study. A written informed consent was obtained from the mothers.Participants of the study were part of the pregnancy surveillance initiated in KDS-HRC Salter’s hang up scale to the nearest 100 g following standard techniques The newborns were weighted naked or in minimal clothing using a In addition, using a questionnaire adapted from relevant research reports Data were entered using Epi data version 2.2 and analyzed using STATA software version 11 Birth weights of babies less than 2500g was labeled as LBW and 2500g and above was considered normal weight During analysis, age was grouped using 10 year interval; maternal education was grouped as illiterate for those who can’t read and write and all others are labeled as literate. The ten rural Kebeles are labeled as rural and the two small towns (Kersa and Weter) are labeled as rural towns.To determine the economic status of families, wealth index was used. The wealth distribution was generated by applying principal components analysis to 33 household variables Travel time to the nearest health facility was used to assess access to health care and it was based on the women’s estimate. Later during analysis, the reports were grouped into less than 40, 40–79 and 80+ minutes. The grouping was made based on local people’s reference that “a woman can walk an estimated distance of 5 km in 40 minutes.”To proxy maternal nutritional status MUAC was used. A MUAC of less than 23cm was considered to be a sign of poor nutrition. MUAC doesn’t vary much during pregnancy and is therefore an appropriate measure of nutritional status than BMI or weight Fertility desire was recorded based the report of the women. All mistimed and unwanted pregnancies were labeled as unintended and otherwise intended. ANC attendance was recorded using woman’s report or report plus ANC card. Women who reported to visit at least once were labeled as ANC attended and those reported not visiting health institution for ANC attendance were labeled as not attended.Before entering the variables in to regression models, missing birth weight was analyzed. Missing birth weight was 26.2%. It was significantly associated with women from poor family, women with MUAC of 23+ cm; experience of physical violence during pregnancy and travel time to the nearest health facility less than 40 minutes. This suggests missed observations were significantly different from those allowed their babies to be measured. Therefore, to curb the problem of missing to the outcome variable (Birth Weight) sample weighting was used for those who allowed their children to be weighted. The purpose of sample weighting was to improve representativeness of the sample in terms of characteristics of the study population Finally data were analyzed using four separate regression models. The first three models were used to see the relative effect of variables each other in block before entering them in the final model. In model one socio demographic and economic variable were entered. In model two parity and maternal nutritional status were entered. In model three variables relating to the index pregnancy were entered. In the final model all variables were entered together. Statistical level of significance was set at alpha 5%.A total of 1295 live births were registered and of these the weights of only 956 babies were measured. This gives a retention rate of 73.8%. The remaining babies were not weighted because the mothers did not let their babies weighted for cultural and traditional reasons. LBW was detected among 271 babies .More LBWs were observed among women whose ages were less than 20 and more than 40, who were illiterate, who were rural resident and who were poorer than their counterparts. LBW was significantly associated with rural residence and poor wealth status (p<0.05 for both) .Compared to parity of 1–4 and 5–6; primipara and parity of 7+ had more LBW babies. Women who had MUAC of less than 23 cm had more LBW babies than those who had MUAC of 23 cm and more. Unadjusted OR also showed a statistically significant association between LBW and MUAC of less than 23 cm .More LBW babies were observed among women who never followed antenatal care than those who attended one or more times. Experience of physical violence during the index pregnancy increased the risk of having LBW, compared to those who experience sexual violence or no violence. Women who reported estimated time to walk to the nearest health facility 40–79 minutes had more LBW babies than those who reported less than 40 minute and more than 80 minutes. Significant associations between LBW and antenatal follow-up, physical violence and 40–79 minutes to walk to health facility were detected .In the first block logistic regression model, LBW was significantly high among rural residence and poor,  and , respectively. In the second model, LBW was significantly associated with MUAC less than 23 cm .In the third model, LBW was significantly associated with not attending ANC and experience of physical violence during the index pregnancy,  and , respectively .In the final mode, LBW was significantly associated to the poor , MUAC less than 23 cm , not attending ANC , experience of physical violence  and time to walk to health facility 40–79 minutes  .In this study the incidence of LBW was 28. 3%. It was found that poor economy, MUAC less than 23 cm, not attending ANC, experience of physical violence and time to walk to health facility 40–79 minutes were strong predictors of LBW.Obtaining birth weight data from the community setting within 24 hour is the major strength of the study. In many other researches, data on birth weight have not been collected within 24 hours due to the difficulty in acquiring information on delivery immediately after it happened In assessing access to health care, time to walk to health facility was used. It was obtained by asking respondents to estimate the time they need to walk to the nearest health facility. In this study most of the respondents were illiterate, and their capacity to estimate time against distance may not be as accurate as the estimate that can be obtained from literate person.In this study gestational age of the babies were not calculated. The main reason for this was majority of the women were unable to tell their last menstrual period. Therefore, it was not possible to control gestational age in the analysis.th day after birth, which means getting the correct birth weight is unlikely The incidence of LBW in this study was higher than studies conducted in Zimbabwe and Ethiopia. The Harare teaching hospital study revealed an incidence of 199 per thousand live births, whereas the study conducted in Black Lion hospital of Ethiopia showed prevalence of 8.8% Poor wealth status was one of the strong predictors LBW. Similar findings were observed in other studies conducted in developing countries, including Sub-Sahara Africa. Failure to achieve adequate weight gain during pregnancy as a result of poor feeding pattern eventually affects the birth weight of the newborn. Reduction in the ability to seek medical care, travel for referrals and purchase of food/hygiene items are directly related to poverty, which indirectly affect the birth weight of the baby The nutritional status of women as proxy by MUAC was also a strong predictor of LBW. Other studies conducted in African countries also reached on similar conclusion Not attending ANC showed a significant association with LBW. Other studies had also identified similar finding Experience of physical violence during pregnancy was found to predict LBW. Violence creates stress and psychological disturbance that affects maternal nutritional balance As time to walk to health facility increased more than forty minutes, the occurrence of low birth weight increased by 63%. In this study, more than 80 minute was not observed to increase low birth weight. This may be explained by the fewer sample size in that category as evidenced by wide confidence interval.The percentage of newborns delivered having LBW was extremely high in this study. The significant predictors of LBW were being poor, not attending ANC, MUAC less than 23 cm, experience of physical violence during the index pregnancy and longer time to walk to health facility. This is significant because LBW is highly associated with neonatal and infant morbidity and mortality.Improving access to health care and improving the nutritional status of women particularity pregnant women would reduce the occurrence of LBW.In Ethiopia, all rural Kebeles are staffed with health extension workers; these health workers are expected to provide house to house health promotion services. In this regard, they should emphasize providing ANC for the pregnant women. Through ANC, it is possible to identity a women who are at risk of having LBW baby. Giving necessary advice such as the need for balanced diet and provision of health promotion intervention such as provision of iron tables for the anemic can be easily facilitated.In addition, empowering women, families and neighborhoods to work on their own health care such as peer discussion to use ANC, would reduce the number of babies borne having LBW. These activities would prevent not only the occurrence of LBW but also problems associated with LBW such as child morbidity and most importantly child mortality thereby helps the country achieve child MDG targets.


