An imbalance between degradation and reconstruction of the aortic wall is one of the leading causes of acute aortic dissection (AAD). Vitamin D seems an intriguing molecule to explore in the field of AAD since it improves endothelial function and protects smooth muscle cells from inflammation-induced remodeling, calcification, and loss of function, all events which are strongly related to the aging process. We quantified 25-hydroxy vitamin D, calcium, parathormone, bone alkaline phosphatase, and osteocalcin levels in 24 elderly AAD patients to identify a potential pathological implication of these molecules in AAD. Median 25-hydroxy vitamin D  and calcium levels  suggested hypovitaminosis D and a moderate hypocalcemia. Thirty-eight percent of AAD patients had severe (<10 ng/mL), 38% moderate (10–20 ng/mL), and 24% mild 25-hydroxy vitamin D deficiency (20–30 ng/mL). A significant inverse correlation was observed between 25OHD and osteocalcin levels. All the other molecules were unchanged. A condition of hypovitaminosis D associated to an increase in osteocalcin levels is present in AAD patients. The identification of these molecules as new factors involved in AAD may be helpful to identify individuals at high risk as well to study preventing strategies. Increasing data suggest that vitamin D, in addition to its role in bone metabolism, may exert important extraskeletal effects. Among different tissues and organs, the cardiovascular system has been regarded as one of the main targets of vitamin D actions. Epidemiological studies indicated that serum 25-hydroxy vitamin D (25OHD) levels are inversely associated with atherosclerosis, hypertension, coronary artery disease, peripheral artery disease, and stroke as well as with most of the main traditional cardiovascular risk factors –7. On thRecent studies suggested the existence of an inverse relationship between 25OHD levels and the presence and size of aortic aneurysms and thoracic aortic dilatations –11. The An imbalance between degradation and reconstruction of the aortic wall is also one of the leading causes of acute aortic dissection (AAD). AAD is an acute event caused by the formation of a tear within the artery wall and the creation of a false lumen within aortic wall between the intima and the media. It is associated with high morbidity and mortality if undiagnosed or not properly treated and is mainly characterized by cystic medial necrosis, elastic layer degradation, smooth muscle cell apoptosis, and inflammation . Any conToday, no studies assessed the vitamin D status in AAD patients. Thus, our aim was to evaluate whether vitamin D and vitamin D-related bone markers, which have also been related to metabolic and cardiovascular diseases, namely, osteocalcin (OC), parathormone (PTH), and bone alkaline phosphatase (BAP) , 21, areThis is a retrospective investigation study of 44 consecutive AAD patients who attended the I.R.C.C.S. Policlinico San Donato from 2005 to 2012 and were included in the International Registry of Aortic Dissection (IRAD) , accordi2) was calculated.AAD were repaired using deep or moderate hypothermic circulatory arrest and selective antegrade cerebral perfusion. All patients were managed with open distal anastomosis. Height, weight, medication intake, and risk factors were recorded. Body mass index . The lower detection limit was 4 ng/mL; the intra- and interassay coefficients of variation (CVs) were 3.7 to 7.7% and 5.8 to 10.9%, respectively. BAP was quantified by a direct, 2-site sandwich-type immunoluminometric assay utilizing two monoclonal antibodies . The lower detection limit was 1.5 μg/L; the intra- and interassay CVs were 3.2 to 4.0% and 6.5 to 8.1%, respectively. A direct, 2-site, sandwich-type immunoluminometric test which utilizes directly coated microparticles was used for OC detection . The lower detection limit was 0.5 ng/mL; the intra- and interassay CVs were 3.0 to 8.0% and 4.0 to 9.0%, respectively. 1–84 PTH was quantified with a 2-site, sandwich-type immunoluminometric test using directly coated microparticles . The lower detection limit was 4.0 pg/mL; the intra- and interassay CVs were 3.0 to 5.9% and 5.5 to 9.0%, respectively.Serum sample was obtained after centrifugation at 1000All analyses were performed using the LIAISON Analyzer (DiaSorin). Calcium was quantified by a colorimetric method using Vitros 5600 System . The lower detection limit was 4.0 mg/dL; the intra- and interassay CVs were 0.04 to 0.12% and 0.9 to 1.9%, respectively.T-test, for normal variables, or Mann–Whitney U-test, for nonparametric variables, was used to compare two groups. Chi-square test was used for categorical outcomes. For multiple comparison (three groups), the Bonferroni correction was used. Pearson  or Spearman  correlation tests were used to test the univariate association between variables. All statistical analyses were performed using GraphPad Prism 5.0 biochemical statistical package . A p value <0.05 was considered significant.The normality of data distribution was assessed by the Kolmogrov-Smirnoff test. Data were expressed as mean ± standard deviation (SD), median (25th–75th percentiles), or number and percentage. The demographic, anthropometric, and clinical characteristics of patients enrolled in the study are reported in p < 0.05 for 25OHD < 10 ng/mL versus 25OHD > 20 ng/mL) at decreasing 25OHD level.Relative to the other vitamin D-related bone turnover markers, OC levels (17.95 ± 8.1 ng/mL) as well as BAP (7.39 ± 3.2 mg/L) and PTH  were within the ranges of normality. After AAD patient classification according to 25OHD status, we observed a significant increase in OC concentrations (p < 0.05), whereas no differences were observed in the levels of all the other markers (data not shown). Univariate correlation analysis clearly showed the existence of an inverse association between 25OHD and OC . No correlation was observed with the other parameters, instead.Differently, statistically significant differences were observed neither in BAP and PTH levels among groups  nor in ap < 0.01 for 25OHD < 10 ng/mL versus 25OHD > 20 ng/mL; p < 0.05 for 10 ng/mL < 25OHD < 20 ng/mL versus 25OHD > 20 ng/mL). Anyway, univariate correlation analyses with 25OHD and the other parameters of calcium metabolism did not reach the statistical significance .Mean calcium levels suggested a mild condition of hypocalcemia in AAD patients. As expected, calcium levels decreased at decreasing 25OHD levels (AAD patients were further classified according to dissection localization. Levels of 25OHD and OC were, respectively, higher and lower in type B compared to type A patients, although just over the limit of statistical significance . None ofThe main observation of our study is that AAD patients