The mean (SD) dietary GI and GL was 54 (7) and 135 (59) respectively and the top 3 contributors to dietary GL were breads (14.4%), cereal-based dishes (10.3%) and breakfast cereals (ready to eat) (6.6%). There were small but significant differences in the GL contribution pattern between the sexes. The findings indicate that the average dietary GI of Australian adults is similar to that of other population groups, with a large proportion of starchy and energy-dense nutrient-poor foods that contribute to a high GL.This study aimed to determine the major food groups contributing to dietary glycaemic load (GL). Plausible food intake data collected using a multiple-pass 24 hour recall from a weighted sample of 6326 adult respondents  of the 2011–2012 Australian Health Survey dataset (AHS) were analysed. The GI of foods was estimated based on a previously published step-wise method. Descriptive statistics were calculated for dietary glycaemic index (GI), GL and contribution to GL by major food groups, stratified by age and sex. Trends across age groups were assessed using linear regression. Pearson’s χ Evidence from recent meta-analyses123456et al.et al.To better design appropriate dietary intervention strategies to lower the dietary GI and GL, it is important to know the current levels and contributors to dietary GI/GL of the target population. Until recently, nationally representative data for Australian adults were scarce. Several Australian population-based cohort studies1011vs. women of the same age.Australia has conducted several national nutrition surveys in the past 40 years14Therefore, using data from the recently released Australian Health Survey (AHS)2, with 37.6% and 22.5% considered overweight and obese, respectively. Extreme low-reporters excluded from the analysis were similar mean age to those included  and had a higher mean BMI of 29.7 ± 6.3 kg/m2 (p < 0.001); whereas extreme high-reporters excluded had a significantly lower age of 40 ± 17 years , and lower BMI of 23.1 ± 4.3 kg/m2 (p < 0.001), compared with participants with plausible dietary intake. Respondents excluded due to missing weight data were significantly older .The characteristics of the participants are summarized in p < 0.001).The dietary intake of the subjects, stratified by age groups, is summarized in per capita basis. The ranking of GL contributions per food group varied considerably among age groups (Bread and bread rolls (14.4%); cereal-based dishes (10.3%); and breakfast cereals (ready to eat) (6.8%) were identified as the top 3 contributors to dietary GL, accounting for nearly 40% of the dietary GL on a e groups .per capita contribution to dietary GL by the top 20 contributors, stratified by age group. Cereal-based dishes; flours, cereals and starches; sweetened beverages; fruit and vegetable juices and drinks; poultry-based dishes; and pastas all contributed less to dietary GL as age increased, while bread and bread rolls; breakfast cereals (ready to eat); sugar, honey and syrup; cake-type desserts; sweet biscuits; dairy milk; tropical and subtropical fruit; pome fruit; savoury biscuits; and frozen milk products contributed more to dietary GL as age increased . Post hoc comparisons  indicated that adults aged 19–30 years had significantly lower proportions of dietary GL from bread and bread rolls, and sugars, honey and syrups; and significantly higher proportions of dietary GL from cereal-based dishes, sweetened beverages and fruits and vegetables juices and cordials, compared with their older counterparts. They also had significantly higher proportions of dietary GL from poultry-based dishes and pasta compared with adults aged 51 years or above, and a significantly lower proportion of dietary GL from dairy milk.p < 0.001) were observed for breakfast cereals (ready to eat) (M: 7.1 ± 11.4% vs. F: 6.1 ± 10.5%); flours, cereals and starches (M: 7.0 ± 16.5% vs. F: 5.6 ± 14.4%); sweetened beverages (M: 5.1 ± 10.3% vs. F: 3.2 ± 8.9%); cake-type desserts (M: 3.1 ± 8.5% vs. F: 4.7 ± 11.2%); tropical and subtropical fruit (M: 1.8 ± 4.5% vs. F: 2.5 ± 5.8%); pome fruit (M: 1.6 ± 3.8% vs. F: 2.0 ± 4.2%); savoury biscuits (M: 1.4 ± 5.2% vs. F: 2.1 ± 6.4%); and chocolates (M: 1.3 ± 4.7% vs. F: 1.9 ± 5.9%). Percentage GL contributed from fancy breads was also marginally different between sexes .When examined by sex , signifiPer consumer analyses, as presented in ptrend < 0.001); while decreasing trends with age were observed for breakfast cereals (ready to eat); flours, cereals and starches; sweetened beverages; and fruit and vegetable juices and drinks . Interestingly, post hoc analysis revealed that these trends across age groups were mostly driven by the significant difference between the 19–30 year olds and their older counterparts.p < 0.001) were observed for flours, cereals and starches (M: 35.5 ± 19.3% vs. F: 30.5 ± 19.1%); sweetened beverages (M: 15.0 ± 12.9% vs. F: 12.9 ± 13.8%); cake-type desserts (M: 19.1 ± 11.6% vs. F: 23.0 ± 14.3%); tropical and subtropical fruit (M: 9.0 ± 6.0% vs. F: 10.5 ± 7.3%); and pome fruit (M: 7.0 ± 4.9% vs. F: 7.9 ± 4.8%). Percentage GL contributed from chocolates was marginally different between sexes .When examined by sex , signifiEnergy- and SEIFA adjustments to the analyses did not materially alter the interpretations and conclusions .This study is the first to report the GI and GL of a nationally representative sample of Australian adults. Overall, the mean GI and GL of participants was 54 (SD 7) and 135 (SD 59), respectively. Both GI and GL decreased with advancing age. Common carbohydrate-rich staple foods including bread and bread rolls, cereal-based dishes and breakfast cereals (ready to eat) were the top contributors to dietary GL in Australian adults.vs. 53.9 ± 6.8; p < 0.001) and dietary GL per MJ , but lower fibre density . We also observed significant differences  in the per consumer percentage GL contribution from different food groups between adults and children and adolescents. Adults had higher proportions of their dietary GL contributed by flours, cereals and starches (6.3% vs. 4.6%); potatoes (5.4% vs. 4.5%); sugars, honey and syrups (4.0% vs. 1.7%); and pastries (2.5% vs. 1.8%); and lower proportion of their dietary GL from fruit and vegetable juices and drinks (2.8% vs. 3.9%); sweet biscuits (2.2% vs. 3.2%); dairy milk (2.1% vs. 2.9%); pome fruit (1.8% vs. 2.6%); savoury biscuits (1.7% vs. 3.0%); and frozen milk products (1.1% vs. 1.5%). Furthermore, in the current study, older adults had a large proportion of their dietary GL in the form of multigrain bread and bread rolls which have a lower GI than conventional breads, as also reported by Barclay et al.Our group had previously reported data on children and adolescent respondents from the same national surveyet al.et al.21When comparing dietary GI and GL data from different international studies, it is important to take the dietary assessment methods used into account. For example, the study by Murakami vs. 54) but lower GL (123 vs. 135) than that assessed by a semi-quantitative FFQ in a sub-population of 2,900 Australian adults in the Blue Mountains regionvs. 115) and lower dietary GI (54 vs. 56) than that assessed using a diet history in a small sample of 459 women in Brisbane Australia. Again, differences in the dietary assessment methods and the different periods of data collection may have contributed to these disparate findings. Clearly, nationally representative data much better inform policy discussion.Our results also imply that older adults (aged 51 years or above) in the Australian population have a similar dietary GI , followed by cereal-based dishes (10.3%), breakfast cereals (ready to eat) (6.6%), flours, cereals and grains (6.3%), potatoes (5.4%) and sweetened beverages (4.2%). To inform public health strategies and policy discussions regarding lowering dietary GI and GL, it is important to look at both the per consumer and per capita data, where the per capita data provide a snapshot of the population, and the per consumer data provide the actual intake level and pattern for those who consume a higher GI/GL diet. The per consumer analysis revealed that flours, cereal and starches were higher contributors (33.2%), followed by cereal-based dishes (29.1%), breads (21.9%), cake-type desserts (21.1%), potatoes (16.3%) and pasta (21.6%). The large differences between the per capita vs. per consumer results suggests that the per capita data were likely skewed to the lower end by a large number of non-consumers. Hence to better target individual-based dietary intervention strategies, the per consumer data should be used. Per capita data should be used for developing population-based strategies.On a It is often assumed that a low GI diet is one with a GI less than 55 given low GI foods are defined as those that have a GI of 55 or less2324et al.While dietary GI represents the quality of carbohydrates in the diet, dietary GL which takes into account both quantity and quality of the dietary carbohydrates was found to better predict postprandial glycaemia and insulinaemiaThe use of a published method27vs. dense multi-grain breads (low GI), and as such we were unable to tell whether the high contribution of breads to dietary GL is due to the choice of high GI breads, of the high consumption of breads in general, or both. However, this limitation likely only affected food groups with considerable within group variability such as breads and cereals. Using the pre-defined food classification allows better comparisons of other secondary analyses of the Australian Health Survey, and the results will be more interpretable by dietitians and public health practitioners who are familiar with the survey.Our study is however limited in several ways. First, we used only the first 24 hour recall, which is unlikely to capture the usual dietary pattern of an individualIn conclusion, the average dietary GI of Australian adults was found to be substantially higher than that associated with reduced risk of chronic disease in other populations. Significant age and sex differences in GL contribution pattern were also observed. Breads remain the top contributor to dietary GL among Australian adults, while younger adults are deriving more of their dietary GL from a wide range of foods, especially processed