Children with attention-deficit/hyperactivity disorder (ADHD) are two to three times more likely to experience sleep problems. The purpose of this study is to determine the relative contributions of circadian preferences and behavioral problems to sleep onset problems experienced by children with ADHD and to test for a moderation effect of ADHD diagnosis on the impact of circadian preferences and externalizing problems on sleep onset problems.After initial screening, parents of children meeting inclusion criteria documented child bedtime over 4 nights, using a sleep log, and completed questionnaires regarding sleep, ADHD and demographics to assess bedtime routine prior to PSG. On the fifth night of the study, sleep was recorded via ambulatory assessment of sleep architecture in the child’s natural sleep environment employing portable polysomnography equipment. Seventy-five children (26 with ADHD and 49 controls) aged 7–11 years  participated in the present study.In both groups of children, externalizing problems yielded significant independent contributions to the explained variance in parental reports of bedtime resistance, whereas an evening circadian tendency contributed both to parental reports of sleep onset delay and to PSG-measured sleep-onset latency. No significant interaction effect of behavioral/circadian tendency with ADHD status was evident.Sleep onset problems in ADHD are related to different etiologies that might require different interventional strategies and can be distinguished using the parental reports on the CSHQ. Attention-deficit/hyperactivity disorder (ADHD) is characterized by impulsivity/hyperactivity and inattention, with symptom onset before 7 years of age and impaired functioning in two or more environmental settings. ADHD isSleep problems presenting in a child with ADHD may stress both the child and family. Furthermore, sleep disturbances can cause excessive daytime fatigue and interfere with several aspects of an individual’s daytime functioning, including mood, attention span, and behavior,13, whicDespite the clinical and scientific relevance of sleep problems to the understanding and management of ADHD, the etiologies of such sleep issues remain unclear. This is particularly important for an accurate evaluation of possible causes related to parental descriptions of sleep onset problems in children with ADHD and in choosing the most effective intervention.Amongst the potential etiologies for sleep onset problems, both behavioral and biological explanations have been advanced within the current literature. The behavioral model posits that prolonged sleep onset and delayed bedtime in children with ADHD are a result of behavioral problems, leading to increased resistance at bedtime and difficulty in settling down in the evening. According to this model, an intervention should target the behavioral problems that lead to delayed bedtime and sleep deprivation in children with ADHD. In contrast, the circadian model suggests that children with ADHD are averse to bedtime because they are sent to bed before feeling the need to sleep. In particular, it is suggested that a delay in the endogenous circadian rhythm delays sleep and waking times. Such diIn addition, it is also unclear if such problems are unique to children with ADHD. It has been suggested that children with ADHD may have a deficit in arousal regulation and further factors associated with sleep or bedtime behavior might, thus, be specific to this population. However, to date, this hypothesis has not been tested. As such, the objectives of this study were two-fold: The first goal was to determine the relative contributions of circadian preferences and behavioral problems in relation to sleep onset issues, specifically – sleep onset insomnia and bedtime resistance – experienced by children with ADHD and in controls. Since inconsistencies in the detection of sleep problems using PSG and parental reports have been documented, we examined and compared both sources of information. The second objective of this study was to test for a moderation effect of diagnosis (ADHD vs. control) on the impact of circadian preferences and externalizing problems on sleep problems. We hypothesized that 1) stronger evening circadian preferences would be associated with longer SOI in both children with ADHD and controls; 2) a greater level of externalizing problems would be associated with elevated bedtime resistance in both groups; and, 3) that a stronger impact of circadian evening preferences and behavioral problems on bedtime behavior would be evident in the ADHD group compared to controls.Participants were recruited from elementary schools in Montreal and surrounding regions. Children with ADHD and controls were recruited by flyers and advertisements through local schools, psychologists, and centers aimed at helping children with ADHD such as the Montreal Children’s Hospital and the Douglas Mental Health University Institute’s ADHD outpatient clinic. The study was approved by the Douglas Mental Health University Institute’s Research Ethics Board. Parents signed informed consent forms and all of the children assented to participation in the study.Data collection occurred in a quiet location within participants’ home environments.Eighty children were screened and invited to participate in the study. Five of these children received a total score above a predetermined cutoff score on initial screening and were subsequently excluded because of the possible confound of SDB (2) or RLS (3), leaving a final study group of 26. Hence, a total of 75 children , aged 7–11 years , participated in the study.i.e., giving positive answers to 33% or more of the 22 questions of either scale) were excluded from the study. We subsequently excluded all children with a symptom complex suggestive of these disorders.The diagnosis of ADHD was determined by criteria from DSM-IV, using the Diagnostic Interview Schedule for Children (DISC-IV); which wSeventeen (65.4%) of the children with ADHD had Combined subtype ADHD, eight (30.8%) were Inattentive subtype, and one (3.8%) was Hyperactive/Impulsive subtype. As well, 33.8% of the children with ADHD had used medication for ADHD symptoms , Concerta (4), or Strattera (1)). Eight (30.8%) of the 26 patients with ADHD received a comorbid diagnosis of oppositional defiant disorder, and 2 (7.7%) received a comorbid diagnosis of conduct disorder. None of the patients with ADHD elicited symptoms suggestive of comorbid diagnosis of depression, anxiety or any other disorder which may interfere with sleep.For study design please see FigureAfter initial screening, eligible children were invited to participate in the study and their parents received a package that included a sleep assessment battery, standard ADHD assessment questionnaires, a demographic questionnaire, and a consent form. Parents were asked to complete and return all questionnaires. The sleep assessment battery sought to provide comprehensive baseline data on sleep-related patterns, behavior, and preferences in children with ADHD, and included the Children’s Sleep Habits Questionnaire (CSHQ), the chie.g., chocolate or cola) for at least 48 hours before evaluation. The “wash-out” period of 48 hours was chosen because the half-life of the stimulant methylphenidate is, on average, 2.4 hours when either of the two dosage forms (long- or short-acting) is employed = 2.32; p < 0.05). Univariate analyses indicated that children with ADHD scored higher on the CSHQ subscales of Sleep Onset Delay, Sleep Duration, Night Waking, Sleep Anxiety, and Daytime Sleepiness. A separate ANCOVA evaluating between-group differences in Total CSHQ scores indicated that children with ADHD also scored higher on this measure.The MANCOVAs conducted to explore between-group differences in reported sleep problems on CSHQ measures revealed a significant main effect of group , suggesting that these two measures, as expected, were examining similar constructs; CBCL externalizing scores were found to be negatively related to age , such that younger children tended to have greater externalizing scores; no significant correlations were found between bedtime resistance and parental reports of sleep-onset delay or PSG-assessed sleep-onset latency or between any of the other predictor variables, suggesting colinearity assumptions were not violated.To explore whether behavioral or circadian tendencies contributed to sleep onset problems in children with ADHD, multiple linear regression analyses were performed on data from 75 participants Table. Inter-cRegression analyses are reported in TableAnalyses revealed that scores on the CMEP contributed above and beyond puberty and age in predicting parental report of sleep onset delay. No interaction effect between diagnostic group and any predictor of outcome measures was evident and no effect was significant, other than for CMEP scores  and biological (circadian) factors previously found to be associated with ADHD symptoms contribute to sleep-onset insomnia and bedtime resistance commonly encountered in children with ADHD. In addition, we tested whether externalizing problems and circadian preferences differentially affect sleep problems in children with ADHD compared to controls. The results showed that each of the two factors  was associated with a different bedtime problem. In both groups of children, externalizing problems yielded significant independent contributions to parental reports of bedtime resistance, whereas an evening circadian tendency contributed both to parental reports of sleep onset delay and to PSG-measured sleep-onset latency. No significant interaction effect of behavioral/circadian tendency with ADHD status was evident.Previous reports found that bedtime struggles and delayed sleep-onset were associated with sleep disorders characteristic of children with ADHD and suggested that what parents perceive to be behavioral problems might sometimes reflect the presence of circadian sleep disorders. Our present findings suggest that parents effectively differentiate between child “unwillingness”  versus “inability” (sleep-onset delay) to fall asleep. We further show that the “mixed bag” of bedtime problems in children with ADHD may have different etiologies.per se that predicted parental descriptions of increased bedtime resistance and struggle. Previous reports have shown that sleep problems and externalizing behaviors often occur together[Consistent with previous reports, we found that children with behavioral problems showed resistance at bedtime and considerable difficulty in settling down in the evening. Howevertogether,45. DefiIn the present study, we found that children with ADHD had a stronger circadian evening preference than did controls. This is in line with previous work showing that the evening increase in endogenous melatonin levels was delayed in medication-free children with ADHD. FurtherConsistent with the results of a number of other studies, we founAlthough no significant between-group difference was found for bedtime resistance, it was noticeable that both groups of children showed levels of resistance previously considered to be characteristic of children with ADHD. Hence, Our findings that bedtime problems associated with ADHD may vary in etiology emphasize the need for careful and accurate diagnosis of the source of sleep problems in children with ADHD. When assessing a child or adolescent with ADHD, it is necessary to distinguish a physiological sleep problem from a behavioral condition of which the sleep problem is but one of the symptoms.To our knowledge, this is the first published study that used CMEP for the assessment of circadian tendencies in children with ADHD when compared with objective PSG measures of sleep. The findings of the present study emphasize the importance of using CMEP scale for the assessment of circadian preference in children with sleep onset problems, especially those with the diagnosis of ADHD.Whereas PSG has been considered to be the “gold standard” for diagnosis of sleep disorders and in recording of sleep, PSG is both costly to implement and burdensome for families. Our present findings suggest that both objective and subjective measures could be used to provide complimentary information regarding potential sleep problems. Whereas parental reporting is a simple, reliable, and inexpensive method for accurately diagnosing sleep onset problems, PSG allows one to obtain objective, physiological information that can be used to examine primary sleep disorders that are common in children with inattention and hyperactivity. Given