displayed hypovitaminosis D and there was an inverse relationship between 25OHD and OC levels. Otherwise, no changes in the concentrations of the other bone-related molecules, according to vitamin D status, have been observed in these patients.Adequate vitamin D levels are first of all important for calcium absorption and thus the regulation of calcium homeostasis. Since low concentration of 25OHD is one of the main cause of hypocalcemia, it is not surprising to observe a trend of decrease in calcium concentration according to the vitamin D status of AAD patients. Otherwise, we did not observe an increase in PTH secretion as a compensatory mechanism to keep calcium concentration normal, as well as any significant correlation between calcium and vitamin D levels. Since PTH secretion is promoted by a decrease in free calcium level, it is possible that, despite the decrease in total calcium level, free calcium did not change. Unfortunately, in the present study, we could measure total but not free calcium.The unchanged levels of the bone turnover marker BAP seemed also to suggest the lack of activation of bone-related compensatory mechanisms in the regulation of calcium homeostasis in AAD patients. BAP is mainly produced by ostoblasts and plays important roles in bone matrix calcification . IncreasIn the present study, we also evaluated another bone matrix protein, OC, a vitamin D-related product which is released by osteoblasts and has potential role not only in osteoblast-osteoclast interaction and bone resorption but also in metabolism. Concerning OC, we observed the existence of an inverse correlation between this molecule and vitamin D. The fact that we could not observe any association between hypovitaminosis D and bone turnover did not exclude the possibility that reduced vitamin D levels may have consequences on vascular and cardiovascular health. Patients evaluated in this study, in fact, displayed an acute vascular event which was the result of inflammation-related processes and alteration in the composition of the media tunica of the vascular wall. Recent reports indicated that OC is associated to arterial diseases, thus being a molecule able to exert important effects also on vascular tissues which is similar to bone tissue in term of regulation of mineral homeostatsis . In addiThis has been mainly observed in the presence of inflammation  that, asIn our study, we also compared the type of AAD according to dissection localization. Usually, type A AAD, that is, at ascending aorta, is associated to worsen prognosis. By comparing the two types, we could not observe any difference in the levels of the evaluated molecules, although in type B patients, levels of 25OHD were higher and OC lower compared to type A, but at the limit of the statistical significance, probably due to the reduced number of type B patients.Our study suggests that in AAD hypovitaminosis D is not associated to changes in bone-related metabolic pathways but is inversely related to OC which could be an interesting molecule able to mediate the effect of inadequate 25OHD level at vascular level. Future studies exploring local vascular mechanisms of vitamin D and OC could be helpful to better highlight the role of these molecules in this disease. Whether maintaining an optimal vitamin D status could be a preventing strategy needs also further analyses.


With improved survivorship, the prevalence of breast cancer-related lymphedema (BCRL) continues to increase, leading to impairment of a patients’ quality of life. While traditional diagnostic methods are limited by an inability to detect BCRL until clinically apparent, bioimpedance spectroscopy (BIS) has been shown to detect subclinical BCRL. The purpose of this study is to evaluate the role of BIS in the early detection of BCRL, as well as assessment of response to BCRL treatment.n = 326) underwent preoperative and postoperative L-Dex measurements. Patients were identified as having subclinical lymphedema if they were asymptomatic and the L-Dex score increased >10 U above baseline and were monitored following treatment. Patients were stratified by lymph node dissection technique [sentinel lymph node biopsy (SLNB) vs. axillary lymph node dissection (ALND)] and receipt of BCRL treatment.A retrospective review of 1,133 patients treated between November 2008 and July 2013 at two surgical practices was performed. Eligible patients (n = 9) and 26.7% for ALND (n = 31). Of those diagnosed with BCRL, 50% resolved following treatment, 27.5% underwent treatment without resolution, and 22.5% had resolution without treatment. The prevalence of persistent, clinical BCRL was 0.5% for SLNB and 8.6% for ALND.The average age of the cohort was 56.2 years old, and mean follow-up was 21.7 months. Of the 326 patients, 210 underwent SLNB and 116 underwent ALND. BCRL was identified by L-Dex in 40 patients (12.3%). The cumulative incidence rate of subclinical lymphedema was 4.3% for SLNB (This study demonstrates both the feasibility and clinical utility of implementing L-Dex measurements in routine breast cancer care. L-Dex identified patients with possible subclinical BCRL and allowed for assessment of response to therapy. Breast cancer represents the most common non-cutaneous cancer among women and as such, treatment paradigms and survivorship strategies continue to evolve . Over thIncidence rates from BCRL vary widely with rates of 0–94% reported in the literature Table  based onAt this time, the importance of early detection and subsequent early intervention has been demonstrated with the publication of a prospective trial as well as several other studies demonstrating improvements in patient outcomes with intervention at earlier phases of BCRL , 21–26. A retrospective chart review of 1,133 breast cancer patients treated at two surgical practices between November 2008 and July 2013 was performed with Institutional Review Board approval given by the Western Institutional Review Board. Inclusion criteria included (1) some form of axillary staging [sentinel lymph node biopsy (SLNB) or axillary lymph node dissection (ALND)], (2) preoperative L-Dex assessment, and (3) a minimum of two subsequent L-Dex assessments. Exclusion criteria included (1) bilateral axillary surgery, (2) previously documented diagnosis of BCRL, (3) pregnancy, and (4) implanted electronic cardiac device, such as a pacemaker. A total of 326 patients meeting such criteria were identified and represent the cohort upon which the analysis was performed. Surgical technique, axillary sampling technique, and L-Dex assessments were available but body mass index (BMI), utilization of systemic and radiation therapy was not.With regard to assessment technique, L-Dex readings were taken using