foods. Dietary interventions aiming at reducing dietary GI and/or GL should hence be age- and sex-specific, where interventions aiming at elderly should focus on swapping high GI staple food with low GI versions, while for younger adults discouraging consumption of high GI processed foods may be more effective.This analysis used data from the National Nutrition and Physical Activity Survey (NNPAS) component of the 2011–2012 AHS. The AHS was conducted by the Australian Bureau of Statistics (ABS) between 2011 and 2012, with a response rate of 77%2); normal weight (18.5–24.99 kg/m2), overweight (25.0–29.99 kg/m2) and obese (≥30.0 kg/m2)Weight and height were measured without shoes and heavy clothing where possible, using a digital scale and a stadiometer, to the nearest 0.1 kg and 0.1 cm, respectively. Body mass index (BMI) was calculated as weight in kg divided by the square of height in meters. Participants were then classified into underweight, normal weight, overweight or obese based on the following BMI cut-offs: underweight  had dietary information collected in a face-to-face interview between 29 May 2011 and 9 June 2012. A phone interview was conducted in ~60% of the participants at least 8 days after the face-to-face interview to collect a second 24 h recall. A two dimensional food model booklet was provided to the respondent to aid portion size estimation27n = 1752). All remaining foods had their GI values assigned based on a step-wise, systematic approach, using either the GI value of an exact match  or a closest match  in one of the 4 commonly used GI databases373839n = 1526), or the median GI of the corresponding food subgroup . A small proportion (n = 60) of foods were assigned a default GI as none of the earlier steps could be used to assign a GI to them.All participants aged 19 to 95 years  × available carbohydrates (in grams) in a serving of that specific food. The daily dietary GL of each subject was then calculated as the sum of GL from all foods consumed in that day. Dietary GI was calculated by the following formula:  × 10016et al.n = 6278), of which 52.4% were male.In this study, only the first 24 h recall collected from a face-to-face interview were used. This is likely to reduce the influence of under-reporting which is common in the follow-up recall via phone interviewper capita and per consumer results were both presented. Per capita analyses included all participants and provide a snapshot of an average individual, whilst per consumer analyses included only the participants who had reported consumption of the specific food group on the day of the food recall. The latter provided information on average serving size. Age groups pre-defined in the AHS were used in the stratified analysis to allow better comparisons across surveys within Australia, and post-hoc comparisons between age groups were performed by using the Bonferroni post-hoc test for one-way ANOVA. Differences between sexes for each age group were tested by one-way ANOVA. Linear regression was used to test for trends across age groups. All main statistical analyses were carried out using SPSS version 22.0 . Additional analyses adjusted for energy and SEIFA were performed using General Linear Model in SPSS Complex Samples. A p value of <0.001 was considered to be statistically significant to minimize type I errorThe AHS had over-sampled males to females, and had a much larger percentage of adults than children. Furthermore, major city residents were over-represented33How to cite this article: Louie, J. C. Y. et al. Dietary glycaemic index and glycaemic load among Australian adults – results from the 2011–2012 Australian Health Survey. Sci. Rep.7, 43882; doi: 10.1038/srep43882 (2017).Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


Enterovirus 71 (EV-A71) shows a potential of rapid death, but the natural history of the infection is poorly known. This study aimed to examine the natural history of EV-A71 infection.st and October 31st, 2012, at three hospitals in Guangdong, China. Subjects with positive EV-A71 RNA laboratory test results were included. Disease progression was documented with MRI, autopsies, and follow-up. Symptoms/signs with potential association with risk of death were analyzed.This was a prospective longitudinal observational study performed between January 1Among the 288 patients, neurologic symptoms and signs were observed . Some of them occurred as initial symptoms. Myoclonic jerks/tremors were observed among >50% of the patients; nearly 40% of patients presented fatigue and 25% were with vomiting. Twenty-eight patients (9.7%) presented poor peripheral perfusion within 53.4 ± 26.1 h; 23 patients (8.0%) presented pulmonary edema and/or hemorrhage within 62.9 ± 28.6 h. Seventeen (5.9%) patients were in a coma. Seven (2.4%) patients died within 62.9 ± 28.6 h. Seventy-seven survivors underwent head and spinal cord MRI and 37.7% (29/77) showed abnormalities. Two fatal cases showed neuronal necrosis, softening, perivascular cuffing, colloid, and neuronophagia phenomenon in the brainstem.Patients with EV-A71 infection showed high complexity of symptoms and onset timing. Death risk may be indicated by autokinetic eyeball, eyeball ataxia, severe coma, respiratory rhythm abnormality, absent pharyngeal reflex, ultrahyperpyrexia, excessive tachycardia, pulmonary edema and/or hemorrhage, and refractory shock and ataxic respiration. Early assessment of these symptoms/signs is important for proper management.The online version of this article (doi:10.1186/s12879-017-2228-9) contains supplementary material, which is available to authorized users. Recently, the enterovirus 71 (EV-A71) has been considered as the second most significant neurotropic enterovirus, next only to poliovirus . During Analysis of the 1998 Taiwan EV-A71 HFMD outbreak showed that complications included encephalitis, pulmonary edema or hemorrhage, aseptic meningitis, myocarditis, and paralysis were associated with EV-A71 infection –6. AnalyTherefore, this prospective observational study was carried out to characterize the various symptoms of EV-A71 infection and to explore the associations between clinical manifestations and their indications for disease severity and progression.http://www.gov.cn/gzdt/2008-05/03/content_960347.htm, which is similar to an English version from Hong Kong: http://www.dh.gov.hk/chs/useful/useful_ld/files/ltod20070523.pdf). This study was approved by the Ethics Committees of all three participating hospitals. Written informed consent was obtained from the legal guardians of the patients.This was a prospective, longitudinal observation aiming to record the clinical manifestations and onset timing of cases with severe EV-A71 infection. A study board consisting of pediatricians, pediatric neurologists, and HFMD specialists of the National Health and Family Planning Commission of The People’s Republic of China, and experts of the National Clinical Research Center for Respiratory Diseases and of the Institute of Neuroscience of Guangzhou Medical University was organized in 2008. By reviewing case records of HFMD from our experience as well as from previous studies –12, 91 sst and October 31st, 2012 at three hospitals: the Guangzhou Women and Children’s Medical Center Affiliated to Guangzhou Medical University and Sun Yat-sen University, the Dongguan Taiping People’s Hospital, and the Zhongshan People’s Hospital, all of which are major treatment centers of HFMD in the coastal cities of the Guangdong province. The three centers are located in the core area of the Zhujiang Delta, where it is dominated by the monsoon climate from the South Asian Ocean. This region presents a high HFMD incidence. Autopsies were performed at the Forensic Medical Identification Center of Sun Yat-Sen University.Cases of EV-A71 were observed between January 1Patients <14 years old with suspected EV-A71 infections  were screened by real-time PCR using a throat swab sample, as previously described . SubjectPatients were triaged in a standard manner, which determined the interval of the clinical assessments. Critical patients were continuously monitored. Emergency patients were assessed at 15-min intervals. Urgent patients were assessed at 15–30 min intervals. There was no non-urgent patients in the present study.MRI was performed using a PHILIPS Achieve 1.5 T . One senior radiologist and one senior neurologist analyzed the MRI scans.t-test was used to detect differences in occurrence timing of each symptom. Correlations were assessed using the Pearson’s test. Statistical analyses were performed using SPSS 22.0 . Two-sided P-values <0.05 were considered significant.Descriptive statistics were used to interpret the patterns of the 91 observational factors and the sequences of occurrence. The Student’s st and October 31st, 2012, 288 inpatients with confirmed severe EV-A71 infection were registered in the three participating hospitals, including seven (2.43%) fatal cases. The patients were classified as severe if they experienced any neurological complications  and/or cardiopulmonary complications ; otherwise, they were categorized as mild cases. There were 188 males and 100 females, with a median age of 25 months ; 161 (55.9%) patients were from urban areas and 127 (44.1%) were from rural areas. Among the 288 patients, 274 (95.1%) were diagnosed with HFMD or herpangina, and the remaining 14 (4.9%) were diagnosed with neither but were positive for EV-A71 RNA and with neurological symptoms.Between January 1Seven (2.43%) patients died within 5 days after onset; these patients were aged 27.7 ± 13.2 months. The median time from onset to death was . There was no difference in gender, age, weight, and demographic distribution between dead and surviving patients. Among the survivors, 258 were fully recovered, and 23 had slight neurological symptoms/signs when they were discharged from the hospital; 19 of them were followed up from 2 to 20 weeks, and none of them was found neurological sequelae.Recorded clinical manifestations and timings are listed in Table Among the dyskinesia indicators, emotional movement disorders were the major symptoms of dyskinesia: 211 patients (73.3%) had startles, which appeared within 30.0 ± 23.0 h as the first single symptom. Anxiety occurred in 39 patients (13.5%) and occurrence time was 