the high prevalence of ADHD and the elevated rates of associated sleep disorders, our finding that parental reporting is accurate is clinically important, as it can be used as the basis for an accurate and feasible assessment method in a clinical/pediatric setting. Pediatricians should be educated on this topic and provided with the scales for integration into practice.Some limitations of this work are apparent. First, although the participants were below the cutoff score on the Pediatric Sleep Questionnaire, had no indication of desaturation, and no paradoxical breathing at the thoracic and abdominal channels, the presence of sleep-disordered breathing cannot be completely excluded because of the lack of nasal cannula or thermistor. Other limitations include the lack of employment of objective circadian measures, such as salivary melatonin level or core body temperature, to accurately determine the extent of the circadian discord. In addition, approximately one-third of the children with ADHD prescribed stimulants to control symptoms had potentially short wash-out periods. This should be noted, because a transient worsening of symptoms (a “rebound”) might have occurred after medication was stopped. Future studies should include a washout period of at least 1 week, or should examine only medication-naïve children when the impact of sleep practices on reported sleep problems is under consideration.In conclusion, our findings indicate that sleep onset problems in children with ADHD may be related to both behavioral factors and circadian preference. Our data further indicate that CSHQ is a useful clinical tool that should be incorporated into sleep evaluation in children with ADHD. Our findings add to the understanding of specific etiologies underlying sleep disorders and ADHD and contribute to closing the gap between data obtained using subjective or objective sleep measures in ADHD children.The authors declare that they have no competing interests.RG has made substantial contributions to the conception and design of the study in addition to analysis and interpretation of data and writing the manuscript. LF, LB, SW have made substantial contributions to the acquisition of data and revision of the manuscript. AR has made substantial contributions to the analysis of data as a statistical consultant. SF has made substantial contributions to analysis of data. JC has made substantial contributions to the design of the study and interpretation of data. All authors have read and approved the final manuscript.This study was supported by grants to Dr. Gruber from the Canadian Institutes of Health Research  and the Fonds de la recherche en santé . The authors of this manuscript declare no conflicts of interest.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-244X/12/212/prepub


Due to the specific structure of surfactants molecules they are applied in different areas of human activity . After using and discharging from wastewater treatment plants as effluent stream, surface active agents (SAAs) are emitted to various elements of the environment , where they can undergo numerous physic-chemical processes  and freely migrate. Additionally, SAAs present in the environment can be accumulated in living organisms (bioaccumulation), what can have a negative effect on biotic elements of ecosystems . They also cause increaseing solubility of organic pollutants in aqueous phase, their migration, and accumulation in different environmental compartments. Moreover, surfactants found in aerosols can affect formation and development of clouds, which is associated with cooling effect in the atmosphere and climate changes. The environmental fate of SAAs is still unknown and recognition of this problem will contribute to protection of living organisms as well as preservation of quality and balance of various ecosystems. This work contains basic information about surfactants and overview of pollution of different ecosystems caused by them . Surface active agents  are a group of compounds with specific chemical composition of their molecules (one part soluble in polar medium: hydrophilic and second in nonpolar medium: hydrophobic). The main classification of surfactants is based on charge of hydrophilic part of their molecules: cationic, anionic, and nonionic compounds .Occurrence of polar and nonpolar parts in the SAAs molecules gives them special properties against different medium. Surfactants are characterized by tendency to absorb at different types of surfaces or interfaces. Another property of the SAAs is ability to association in solution and formation of micelles . During wetting/waterproofing,de- or foaming,de- or emulsification,dispersion or flocculation of solids particles in liquid phases,solubilization of non-/sparingly soluble reagents in solvents,increase or decrease of viscosity of solution phases .Due to the specific chemical structure of surfactants molecules they are applied in different areas of human activity. During formulation of households or industrial products compounds from the group of surfactants are used because their presence leads to improving efficiency of the following processes:In In 2008 the annual global production of surface active agents was about 13 million metric tons. The expected growth of production of that class of compounds is forecasted as 2.8% annually to 2012. Approximately 65% of total production corresponds to compounds classified as anionic surfactants [After use, compounds from the group of surfactants are emitted to various elements of the environment , where they can undergo numerous physical and chemical processes. Therefore, the specific properties of those chemical compounds cause increasing their mobility and unrestrained circulation in the environment. Those processes might significantly contribute to disrupting the water cycle within various ecosystems; hence, it is essential to obtain answers to what levels of concentrations surfactants are present in the environment. Investigation of environmental fate of surface active agents can help increase level of knowledge about pollutants migration pathways and better protect living organisms or different ecosystems. In recent years the interest of SAA has increased and scientists have started to estimate the possible effects of those compounds on the environmental balance , 6.In this paper, basic information about surface active agents  and their fate after discharging to wastewater treatment plants are mentioned. The brief review of sorption and degradation processes of surfactants in water systems is presented. Such information can be useful during investigation of their presence and behavior in environment. Moreover, the analytical protocols used for determining total concentration of surfactants or individual analytes from particular group of SAA in environmental samples are described. This work contains overview of contamination of different ecosystems caused by surfactants . Possible impact of compounds from the group of SAA on biotic and abiotic elements of the environment  is presented.Surface active agents are one of the most common applied compounds in industrial, agricultural, and household activities and after use a huge number of surfactants (and/or their degradation products) are discarded to wastewater-treatment plants (WWTP). In nonurban areas (there is no WWTP) wastewaters that contained various classes of surface active agents are discarded directly to surface waters and they might be dispersed into different elements of environment. In wastewater treatment plants compounds from group of SAAs are completely or partially removed by a combination of different processes  and their degradation pathways were investigated , 8.Chemical compounds from this group are degraded during secondary treatment processes and under optimized conditions about 90–95% of initial SAAs concentration contained in influent streams can be eliminated (depending on efficiency of WWTPs) , 11. Morμg/L or g/kg) can occur, are discharged into surface waters or used as fertilizer in agricultural areas, respectively. Such practices lead to emission of surfactants and their metabolites into different parts of the environment  [After appropriate processes in WWTP effluents and sewage sludge, in which different types of surfactants or their degradation products  , 12. In If compounds from the group of SAAs are present in water ecosystems, they can also undergo sorption and aerobic/anaerobic degradation processes. Those processes are responsible for elimination and transport of pollutants to water systems. Sorption processes inhibit also degradation of chemical compounds because their bioavailability can be reduced. In 13LAS, NP, and NPE1-2O) were observed in sediment or suspended solid samples. The higher concentrations of more polar compounds  were observed in the dissolved form [more polar anionic SAAs were estimated in the dissolved phases;less polar cationic and nonionic SAAs were estimated in the particulate phases (their transport will be associated with suspended solids) .Generally, the higher concentrations of less polar compounds from the group of surfactants  is the primary transformation taking place in different ecosystems to reduce impact on living organisms. Biodegradation is an important process not only in wastewater treatment plants but also in the environment. During this process microorganisms are able to utilize surfactants as substrates to produce energy and nutrients or cometabolize them by microbial metabolic reactions. There are many factors  that affect the efficiency of biodegradation of compounds from the group of surfactant in the environment. Most of compounds from this group can be rapidly degraded by microorganisms in ecosystems in the presence of oxygen (according to the current legislation), while some of them  may be persistent under anaerobic conditions , 15, 17.4N+ < R3MeN+ < R2Me2N+ < RMe3N+ < Me4N+; Me—methyl group) and substitue them with a benzyl group [Cationic surfactants are aerobic biodegradable compounds that characterized different degradation mechanisms. The degradation pathway for alkyl trimethyl or dimethyl ammonium compounds (TMAC and DMAC) is initialed by N-dealkylation and followed by N-demethylation reaction  . Cationiyl group , 21. Theyl group .ω-oxidation of the alkyl chain terminal carbon, successive β-oxidation, and further desuphonation [Analytes from the group of LAS contained longer alkyl chains and benzene group in external position is more susceptible to biological degradation. Mono- and dicarboxylic sulfophenyl acids (SPC), LAS biodegradation intermediates having an alkyl chain length of 4 to 13, are formed during the following stages: honation , 23–26.