the L-Dex U400. Measurements were taken with patients lying supine on a non-metallic surface utilizing a standardized technique as demonstrated in Figure Patients were prospectively monitored preoperatively and assessed every 3 months within the first 2 years after surgery, in accordance with recommendations from Stout Gergich et al. , unless A total of 326 women met the selection criteria and were evaluated for this study. The mean age of the cohort was 56.2 years old with a mean time from preoperative L-Dex assessment to first postoperative assessment of 5.4 months. Mean overall follow-up was 21.7 months (range: 3.7–54.0 months). For the entire cohort, 155 patients (47.5%) underwent lumpectomy and 171 (52.5%) underwent mastectomy. Two hundred ten patients underwent SLNB  and 116 ALND . Table n = 40) with a rate of 4.3% (n = 9) for those undergoing SLNB and 26.7% (n = 31) for those undergoing ALND. For those undergoing SLNB, the mean time to positive L-Dex assessment was 5.6 months (2.3–13.6 months) from preoperative assessment and mean follow-up from subclinical BCRL diagnosis of 15.7 months (0.6–43.7 months). For those undergoing ALND, the mean time to positive assessment was 7.5 months (1.3–26.1 months) from preoperative assessment and mean follow-up from subclinical BCRL diagnosis of 13.4 months (0–18.1 months). Among the 286 patients not diagnosed with BCRL, 30 patients used prophylactic garments when performing perceived high risk activities, such as air travel with no patient progressing to BCRL.Breast cancer-related lymphedema status based on L-Dex score, treatment, and outcomes, stratified by axillary surgery are displayed in Table Of the 31 patients diagnosed with BCRL who underwent treatment , 20 (64.5%) had resolution of their BCRL at the end of the study. Nine patients  diagnosed with BCRL did not undergo treatment with all demonstrating resolution of their BCRL. Seventeen SLNB patients and 13 ALND patients, whose L-Dex scores were considered normal but showed an increasing trend coincidently with self-report of minor symptoms, were treated conservatively with a compression sleeve with no patient progressing to BCRL at last follow-up. Nine (seven ALND and two SLNB) patients had their L-Dex score resolve spontaneously without treatment . Further analysis of these subjects shows that for eight of these patients, positive assessment occurred within 9.1 months of initial surgery (mean = 4.3 months) and was resolved by the next visit .The results of this study demonstrate several key findings: 1) L-Dex was incorporated into routine breast cancer clinical practice and was used as part of routine follow-up care, (2) L-Dex scores identified patients in need of BCRL intervention, and (3) L-Dex was able to identify an improvement in BCRL following treatment. The incidence of BCRL for all patients in the present study, regardless of treatment type, was 12.3% . The incidence rates reported in the literature for all cancer treatments varies from 0 to 94%. These rates are primarily based on clinically evident BCRL measurable by tape circumference or water displacement. The cumulative incidence of BCRL in this cohort is lower than that expected from the literature . Additio L-Dex waThe changing management paradigm for BCRL is based on earlier detection and early intervention in order to prevent the chronic sequelae of BCRL that are irreversible , 5. As sThe low chronic incidence rates demonstrated in the present study suggest that early detection is integral in the management of patients at risk of BCRL. By comparing ALND patients monitored with L-Dex technology with a control group monitored with tape measurements only, Soran et al. showed the importance of using L-Dex technology in early subclinical detection of BCRL and early intervention . The incOur result demonstrated that a subset of patients had spontaneous resolution of BCRL without treatment. This is consistent with Kilbreath et al. who found swelling in the first year after breast cancer treatment could possibly be transient due to the effects of exercise, surgery, anesthesia, and/or taxane therapy . CurrentThere are limitations to the current analysis. This was a retrospective review and therefore, subject to the limitations of such an analysis. While the initial cohort was large, due to the small number of events, further data are required to validate these findings. Additionally, we were unable to evaluate other factors associated with an increased risk of lymphedema  due to limits on the data available. Finally, because intervention was based on clinician discretion, no cut point for beginning intervention could be determined at this time. However, this study represents one of the few studies available that demonstrate the ability to use L-Dex as part of routine clinical breast care to identify subclinical BCRL and allow early intervention to prevent long-term chronic BCRL.The results of this retrospective study demonstrate that L-Dex assessments can be incorporated into routine breast cancer programs as part of follow-up. This is critically important given the recent changes in the NCCN survivorship guidelines for post-treatment follow-up care for breast cancer patients establishing that health-care providers “educate, monitor, and refer for lymphedema management.” Additionally, the analyses suggest that L-Dex assessments can identify subclinical BCRL and subsequently monitor the return to baseline following conservative interventions. Further studies are required to demonstrate the long-term benefits of early detection and subsequently early intervention predicated upon subclinical detection of BCRL.AL contributed a portion of the retrospective patient data from her patient pool, analyzed all data, formulated a hypothesis, and was the primary author of this manuscript. BA contributed the remaining retrospective patient data from her patient population, assisted in formulating the hypothesis, and contributed significantly to the writing of this manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Enterococcus durans (50), En. faecium (15), En. faecalis (6), En. italicus (3), and En. hirae (1) were evaluated. The results revealed that ciprofloxacin resistance and biofilm formation are correlated with isolates originated from Golija mountain (Serbia), while gelatinase activity was more common in isolates from Prigorje region (Croatia), pointing to uncontrolled use of antibiotics and anthropogenic impact on dairy products' microbiota in these regions. The virulence genes were sporadically present in 13 selected dairy enterococci isolates. Interestingly, biofilm formation was correlated with higher ability of strains to reduce the adhesion of E. coli and Salmonella Enteritidis to HT29-MTX