40.1 ± 31.1 h. Dysphoria was observed in 17 patients, 14 with fright, two with gibberish, and one with mania. Muscle strength and muscle tone disorders were the most common symptoms of limb/muscle movement disorders. There were 162 patients (56.3%) with myoclonic jerks or tremors occurring within 33.3 ± 23.0 h, including one case of galloping tongue. There were 22 patients with acute flaccid paralysis, 12 of them involving all extremities, and 10 in the upper or lower limbs. For abnormal eyeball movement indicators, nystagmus was presents in 16 patients (5.6%) and appeared within 51.8 ± 28.5 h; 10 patients showed eyeball ataxia occurring within 57.6 ± 31.7 h; eight patients showed autokinetic eyeball within 43.8 ± 18.3 h after onset. In abnormal pupil movement indicators, abnormal light reflex was found in five cases. Pharyngeal reflex was absent in eight patients (2.8%) within 63.9 ± 34.7 h.More than half of the patients had various levels of disturbance of consciousness. Fatigue/sleepiness  was found in 115 patients (39.9%), occurring within 44.0 ± 26.5 h, most of them complaining of sleepiness and less movements; somnolence was present in 25 patients (8.7%). There were 17 patients with different levels of coma.P < 0.05). Twenty–eight patients (9.7%) presented poor peripheral perfusion within 53.4 ± 26.1 h, including capillary refill time (CRT) extension occurring in 27 patients (9.4%) within 52.9 ± 25.4 h; 25 patients showed clammy skin, pale skin in 13, and mottled skin in 12. In the seven fatal cases, poor peripheral perfusion occurred in the early stages . Refractory shock was observed in nine patients (3.1%), and occurred within 59.9 ± 23.7 h (P < 0.001).For the circulatory system, 15 patients (5.2%) had different kinds of arrhythmia within 57.8 ± 38.8 h. There were 12 patients (4.2%) with tachycardia within 49.6 ± 26.3 h, and seven had excessive tachycardia within 58.1 ± 23.2 h; five of them died (P = 0.001). Sixteen patients presented tachypnea within 49.1 ± 27.7 h. Eight patients were with hyperventilation within 57.5 ± 35.1 h. Pulmonary edema and/or hemorrhage was observed in 23 patients (8.0%), which appeared within 62.9 ± 28.6 h; seven of them died (P = 0.001).For the respiratory system, irregular respiratory rhythm was observed in 19 patients (6.6%) within 60.1 ± 27.1 h, and seven of them died (P = 0.512), persistent hyperpyrexia and anxiety (P = 0.997), persistent hyperpyrexia and fatigue/sleepiness (P = 0.904), fatigue/sleepiness and fright (P = 0.216), fright and dysphoria (P = 0.567), or nystagmus and CRT extension (P = 0.543).Exceped the prodromal symptoms, the 50 symptoms/signs with the highest frequency are shown in Table P < 0.05). Among the 42 symptoms/signs . It could be summarized that the usual progression was fever, fatigue/sleepiness, poor peripheral perfusion, abnormal respiratory rhythm, pulmonary edema and/or hemorrhage, refractory shock, and death (Table Among the seven non-surviving cases 2.43%), six were from rural areas, and four were male. The median age of death was 29 months  and the patients weighed 12.14 ± 2.12 kg . The median time from onset to death was 77 h . All dead patients had neurological symptoms/signs , eyeball ataxia , severe coma , respiratory rhythm abnormality , decreased or absent pharyngeal reflex , ultrahyperpyrexia , excessive tachycardia , pulmonary edema and/or hemorrhage , refractory shock , and ataxic respiration    and died 3–4.4 days after illness onset; three of them experienced excessive tachycardia and hypertension recurring 5 to 23 h until death. It should be noticed that five of the surviving patients and one non-surviving patient were not in a coma before pulmonary edema. The mortality from pulmonary edema was the last cause in the FNS in our study.P < 0.05). This suggests that patients who developed these symptoms/signs were prone to develop the symptoms/signs of the FNS. Table In order to determine if some factors could indicate the eventual progression to the FNS, time-correlations between the occurrence of the FNS (any component) and other symptoms/signs were analyzed using the Pearson correlation analysis. CRT extension, fatigue/sleepiness, tachycardia, anxiety, hyperventilation, vomiting, persistent hyperpyrexia, myoclonic jerks/tremors, nystagmus, startles, dysphoria, and fright were time-correlated with FNS  of patients. The affected neural structures included the thalamus, basal ganglia, midbrain and rhombencephalon, and spinal cord Table . Among t9/77 of pAmong patients with telencephalon/meningeal changes, five cases showed startles and fatigue/sleepiness, four with vomiting, four with myoclonic jerks/tremors, three with persistent hyperpyrexia, three with mental disorders, and even disturbance of consciousness and central respiratory disorders.Among the cases with MRI changes in the brain stem, disturbance of consciousness or mental disorders were not found, but startles, myoclonic jerks/tremors, decreased muscle tension, decreased muscle strength and acute flaccid paralysis were observed.Two fatal cases revealed neuronal necrosis, softening, perivascular cuffing, colloid, and the neuronophagia phenomenon in the brainstem Figs.  and 5. TAmong the severe cases of EV-A71 infection identified in the present study, neurological symptoms and signs occurred at each stage, which confirmed the neurotropic nature of EV-A71. Some of the symptoms such as myoclonic jerks/tremors and vomiting related to EV-A71 infection could not be related to damages in specific neural region –16. HoweIn the present study, the age distribution  was not only similar to a large study from China , 7. WhetIt has been suggested that the duration from onset to present neurological symptoms also depends on the velocity of reverse axonal transportation, the length of motor nerve and the characteristics of the virus including its virulence, mode of spread and transmission. In the present study, seven patients died within 5 days after onset with autonomic dysfunction. All of them developed subsequent irregular respiration, pulmonary edema/hemorrhage, and refractory shock. The rapid progress of fatal cases might be explained by the direct EV-A71 invasion of the medulla oblongata via the cranial nerve (CN). The autopsy showed that the cause of death was acute central nervous failure after brain stem encephalitis. Two dead patients showed neuronal necrosis, softening, perivascular cuffing, colloid, and neuronophagia phenomenon in the brainstem. The brain and cerebellum lesions were very similar. Pulmonary necrosis and inflammatory cell infiltration were not observed, suggesting that they had pulmonary hemorrhage without inflammation. Cardiac specimens showed breakage or undulatory array of some myocardial fibers without inflammation, necrosis, or hemorrhage. There were no pathological changes in other organs. Therefore, the pathologists confirmed that these patients died of acute central nervous system dysfunction caused by brainstem encephalitis.The occurrence timing of some neurological symptoms or signs that indicate nerve lesions was not different between the survivors and non-survivors. Therefore, it may be hypothesized that the virus invasion path may go through multiple neural pathways or different cranial nerve fibers, but this will have to be confirmed. Six cases presented pulmonary edema/hemorrhage without coma, and one of them had abnormal MRI signals in the spinal cord and spinal roots without abnormal change in brain. This phenomenon might be related to the direct attack on the medulla oblongata, spinal cord or sympathetic trunk, without involving awakening centers . The posData of the present study showed that when FNS occurred, patients could die in a short time. Clinicians need to pay great attention to these symptoms/signs. FNS are mainly symptoms caused by damage to the medulla oblongata, including autokinetic eyeball, eyeball ataxia, severe coma, respiratory rhythm abnormality, decreased or absent pharyngeal reflex, ultrahyperpyrexia, excessive tachycardia, pulmonary edema and/or hemorrhage, refractory shock, and ataxic respiratory.Among the patients that underwent MRI examination in the acute phase, patients with positive MRI results showed some neurological symptoms and signs. On the other hand, in the present study, 62.3% of patients had normal MRI results, while patients with encephalomyelitis may have lesions of both the brainstem and the spinal cord. This delay could be associated with MRI time lag in acute neurologic injury or necrosis –44, i.e.The severe EV-A71 infected patients might deteriorate and even die in a short time, therefore hindering practice of neuroimaging. In addition, some uncertain factors affected the implementation of MRI in early phase, such as parent’s wishes, equipment condition, patient tolerance and checking-time selection. Primary assessments based on symptoms and signs are advantageous in these circumstances . BecauseThe results of the present study could be used to propose a staging system for severe EV-A71 infection. We regarded patients with FNS as Stage 2c Fig. , which rTaiwan summarized the epidemiology and clinical features of EV-A71 infection and showed that nervous system involvement was an important factor in critically ill patients . China mThis study has some limitations. First, the cases were only from coastal cities in the Guangdong Province, and the generalisability might be challengeable. In addition, the sample size was small, precluding any reliable calculation of sensitivity and specificity. Secondly, we only interpreted the observation and analysis of the clinical facts. The present facts supported that neurological symptoms were valid for early assessment. Thirdly, due to ethical reasons, autopsy only confirmed the major cause of death, and we could not perform a comprehensive and exploratory autopsy; thus, brainstem lesions could not be explained as an independent reason for fatality. There could be a slight difference between the time of symptom onset and the time of symptom observation, but this difference was of maximum 15–30 min.Severely affected patients may die from EV-A71 infection. Close attention and early assessment of certain symptoms/signs is important for severe infection administration. Symptomatic treatment is warranted in all patients.