β-oxidation pathway) [For example, it can be observed that in surface waters primary degradation  of compounds from the group of LAS is completed after 4 days; the average half-life of some analytes is 10–24 h and 56–90% of mineralization can be finished from 7 days up to 30 days . Compounpathway) . Biodegrpathway) . Removalpathway) , 30. Forpathway) .2C [ω-oxidation and later α, β-oxidation reaction of the alkyl chain [1-2O, and NPEC) [ω-oxidation and later α, β-oxidation reaction of the alkyl chain [ω-oxidation and successive α, β-oxidation reactions of the alkyl chain [Degradation process of nonionic surfactants was investigated mainly using compounds from the group of nonylphenol ethoxylates. Their primary biodegradation is relatively fast (from 4 to 24 days) and their mineralization is typically from 50 to 80% –34. Degr2C –35. Alkynd NPEC) , 38. Degyl chain , 40. Braβ-oxidation reactions [Therefore, surfactants occurred in water systems can be degraded easily  depending on their properties and environmental parameters. Compounds from the group of SAA can undergo such processes like attachment to suspended solids and accumulation in sediments. In conditions with lack of oxygen (starting in depth of few cm) surfactants can be only degraded by anaerobic pathways. Generally, in anaerobic condition processes are slower or they are not observed  and pollutants occurred longer in sediment , 13. Howeactions –43.the complex composition of environmental samples—interfering components which increased or reduced the identified levels of analytes,the low concentrations of individual surfactants in such samples,the diverse chemical structures of surfactants—moreover commercially available surfactants are mixtures containing twenty or more individual compounds,the amphiphilic nature of surfactants ,the limited availability of commercial standard solutions of surfactants .For different reasons, it is important to detect, identify, and monitor levels of surfactants in aquatic environments. Prior analytics of pollutants from the group of SAAs in environmental samples pose new challenges. The searching for new tools and new sources for obtaining information about degree of pollution different environmental compartments is dictated by toxicological considerations, the desire to increase the accuracy of the description of the environment, and the study and protection of aquatic ecosystems balance. It is imperative to have the appropriate analytical tools  in order to monitor the presence of surface active agents in various environmental samples. The determination of SAAs in such samples causes a lot of problems, mainly because of , 6the coThe complex and frequently variable matrix composition of environmental samples and the trace levels of SAAs mean that suitable isolation and/or preconcentration techniques have to be applied at the sample preparation stage. As a consequence of amphiphilic nature of surfactants molecules during preparation stage an internal standard has to be added to the sample before the solvent extraction , which problematic because of lack of available commercial standards. Moreover, the analytical methodologies enabling the determination of a wide range of SAAs present at different levels in environmental samples should be validated against certified reference materials. Nowadays, only liquid reference materials are available on the market; they can be used to validate methodologies for determining total contents of ionic (cationic or anionic) and nonionic surfactants. On the other hand, there are no reference materials suitable for validating entire analytical procedures. Those problems have influence on quality control and quality assurance of measurement results and they might cause difficulties in obtainment of the reliable analytical information , 45. CurThe determination of total concentration of ionic and nonionic surfactants in different types of liquid/solid samples can be carried with use of standard analytical methodologies . Cationic surfactants can be determined as sum of substances that form ion pairs with disulfine blue (DiSB) and isolated with use of appropriated extraction technique. Next, the absorbance of the organic extract is measured spectrophotometrically . Total cFor determining the individual surfactants belonging to different classes of chemical compounds should be applied modification of the available analytical procedures with use of isolation or/and preconcentration techniques at sample preparation stage  and chromatographic techniques coupled with different detection systems  at the final determination stage. Intensive researches in this direction are carried out in a few research centers located only in Spain, Germany, USA, China, and so forth. Gas chromatography (GC) is limited to volatile analytes and this requirement meets only low molecular mass nonionic surfactants (contain low number of ethoxylated groups). This technique is suitable during determination of concentration of other nonionic and anionic after derivatization processes with specific agents. In the literature data not mentioned about application of GC technique to separate cationic SAAs. Presently, high-performance liquid chromatography (HPLC) is the most often used analytical technique during analysis of surface active agents from all classes compounds  in environmental samples. In most cases, derivatization processes of analytes are not necessary, because LC technique is suitable for determining level of low-volatility analytes with large molecules. It gives the possibility of excluding this operation from analytical procedures and simplifies them—due to the Green Analytical Chemistry (GAC) concept. An undoubted advantage of HPLC technique is that surfactants levels can be measured in a very short time. For determining the concentration of individual analytes from group of surfactants in appropriate prepared solvent extracts the following types of detectors can be used: fluorescence (FLD), ultraviolet (UV), conductometric (CD), mass spectrometry (MS) or their combinations , 6. MassAnalysis of the literature data confirmed that surface active agents are presented in various elements of the environment, but researchers focused mainly on determination of levels of anionic and nonionic analytes. Thus, due to the very limited literature data on the determination of cationic SAAs, which are characterized by higher toxicity to living organisms and undergo sorption onto solid surfaces. Additionally, commercial available surfactants are mixtures of various homologues and/or isomers and their determination in environmental samples becomes more problematic. Those aspects indicate the need to develop new methodologies with the use of more selective and specific analytical tools.The ranges of concentrations or mean values of compounds from different groups of surfactants determined in solid and liquid environmental samples are presented in Accordingly, for the first time the total concentration of various groups of SAAs has been determined in atmospheric deposits samples collected during different seasons on Polish territory  , 54.A fraction of surfactant from influent stream can be emitted to the environment via wastewater treatment plant effluent discharge into surface waters. In aquatic ecosystems amount of compounds from the group of SAAs can be reduced by different processes . Cationic surface active agents exhibit specific properties that may prevent (or retard) the growth or cause mortality of microorganisms. These properties allows for appling them as disinfectants and antiseptic agents in different products. On the other hand, the occurrence of cationic surfactants in aquatic ecosystems is very dangereous for aquatic organisms or, in case of humans, may cause irritation or burns to the skin, eyes and respiratory system , 56. AniSome compounds from the group of surfactants have a low biodegradability or their degradation products are more toxic than initial compounds or even become endocrine disruptor compounds (EDC). The toxicity properties of compounds from the group of surfactants can be used to estimate their environmental risks. In p-n-alkylphenol were published. Further research data shows that compounds from OP, NP, NPnEO , and NP1EC have estrogenic activities against different living organisms [1EC, NP2EC, and NP2EO) were observed positive results [Chemical compounds (EDC) that occurred in the environment can present disruption properties of normal functioning of hormonal system in living organism . In 1938rganisms –67. In i results .In previous section of article information about sorption and degradation processes of surfactants in wastewater treatment plants or in the environment is presented. Herein, other important aspects of impact of compounds from the group of surfactants on different part of abiotic environment will be reviewed. So far, an atmospheric input of compounds from the group of surfactants to aquatic environment has not been considered yet. However, evaporation processes of semivolatile compounds (anionic and nonionic surfactants or product of their degradation) from surface waters, soils and vegetation have been recognized as a significant source for such contaminants in the atmosphere .Compounds from the group of SAAs are able to interact on processes that occurred at different interfaces. They can also have influence on processes between air and water interfaces . Rain droplets contain surfactants monolayer is one of the example mentioned type of interfaces. But processes in this area are still under investigation. Nowadays, scientists believe that there are two mechanisms of surfactant amount reduction on a water surface. First, rain drop can split into many small drops covered with surface active agents. These drops can be transported away from the surface of water by moving air masses. Secondly, bubbles can be formed during rain drop impacts and they will move to water surface, burst, and lead to transporting pollutants to the air –74.Moreover, the occurrence of surfactants in abiotic environment might disturb equilibrium of different compartments. Surfactants are able to form films on aqueous surfaces decreasing the surface tension. It potentially hinders water evaporation and gaseous transportation across the aqueous interface. Those compounds can also increase the solubility of organic compounds in the aqueous phase (increasing mobility of toxic agents in different ecosystems). In the environment this specific system can be observed as a thin boundary between the water basin  and the atmosphere, named as sea-surface microlayer (SML). The formation of SML is complicated and it is unknown which physical and chemical processes have influence on migration of chemical compounds in this boundary. For several times occurrence of surface active agents in SML has been proved. Research in this area has shown that SAAs (consist of low-molecular-weight carbonyl compounds) can be also generated in microlayer by microorganisms. In surface water those compounds are produced photochemically from the degradation of refractory dissolved organic matter  and they are taken up quickly by microorganisms –77.Accumulation of surfactants at surface of water can be toxic for marine and freshwater organisms. Some of compounds from the group of surfactants can lead to chronic toxicity or estrogenic responses towards aquatic species , 78–82. The early studies of surfactants in rain water and atmospheric aerosols found that their concentrations were too low to have any effect upon the cloud physical process at high dilution . More coAs a result of the intensification of certain types of human activity an upward trend is observed in the content of surfactants in the environmental compartments. Due to their widespread use and freely migration between phases, surfactants and products of their degradation have been detected at various concentrations in different part of abiotic and biotic environment. The occurrence of surface active agents was confirmed in atmospheric precipitation and deposits, surface waters, sediments, soils, and living organisms. Compounds from the group of surfactants have been detected in samples  from the areas of residence and economic exploitation by humans and from remote regions like the Antarctic as well. Antarctic ecosystems are sensitive to anthropogenic modifications and highly susceptible to human impact. The global source refers to long-range atmospheric transportation of pollutants  from lower latitude area, but their transport pathways are not well understood .Thus, it is widely accepted that SAAs play an important role of anthropogenic pollutant emission, having versatile environmental consequences. On the other hand, review of the literature also shows that we are far from understanding the environmental fate of surfactants. There is lack of data on composition, properties, and behavior of the organic material  in the atmosphere or other environmental compartments , 92. TheCompounds from the group of surfactants are widely applied in different areas of human activities and after use they are discharging to wastewater treatment plant. After appropriated processes , they can be emitted to surface waters with effluent streams. In aquatic ecosystems surfactants and their degradation products undergo similar processes as in WWTPs. Compounds from the group of SAAs can also affect living organisms and abiotic parts of the environment. They can be toxic for different types of organisms or disturb their endocrine balance. Moreover, they can interact with different interfaces  and change natural processes in those systems.During last decades in environmental samples different types of surfactant were determined using analytical protocols depending on what information is required . But it is still imperative to develop new analytical procedure for investigation analytes from the group of SAAs that occurred in ecosystems to make them easier, less cost-consuming, and more safe for biotic and abiotic elements of the environment. Research data confirmed that they are able to spread between waters, soils, atmosphere, and living organisms from different geographic regions. But we are far from understanding migration pathways and their behavior of those pollutants, their impact on different ecosystems and living organisms. There is a need to investigate those processes in environment to protect abiotic and biotic part of the environment.