cells. To our knowledge this is the first study reporting the presence of the esp gene (previously correlated with pathogenesis) in dairy enterococci isolates, mostly associated with the genes involved in adhesion property. Hence, the results of this study revealed that the virulence genes are sporadically present in dairy isolates and more correlated to adhesion properties and biofilm formation, implicating their role in gut colonization rather than to the virulence traits.Enterococci have controversial status due to their emerging role in nosocomial infections and transmission of antibiotic resistance genes, while some enterococci strains are used as probiotics for humans and animals and starter cultures in dairy industry. In order to improve our understanding of factors involved in the safe use of enterococci as potential probiotics, the antibiotic susceptibility, virulence and probiotic traits of 75 dairy enterococci isolates belonging to  Enterococcus faecalis and En. faecium are found to be the most frequent enterococci species in human infections and represent the third most frequently isolated multidrug resistant nosocomial pathogens from bacteraemia . On the other hand, enterococci are commensals in gastrointestinal tract (GIT) as well as important constituents of traditional Mediterranean food of animal origin and contribute to their characteristic taste and flavor  suggested the anthropogenic impact on the spread of antibiotic resistance genes among enterococci in urban surface waters that is most likely attributed to the uncontrolled use of antibiotics  containing 5% (v/v) defibrinated horse blood after 48 h of incubation at 37°C , gentamicin (16–32 μg/ml), kanamycin (512–1024 μg/ml), streptomycin (64–128 μg/ml), clindamycin (2–4 μg/ml) and tylosine (2–4 μg/ml). Microdilution tests were performed in GM17 . Cell density was monitored after 24 h incubation at 37°C at 595 nm using Plate Reader Infinite 200 pro (MTX Lab Systems).Minimal inhibitory concentrations (MICs) were determined by microdilution testing following CLSI,  criteriaThe total DNA of 13 antibiotic-susceptible and without haemolytic and gelatinase activity enterococci strains was used in PCR reactions to detect the presence or absence of genes for virulence determinants. The primer sequences of the target genes, the expected amplicon sizes and annealing temperatures are given in Table Production of antimicrobial compounds produced by enterococci was tested by the deferred antagonism method using various indicator strains  in 0.01% HCl. The wells were then incubated at 37°C overnight. Adsorption of the dissolved formazan crystals was measured using Plate Reader Infinite 200 pro at 570 nm. The results were expressed as the percentage of optical density (metabolic activity) compared to the control (cultures of non-treated cells), used as 100% as follows:The cytotoxicity of enterococci on HT29-MTX cell line was evaluated through a microculture tetrazolium  assay Mosmann, . The colMetabolic activity (%) = (O.D. of treated cells/(O.D. of non-treated cells) × 100.in vitro test as described previously  was washed and suspended in PBS (pH 7.0) and added to the wells. Plates were incubated at 37°C for 2 h. After incubation, wells were washed three times with 200 μl PBS to remove unbound bacteria. Another 200 μl of 0.5% (v/v) Triton X-100 (Sigma) was then added to isolate attached bacteria. The viable cell count expressed as CFU/ml was determined in all cases by plating on GM17. Percentage adhesion was calculated from the viable counts adherent to the mucin with respect to the initial counts (%) =  × 100.Mucin adhesion assays were performed in 96-well polystyrene microtitre plates  according to the method of  = (CFU/mL adhered bacteria/CFU/mL added bacteria) × 100.For adhesion experiments, 13 ± 1 day-old cellular monolayers of HT29-MTX were used according to Živković et al. . CellulaE. coli ATCC25922 and Salmonella Enteritidis 654/7E to adhere to the intestinal epithelium in the presence and absence of enterococci was tested according to Živković et al. (E. coli or a combination of E. coli and enterococci (ratio 1:1) and Salmonella Enteritidis or a combination of Salmonella Enteritidis and enterococci (ratio 1:1) were independently added to the HT29-MTX monolayers at a ratio of 10:1, (bacteria: eukaryotic cell) and incubated at 37°C, with 5% CO2 for 1 h. The percentage of adhesion was calculated as follows: 100 × CFU/mL bacteria adhered to HT29-MTX monolayers / total CFU/mL bacteria added (adjusted for dilution). Numbers of bacteria were determined via viable cell count expressed as CFU/ml determined by plating on GM17. To determine the capability of the enterococci to inhibit the adhesion of E. coli and Salmonella Enteritidis to HT29-MTX monolayers, data were referred to that obtained with the E. coli and Salmonella Enteritidis alone .The capability of reference strains ć et al. . The bacp ≤ 0.05 was here considered statistically significant. Data were presented as the mean ± standard deviation of three independent replicates. Rules of association between geographic origin of strains and different phenotypic characteristics were mined as described in Hahsler et al.  and En. italicus (1 out of 3), 20% En. durans (10 out of 50), and 6.7% En. faecium (1 out of 15). En. hirae strain did not exhibit haemolytic activity on blood agar plates after 48 h incubation , collagen adhesin (ace), cytolysin (cylA), enterococcal surface protein (esp), cell wall adhesins , gelatinase (gelE), hyaluronidase (hylN), serine protease (sprE), response regulator (fsrA), signaling peptide (fsrB), and histidine kinase (fsrC) (Table efaAfs gene and nine strains (69.2%) were positive for the agg gene. Seven strains (53.8%) were positive for the sprE gene, while six strains (46.2%) were positive for esp and efaAfm genes. Three strains (23.1%) were positive for the fsrA gene, while two strains (15.4%) were positive for the gelE gene. One strain (7.7%) was positive for fsrC gene, while fsrB gene was not detected.The 13 antibiotic susceptible strains were further tested for presence of the virulence genes encoding aggregation factor , bacterial toxin with β-haemolytic properties in humans, as well as the hylN gene, encoding hyaluronidase, degradative enzyme associated with tissue damage  locus  and Salmonella Enteritidis 654/7E (63.9%) , the agg gene (69.2%), the esp gene (46.2%), and the efaAfm gene (46.2%), and associated with the adhesion ability. Aggregation substance, encoded by the agg gene, is a pheromone-inducible surface glycoprotein involved in bacterial conjugation and facilitating plasmid transfer, as well as adhesion to an array of eukaryotic surfaces . Interestingly, the esp gene in these strains was