Nature Communications 10.1038/s41467-017-01045-x, published online 31 October 2017Correction to: 2), which is stronger than in previous reports’, should have read ‘Intensity ranged from 0.5 to 2.5 mA peak to peak (peak current density 0.0625–0.3125 mA/cm2), which is stronger than in previous reports.’ These errors do not affect any of the Article’s conclusions. Correct versions of Fig. 2 and Supplementary Fig. 11 are presented below as Figs. 1, 2.It has come to our attention that we did not specify whether the stimulation magnitudes we report in this Article are peak amplitudes or peak-to-peak. All references to intensity given in mA in the manuscript refer to peak-to-peak amplitudes, except in Fig. 2, where the model is calibrated to 1 mA peak amplitude, as stated. In the original version of the paper we incorrectly calibrated the computational models to 1 mA peak-to-peak, rather than 1 mA peak amplitude. This means that we divided by a value twice as large as we should have. The correct estimated fields are therefore twice as large as shown in the original Fig. 2 and Supplementary Fig. 11. The corrected figures are now properly calibrated to 1mA peak amplitude. Furthermore, the sentence in the first paragraph of the Results section ‘Intensity ranged from 0.5 to 2.5 mA (current density 0.125–0.625 mA mA/cmFig. 1Fig. 2


Helicobacter pylori increases 10 times the risk of developing gastric cancer. Juglone, a natural occurring 1,4-naphthoquinone, prevents H. pylori growth by interfering with some of its critical metabolic pathways. Here, we report the design, synthesis, and in vitro evaluation of a series of juglone derivatives, namely, 2/3-phenylaminojuglones, as potential H. pylori growth inhibitors. Results show that 5 out of 12 phenylaminojuglones (at 1.5 μg/mL) were 1.5–2.2-fold more active than juglone. Interestingly, most of the phenylaminojuglones (10 out of 12) were 1.1–2.8 fold more active than metronidazole, a known H. pylori growth inhibitor. The most active compound, namely, 2-amino)-5-hydroxynaphthalene-1,4-dione 7, showed significant higher halo of growth inhibitions (HGI = 32.25 mm) to that of juglone and metronidazole (HGI = 14.50 and 11.67 mm). Structural activity relationships of the series suggest that the nature and location of the nitrogen substituents in the juglone scaffold, likely due in part to their redox potential, may influence the antibacterial activity of the series.Infection by  Helicobacter pylori (H. pylori) is a Gram-negative bacillary spiral-shaped bacterium that colonizes the human stomach [H. pylori belongs to the group 1 of carcinogens according to the International Agency for Research on Cancer (IARC) [H. pylori express high levels of urease, converting urea into ammonium and carbonic anhydride. This creates an alkaline local medium that allows the survival of H. pylori in the acidic environment of the stomach, facilitating the colonization of the gastric mucosa [H. pylori infection is acquired during childhood, and if it is not treated, it may remain throughout the entire patient life [H. pylori [H. pylori infection [ stomach  and is a stomach , 3. Due r (IARC) , 5. In tc mucosa –8. Frequent life . Approxi. pylori –12 but m. pylori , 14. In H. pylori includes a double antibiotic therapy plus a proton pump inhibitor. This high-cost treatment regimen is often problematic (failure rates between 20 and 40%), with undesirable side effects that limit patient compliance and lead to the selection of antibiotic-resistant bacteria [H. pylori has been associated with the consumption of many food of vegetal origin, including wine and green tea which are rich in phytochemicals such as flavones, isoflavones, flavo- and flavanols, anthocyanidins, tannins, and stilbene derivatives [H. pylori infection and prevent gastric cancer [Currently, the eradication treatment of bacteria –17. Loweivatives –21. Takec cancer .Quinones represent an important class of naturally occurring compounds that are widely found in animals, plants, and microorganisms . These cH. pylori growth by different mechanisms to those reported for standard antibiotic drugs and could be used as an alternative approach to avoid the development of bacterial resistance. Regarding the antimicrobial effects mediated by quinones, they act on cell surface-exposed molecules, cell wall polypeptides, and membrane-bound enzymes of H. pylori. For instance, juglone is a promising inhibitor of H. pylori growth because of its capacity to interfere with essential processes such as inhibition of 3 key H. pylori enzyme activities: cystathionine γ-synthase (HpCGS), malonyl-CoA acyl carrier protein transacylase (HpFabD), and β-hydroxyacyl-ACPdehydratase (HpFabZ) [H. pylori activity of several quinones, including juglone, menadione, and plumbagin, has been shown by MIC90 values around 0.8–25 μg/mL [Wolinella succinogenes, a target closely related to QFRs from H. pylori [H. pylori activity [H. pylori by acting on bacterial thymidylate synthase [Phytochemicals that display antimicrobial activity may inhibit (HpFabZ) . The ant25 μg/mL . Meanwhi. pylori . Moreoveactivity , 33. Finsynthase .H. pylori agents. To this end, a series of 2- and 3-phenylaminojuglone-based substances was prepared from juglone to assess their anti-H. pylori activity. In addition, we evaluated the influence of stereoelectronic and hydrophobic parameters of these compounds on the anti-H. pylori activity.The aim of the study was to design new anti-Suspensions of 1,5-dihydroxynaphthalene , rose bengal , and water (150 mL) were exposed to green LEDs for 5 h while a gentle stream of air was bubbled through the solution. Thereafter, phenylamines 3 (1.5 mmol) were added and the solutions were stirred for 4 h at room temperature (RT). Work-up of the reaction mixtures followed by column chromatography over silica gel (3 : 1 petroleum ether/ethyl acetate) provided pure compounds 4–15 .−1. Proton nuclear magnetic resonance (1H NMR) spectra were measured at 400 and 300 MHz in a Bruker AM-400 and Ultrashield-300 spectrometers. Chemical shifts are expressed in ppm using TMS as an internal reference , and (J) coupling constants are expressed in hertz (Hz). Carbon-13 nuclear magnetic resonance (13C NMR) spectra were measured at 100 and 75 MHz in a Bruker AM-400 and Ultrashield-300, spectrometers. Silica gel (70–230 and 230–400 mesh) and TLC on aluminum foil 60 F254-supported silica  were used for the chromatography analytical columns and TLC, respectively.All reagents were of commercial quality and used without further purification. The melting points were measured in a Stuart Scientific SMP3 equipment. The IR spectra were obtained in a vector 22-FT Bruker spectrophotometer using KBr disks, and wavelengths are expressed in cm+. The first and the second halfwave potential values (EI1/2) of juglone and phenylaminojuglones, evaluated from the voltammograms obtained at a sweep rate of 100 mV s−1, are summarized in Calculation of lipophilicity (ClogP) and molar refractivity (CMR) was assessed by using the ChemBioDraw Ultra 11.0 software and the obtained values are shown in Cellulose acetate filters, sodium chloride, and bacto agar were purchased at Asahi Glass,  and JT Baker (Mexico), respectively. Metronidazole was from Sigma Aldrich . All other chemicals were ACS reagent grade. Stock solutions of juglone and its analogs were prepared by dissolving 50 mg of the compound in 1 mL of 100% DMSO. Solutions were sterilized by filtration through cellulose acetate filters .H. pylori 26695 (ATCC 700392), isolated from a United Kingdom patient with gastritis, was obtained from the American Type Culture Collection . Frozen stocks of H. pylori were recovered and routinely grown for 48 h at 37°C, 5.5% CO2, and 70 to 80% relative humidity on Trypticase soy agar plates (TSA) from Becton Dickinson  supplemented with 0.4% H. pylori selective supplement Dent , 0.3% IsoVitalex (Oxoid), and 5% horse serum from Thermo Fisher Scientific HyClone  [600) at pH 7.0 and subsequently diluted to a starting OD600 of 0.05. To measure the growth of H. pylori in liquid medium, a serial dilution was prepared, aliquots of the various dilutions were plated on Trypticase soy agar plates, and the number colony-forming units (CFU) was determined [21, USA) , 39. Fortermined .H. pylori (3 × 107 cells/mL) were inoculated in 5 mL of TSB and supplemented with a range of concentrations (0.0 to 1.0 μg/mL) of juglone or a derivative compound. After incubation at 37°C for 48 h with constant shaking at 250 rpm in a controlled atmosphere (5.5% CO2 and 70% relative humidity), bacterial growth was determined by turbidimetry at 600 nm or by counting colony-forming units on TSA plates [A plates , 42.μL aliquots were taken at the end of the incubation period to prepare serial dilutions in PBS. Aliquots of 10 μL from each of these dilutions were plated on TSA and incubated for 48 h at 37°C [From each of the experimental culture tubes described in the previous section, 100  at 37°C . The numμL of H. pylori suspension containing 3 × 107 cells/mL was evenly spread over the TSA plates with a metal handle loop. Then, three-millimeter diameter wells were made in the plates and 30 μL of a series of compound solutions was deposited in the wells (corresponding to 0 to 1 mg/well). After 48 h of incubation at 37°C, the diameter of the growth inhibition halos was determined.The procedure was performed as described by Rodríguez et al. . One hunμM of MDA equivalents. The prooxidant activity of some selected quinones is shown in The assay was based on TBARS method according to Halliwell et al. . Brieflyp value < 0.05 was set as statistically significant.All experiments were performed at least 3 times and groups were compared by ANOVA test using GraphPad Prism software . Two-way ANOVA test was used to analyze the dose-response curves. A 1H-NMR and 13C-NMR) and high-resolution mass spectrometry (HRMS). The location of the phenylamino substituents at the quinone nucleus in compounds 4–7 and 8–15 was determined by bidimensional nuclear magnetic resonance (2D-NMR) . Compared to the antibacterial effect mediated by juglone (HGI: 14.50 mm), 5 out of 12 phenylaminojuglones were more efficient than juglone with HGI values ranging from 22.25 to 32.25 mm. A clear representation of this inhibitory effect is unveiled when the antimicrobial activity of compounds based on molar amounts was compared. For instance, the HGI of 33.25 mm of 7 was obtained at 4.5 μM while the HGI of juglone (14.50 mm) and metronidazole (11.67 mm) were obtained at 9.2 and 9.35 μM, respectively. In other words, 7 reached a high inhibitory effect on H. pylori growth at half of the doses required by juglone and metronidazole whose effects were by far lower than 7.Juglone and most of its analogs (except 5 and 9) were more active on The C-H functionalization in the 1,4-naphthoquinone scaffold at either C-2 or C-3, like in the pairs 4/8, 5/9, 6/12, and 7/13, resulted in similar antibacterial activities as shown by their halo of inhibition. For instance, 4 and 8 have an HGI of 13.25 and 13.75 mm, respectively, and 7 and 13 have an HGI of 32.25 and 28.50 mm, respectively.2NPhSO2Ph-4′-NH2), displayed higher inhibitory activity on H. pylori growth than juglone at all the tested doses. Since dapsone may act against bacteria by inhibiting the synthesis of dihydrofolic acid [pylori activity of 14. Finally, arylaminojuglone 15 derived from 2 and benzidine (4-H2NPh-Ph-4′-NH2) showed a lower range of activity than juglone. It should be noted that amines 3 phenylamine, 2-methylphenylamine, 3-methoxyphenylamine, 4-methoxyphenylamine, 4-hydroxyphenylamine, 3,4,5-trimethoxyphenylamine, and benzidine were devoid of anti-pylori activity when added in the absence of juglone (data not shown).Compound 14, obtained by oxidative amination of 2 with dapsone (4-Hlic acid  it is li7) on H. pylori viability in liquid medium. To this end, TSB medium was supplemented with increasing concentrations of each compound and incubated for 48 h. Next, aliquots were removed and colony- forming units (CFU) were counted. Bacteria viability results are expressed as CFU/mL.Once determining the effect of quinone-derived compounds in solid medium , we investigated the effect of juglone and the more active phenylaminojuglone , compound 7 reduced dramatically the viability of H. pylori while juglone, at the same concentration, did not affect significantly the bacteria viability. Second, it is required to use 0.6 μg/mL of juglone in order to reach the inhibitory effect of 7 (0.2 μg/mL) on the growth of H. pylori (3-fold increase). In terms of molarity, as we have previously shown, such difference is even higher in favor to 7. Indeed, 0.2 μg/mL of 7 corresponds to 0.56 μM, while 0.6 μg/mL of juglone corresponds to 3.44 μM, a difference of 6-fold to obtain similar inhibitory effects.