Inattentiveness, impulsivity and hyperactivity are the primary behaviors associated with attention-deficit hyperactivity disorder (ADHD). Previous studies showed that peripheral blood gene expression signatures can mirror central nervous system disease. Tourette syndrome (TS) is associated with inattention (IA) and hyperactivity/impulsivity (HI) symptoms over 50% of the time. This study determined if gene expression in blood correlated significantly with IA and/or HI rating scale scores in participants with TS.RNA was isolated from the blood of 21 participants with TS, and gene expression measured on Affymetrix human U133 Plus 2.0 arrays. To identify the genes that correlated with Conners’ Parents Ratings of IA and HI ratings of symptoms, an analysis of covariance (ANCOVA) was performed, controlling for age, gender and batch.P<0.05, |Partial correlation (rp)|>0.4). Immune, catecholamine and other neurotransmitter pathways were associated with IA and HI behaviors. A number of the identified genes (n=27) have previously been reported in ADHD genetic studies. Many more genes correlated with either IA or HI scales alone compared to those that correlated with both IA and HI scales.There were 1201 gene probesets that correlated with IA scales, 1625 that correlated with HI scales, and 262 that correlated with both IA and HI scale scores (These findings support the concept that the pathophysiology of ADHD and/or its subtypes in TS may involve the interaction of multiple genes. These preliminary data also suggest gene expression may be useful for studying IA and HI symptoms that relate to ADHD in TS and perhaps non-TS participants. These results will need to be confirmed in future studies. Inattentiveness, impulsivity and hyperactivity are common behaviors seen in children. When pronounced, these behaviors may lead to the diagnosis of attention-deficit hyperactivity disorder (ADHD) . ADHD isTourette syndrome (TS), characterized by motor and vocal tics, is often associated with ADHD symptoms. TS is a heritable, complex genetic disorder where multiple genes, each with a modest effect, are postulated to interact with unknown environmental factors to produce the phenotype ,6. PatieRecent studies suggest that ADHD symptoms might best be considered as continuous quantitative traits rather than diagnostic categories . This haThus, the current study quantified IA and HI behaviors using the Conners’ Parent Rating Scales-Revised (CPRS-R) in a group of participants with TS. The well-validated Conners’ scale is widely used in research and clinical practice to diagnose ADHD and evaluate treatment effects in the disorder . Gene exAll participants with TS were recruited via the Tourette Syndrome Association, clinical referrals, local advertisements, physician referrals, and through the University of California at Davis. The participants were recruited as part of a functional magnetic resonance imaging study of tic severity and cognitive control conducted by Dr. S. Bunge and colleagues . All of Blood sample collection and RNA isolation were performed as described previously . Whole bA260/A280 absorbance ratio greater than 2.0 and a 28s/18s rRNA ratio equal to or exceeding 1.8 was utilized.Total RNA was isolated using the PAXgene Blood RNA Kit (Qiagen) according to the manufacturer’s protocol. RNA quality was assessed using the Agilent 2100 Bioanalyzer  and quantified using fiberoptic spectrophotometry . RNA yielding both an Human Genome U133 Plus 2.0 microarray processing was performed according to the manufacturer’s protocol. The Ovation RNA Amplification System V2 kit and the Ovation® WB Reagent kit  were used to optimize whole blood amplification starting with 50 ng total RNA, the amplified cDNA was fragmented and labeled using NuGEN’s FL-Ovation™ cDNA Biotin Module V2 . Hybridization, washing and scanning were performed according to the Affymetrix Human U133 Plus 2.0 protocols .2-transformation and median polish probe set summarization. Principal Components Analysis (PCA) was employed to detect outliers because outliers can have a profound influence on correlation coefficients.We deposited the raw data at GEO under accession number GSE30470 and can confirm all details are MIAME (Minimum Information About a Microarray Experiment) compliant. Raw data (Affymetrix.CEL files) were imported into Partek Genomics Suite 6.4 . Probe summarization and probe-set normalization were performed using Robust Multi-Chip Average (RMA), which included background correction, quantile normalization, logP<0.05 and |rp| >0.4, supplemented with a pathway and network over-representation approach. Ingenuity Pathways Analysis  was used to identify statistically significant functional categories in the data set using a modified Fisher Exact test, with P<0.05 considered significant. To further support the pathway-related ADHD genes, those involved in the significant pathways were subjected to a co-expression analysis by first performing gene-gene correlation in Partek, and then hierarchical clustering based on gene-gene correlation coefficients by Genesis (Gene Expression Similarity Investigation Suit) software. Chromosome over-representation was identified using the NIAID/NIH DAVID Bioinformatics Resources (http://david.abcc.ncifcrf.gov).To identify the genes that correlated with the CPRS-R inattention or hyperactivity/impulsivity scales, an analysis of covariance (ANCOVA) was performed, controlling for the effects of age, gender and batch (random effect). Of the ~54,000 probesets on the Affymetrix U133 plus 2.0 array, about 36,000 were analyzed after filtering out the probesets targeting non-annotated transcripts, opening reading frames and hypothetical genes. No probesets met the high stringency of a false-discovery correction for multiple comparisons. Thus, we initially considered a main effect of probesets meeting criteria P=0.13 for HI, P=0.48 for IA). All of the participants were medication naive as per parental reports, except for two participants who had previously taken atomoxetine  to treat ADHD symptoms. One of these participants ended medication approximately 1 month before participation in the study. The other stopped taking medication 40 hours before participation.The mean age of the 21 participants with TS in this study was 10.5 years . There were 17 males (81.9%) and 4 females (18.1%), including 15 persons identifying themselves as Caucasian (71.5%), 2 Hispanic (9.5%), and 4 as Other ethnic category (19.0%). The mean tic severity was 23.4 . The mean HI rating score was 66.1 , and the average IA rating score was 63.1 . The HI and IA scores had a normal distribution     correlated with IA scores  and glutamate receptor, ionotropic, N-methyl D-aspartate 2B (GRIN2B) , dopamine receptor D2 (DRD2), monoamine oxidase A (MAOA), and solute carrier family 6 , member 4 (SLC6A4) – all having previously been associated with ADHD  in regulation of the immune response, growth hormone and natural killer cell signaling (Table MOBP), dopamine receptor D1 (DRD1), forkhead box P1 (FOXP1) and fatty acid desaturase 2 (FADS2)  may provide a molecular correlate of the combined symptoms in ADHD, as well as facilitate an understanding of the association between IA and HI symptoms. Given the small number of participants, the results are preliminary and will need to be confirmed in subsequent studies. This study did not test whether the genes identified could be used to distinguish individuals with ADHD of the predominantly IA, predominantly HI, or combined types. The current study identified genes that correlated with IA, HI scales or both across all of the participants with TS. These genes might be useful in identifying ADHD phenotypes but future studies with a much larger cohort would be needed to address this question.One of the questions this study raises is how RNA expression in peripheral blood cells might correlate with IA or HI symptoms that are thought to be mediated by central nervous system pathways. First, many of the neurotransmitters and receptors expressed in brain are also expressed in peripheral leukocytes ,10-12. FMost of the top pathways associated with the common IA-HI genes in participants with TS were immune-related including IL-4 Signaling, B cell receptor signaling, T cell receptor signaling, and glucocorticoid receptor signaling. Glucocorticoid release, which is mediated by the hypothalamic-pituitary-adrenal axis, could affect IA and HI symptoms and gene expression of leukocytes . NetworkSLC6A2 and GRIN2B observed in the common IA-HI gene list have been associated with ADHD. SLC6A2 is a norepinephrine transporter that has been studied in ADHD due to the fact that drugs that block the norepinephrine transporter are efficacious in treating ADHD [SLC6A2 gene have been associated with ADHD [GRIN2B gene were associated with ADHD, and quantitative trait analyses showed associations of these markers with both the IA and HI symptom dimensions of ADHD. Disruption of GRIN1 (2A-D), another glutamate receptor subunit gene, leads to significant alterations in cognitive and/or locomotor behavior including impairments in latent learning, spatial memory tasks and hyperactivity [The neurotransmitter genes ing ADHD ,28. SNPsith ADHD . Glutamaactivity .One of the top canonical pathways over-represented in HI-candidate genes was the role of NFAT in the regulation of the immune response and natural killer cell signaling. This is consistent with a previous report of natural killer cell genes being differentially expressed in TS patients diagnosed with ADHD . Other HCOMT, DRD2, MAOA and SLC6A4 were also included in the HI-candidate gene list and have been previously associated with ADHD [DRD2, COMT and MAOA are catecholaminergic genes. SLC6A4 is a serotonin transporter that transports the neurotransmitter serotonin from synaptic clefts into presynaptic neurons. MAOA is a mitochondrial enzyme which degrades norepinephrine, dopamine and serotonin [COMT also catalyzes degradation of catecholamines including dopamine, norepinephrine and epinephrine. The DRD2 dopamine receptors mediate the effects of dopamine in the indirect basal ganglia pathway. The density of DRD2 receptors is highest in the basal ganglia, and HI is related to excessive dopamine activity in the basal ganglia [The neurotransmitter-related genes ith ADHD ,29 . DRDerotonin . COMT al ganglia .DRD1, MOBP, FOXP and FADS2. DRD1 is most abundant in the prefrontal cortex (PFC) which is believed to be critical for regulating attention, motivational behavior and emotion. Either too little or too much DRD1 receptor stimulation impairs PFC function [DRD1 with the ADHD IA symptoms in particular [Genes expressed in blood that correlated with IA symptoms and have been previously associated with ADHD included function . In addirticular .FOXP1 and MOBP genes are associated with ADHD [FOXP1 is a FOX transcription factor family member. FOX transcription factors regulate tissue- and cell type-specific gene transcription during both development and adulthood. Another family member FOXP2 is involved in developmental speech and language disorders and directly regulates targets related to neural development and synaptic plasticity and developmental disorders like autism and schizophrenia [GWAS have suggested that SNPs in the ith ADHD . FOXP1 iophrenia .This study only addressed gene expression correlated with the ADHD symptoms (IA and HI) in participants with TS, and did not consider other co-morbidities like tic severity or obsessive-compulsive symptom severity. It is not known if the genes associated with IA and HI symptoms in the TS subjects could be replicated in general populations of children with ADHD. Given that many genes overlapped between IA and HI symptoms in subjects with TS, some of these might also overlap in subjects with ADHD without TS.Two participants who had been previously prescribed medication were included in the current study. To determine if these subjects might have biased the results, our Principal Components Analysis (not shown) revealed that there were no outliers in the gene expression data, suggesting these two individuals did not significantly bias the correlations observed. Moreover, our previous studies including these individuals did not show them to be outliers with regard to fMRI findings or alternative splicing ,13. NeveThe largest limitation of the study is that, in spite of many genes being correlated with HI and/or IA symptoms, no gene passed multiple comparison correction testing using the Benjamini-Hochberg False Discovery rate (FDR<5%), and none of the genes were confirmed using an independent method such as RT-PCR. Thus, a future confirmatory study likely including RT-PCR and possibly corrections for blood cell types in a much a larger sample size will be needed to validate the genes reported here.Genetic studies have shown that of the many genes involved in ADHD, a given gene may only contribute a small percent to the symptoms ,6,17. ThThese findings support the concept that the pathophysiology of ADHD and/or its subtypes in TS may involve the interaction of multiple genes. Even with limitations, the results suggest a gene expression approach may be useful for defining molecular correlates of IA and HI symptoms in ADHD phenotypes in subjects with TS. A similar approach might be useful