mostly correlated with the agg, efaAfs and efaAfm genes involved in adhesion property of the strains and the sprE gene. Moreover, in En. durans strains BGPT2-84 and BGAL3-19 the presence of the esp gene was correlated with biofilm formation ability. The ace gene encoding collagen adhesin was not detected in any of the tested isolates.Although adhesion properties of pathogenic bacteria are more intensively studied, various cell-surface proteins in probiotic lactic acid bacteria (LAB) and bifidobacteria are shown to have adhesion properties and they have been associated with probiotic activity  Figure . InteresEn. italicus (66.7%), three out of six En. faecalis (50%), six out of 15 En. faecium (40%), and 19 out of 50 En. durans (38%) strains, respectively, while En. hirae strain was not able to form biofilm .On the other hand, while numerous studies reported the pathogen exclusion by lactobacilli probiotic strains , p = 9.4 × 10−8  and p < 0.001 (t-distribution).The correlation between the two trees (phylogenetic and phenotypic) is statistically significant based on Bakers gamma correlation coefficient (0.1039904), which suggests that clustering in phenotypic groups is still under strong influence of phylogenetic relatedness. Depending on the method used p values are In conclusion, to our knowledge, this is the first study combining the data on virulence genes, gelatinase production, haemolytic activity, antibiotic resistance, biofilm production and probiotic features of dairy enterococci isolates, in order to improve our understanding of factors involved in virulence vs. probiotic properties of dairy enterococci. Our results indicate that the virulence genes are sporadically present in tested dairy enterococci isolates and that are not correlated with cytotoxic activity, suggesting that adhesion and biofilm formation are more associated with gut colonization.KV, NG–conception and design of the study; NP–performed the main work; KV, AT-V, and JD–participated in the research—virulence traits, antibiotic susceptibility and cytotoxic effect, antimicrobial activity; KV, JD, and MD–participated in the research—gut colonization, adhesion properties and biofilm formation; SM, MT, SB, and NG–participated in the research—relationship between isolates' origins, phylogenetic relatedness and virulence traits; NG, KV, SM, JD, and MT–analyzed, interpreted and critically revised the data; NP, KV, NG, and SM–prepared the manuscript for submission. All authors finally approved the version to be published.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Sleep apnea is common in patients with atrial fibrillation, but the effect of the cardioversion of atrial fibrillation to sinus rhythm on central and obstructive apneas is mainly unknown. The primary aim of the study was to analyze the association between cardioversion of atrial fibrillation and sleep apneas, to investigate whether obstructive or central sleep apneas are reduced following cardioversion. A secondary objective was to study the effect on sleep quality.Twenty-three patients with atrial fibrillation were investigated using overnight polysomnography, including esophagus pressure monitoring and ECG, before and after the cardioversion of persistent atrial fibrillation.Obstructive sleep apnea occurred in 17/23 patients (74%), and central sleep apnea in 6/23 patients (26%). Five patients had both obstructive and central sleep apnea. Sinus rhythm at follow-up was achieved in 16 patients. The obstructive apnea-hypopnea index, central apnea-hypopnea index, and the number of patients with obstructive or central sleep apnea did not differ before and after restoration of sinus rhythm. Sleep time, sleep efficiency, time in different sleep stages, and subjective daytime sleepiness were normal and unaffected by cardioversion.Both obstructive and central sleep apneas are highly prevalent in patients with persistent atrial fibrillation. Obstructive sleep apneas are unaffected by the cardioversion of atrial fibrillation to sinus rhythm. The sleep pattern is normal and unaffected by cardioversion in patients with atrial fibrillation.Trial number NCT00429884. Atrial fibrillation is common and increases with age. It occurs in about 6% after the age of 55 and in 14% after the age of 80 ,3. ObstrThe effect of cardioversion of atrial fibrillation on obstructive and central sleep apnea is, however, mainly unknown. The primary aim of the study was to analyze the association between cardioversion of atrial fibrillation and sleep apneas, to investigate whether obstructive or central sleep apneas are reduced following cardioversion. A secondary objective was to study the effect on sleep quality.The study comprised 28 patients scheduled for elective cardioversion of persistent atrial fibrillation at the Department of Cardiology, Umeå University Hospital, without previously diagnosed sleep apnea. The Regional Ethics Committee at the medical faculty at Umeå University approved the study protocol, and written informed consent was obtained from all the participating patients. Five patients were subsequently excluded, due to severe heart failure, unstable angina pectoris, or withdrawal of consent.The patients were included two to four weeks prior to the cardioversion. A detailed medical history was obtained, a 12-lead electrocardiogram (ECG) was performed, and the patients underwent an echocardiography. Overnight polysomnography was performed within one week prior to the cardioversion. A follow-up polysomnography and ECG was done within one week after cardioversion.®) 1 mg/kg i.v. administered by an anesthesiologist. R-wave-synchronized mono- or biphasic anterio-posterior shocks were given in a step-up protocol at 200 J, 300 J, and 360 J to achieve sinus rhythm.Cardioversion was performed on an elective outpatient basis. Sedation was induced by propofol  was performed in hospital at 20 °C/68 °F and included continuous recordings of electroencephalograms , electro-oculograms, submental electromyograms, airflow with a three-port oro-nasal thermistor, respiratory effort from continuous esophageal pressure  and piezo-electric belts , finger pulsoximetry (Embla A10 flex Sensor), electrocardiograms (V5), and a body position sensor.Arterial samples for the analysis of blood gas parameters were taken in the supine position.All the recordings were scored manually. An obstructive apnea was defined as the cessation of airflow for at least 10 seconds with continuing abdominal and thoracic movements, according to the American Academy of Sleep Medicine . An obstThe Epworth Sleepiness Scale (ESS) was used to assess the degree of daytime sleepiness. The ESS is a self-administered, validated questionnaire with eight questions relating to the risk of falling