μg/mL. Compound 7 provoked a rapid and strong inhibition of H. pylori growth decreasing the CFU values from 4.95 × 106 to 5.15 × 102 30 minutes after incubation. In contrast, metronidazole slightly decreased the CFU values from 5.3 × 106 at the beginning of the incubation to 3.23 × 106 after 30 min. At the end of the 180 min of incubation, the CFU value for metronidazole was still high reaching 4.80 × 105 whereas 7 practically causes a total loss of the bacteria. It should be noted that in terms of molarity, 7 was tested at 2.25 μM while metronidazole was used at 4.7 μM, highlighting the ability of 7 as a potential anti-pylori molecule.H. pylori biological activity. Among the members of the series, five congeners were found 1.9- to 2.8-times more active than one standard therapeutic drug , a currently standard anti-pylori drug [Traditional medicine used by ancient cultures relies on the use of natural compounds with biological activity, which can be used as starting molecules to modify their structures for improving their pharmacological properties. The aim of this work was to synthesize a series of phenylaminojuglones with anti-ori drug , 50–52.pylori activity depends on the nature and location of the nitrogen substituents at the quinone nucleus of the juglone scaffold. Thus, insertion of the 3,4,5-trimethoxyphenylamino group at the 2 position in juglone, as in compound 7 (HGI: 32.25 mm), induced a strong effect on the antibacterial activity of the juglone scaffold (HGI: 14.50 mm). Conversely, the insertion of the phenylamino, 2-phenylamino, and 4-methoxyphenylamino groups, as in compounds 4 (HGI: 13.25 mm), 5 (HGI: 11.00 mm) and 6 (13.50 mm), causes decreasing effects on the antibacterial activity of the juglone scaffold. Inspection of μg/mL) correspond to 6 μM of compound 4, 4.5 μM of compound 7, and 9.2 μM of juglone, strengthening the assumption about the efficacy of compound 7.Even though the discovery of molecular mechanisms underlying the antibacterial effects of the phenylaminojuglones was beyond our objectives, we noted that their anti-pylori activity: HGI: 10.75 and 28.50 mm, respectively. It appears then that compounds with a significant hydrophilic degree will have a more pronounced antibacterial activity. Interestingly, it has been reported that the membrane surface of H. pylori is rather hydrophilic and it is negatively charged [By comparing data from  charged . This prE1/2 values for the first one electron transfer, corresponding to the formation of the radical-anions of compounds 4–15, are spread into a broad potential range from −790 to −450 mV. In addition, we noted that 7 out of 12 phenylaminojuglones have higher redox potential values (from −450 to −510 mV) than juglone (−517 mV). It is tempting to assume that the effect on redox potential by the insertion of nitrogen groups, such as PhNH-, in the 5-hydroxy-1,4-naphthoquinone (juglone) scaffold may be clearly predicted, but the situation is a little bit more complex. Indeed, it is reasonable to assume that the electron acceptor ability of these phenylaminojuglone will depend, in part, on the location of the nitrogen donor in the quinone core and on the extent of the conjugative effect of this group to the intramolecular hydrogen bond of the molecule. A similar situation by taking the 1,4-naphthoquinone scaffold has been discussed by Aguilar-Martinez et al. [It is to be expected that the redox status of the cellular system would be modulated by ROS. Since the ease of ROS generation through reduction of a quinonoid would depend on its electrochemical parameters, the redox potential of a quinone would influence its overall biological profile, which encompasses the functional, toxicological, mutagenic, and antitumor activities. With this view, the redox potential of these naphthoquinones was determined by cyclic voltammetry using acetonitrile, an aprotic solvent, which mimics the environment of the cell membrane . Figure z et al. .pylori activity. Indeed, when compared compounds 5 and 14, they have the same redox potential values (−495 mV) and similar lipophilia (1.65 versus 1.25) but different polarizability values. Accordingly, compound 14 with a high molar refractivity (115.63), it has a high anti-pylori activity (HGI: 22.25 mm). However, compound 7 has less lipophilicity, high polarizability and low redox potential compared to 12, and their HGI values were markedly different: 32.25 and 12.75 mm, respectively. All these results illustrate how difficult is to attribute a biological response to a given molecular descriptor. Interestingly, H. pylori infection, it should be noted that the administration of coenzyme q10 decreases mucosal inflammation in such patients [Regarding the influence of other molecular descriptors such as molar refractivity, it seems that high polarizability values enhance the anti-patients .pylori compound already active as soon as 30 min of incubation at a very low concentration (0.56 μM). When using at 4.5 μM (1.6 μg/mL), the calculated halo of growth inhibition for 7 was 32.25 mm. These preliminary results make 7 an interesting lead molecule modulated by other substituting groups and to conduct further assays.In conclusion, compound 7 is a promissory anti-


To compare the effects of baked flaxseed versus those who received a placebo on constipation symptom scores, weight, glycemic and lipid control in constipated patients with type 2 diabetes (T2D).2 received either 10 g of flaxseed pre-mixed in cookies twice per day or placebo cookies for 12 weeks. The constipation symptom scores, BMI, fasting plasma glucose (FPG), glycosylated hemoglobin (HbA1c), and lipid profile were determined at the beginning and end of 4, 8, and 12-week period. Constipation was evaluated with a stool diary (ROME III).In a single-blinded, randomized controlled trial, 53 constipated patients with T2D with body mass index (BMI) 20.5–48.9 kg/m2), FPG (− 26.7 mg/dl), cholesterol (− 37.3 mg/dl), triglycerides (− 10.4 mg/dl), LDLC (− 21.0 mg/dl), HDLC (4.7 mg/dl), cholesterol/ HDLC ratio (− 1.4 mg/dl) significantly decreased from baseline in the flaxseed group . The differences of absolute change of constipation symptom scores (2.46 vs. 0.41), weight (− 3.8 vs. 0.0 kg), BMI (− 1.5 vs.-0.1 kg/m2), FPG (− 26.7 vs.-1.9 mg/dl), >HbA1c (− 0.8 vs. 1.0%), cholesterol (− 37.3 vs. -10.4 mg/dl), LDLC (− 21.0 vs. -4.3 mg/dl), and HDLC (4.7 vs. -4.4 mg/dl) between the flaxseed and placebo groups were statistically significant . The compliance was good and no adverse effects were observed.After the 12-week intervention, constipation symptom scores (2.46), weight (− 3.8 kg), BMI (− 1.5 kg/mIn constipated patients with T2D, flaxseed cookies used as a snack may be a useful tool for decreasing constipation symptoms, weight, glycemic and lipid levels.IRCT20110416006202N2.irct.ir:  Type 2 diabetes (T2D) is an increasingly common chronic disease and a major health concern that has been expected to continue far into the future . ConstipFlaxseed (linseed) is a functional food that is a rich source of polyunsaturated fatty acid, mainly alpha linolenic acid, an omega-3 fatty acid, as well as soluble fiber, lignan precursors, and other substances that may have health benefits, whose mechanism of action has remained elusive , 12. HowTherefore, this single-blinded, randomized, placebo-controlled trial was designed to test the hypothesis that the 10 g flaxseed consumption per day, as compared to a placebo, would reduce constipation symptoms, body weight and improve the glycemic and lipid levels in constipated patients with T2D. The objective of this trial was to assess the beneficial effects of adding flaxseed to the normal diet for 12 weeks of intervention among patients with T2D and chronic constipation.The study was approved by the Isfahan University of Medical Sciences ethics committee , and was conducted in accordance with Good Clinical practice. The study protocol was registered at irct.ir as IRCT20110416006202N2.This is a single-blinded, parallel-design, randomized, placebo-controlled trial, conducted for 12 weeks. Fifty-four consecutive constipated patients with T2D attending outpatient clinics in Isfahan Endocrine and Metabolism Research Center affiliated to Isfahan University of Medical Sciences, Iran from 20 Jan. to 15 Oct. 2017 were included. Constipation was diagnosed by the Rome III criteria . A one-wThe ROME III  definitiA total of 375 patients with T2D were screened and 60 patients were recruited. Four patients declined to participate, and 2 patients had medical comorbidities and did not meet our study criteria. The 54 participants (10 (18.5%) men, 44 (81.5%) women) were assigned randomly and equally to one of two treatment groups. Of those randomized, one patient in the flaxseed group lost to follow-up and were not evaluated  who was not involved with the study conduct. Only one author (NS) was not blinded to subject allocation and did not participate in data analysis.2) was calculated as weight (kg) divided by the squared of height (m2). The physician defined T2D  (measured using the glucose oxidase method), glycosylated hemoglobin (HbA1c) (measured by ion-exchange chromatography), total cholesterol (TC), high-density lipoprotein cholesterol (HDLC), triglyceride (TG) (measured using standardized procedures), and low-density lipoprotein cholesterol (LDLC) , were cociation ) and allParticipants in both groups maintained a constipation symptom diary for 12 weeks of intervention and for 1 week run-in phase and a 4-week without treatment. Constipation was assessed with a 5-point Likert Scale  included bowel movement frequency, feelings of complete evacuation, use of digital maneuvers, stool consistency , straining during bowel movement, pain during bowel movement, and overall feeling of constipation by a previously validated stool and symptom diary . In addip-values. Paired-Student’s t-test was conducted to analyze the difference between the baseline and week 12 for each of the two treatment groups, flaxseed and placebo. We used the chi-squared or Fisher’s exact test to compare proportions. To adjust for baseline characteristics in the analysis to account for the imbalance of the two groups, a general linear model was used to compare age, gender, LDLC, and constipation symptoms-adjusted means. The results are expressed as the mean (SD), and P < 0.05 was considered statistically significant. All statistical tests were two-sided, and all analyses were done with SPSS software for Windows .Primary outcome measures included analysis of numerical values of constipation intensity according to global constipation symptom score at the beginning, 4, 8, 12 and 16 weeks after entry in flaxseed and placebo groups. Secondary outcome measures included analysis of the body weight, glycemic, and lipid control in the beginning, 4, 8, 12 and 16 weeks after entry in both groups. The sample size was calculated when the study was designed and was based on the comparison of two means. Assuming an SD for the treatment differences in global constipation symptom score of 2.5, as observed in other studies , we calcAll 53  patients who completed treatment were available for follow-up at 4, 8, 12, and 16 weeks. Except for the higher LDLC and some constipation symptoms in the flaxseed group than in the placebo group at baseline, the two treatment groups were generally matched with regard to age, gender, diabetes duration, weight, BMI, FPG, HbA1c, triglyceride, HDLC, and constipation symptom score (Tables P < 0.001 for flaxseed and P = 0.003 for placebo). Mean global constipation symptom score changes was different between flaxseed and the placebo group  .On the global constipation symptom survey, 2 (7.4%) patients who received placebo cookies and 11 (42.3%) patients who received flaxseed reported improvement of symptoms and rated their improvement as at least somewhat better (+ 2) when compared to baseline symptoms. The mean (SD) global constipation symptom scores after 12 week treatment were − 0.59 (1.4) and 1.08 (1.4) for placebo and flaxseed, respectively (P < 0.05). Comparing week 12 vs. baseline for placebo and flaxseed, stool consistency in flaxseed groups improved (P < 0.001), but not in placebo group (P = 0.307).When comparing placebo vs. flaxseed the stool consistency were different at 4, 8, and 12 weeks (P = 0.947 and P = 0.930). Mean weight loss and BMI changes from baseline was different between the flaxseed and the placebo group  (Table A significant decrease in body weight and BMI observed in the flaxseed group (P < 0.001), whereas no significant change was observed in the placebo group (P = 0.137). Mean FPG and HbA1c changes from baseline were different between flaxseed and the placebo group .FPG and HbA1c were reduced in the flaxseed group while no changes observed in the placebo group. In the flaxseed group, the HbA1c shows reduced about 0.8%, in which this reduction was not significant (P < 0.001), but not in the placebo group (P = 0.311). Mean cholesterol changes from baseline were different between flaxseed and the placebo group .Cholesterol levels decreased throughout the study period in the flaxseed group (P = 0.045), change from baseline were not different between the two groups .Although within-group analysis showed a significant decrease in the mean of triglyceride in both groups (P = 0.482). Mean changes from baseline in LDLC differed between the two groups .While LDLC was decreased in the flaxseed group , no changes observed in the placebo group (P = 0.011).While HDLC was increased in the flaxseed group (non-significant), no changes observed in the placebo group. Mean changes from baseline in HDLC differed between the two groups , but not in the placebo group (P = 0.332), change from baseline were not different between the two groups  . So, it could be argued that the treatment group “regressed to the mean”. However, we adjusted for baseline covariates to account for the imbalance of the two groups.Based on the partial therapeutic benefit obtained with flaxseed cookies, its ease of administration, and lack of major side-effects, these results suggest that flaxseed in the form of cookies is useful in the treatment of constipation symptoms as well as the control of weight, glycemia, and lipids in people with T2D. Accordingly; none of our studied patients had any complaints about the application of this therapy.In constipated patients with T2D, flaxseed consumption may relieve constipation symptoms as well as decrease body weight, glycemic and lipid levels. Patients with T2D should be encouraged to increase the fiber content of their diet. Flaxseed consumption may be a valuable dietary approach for the prevention and treatment of T2D. The encouraging results obtained in this trial highlight the need for a larger sample size and longer follow-up period, probably blinded, to confirm these findings and better understand the mechanisms by which flaxseed may relieve constipation in patients with T2D.