in ADHD phenotypes in subjects without TS.rp: Partial correlation; TS: Tourette syndrome; DSM-IV: Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition; CPRS-R: Conners’ Parent Rating Scales-Revised; RMA: Robust Multi-Chip Average; SLC6A2: Solute carrier family 6 member 2; GRIN2B: Lutamate receptor, ionotropic, N-methyl D-aspartate 2B; NFAT: Nuclear factor of activated T-cells; COMT: Catechol-O-methyltransferase; DRD1: Dopamine receptor D1; DRD2: Dopamine receptor D2; MAOA: Monoamine oxidase A; SLC6A4: Solute carrier family 6, member 4; MOBP: Myelin-associated oligodendrocyte basic protein; FOXP1: Forkhead box P1; FADS2: Fatty acid desaturase 2; ChAT: Choline acetyltransferase; Ach: Acetylcholine; GWAS: Genome wide association studies; GHRs: Transmembrane receptors; JAK2: Janus kinase 2; MAPKs: Mitogen-activated protein kinases; PI3K: Phosphatidylinositol 3-kinases; PKC: Protein kinase C; STATs: Signal transducer and activator of transcriptions; OCD: Obsessive–compulsive disorder.ADHD: Attention-deficit hyperactivity disorder; ANCOVA: Analysis of covariance; The authors declare that they have no competing interests.All the authors contributed substantially to the conception and design of the study. YT, JG, BS, BC, BA, CB, NB, PH contributed to the acquisition of data. YT, BS and GJ analyzed and interpreted data. YT,JBS and FRS drafted the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1755-8794/5/49/prepubTable S1. ADHD symptom related genes.Click here for fileTable S2-1. Canonical pathways with significant over-representation of genes that correlated with CPRS for IA, HI or both . Table S2-2. Chromosomes significantly over-enriched with genes that correlated with CPRS IA, HI scales or both .Click here for fileFigure S1. Co-expression analysis results of 24 common Inattention/ Hyperactivity/ Impulsivity (IA-HI) pathway-related probesets by using two-way clustering of gene-gene correlation data.Click here for fileFigure S2. Co-expression analysis results of 106 Hyperactivity/ Impulsivity (HI) pathway-related probesets by using two-way clustering of gene-gene correlation data.Click here for fileFigure S3. Co-expression analysis results of 48 Inattention (IA) pathway-related probesets by using two-way clustering of gene-gene correlation data.Click here for fileTable S3. The multiple probesets targted to a specific gene were highly correalted each other.Click here for file


The influence of comorbid symptomatology is unclear. Here we sought to address the question of gray matter abnormalities in GTS patients In GTS we detected a cluster of decreased gray matter volume in the left inferior frontal gyrus (IFG), but no regions demonstrating volume increases. By comparing subgroups of GTS with comorbid ADHD to the subgroup with comorbid OCD, we found a left-sided amygdalar volume increase.From our results it is suggested that the left IFG may constitute a common underlying structural correlate of GTS with co-morbid OCD/ADHD. A volume reduction in this brain region that has been previously identified as a key region in OCD and was associated with the active inhibition of attentional processes may reflect the failure to control behavior. Amygdala volume increase is discussed on the background of a linkage of this structure with ADHD symptomatology. Correlations with clinical data revealed gray matter volume changes in specific brain areas that have been described in these conditions each. Gilles de la Tourette syndrome (GTS) is a complex chronic motor and vocal tic disorder with childhood onset. However, the majority of patients, in addition, suffer from different comorbid disorders, most often obsessive-compulsive disorder (OCD) and attention deficit hyperactivity disorder (ADHD) ,2. On avwith comorbid OCD and/or ADHD compared to healthy controls using VBM. More specifically, we applied a sophisticated computational image analysis approach both to compare regional volumes of gray matter throughout the brain [VBM has been proven to be a powerful method for the in vivo study of human brain structures ,26. Thishe brain  and to tp = 0.03). However, none of the patients had an IQ below 92. Results of clinical assessments regarding severity of tics, OCD, ADHD, depression, and anxiety differed significantly between GTS patients and normal controls  was found significantly lower in GTS patients compared to healthy controls  in GTS patients compared to healthy controls  are significantly different compared to the GTS group without ADHD comorbidity  in regard to gray matter volumes, we directly compared these groups. By comparing the subgroups with comorbid ADHD (GTS + ADHD and GTS "plus") with patients suffering from Tourette with OCD symptomatology only (GTS + OCD), we found an increase of gray matter density in the left amygdala. This cluster was also significant in comparison to healthy controls, indicating differences due to pathology. We found no significant clusters of volume reductions here.p < 0.001, and represent multiple regression models best predicting GM values of GTS patients.Correlations between clinical scores and cortical gray matter volumes in patients suffering from GTS are presented in Figure Tic severity measured with the RVTRS correlated negatively with the volume of the right superior frontal gyrus (r = -0.65) and the left insula (r = -0.63). No regions showed a positive correlation with the RVTRS.The left postcentral gyrus showed a negative correlation with the Y-BOCS scores assessing obsessions (r = -0.63) and compulsions (r = -0.69) in patients with GTS. No regions showed a positive correlation with the Y-BOCS.By conducting multiple regression analyses including ADHD scales , we found a positive correlation with gray matter volumes of the left  and right putamen  while no negative correlations could be observed.The present study identified a common pattern of gray matter brain tissue alteration in actually unmedicated adult patients with GTS and comorbid OCD and/or ADHD characterized by left inferior frontal gyrus decreases of GM volumes. There were no significant differences between healthy controls and GTS patients concerning a GM volume increase. Based on our results, we suggest that the left IFG may constitute a common underlying neurological correlate of GTS with comorbid OCD and/or ADHD. As can be seen in Figure But are there any functional changes of the left IFG in GTS? We are aware of only a single PET-study  reportinOur results of a comparison of the GTS subgroups with ADHD comorbidity to the GTS group without ADHD comorbidity (GTS and OCD) revealed a volume increase of the left amygdala which could also be observed in comparison to healthy controls. Structural alterations of the amygdala -gray matter volume increase in particular  - have bVBM analyses reflect a number of anatomical features, including gray matter alterations and shifts in gyral or sulcal anatomy. Therefore, it can be speculated that GM volume decreases observed in this study might be the consequence of malformated cortical development, such as abnormal neuronal migration. Future studies using different volumetric measurements such as an estimation of cortical thickness may further contribute to the underlying structural differences in GTS ,41.We failed to detect group differences in regions previously associated with GTS pathology including the basal ganglia -10 and tA limitation of the study was the small subgroup of only 4 patients with comorbid ADHD (without OCD). When excluding this subgroup from additional analyses, differences between GTS patients and controls even increase strengthening our assumption that GM volume reduction in the inferior frontal gyrus are mainly based on the presence of comorbid OCD. However, future structural studies including larger samples of patients with comorbid ADHD are needed to better clarify the impact of comorbid ADHD.Although the group of patients and the control group were not comparable in regard to IQ, we do not think that group differences can be explained by these small IQ differences, since - to the best of our knowledge - there are no VBM studies available reporting morphological alterations on the basis of small group differences in intelligence. Our results are in line with recent findings in children demonstrating that IQ scores in patients with GTS "plus" (with comorbidities) are slightly reduced compared to healthy .Although all patients included in this study were unmedicated for at least 6 months prior to MRI imaging, some patients were medicated with typical or atypical antipsychotics before. In patients with Tourette syndrome, it has been demonstrated that neuroleptic medication increases both caudate and globus pallidus volumes . BecauseWe found several significant correlations between GM volumes and different clinical scores. Tic severity (RVTRS) correlated negatively with GM volumes of both the left insula and the right premotor cortex. Although, video-based tic ratings represent only a short period of time, they are regarded as the most objective tic measurement. Since in adult patients, spontaneous tic fluctuations are less marked compared to children, it can be speculated that the RVTRS indeed represents tic severity in an individual patient.This result is in line with recent morphological  and funcIn line with findings of a recent study in patients with "pure" OCD , we founIn contrast, we found a positive correlation between the severity of ADHD  and bilateral GM volumes in the putamen. This finding is in line with results obtained from recent MRI studies in children with "pure" ADHD consistently demonstrating an involvement of the putamen ,58 with By means of VBM we found a gray matter reduction in the left IFG in GTS patients with comorbid OCD/ADHD compared to healthy controls. Furthermore, the volume of the left amygdala was found to be increased in the subgroup of GTS patients with ADHD symptomatology. From our data it is suggested that the coexistence of tics and ADHD and/or OCD represents an additive model, since in patients with GTS "plus" severity of tics, ADHD, and OCD each correlated with those brain regions  that are also involved in "pure" GTS, "pure" ADHD and "pure" OCD.Forty-one adult (≥ 18 years) male patients with the diagnosis of GTS according to DSM IV-TR criteria were scanned for this study. All patients suffered either from comorbid OCD, ADHD, or both. All patients were actually unmedicated for at least six months before entering the study. For the diagnosis of ADHD we used both, a psychiatric examination and different self-rating scales including the DSM-IV symptom check-list for ADHD  consistiNone of the patients had a history of head trauma, epilepsy, brain surgery, systemic illness, drug or alcohol abuse, or any other significant comorbid disorder. In all patients a neurological and psychiatric examination was performed by one of the authors (KR MV) who is experienced in the diagnosis of TS, OCD, and ADHD. Twenty-five healthy control subjects individually matched for age and gender were enrolled in this study. Exclusion criteria were the same as for the patients' group. Healthy controls were interviewed and examined in the same way as patients. Handedness for all participants was assessed with the Edinburgh Handedness Inventory . The stuand ADHD, n = 8), (2) GTS-OCD (n = 17), and (3) GTS-ADHD (n = 4).Of 66 participants included in the study, data sets of 53  participants were used for further analyses. Data sets of 13 participants did not survive our strict image quality control due to different reason: poor image quality due to excessive movement artifacts (n = 9 GTS patients), anatomical abnormalities that prevent reliable and accurate spatial normalization , and claustrophobia leading to incomplete measurements (n = 3 GTS patients). Demographic and clinical parameters are summarized in Table 3).All MRI scans were obtained at 1.5 Tesla General Electric Signa Horizon LX . A high-resolution three-dimensional T1-weighted spoiled gradient recalled echo (SPGR) sequence generated 124 contiguous sagittal slices  http://dbm.neuro.uni-jena.de/vbm.html running under the MATLAB 7.10  environment. Voxel-based morphometry is a whole-brain, unbiased, semi-automated technique for characterizing regional cerebral differences in structural magnetic resonance images [p < 0.001. This method is implemented in SPM8 and refers to the estimated smoothness of the images. After determining of the number of resels, the expected Euler characteristic is calculated. This is used to give the correct threshold (number of voxels) that is required to control for false positive results.Data were processed and examined using the SPM8 software (Wellcome Department of Imaging Neuroscience Group, London, UK; e images . Before e images . Images http://web.mit.edu/swg/software.htm.Resulting parameter estimates were extracted from significant clusters and scaled to the global mean with the REX toolbox MW performed the analyses and drafted the manuscript. SB participated in the design of the study and the collection of data. TP and JG participated in data collection, and reviewed the manuscript. AG, NB, HB, and RD reviewed the manuscript. KRM-V participated in study design and coordination and edited the manuscript. All authors read and approved the final manuscript.