asleep in different situation, with answers scoring from 0 to 3. Possible scores ranged from 0 to 24, and excessive daytime sleepiness was defined as an ESS score of ≥11 .t test was used for comparing normally distributed variables, and Wilcoxon’s signed rank test was used when comparing non-normally distributed variables. Proportions were compared using Fisher’s exact test or McNemar’s test for paired proportions. A P value of <0.05 was considered significant. Statistical calculations were performed with SPSS v 22 .Continuous variables were given as the means ± standard deviation or medians and interquartile range (IQR). Categorical variables were presented as percentages. A paired 2, and 19 patients (83%) had sleep apnea (AHI ≥5 events/h). Obstructive sleep apnea (obstructive AHI ≥5 events/h) occurred in 17/23 patients (74%) and central sleep apnea  in 6/23 patients (26%). Five patients had both obstructive and central sleep apnea. Twenty-two patients had a normal or slightly impaired left ventricular function, with a left ventricular ejection fraction above 45%.Twenty-three patients, 14 men and 9 women, were investigated before and after the cardioversion of atrial fibrillation . They weP = 1.0) (P = 0.5). Central sleep apnea occurred in 2/16 (12%) patients before cardioversion, and in 3/16 (19%) who were in sinus rhythm at follow-up (P = 1.0). Neither the obstructive apnea-hypopnea index  nor the central apnea-hypopnea index  changed when sinus rhythm was restored among patients who were in sinus rhythm at follow-up (P = 0.045).Twenty-one patients were converted to sinus rhythm, and 16 of the 23 patients (70%) were in sinus rhythm at follow-up. Among the 16 patients who were in sinus rhythm at follow-up the first polysomnography was performed median 5.5 days (IQR 2–11.2) before cardioversion and the second polysomnography median 4 days (IQR 2–9.2) after cardioversion, and sleep apnea occurred in 13 (81%) before cardioversion and in 14 (88%) after cardioversion (P = 1.0) . Obstrucollow-up . The proTotal sleep time, sleep in different sleep stages, sleep efficiency, sleep in a supine position, and daytime sleepiness according to the Epworth Sleepiness Scale did not change when sinus rhythm was restored .Sleep apnea was common in the present patients with atrial fibrillation, and 83% of the included patients had sleep apnea, although none of them had been diagnosed with sleep apnea previously. Obstructive sleep apnea occurred in 74% and central sleep apnea in 26% of our patients. At baseline, central sleep apnea was more common among patients with recurrence of atrial fibrillation at follow-up. The prevalence and the degree of sleep apnea were not affected by cardioversion. Sleep quality was normal at baseline and remained unaffected after the cardioversion.The high prevalence of obstructive sleep apnea in the present patients with atrial fibrillation is close to the findings by Braga et al. and Albuquerque et al. ,16. ObstCentral sleep apnea also occurs in patients with atrial fibrillation ,28. In tFox et al. recently reported that sleep apnea was reduced after restoration of sinus rhythm in 116 patients with atrial fibrillation . This reOther studies have addressed the question of whether restoring sinus rhythm reduces sleep apnea or not. Naruse et al. reported a decrease in the obstructive apnea-hypopnea index among 25 patients after radiofrequency catheter ablation of atrial fibrillation . They, hA strength of the present study was the use of polysomnography including EEG, and esophageal pressure monitoring, the gold standard to distinguish central from obstructive apneas and hypopneas. A limitation was the low number of included patients. A major limitation was the even lower number of patients with central sleep apnea, and we can therefore not exclude an effect of cardioversion on central apneas.In conclusion, both central and obstructive sleep apneas are common in patients with persistent atrial fibrillation. Obstructive sleep apneas are unaffected by the cardioversion of atrial fibrillation. The sleep pattern is normal and unaffected by cardioversion in patients with atrial fibrillation.


Obesity is known to be related to the development of type 2 diabetes mellitus (T2D). The most commonly used anthropometric indicator (body mass index [BMI]) presents several limitations such as the lack of possibility to distinguish adipose tissue distribution. Thus, this study examines the suitability of a body shape index (ABSI) for prediction of body composition and sarcopenic obesity in obese or overweight T2D subjects.1c], mean blood glucose, fasting plasma glucose [FPG], high-density-lipoprotein cholesterol [HDL], low-density-lipoprotein cholesterol, total cholesterol, and triglycerides [TG] levels; the ratio TG/HDL was also calculated as a surrogate marker for insulin resistance).Cross-sectional study in 199 overweight/obese T2D adults. Anthropometric  and body composition  data was collected, as well as metabolic parameters . Moreover, a significant association between ABSI and FPG was found in men.ABSI was significantly associated with age and waist circumference. It showed a statistically significant correlation with BMI exclusively in women. Regarding body composition, in men, ABSI was associated with FM (%), while in women it was associated with both FM and FFM. Both males and females groups with high ABSI scores were significantly older  and showed lower FFM values  compared with low-ABSI groups. Multiple linear regression revealed that ABSI independently predict FMI and the FM/FFM ratio in women. Sarcopenic obesity was identified in 70 (36.5%) individuals according to the FM/FFM ratio. The AUROC of ABSI was 63.1%  and an ABSI value of 0.083 mABSI could be useful to identify visceral and sarcopenic obesity in overweight/obese adults with T2D, adding some relevant clinical information to traditional anthropometric measures.The online version of this article (10.1186/s13098-018-0323-8) contains supplementary material, which is available to authorized users. According to the global trends, the prevalence of diabetes mellitus is expected to increase considerably , 2. ObesSome anthropometric measurements considered surrogates for visceral obesity have long been used in medical settings for obesity-associated health risk evaluation. A commonly used measure is the body mass index (BMI). However, it does not serve to distinguish between muscle and fat accumulation and gives no indication of body shape . An incrWaist circumference (WC) is widely seen as a measure of central adiposity. Its association with insulin resistance has been reported to be better than that of BMI –11. RelaKrakauer et al.  developeAdditionally, ageing is usually associated with