Matricaria chamomilla L. extract. The flow rate was set at 1.0 ml/min and the detection wavelength was kept at 350 nm. The calibration curves were linear in the range of 0.11–11.36 μg/ml for quercetin, 0.11–11.20 μg/ml for luteolin, and 0.11–10.60 μg/ml for apigenin, respectively. The intraday and interday precisions (RSD) were less than 8.32 and 8.81%, respectively. The lower limits of quantification (LLOQ) of the three compounds were 0.11 μg/ml. The mean recoveries for quercetin, luteolin, and apigenin were 99.11, 95.62, and 95.21%, respectively. Stability studies demonstrated that the three compounds were stable in the preparation and analytical process. The maximum plasma concentration (Cmax) was 0.29 ± 0.06, 3.04 ± 0.60, and 0.42 ± 0.10 μg/ml, respectively. The time to reach the maximum plasma concentration (Tmax) was 0.79 ± 0.25, 0.42 ± 0.09, and 0.51 ± 0.13 h, respectively. The validated method was successfully applied to investigate the pharmacokinetics study of quercetin, luteolin, and apigenin in rat plasma after oral administration of M. chamomilla extract.A simple and sensitive HPLC-UV method has been developed for the simultaneous determination of quercetin, luteolin, and apigenin in rat plasma after oral administration of Matricaria chamomilla L. (German chamomile), a member of the Asteraceae family, is native to southern and eastern Europe and cultivated also in countries of America and Asia [ M. chamomilla have led to the isolation of essential oil, flavonoids, terpenoids, and coumarins [ M. chamomilla extracts.and Asia –3. It haand Asia , 5. A diand Asia , 7, antiand Asia , anticanand Asia , analgesand Asia , antipruand Asia , antiulcand Asia , and acaand Asia . These pand Asia . Previououmarins –19. Amonoumarins , 21. Sevoumarins –25. Howe M. chamomilla extracts. It was expected that the results of this study would provide some references to the further pharmacological study of M. chamomilla.In this study, a simple and sensitive HPLC-UV method was first developed and validated for simultaneous determination of quercetin, luteolin, and apigenin in rat plasma. A compound, kaempferol, was selected as the internal standard . The met M. chamomilla was provided by the Xinjiang Medical University and authenticated by Prof. Chunsheng Liu (Beijing University of Chinese Medicine). All other reagents were of analytical grade.Reference standards of quercetin , luteolin , and apigenin  were purchased from Shanghai Standard Biotech Co., Ltd. . Kaempferol  was obtained from National Institutes for Food and Drug Control  and used as an internal standard (IS). Methanol (HPLC-grade) was obtained from Fisher (USA). HPLC-quality water was obtained using a Cascada™ IX-water Purification System .18 column  with a mobile phase consisting of methanol and 0.2% phosphoric acid  with a constant rate of 1.0 ml/min. The injection volume was 10 μl and the column temperature was set at 25°C. The wavelength was set at 350 nm for quantitative analysis; the proportion of components in the mobile phase was optimized to obtain a well separation of quercetin, luteolin, apigenin, and kaempferol (IS).All analyses were performed on a Shimadzu HPLC system, equipped with LC-20AT pump, a Shimadzu SCL-10A system controller, and a SPD-20A DAD-UV detector. The HPLC analysis was performed on an Agilent Eclipse XDB-Cμg/ml of IS. All solutions were protected from light and stored at 4°C.The concentrated stock solutions of quercetin, luteolin, and apigenin were prepared by dissolving the reference standards in methanol to final concentration of 0.11 mg/ml. For the assay of plasma samples, working solutions were prepared by appropriate dilution of the stock solution with methanol. A stock solution of IS was prepared in methanol and then further diluted with methanol to prepare the working internal standard solution containing 22.52 μg/ml, luteolin at 0.56, 2.80, and 8.96 μg/ml, and apigenin at 0.53, 2.65, and 8.48 μg/ml, respectively.QC samples were prepared in the same way as the calibration samples, representing three different level concentrations  of quercetin in plasma at 0.57, 2.84, and 9.09 μL rat plasma was transferred to a 1.5 mL plastic tube to which was added 10 μL IS (22.52 μg/ml) for processing. After being vortex-mixed for 30 seconds, the plasma sample was then extracted with 800 μl of ethyl acetate and centrifuged at 18000g for 10 min, where ethyl acetate acted as a deproteinization-extraction agent. The procedure of extraction was performed two times. The organic layer was dried at 40°C under nitrogen stream. The residue was reconstituted in 100 μL of methanol solution, vortexed for 1 min, and centrifuged at 18000g for 10 min. The supernatant transferred to a glass insert, and 10 μL of the sample was injected into the HPLC column for analysis. The method of sample preparation was also applied for determination of accuracy, precision, and recovery.An aliquot of 100 The method was developed and conducted according to FDA guidelines with respect to specificity, linearity, LLOQ, accuracy, extract recovery, and stability. M. chamomilla extract.The specificity study was to investigate whether endogenous constituents and other substances existing in samples will interfere with the detection of the analytes and IS. The specificity of this method was ascertained by comparatively analyzing blank plasma samples from six different sources of rats, corresponding to blank plasma spiked with the three analytes and IS and the plasma samples from the rats after oral administration of theY) of analytes to the IS versus the spiked concentrations (X) of analytes at least six-point calibration curves. The acceptance criterion for a calibration curve was a correlation coefficient (r) of 0.99 or better. The LLOQ, which was defined as the lowest concentration in the calibration curve with acceptable the relative standard deviation (RSD), was within ±20% and accuracy within 100 ± 20%.The linearity of each calibration curve was determined by plotting the peak area ratio  and thaw (room temperature) cycles, stored at −20°C for 30 days. They were considered stable when 85–115% of the initial concentrations were got. All stability testing QC samples were determined by using the calibration curve of freshly prepared standard samples.Male Sprague-Dawley rats (body weight 160 ± 20 g) were purchased from Beijing Vital River Laboratory Animal Technology . Rats were housed in a temperature- and humidity-controlled environment  and maintained on a 12-hour light/dark cycle for 3 days with free access to food and water before starting the experiment. The animal experiments were performed in accordance with the principles of the International Guide for the Care and Use of Laboratory Animals and were approved by the Committee on Animal Care and Usage of the Beijing University of Chinese Medicine (number SYXK 2011-0024). M. chamomilla  freshly prepared in 0.5% CMC-Na solution were orally administered to six rats. Blood samples (0.5 mL) were collected from the ocular fundus veins of rats before administration and after 10, 20, 30, and 45 min and 1, 1.5, 2, 4, 6, 8, and 12 h after oral administration of M. chamomilla extract. The plasma was immediately centrifuged at 3200g for 10 min to separate out plasma and then stored frozen at −20°C until analysis.The extracts ofThe pharmacokinetic parameters of quercetin, luteolin, and apigenin were calculated by Kinetica 4.4 software . Noncompartmental analysis was used to determine standard pharmacokinetic parameters of analytes. All the results were expressed as means ± standard deviation (SD) of six replicates.An appropriate IS will control variability in extraction and HPLC injection. In this study, several substances, such as chloromycetin, naringenin, genkwanin, and kaempferol were tested as internal standards. Among these, kaempferol has been chosen to be the most appropriate in the present analysis because it is stable and does not exist endogenously in plasma. Moreover, its retention time was suitable and it was well separated from quercetin, luteolin, and apigenin. Therefore, kaempferol was finally selected as the IS due to the relatively equal recovery, and similar polarity and retention time to the analytes.18 column  was investigated to give an adequate separation of quercetin, luteolin, apigenin, and IS. Different compositions of mobile phase systems  were examined and compared in order to obtain good chromatographic behavior. Finally, methanol and 0.2% phosphoric acid  were chosen, which had a satisfactory separation.An Agilent Eclipse XDB-CIn our experiment, liquid-liquid extraction (LLE) and protein precipitation were compared for the sample preparation. Initially, we used the protein precipitation method; 1 mL methanol (or acetonitrile) was added for protein precipitation. However, the results showed that the aimed compounds could not be isolated completely from the proteins and interfering peaks in rat plasma by protein precipitation, but could be isolated by LLE. Therefore, liquid-liquid extraction by ethyl acetate was chosen, which considerably reduced the sample processing time. This method is rapid, and extraction using ethyl acetate is simple without any loss of analytes.The developed method was validated according to FDA guidelines for the following parameters: specificity, linearity, precisions, accuracy, recovery, and stability. M. chamomilla extract are shown in This study confirms the specificity of this method. The specificity of the method towards endogenous plasma matrix was evaluated with plasma from six rats. Typical chromatograms obtained from a blank, a spiked plasma sample with the three analytes and IS, and 20 min plasma sample after an oral administration ofY is the peak-area ratio of analytes to IS and X is the plasma concentration of analytes, respectively. Three calibration curves for quercetin, luteolin, and apigenin were established at six concentrations over the range of 0.11–11.36, 0.11–11.20, and 0.11–10.60 μg/ml, respectively. The LLOQ for three compounds was 0.11 μg/ml, which are sensitive enough for the pharmacokinetic studies of these compounds in rats.The excellent linear relationships are shown in The intra- and interday precision and accuracy are demonstrated