The interplay between olefin metathesis and dendrimers and other nano systems is addressed in this mini review mostly based on the authors’ own contributions over the last decade. Two subjects are presented and discussed: (i) The catalysis of olefin metathesis by dendritic nano-catalysts via either covalent attachment (ROMP) or, more usefully, dendrimer encapsulation – ring closing metathesis (RCM), cross metathesis (CM), enyne metathesis reactions (EYM) – for reactions in water without a co-solvent and (ii) construction and functionalization of dendrimers by CM reactions. Olefin metathesis reactions –7 have btrans-bis-phosphine complexes [cis-bis-phosphine ruthenium benzylidene catalysts. We therefore used Reetz’s bis-phosphines derived from the commercial polyamine DSM dendrimers [3(CO)12] catalyzed by [FeICp(η6-C6Me6)] leading to the substitution of a carbonyl of the [Ru3(CO)12] by a dendritic phosphine on each tether , 3. Activation of mesitylene by the CpFe+ moiety in 4, followed by a one-pot perallylation yielded [CpFe+][PF6-], 5, from which the free arene derivative 6 was obtained on visible-light photolytic decomplexation , 7,  –33. Thereactions –35. When[+ group –38. Thesticinium –41. The lexation –3542. Fictive CM  43–44].–44.+ grend generation Grubbs metathesis catalyst 7 proceeded selectively to produce dendrimers whose tethers were terminated by carboxylic acid groups 3 and iterations , generated in the catalytic cycle of RCM reactions involving terminal olefins, is usually highly susceptible to dimerization and decomposition in CH2Cl2 or C6H6 [11 is used for the RCM reactions. This strongly argues in favor of dendritic protection (probably by encapsulation) of the reactive species. RCM reactions need less catalyst 7 in organic solvents [11. Thus the hydrophobic dendrimer interior should indeed favor the protection this intermediate ruthenium-methylene species from side reactions occurring in the presence of water.Another key feature of the system is that the aqueous solution of the water-soluble dendrimer Olefin metathesis reactions are powerful methods that can be used for the construction of dendrimers and their functionalization with water-solubilizing carboxylate groups and other termini. In turn, water-soluble dendrimers can be used as molecular micelles as exemplified here. The implication of dendrimers in olefin metathesis reactions has mainly been focused on recovering the catalyst by loading the dendrimer with a functionalized catalyst. This strategy has been of very little success, because the % of catalyst used in metathesis reactions was rather high. This is due to the reactivity of methylene-metal intermediates that leads to side reactions. Consequently, another strategy involves protecting the catalytic intermediate in nanoreactors. Dendrimers are shown here to be excellent reactors achieving the goal of decreasing the catalytic amount when water is used as solvent. The success of using water as a reaction medium, even without co-solvent, is important in avoiding polluting organic solvents. Moreover, a very low Ru catalyst loading is possible in RCM with the fully recyclable water solution of the dendritic nanoreactor.


This review incorporates a large number of chromatographic systems modified by the surfactants. A large number of solvent systems and stationary phases are summarized in this paper. Three different kinds of surfactants  are used as modifiers for stationary phases as well as solvent systems. Surfactants are used at all the three different concentration levels  where surfactants behave differently. Modifications of both stationary phases and solvent systems by surfactants produced a new generation of chromatographic systems. Microemulsion solvent systems are also incorporated in this paper. Microemulsion thin-layer chromatography is a new approach in the field of chromatography. It is a well-known fact that amino acids are building blocks of proteins. Amino acids are also essential ingredients of diets of all living beings. Amino acids are biologically important biochemical molecules commonly used in nutritional supplements such as glutamic acid as flavor enhancer  and aspaSurfactants are the promising compounds for the modification of chromatographic data. The unusual results are obtained by using surfactants in chromatographic system as compared to other chemical compounds. Surfactants are the class of compounds which entirely modify the efficiencies of different phases involved in the chromatography. Surfactants expand the potentialities of TLC by resolving complex mixtures especially those containing neutral and charged compounds. Sumina et al.  discusseAccording to the literature surfactants provided important separations of amino acids. Some of the separations are very unique and scientifically important. On the other hand micellar thin-layer chromatography (MTLC) has got many advantages over conventional TLC. The most exciting features found in MTLC as compared to conventional thin-layer chromatography that leads to new generation of chromatographic systems are (a) double solvent front that is observed in chromatogram, (b) modifications of adsorbent surfaces, and (c) a change in elution order of compounds . The useThe following chromatographic systems Tables  and 2 weTLC plates are prepared by mixing silica gel “G” or alumina with double distilled water in 1 : 3 volume ratios with constant shaking for 5 min until homogeneous slurry is obtained. The resultant slurry is coated on the glass plates. The plates are first air-dried at room temperature and then activated by heating at 100°C for 1 h. The activated silica gel plates are then impregnated with desired concentrations of surfactants by developing plates in aqueous solution of impregnant, followed by drying of the plates at 100°C in an electrically controlled oven for 1 h , 18.Precoated plates are impregnated by directly developing in an aqueous solution of surfactants after development plates are activated in an oven at 100°C for 1 hr.Impregnated TLC plates are also prepared by directly mixing silica gel “G” or alumina with aqueous solutions of surfactants in 1 : 3 volume ratios with constant shaking for 5 min until homogeneous slurry is obtained. The resultant slurry is coated on the glass plates. The plates are first air-dried at room temperature and then activated by heating at 100°C for 1 h.A number of sorbent layers have been used for the identification, separation, and quantification of amino acids. The most commonly used material is the silica gel. The silica surface contains silanol (Si-OH) groups as active centers. These groups are active in chemical transformations and provide some unique properties on the surface.The adsorption of monomers of micelles on surfaces takes place by two different ways, that is, hydrophobic adsorption and silanophilic adsorption. The hydrophobic adsorption involves adsorption of alkyl tail of monomer to the alkyl group of stationary phase, while silanophilic adsorption involves adsorption of ionic head group onto the free silanol groups on silica gel .It is necessary to provide an idea about the nature of adsorption of different kinds of surfactants on different surfaces. It is a well-known fact that most of the natural surfaces are negatively charged. Therefore, if a chromatographer needs a hydrophobic surface, a cationic surfactant should be used. Cationic surfactant adsorbs on the surface by orienting positively charged hydrophilic groups towards the negatively charged surface and the hydrophobic group is oriented away from the surface, whereas hydrophilic surface is generated by using an anionic surfactant on the negatively charged surface. In case of positively charged surfaces an anionic surfactant should be used to generate hydrophobic surface, while a cationic surfactant should be used to produce a hydrophilic surface. This could be interpreted on the basis of formation of ad micelles and hemimicelles by surfactants on the appositively charged surfaces. These assemblies are formed on the surface as a result of the charged group of ionic surfactants attached to surfaces of opposite charge (columbic forces) and surfactant chain-chain interactions (noncolumbic).But, if we take the case of nonionic surfactants, it is obvious that these materials could adsorb onto the surface either by hydrophilic or hydrophilic group oriented towards the surface. The nature of these adsorptions could be predicted on the basis of the type of the surface used for the adsorption. As compared to cationic and anionic surfactants, zwitterionic surfactant possesses both positive and negative charges and hence does not change the charge on the surface significantly .In order to achieve some unique separations of amino acids, various surfactants of different kinds such as cationic, anionic, and nonionic have been used as surface modifiers , 20–23. A series of amino acids were tested on different impregnated layers. Chromatographers have used three different kinds of surfactants for the study of mobility pattern and separation of various mixtures of amino acids. Both silica and alumina surfaces were modified by different kinds of surfactants. Mohammad and Zehra , used diIn another study, Mohammad and Haq  have useThe main theme behind this review is to provide modern chromatographers a list of new generation surfaces for the chromatography of amino acids. A lot of information could be generated from This section deals with the