an increase in FM coexisting with a decrease in FFM, leading to sarcopenia . SarcopeThe association of ABSI and body composition measurements has been reported in a geriatric study . OverweiHere we assess ABSI in an observational study of Caucasian obese or overweight subjects with T2D. Our findings suggest that ABSI is associated with body composition and sarcopenic obesity.This cross-sectional observational study was carried out at the Hospital General de Segovia  in accordance with the Declaration of Helsinki, including all amendments, and was approved by the corresponding Independent Research Ethics Committee. All participants provided written informed consent to use their data.2) adult subjects with T2D. A glomerular filtration rate over 60 ml per minute per 1.73 m2 of body surface area  and nor2). Waist circumference (WC) was measured at the uppermost border of the iliac crests in standing positions with a non-stretchable tape (cm). ABSI was calculated as WC/(BMI2/3 * height1/2), expressed in m11/6 kg−2/3. Men and women were each divided into two groups by using the median of individual ABSI measurements as the threshold value. Subjects with ABSI lower than the median values were assigned to the “lower-ABSI” group and subjects with greater ABSI than the median value were assigned to the “higher-ABSI” group.Height (m) and weight (kg) were measured according to standard methods, and the BMI was then calculated as weight/height squared (kg/m2), respectively. The ratio between FM and FFM (FM/FFM) was calculated as an index of sarcopenic obesity. As per Prado et al. [Fat mass (FM) and fat-free mass (FFM) were clinically determined by bioelectrical impedance analysis and calculated through the software supplied by the manufacturers and expressed in kg . These mo et al.  we used The collection of metabolic parameters included glycated haemoglobin (HbA1c), mean blood glucose (MBG), fasting plasma glucose (FPG), high-density lipoprotein cholesterol (HDL), low-density lipoprotein cholesterol (LDL), total cholesterol (TC), and triglycerides (TG) levels. The ratio between TG and HDL (TG/HDL) was also calculated as a surrogate marker for insulin resistance, as is suggested by Cordero et al. .To control for the well-known sexual dimorphism in body composition, all analyses were stratified by gender. Values were expressed as mean ± standard deviation (SD) for quantitative variables and as number and percentages for categorical variables. The Student’s t test or the Mann–Whitney U test for quantitative variables and the Fisher’s exact test or the Chi square test for qualitative variables were used to compare mean values between groups. Correlation analyses between anthropometric measures and body composition and metabolic parameters were evaluated with Pearson’s/Spearman correlation analysis. To determine if ABSI and BMI were independently associated with FM, FFM, FMI, FFMI, FM/FFM ratio, and TG/HDL index, linear regression analyses were performed. Correlations between anthropometric parameters and body composition measurements were adjusted for age, due to the documented relationship between the ageing process and changes in body composition, often without concomitant changes in body weight or BMI . The areBetween March 2012 and April 2016, we identified 203 subjects who met the initial selection criteria; of these, four subjects were excluded due to screening failures (n = 2) and incomplete data (n = 2). Thus, the evaluable population comprised 199 subjects (100 women and 99 men), whose characteristics are shown in Table According to the FM/FFM ratio, 70 (36.5%) subjects had sarcopenic obesity, as defined by an FM/FFM ratio > 0.8. Only 9 of the male subjects showed sarcopenic obesity.Pearson correlations between different variables are shown in Table 11/6 kg−2/3 . For BMI, the cut-off value was 37,45 kg/m2, with sensitivity of 81% and specificity of 77%.The ROC curves of ABSI and BMI for predicting sarcopenia are shown in Fig. 11/6 kg−2/3). Higher-ABSI men had a mean WC significantly greater than the lower-ABSI men group . The higher-ABSI male and female groups were significantly older . With higher-ABSI vs. lower-ABSI FFM was lower , demonstrating the ability of ABSI to identify sarcopenia. Among women, BMI, FM, FMI, FFM, and FFMI exhibited significantly greater values in the lower-ABSI group than the higher-ABSI group , in both males and females. The observed lack of correlation between ABSI and BMI in the general population was confirmed in the male cohort, while there was a modest correlation in the female cohort. This is in line with the recent study conducted by Hardy et al.  that aimABSI was positively correlated with WC both in males and females. A higher ABSI-value is an indication of a higher abdominal fat deposition  that leaThe higher-ABSI females and males showed significantly lower FFM than the lower-ABSI groups, with comparable BMI in men and lower BMI in women. These findings could support the hypothesis that abdominal fat deposition may be associated with a loss of skeletal muscle mass in some subjects, i.e. sarcopenic obesity. Obesity-related sarcopenia, is a syndrome of progressive and generalised loss of skeletal muscle mass and function, characterized by a higher FM in relation to FFM . Our stu11/6 kg−2/3) exactly agreed with the finding of a previous study [The AUROC analysis supports that ABSI showed ability for predicting the presence of sarcopenic obesity . Even tus study , where AThe data presented add to the emerging, growing body of literature regarding different aspects of ABSI. A recent longitudinal study conducted with Australian adults showed a positive association between ABSI and mortality, suggesting that this measure could be a useful predictor of mortality hazard in different populations . In thisThe authors recognise some limitations in the study that should be considered when interpreting the results. First of all, this is an observational study with a cross-sectional design; therefore, there is no patient follow-up, which would be of importance to elucidate the correlation between body composition and ABSI; besides, we cannot rule out that some degree of bias may have been introduced. Secondly, as this study was performed at a single centre, findings in other centres may be different and, certainly, further confirmation is needed. Nonetheless, all these findings should guide further prospective studies with some broader samples .In conclusion, ABSI identifies visceral obesity and sarcopenia in overweight or obese subjects with T2D, adding some relevant clinical information to traditional anthropometric measures.Additional file 1: Table S1. Linear regression analysis of ABSI and BMI with body composition measurements.