in The mean extraction recoveries and RSD for quercetin, luteolin, and apigenin from rat plasma were determined at low, medium, and high concentrations in Stability study showed that the concentrations of quercetin, luteolin, and apigenin were stable in plasma stored at 20°C for 24 h, −20°C for 30 days, and after three freeze-thaw cycles at low, medium, and high concentrations, respectively. All the data are summarized in Pharmacokinetics Study. In this experiment, the method described above was applied to measure the plasma concentrations of quercetin, luteolin, and apigenin in rats administered orally with M. chamomilla extract. The mean plasma concentration-time curves (n = 6) of the analytes are shown in t1/2), maximum plasma concentration (Cmax), time to reach the maximum concentrations (Tmax), and area under concentration-time curve (AUCt0– and AUC∞0–) calculated by the noncompartment model are presented in Cmax of 0.29 μg/ml. The area under the curve (AUC∞0–) was 3.88 μg h/ml. The plasma concentration of luteolin reached a maximum at 0.42 h after administration with an average Cmax of 3.04 μg/ml. The area under the curve (AUC∞0–) was 19.89 μg h/ml. The plasma concentration of apigenin reached a maximum at 0.51 h after administration with an average Cmax of 0.42 μg/ml. The area under the curve (AUC∞0–) was 5.03 μg h/ml.The plasma concentration of quercetin reached a maximum at 0.79 h after administration with an average  M. chamomilla extract, which exhibited anxiolytic and antidepressant activity. However, little data were available regarding the pharmacokinetic characterization of quercetin, luteolin, and apigenin in rats after oral administration of M. chamomilla extract. This is the first report of HPLC-UV quantitative assay for investigating the pharmacokinetics of M. chamomilla extract. There were no interferences from endogenous substances. Pharmacokinetic results showed that the three compounds reached a maximum at about 0.79 h, 0.42 h, and 0.51 h, respectively, indicating that they may be absorbed quickly after oral administration. However, the elimination half-life (T1/2) was about 13.60 h, 4.43 h, and 8.82 h, respectively, suggesting that the elimination of them may be slow in rats after oral administration f M. chamomilla extract.Quercetin, luteolin, and apigenin are the main active constituents present in the M. chamomilla extract. The pharmacokinetic results would be a suitable reference in clinical application of M. chamomilla.The present study described a simple, specific, and reliable HPLC-UV with IS method for the simultaneous determination of three flavonoids in blood samples. This is the first validated HPLC method for analysis of quercetin, luteolin, and apigenin in rat plasma, which was highly sensitive and accurate. The method has been successfully applied to pharmacokinetic studies of quercetin, luteolin, and apigenin in rats after oral administration of


L. monocytogenes growth on rocket and melon under different packaging conditions and storage temperature. For that purpose, rocket and melon were placed on Expanded Polystyrene (EPS) trays, sprayed with L. monocytogenes to a population of 4.5–5.0 log CFU·g−1, packaged using microperforated Oriented Polypropylene (OPP) film in either air or Microperforated Active Modified Atmosphere (MAMA)  including a Whatman paper containing the essential oil, without contact with the product, and stored at 0, 5, 10, and 15 °C. Application of lemongrass exhibited a bactericidal effect on enterococci and a fungistatic effect on yeast-mould populations but only during air storage of rocket. The former took place at all temperatures and the latter only at 10 and 15 °C. No effect on shelf life of both products was recorded. However, an important effect on the sensorial properties was observed; during the first 4–5 days of storage both products were organoleptically unacceptable. Regarding MAMA packaging, it affected only Pseudomonas spp. population resulting in a reduction of 1–2 log CFU·g−1 in both products.The aim of the present study was to examine the effect of lemongrass essential oil vapors on the dynamics of surface microbiota and  Listeria monocytogenes has been implicated as the causative agent in several foodborne outbreaks after consumption of ready-to-eat fruits and vegetables. Contamination of fresh produce is facilitated by the ubiquitous nature of the pathogen as it is widely distributed in soil surface, decaying vegetation, soil sewage, animal feces, river and canal waters, fertilizers, plants, and animals and it is known to survive in plant material for 10–12 years [12 years .Protection of fresh produce against pathogenic microorganisms needs to be carried out through treatments that do not preclude the food from bearing the term “fresh”, which suggests or implies that the food is unprocessed, it is in a raw state and has not been frozen or subjected to any form of thermal processing or any other form of preservation . TherefoThe inherent aroma and antimicrobial activity of EO’s are usually related to the concentration and chemical structure of their components and also to the interactions among the components that are affecting their bioactive properties . Their mCymbopogon citratus) is a tall perennial grass, widely cultivated in warm tropical and subtropical regions [Lemongrass  is widely used for fresh-cut produce preservation. The factors that determine the gas composition of a MAP include the respiration rate and the respiring area of the product, the storage temperature, Oals etc. ,29,30. Tals etc. .L. monocytogenes growth on rocket salad and melon packaged in air and MAMA conditions and stored at 0, 5, 10, and 15 °C.However, there is a lack in the literature concerning the effect of this type of packaging on the surface microbiota as well as foodborne pathogen dynamics, especially in the presence of essential oils that exert antimicrobial action. Therefore, the aim of the present study was to examine the effect of lemongrass oil vapors on surface microbiota and L. monocytogenes strain LQC 15257; belonging to serotype 4b, previously isolated from a strawberry sample was used throughout this study. The strain was in vitro sensitive to the vapors of lemongrass essential oil. Long-term storage took place at −20 °C in nutrient broth supplemented with 50% glycerol. Before experimental use, the strain was grown twice in Brain Heart Infusion broth  at 37 °C for 24 h. Inoculum preparation took place as follows: overnight culture (9 log CFU·mL−1) was centrifuged , washed twice with sterile saline, resuspended in the same diluent and used to inoculate rocket and melon samples at 4.5–5.0 log CFU·g–1 through serial dilutions.Cymbopogon citratus) was purchased by Kokkinakis Essans E.P.E. .The essential oil (EO) of lemongrass (Eruca sativa) and melon (Cucumis melo cultivar Honeydew) cubes (approx. dimensions 3 × 3 × 4 cm) were packaged as follows: 30 g of rocket or 50 g of melon were placed on Expanded Polystyrene (EPS) trays, sprayed or not with 0.5 mL of appropriately diluted pathogen population to obtain the desired final population (4.5–5.0 log CFU·g−1). Holes were punched in the EPS tray in order to allow the essential oil volatiles to reach as much surface of the product as possible. Then 100 μL of the essential oil was applied on a Whatman paper that was then placed outside of the tray, i.e. without any contact with the product. Packaging took place using microperforated, Oriented Polypropylene (OPP) film (O2 permeability: 8000 cm3/m2·24 h·atm) in either air or modified atmosphere  and stored at 0, 5, 10, and 15 °C.Rocket (2 & CO2) within the packaging was measured using a CheckMate 9900 O2/CO2, . Total aerobic mesophilic, yeasts-molds, Enterobacteriaceae, enterococci, Pseudomonas spp., lactic acid bacteria and L. monocytogenes counts were determined according to Paramithiotis et al. [Sampling was performed every 24 h. The atmosphere  was used to statistically assess the effect of MAMA packaging and lemongrass essential oil’s addition to shelf life and microbial population dynamics.L. monocytogenes are presented.In 2 and 0.0% CO2. After the first day of storage, equilibration took place and the atmosphere within the packaging consisted of 18%–20% O2 and 0.00%–2.65% CO2 and was stable throughout storage. During storage in MAMA, rocket reached the limit of marketability on average after 2.5, 4, 9 and 12.5 days at 15, 10, 5, and 0 °C, respectively. The initial atmosphere within the packaging consisted of 5% O2 and 10% CO2. After the first day of storage, equilibration took place and the atmosphere within the packaging consisted of 17.65%–20.80% O2 and 0.15%–3.30% CO2 and was stable throughout storage. Application of lemongrass essential oil seemed to have no effect on the shelf life of the product stored in both atmosphere conditions.Rocket salad reached the limit of marketability on average after 2, 3, 10, and 10.5 days of storage in air at 15, 10, 5, and 0 °C, respectively. The initial atmosphere within the packaging consisted of 20.8% O−1. Enterococci population ranged between below enumeration limit to 3.20 log CFU·g−1, while Enterobacteriaceae and Pseudomonas spp. were enumerated at 3.20–4.40 and 3.90–5.00 log CFU·g−1, respectively. Lactic acid bacteria were below the enumeration limit and absence of L. monocytogenes was verified. At the end of storage period Pseudomonas spp. prevailed the microecosystem of rocket salads stored in air reaching populations of 7.97, 8.40, 8.28, and 8.98 log CFU·g−1 at 0, 5, 10, and 15 °C, respectively. On the contrary, Pseudomonas spp. co-dominated with yeasts during storage at MAMA mostly due to decrease in the Pseudomonas spp. population. More accurately, in all temperatures MAMA packaging resulted in a ca. 1 log CFU·g−1 decrease of the Pseudomonas spp. population.The initial microbiota of the rocket salad was dominated by yeasts-molds that ranged between 5.07 and 6.36 log CFU·g−1 without application of lemongrass essential oil. The same effect was observed regarding yeasts-molds growth at 10 and 15 °C; without lemongrass essential oil application the population reached 7 log CFU·g−1 but when the essential oil was applied the population would not exceed 5.5 log