different kinds of solvent systems generated from the surfactants. Armstrong and Nome  devised Mohammad and Hena  have sep31–M43] in various mixtures. A mixed combination of Tx-100 + SDS + acetone  has been proved to be useful for the selective separation of L-lysine from other essential amino acids. This method was also utilized for the identification of L-lysine in pharmaceutical formulations. In another study, Mohammad and Gupta [One of the most interesting ideas was to use mixed micelles in the separation of amino acids for the separation of amino acids. Usually nonionic and ionic surfactants are mixed together to produce mixed micelles. It is reported that mixed surfactant solution behaves differently than individual surfactants because of the increased ion-dipole interaction between ionic and nonionic surfactants . The mixnd Gupta  utilizedMicroemulsions are thermodynamically stable, single optically isotropic solutions containing oil, water, surfactant, and a cosurfactant  . MicroemRf value could be due to the influence of several factors such as adsorption, distribution, static electricity, hydrophobic force, steric barriers, extraction, and back extraction. Tian and Xie [Tian and Xie  have ext and Xie  have suc and Xie  have proThin-layer chromatography is also a useful analytical tool for the study of various interactions or binding of various compounds. Very few articles are published on the interaction studies of aminoacids with surfactants by TLC.  These interactions are biologically very important and are useful in various pharmaceutical and agrochemical formulations. These interactions have proved to enhance the efficiency of the active ingredients of various formulations. Forgacs  studied


Bupleurum falcatum L. has been used traditionally as a medicinal herb in Korean medicine. The hexane fraction of BF (HFBF), which was profiled with Direct Analysis in Real Time-Mass Spectrometry (DART-MS), activates the secretion of glucagon-like peptide-1 (GLP-1) in NCI-H716 cells significantly. We performed a microarray analysis and GLP-1 ELISA assay, as well as calcium imaging experiments with inhibitors, to investigate the mechanism of action of the HFBF. Through the microarray analysis, it was found that the ITPR2 gene that encodes the inositol 1,4,5-trisphosphate (IP3) receptor is up-regulated and the HFBF induces cell depolarization by inhibiting the voltage-gated channel expression in NCI-H716 cells. In addition, we found that the intracellular calcium in NCI-H716 cells, with Gallein, U73122, and 2APB as inhibitors, was decreased. These results suggest that the HFBF activates the GLP-1 secretion through the Gβγ pathways in the enteroendocrine L cells after treatment with the HFBF. Bupleurum falcatum L. (BF) has been used traditionally as a medical herb  was measured using fura-2 fluorescence dye. NCI-H716 cells grown on a matrigel-coated cover-slide bottom dish were washed three times with PBS and incubated in the dark for 30 min at room temperature with fura-2AM  in PBS. The cells were again washed with PBS three times and analyzed by being illuminated with alternating light of 340 and 380 nm from a rotating filter wheel.Differentiated NCI-H716 cells grown on cover glass were incubated overnight. Cytosolic free calcium [Caμg/mL of the HFBF. Three days before the experiments, the cells were seeded at 1 × 107 cells/well into 6-well culture plates precoated with Matrigel . Other procedures were performed in the same way as described in the GLP-1 assay. Total RNA was extracted by using Hybrid-RTM  according to the manufacturer's instructions. The microarray of each total RNA sample (200 ng) was labeled and amplified using a Low Input Quick Amp Labeling Kit  [μL of hybridization solution . After the aRNAs were placed on an Agilent SurePrint G3 Human GE 8 × 60 K Array  and covered by a Gasket 8-plex slide , the slides were hybridized for 17 hours in a 65°C oven. The hybridized slides were washed in 2 × saline-sodium citrate (SSC), 0.1% sodium dodecyl sulfate (SDS) for 2 min, 1 × SSC for 3 min, and then 0.2 × SSC for 2 min at room temperature. The slides were centrifuged at 3000 rpm for 20 sec to dry.Experiments were carried out in similar GLP-1 secretion study processes and we used 100 μL of hybridization solution . After the labeled aRNAs were placed on an Agilent SurePrint G3 Human GE 8 × 60 K array  and covered by a Gasket 8-plex slide , the slides were hybridized for 17 hours in a 65°C oven. The hybridized slides were washed in 2 × saline-sodium citrate (SSC), 0.1% sodium dodecyl sulfate (SDS) for 2 min, 1 × SSC for 3 min, and then 0.2 × SSC for 2 min at room temperature. The slides were centrifuged at 3000 rpm for 20 sec to dry.Each total RNA sample (200 ng) was labeled and amplified using a Low Input Quick Amp Labeling Kit . The Cy3-labeled aRNAs were resuspended in 50 The arrays were analyzed through the use of an Agilent scanner and the associated software and gene expression levels were estimated with the Feature Extraction v10.7.3.1 . Relative signal intensities for each gene were generated using the Robust Multiarray Average Algorithm. The data were processed according to a quantile normalization method using the GeneSpring GX 11.5.1 . This normalization method used for making the distribution of intensities for each array in the set of arrays was the same. The normalization and log transformed intensity values were analyzed using the GeneSpring GX 11.5.1 . Fold change filters included the requirement that the genes are present in at least 2-fold of the controls for upregulated genes and less than 0.5-fold of the controls for downregulated genes.db/db mouse was purchased from DBL (Korea). The db/db mouse (n = 5) was fasted for 16 hours and then orally administered at the volume of 5 g/kg glucose solution [The solution . HFBF wasolution .Each ELISA and calcium imaging set of data represents at least two separate experiments and each experiment was performed in triplicate. The significance of the data was analyzed with Prism 5 software with one-way ANOVA and Bonferroni tests to compare each set of data. Bars show the SEMs of the means of the three assays.+ = 373.11704). DART-MS results show isosinensetin is a candidate for the active compound of the HFBF which contributes to the GLP-1 secretion effect. A study using isosinensetin may increase the potential of the HFBF as a therapeutic agent for type 2 diabetes mellitus.DART-MS was performed to profile the HFBF . Among tTo confirm the GLP-1 secretion effect due to the HFBF treatment, a GLP-1 assay was conducted. The extracts were diluted with distilled water and were successively partitioned with organic solvents of different polarities as described in the Materials and Methods section. For GLP-1, an ELISA was conducted using a BF fraction including butanol, ethyl acetate, dichloromethane, and hexane. The HFBF activated GLP-1 secretion at higher levels compared to the other extracts (data not shown). As shown in α, β, and γ subunits. Gα and Gβγ act independently through different signaling pathways. As shown in ADCY1 gene that encodes AC is downregulated and PDE-related genes including PDE8B, PDE8A, PDE7B, PDE4D, PDE4D, PDE4D, PDE4C, PDE4B, PDE4A, and PDE2A are also downregulated. The ITPR2 that encodes the inositol 1,4,5-trisphosphate receptor is upregulated  is an incretin hormone which regulates insulin secretion, appetite, and gut motility [β-cells. The major source of GLP-1 is the intestinal L cell that secretes GLP-1 as a gut hormone. In this investigation, HFBF shows the antidiabetic effect in cellular level by secreting GLP-1 secretion. HFBF was treated to NCI-H716 cells with three concentrations: 100, 200, and 500 μg/mL. GLP-1 secretion was increased dose dependently. All concentrations of HFBF increased GLP-1 secretion and had significance. We supposed that the HFBF increases the GLP-1 secretion through sweet taste or bitter taste receptor signal pathway in enteroendocrine NCI-H716 cells. To verify this hypothesis. Lactisole was used as an inhibitor of sweet taste receptor, exactly, human T1R3. Sweet taste receptor is the heterodimer consisting of T1R2 and T1R3. After treatment of HFBF with lactisole, there is no significant effect comparing to the only HFBF treatment (βγ subunit that is related to phospholipase C β2 (PLC β2), diacyl glycerol (DAG), and inositol 1, 4, 5-triphosphate (IP3) and increases intracellular calcium. Gallein, 2APB, and U73122 are inhibitors of Gβγ pathways [βγ subunit, PLC β2, and IP3 receptor. The activated Gβγ activates PLC β2 and then divides PIP2 into DAG and IP3. DAG regulates protein kinase C (PKC) and triggers phosphorylation of PKC. IP3 is recruited to IP3 receptor in ER membrane and, as a result, releases calcium flow into cytoplasm. In βγ-mediated pathway, for the release of GLP-1. GLP-1 is one of target molecules for the treatment of Type 2 diabetes mellitus because it activates the GLP-1 receptor on the cell and then mediates insulin secretion. HFBF also showed regulatory effect of blood glucose level in db/db mouse OGTT experiments. This investigation suggests that HFBF stimulates GLP-1 secretion through Gβγ-mediated signaling pathway. The overall conclusion from the results is that the HFBF could be used as a possible medication for Type 2 diabetes mellitus patients via activation of Gβγ-mediated pathway.This investigation suggests that HFBF stimulates GLP-1 secretion through the Gmotility . Lately,reatment . G protepathways , 20. The