CFU·g−1. However, this was not the case regarding the application of the essential oil in MAMA packaged rocket salads; enterococci as well as yeast-mold populations were not affected. No significant effect on the growth of Enterobacteriaceae and Pseudomonas spp. in both air and MAMA packaged rocket salads was also observed when lemongrass essential oil was applied.Application of lemongrass in air packaged rocket salads seemed to have a bactericidal effect against enterococci. Indeed, when lemongrass essential oil was applied enterococci populations were below the enumeration limit; on the contrary, they reached a population range of 2–5 log CFU·gL. monocytogenes population remained stable at all temperatures throughout storage time.Finally, 2 and 0.0% CO2. After the first day of storage, equilibration took place and the atmosphere within the packaging consisted of 18.40%–19.90% O2 and 1.15%–2.60% CO2, during storage at 15 °C and 20.05%–20.80% O2 and 0.00%–0.65% CO2, during storage at 0, 5 and 10 °C that were stable throughout storage. During storage in MAMA, melon reached the limit of marketability on average after 4, 7, 9, and 15 days at 15, 10, 5, and 0 °C, respectively. The initial atmosphere within the packaging consisted of 5% O2 and 10% CO2. After the first day of storage, equilibration took place and the atmosphere within the packaging consisted of 18.45%–19.80% O2 and 1.10%–2.75% CO2, during storage at 15 °C and 19.10%–20.80% O2 and 0.00%–0.85% CO2, during storage at 0, 5 and 10 °C that were stable throughout storage. As in the case of rocket salad, application of lemongrass essential oil seemed to have no effect on shelf life of the product.Melon reached the limit of marketability on average after 4, 7, 9, and 13 days of storage in air at 15, 10, 5, and 0 °C, respectively. The initial atmosphere within the packaging consisted of 20.8% OPseudomonas spp. that ranged between 3.54–3.95 and 3.00–3.61 log CFU·g−1, respectively. Enterobacteriaceae, enterococci and lactic acid bacteria were below the enumeration limit and absence of L. monocytogenes was verified. At the end of storage period, Pseudomonas spp. prevailed with the microecosystem of melon stored in air reaching populations of 5.65, 7.50, 7.66, and 7.52 log CFU·g−1 at 0, 5, 10 and 15 °C, respectively. On the contrary, Pseudomonas spp. co-dominated with yeasts during storage at MAMA mostly due to a decrease in the Pseudomonas spp. population. More accurately, in all temperatures MAMA packaging resulted in a ca. 1 log CFU·g−1 decrease of the Pseudomonas spp. population.Melon initial microbiota was dominated by yeasts-molds and L. monocytogenes remained stable between 3.6 and 4.3 log CFU·g−1 during storage at 0 °C. However, at 5, 10, and 15 °C growth was observed and population of the pathogen ranged between 5.2 and 6.9 logCFU·g−1. In general, application of lemongrass essential oil seemed to have no effect in the microbial categories studied  and low (1900 cm3/m2 24 h·atm) oxygen transmission rate (OTR) air packaging on the freshness and sensory quality of wild rocket. It was concluded that high OTR films should be selected when storage temperature cannot be controlled as it allows sufficient aerobic respiration and prevents loss of leaf integrity and texture. In the present study, medium OTR was selected fulfilling this requirement, as indicated by the measurements of the atmosphere in the packaging. Both packaging conditions, i.e., air and initial displacement with modified atmosphere  resulted in comparable shelf life of the products under study, most probably due to the equilibration reached already from the first day of storage at all temperatures assessed. However, this displacement significantly affected the dominating surface microbiota as well as the effect of the lemongrass essential oil. In the first case, Pseudomonas spp. dominated the surface microbiota when air was used in both rocket and melon. On the contrary, the initial displacement resulted in co-domination with yeasts-molds. This co-domination resulted from the 1–2 log CFU·g−1 reduction of the Pseudomonas spp. population, compared to the respective in air. This reduction can be attributed to the initial displacement of air with increased CO2 concentration since Pseudomonas spp. are known to be among the most sensitive microorganisms to CO2 [Only a limited amount of studies currently exist regarding the effect of MAMA packaging on the shelf life, the dynamics of the surface microbiota as well as the survival of foodborne pathogens in fresh cut fruits and vegetables and, to the best of our knowledge, the first time that such study is performed in e et al.  studied s to CO2 .Acinetobacter baumanii, Aeromonas veronii, Aspergillus niger, Bacillus cereus, B. subtilis, Botrytis cinerea, Colletotrichum coccodes, Corynebacterium equii, Cladosporium herbarum, Enterococcus faecalis, Enterobacter aerogenes, Escherichia coli, Fusarium verticillioides, Klebsiella pneumoniae, Proteus vulgaris, Rhizopus stolonifer, Salmonella Typhimurium, Serratia marcesens, and Staphylococcus aureus, either by agar diffusion method or/and broth dilution [Fusarium verticillioides growth was not affected when lemongrass oil was added in culture medium. Moreover, the essential oil of C. citratus was reported as more effective than synthetic fungicides like Agrosan GN, Dithane M-43, and copper oxychloride [Several studies have reported antimicrobial activity of lemongrass against fungi and bacteria such as dilution ,43,44,45dilution  reportedchloride ,46.per se, there are much fewer reports on essential oil vapors and particularly on lemongrass vapors. In vitro studies on tomato fruits have indicated complete growth inhibition of Bacillus cinerea and Alternaria arborescens by lemongrass vapors. Moreover Geotrichum candidum was reported as more sensitive to citral and citral-containing oil vapors than to thyme and oregano oils. On the other hand, no inhibition was reported for Rhizopus stolonifer [Although there have been many reports of the antifungal and antibacterial effects of essential oils olonifer .in vitro studies do not necessarily indicate how effective an EO treatment will be when applied in food systems [et al. [Bacillus amyloliquefaciens in combination with lemongrass oil in a pad delivery system within a biodegradable MAP. By combining B. amyloliquefaciens with lemongrass oil, a reduction of the amount of the latter in the delivery system was managed, also reducing the unpleasant odor and taste resulting from the oil.The main concern about essential oils’ use is their effect on the sensorial properties of the treated products. Despite the fact that MICs (minimum inhibitory concentrations) of the most active EOs are very low, they can still alter the organoleptic properties of the treated foods . In addi systems . Indeed, systems . This is systems  and shou [et al.  managed Listeria innocua, E. coli and Salmonella Enteritidis in apple, pear and melon juices at 35 °C [−1 in apple and pear juices and by 5 μL·mL−1 in melon and tryptone soy broth, highlighting the effect of the food matrix. Azarakhsh et al. [Lemongrass oil has also been found effective against at 35 °C . In the h et al.  studied L. monocytogenes population remained stable during storage in air packaged salads at 0, 5, 10, and 15 °C and MAMA packaged rocket salads stored at 0 °C. However, at 5, 10, and 15 °C growth of the pathogen was observed. This may be primarily attributed to the microperforated packaging that allowed the escape of essential oil vapors but was necessary in order to allow sufficient plant tissue respiration. In conclusion, the antimicrobial activity of lemongrass essential oil seemed to be affected by the food matrix and the storage conditions. Moreover, no effect on shelf life of rocket or melon was observed.In the present study, application of lemongrass in air packaged rocket salads seemed to have a significant bactericidal effect on growth of yeast-molds at 10–15 °C and in enterococci populations at all temperatures. Moreover, application of lemongrass essential oil seemed to have no effect in the microbial categories studied in rocket salad packaged in MAMA conditions and in melon packaged in both air and MAMA conditions.


CT scan with delayed phase protocol. Management is usually conservative and generally depends on the etiology, urinomas size, and the presence or absence of kidney failure or infections.Renal collecting system rupture is a rare manifestation of obstructive uropathy. The majority of cases are attributed to ureteric calculi and extrinsic ureteric compression. The optimal diagnostic image is contrast‐enhanced  Klebsiella pneumoniae. She underwent right double J‐stent insertion and had successful lithotripsy 2 weeks later.A 52‐year‐old Caucasian woman presented to the emergency department with acute‐onset right lower abdominal pain for 1 day. She reported a few days history of fever, chills, urinary frequency, and dysuria. Vital signs were noticeable for a temperature of 38.7°C, regular heart rate of 120 beats/min, blood pressure of 104/62, and respiratory rate of 19. Abdominal examination elicited tenderness to the right costovertebral angle. Physical examination otherwise was unremarkable. Laboratory investigations showed a white cell count (WCC) of 19.6 (Ref: 4.0–10.8 K/UL), neutrophil count of 96% (Ref: 36–75%), hemoglobin of 14.9 (Ref: 12.0–16.0 G/DL), normal kidney and liver function tests, as well as a negative pregnancy test. Urinalysis was positive for nitrite, leukocyte esterase (2+), and blood (3+). Urine microscopy showed WBC of 5–10/high‐power field and RBC of 40–60/HPF. Pelvic ultrasound showed normal appearance of uterus, endometrium, and both ovaries. However, a computed tomography (CT) scan of the abdomen showed a 7 mm calculus at the ureterovesical junction causing right hydroureteronephrosis and ruptured calyx (Fig. Calyceal rupture is a rare manifestation of obstructive uropathy None declared.BK: designed, planned, and wrote the manuscript and did the literature review. IK: designed, planned, and revised manuscript. SM: designed, planned, and revised the manuscript, and prepared the photographs. EA: designed, planned, and revised the manuscript and selected the photographs.