We observed a significantly increased frequency of the TT genotype of rs1234315 in BD patients . The frequency of the TT genotype of rs12039904 was significantly higher in patients with VKH disease as compared to controls . Analysis of clinical manifestations in VKH disease and BD did not show an association with the TNFSF4 gene polymorphisms. The study suggests that the TNFSF4 gene may be involved in the susceptibility to VKH disease and BD in Han Chinese.To investigate whether single nucleotide polymorphisms (SNPs) of the Tumor Necrosis Factor Superfamily 4 (TNFSF4) gene are associated with Vogt–Koyanagi–Harada (VKH) and Behcet’s disease (BD) in a Chinese Han population. A two-stage case control study was carried out in 1331 VKH, 938 BD and 1752 healthy controls. Ten TNFSF4 SNPs, including rs1234314, rs1234315, rs2205960, rs704840, rs2795288, rs844654, rs12039904, rs10912580, rs844665, and rs844644, were genotyped using the PCR-restriction fragment length polymorphism method. Genotype and allele frequencies were analyzed between cases and healthy controls using the X Vogt-Koyanagi-Harada (VKH) disease and Behcet’s disease (BD) are two of the most common uveitis entities in China, manifesting as bilateral panuveitis and can lead to serious visual impairment1245789101112Recently, several SNPs in a Tumor necrosis factor superfamily-4 (TNFSF4) have been shown to be shared risk factors for several immune-related diseases, including systemic lupus erythematosus (SLE), primary Sjögren’s syndrome (SS) and systemic sclerosis (SS)1415161718192021Previous studies have shown the important role of TNFSF4 in the development of autoimmune diseases. The association of TNFSF4 polymorphisms with BD and VKH disease has however not been investigated and was therefore the subject of the study presented here. Our results show that polymorphisms of rs1234315 of TNFSF4 are associated with susceptibility to ocular BD, whereas rs12039904 is associated with susceptibility to VKH disease in a Han Chinese population.The demographics and clinical characteristics of the enrolled BD and VKH disease patients in the study are displayed in a = 1.68 × 10−4, OR = 0.484, 95% CI = 0.352–0.664; Pca = 5.91 × 10−3, OR = 0.676, 95% CI = 0.564–0.809; respectively)  as compared to the control group   . To confol group . The com6–0.780) . We coul6–0.780) .b = 4.74 × 10−3, OR = 2.494, 95% CI = 1.532–4.060) was significantly higher than that in normal controls  as compared to normal controls  and the First Affiliated Hospital of Chongqing Medical University  were enrolled from April 2005 to June 2014 and were all Han Chinese. The investigated patients and healthy controls gave a written informed consent before collection of blood. This study was approved by the local ethics research committee at the Chongqing Medical University and was conducted according to the Declaration of Helsinki Principles.In the first-stage study of this case-control study, 394 BD patients, 395 VKH patients and 606 healthy controls were enrolled to identify susceptible SNPs (Pc < 0.05) in the TNFSF4 gene. In the second-stage, another 544 BD and 413 healthy individual, and another 936 VKH and 1166 controls were added to replicate the susceptible SNPs found in the first-stage study. The diagnosis of BD was based on the criteria of the International Study Group for BDGenomic DNA was extracted from blood samples by using the QIAamp DNA Blood Mini Kit .The proper primers of rs1234314, rs1234315, rs2205960, rs704840, rs2795288, rs844654, rs12039904, rs10912580, rs844665, and rs844644 for amplifying target DNA sequence by polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) are depicted in 2 test was used to check whether the data conformed to the Hardy-Weinberg (HWE) principle. Statistical analysis was performed using SPSS 17.0 software . The frequencies of genotypes and alleles were evaluated using a case-control study design and significance was tested using the Pearson X2 test. The patterns of linkage disequilibrium of the SNPs of TNFSF4 were compared using Haploview 4.0 . P values were corrected for multiple comparisons using the Bonferroni correction by multiplying obtained p values with the number of analyses performed. A Bonferroni corrected P values <0.05 was considered to be statistically significant.The XHow to cite this article: Lu, S. et al. Association of TNFSF4 Polymorphisms with Vogt-Koyanagi-Harada and Behcet’s Disease in Han Chinese. Sci. Rep.6, 37257; doi: 10.1038/srep37257 (2016).Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


Ocimum basilicum L. cv “Ceasar”) grown in controlled environments. This culinary herb produces an aroma highly appreciated by consumers, primarily composed of terpenes/terpenoids, phenylpropanoids, and fatty-acid- derived volatile molecules. Basil plants were grown under narrow-bandwidth light conditions, and leaf area, height, mass, antioxidant capacity and volatile emissions were measured at various time points. The results indicate reproducible significant differences in specific volatiles, and in biochemical classes of volatiles, compared to greenhouse grown plants. For example, basil plants grown under blue/red/yellow or blue/red/green wavelengths emit higher levels of a subset of monoterpenoid volatiles, while a blue/red/far-red treatment leads to higher levels of most sesquiterpenoid volatile molecules. Specific light treatments increase volatile content, mass, and antioxidant capacity. The results show that narrow-bandwidth illumination can induce discrete suites of volatile classes that affect sensory quality in commercial herbs, and may be a useful tool in improving commercial production.Narrow-bandwidth light treatments may be used to manipulate plant growth, development and metabolism. In this report LED-based light treatments were used to affect yield and metabolic content of sweet basil ( Plants constantly adapt their growth and physiology to best match ambient conditions. Light modulates plant growth throughout development, with effects on seed germination, seedling establishment, transition to flowering, and adaptation to stress responses . Basil is a highly valued horticultural crop that presents a complex aroma profile. It stands as an excellent system to test the hypothesis that spectral quality can control the critical sensory attributes of herbs, and generate new questions about how broad suites of volatiles may be affected by particular light qualities.This report details how specific light treatments can affect relevant plant traits, such as aroma, in sweet basil (Ocimum basilicum) “Ceasar” were sown in soil and allowed to germinate in greenhouse or LED-illuminated chambers. Greenhouse plants were grown for 7 days under intermittent mist, and then transferred to a glass-covered greenhouse. Under LEDs seedlings were covered with a transparent plastic lid for 7 days and then were grown for an additional 2 weeks. After the first week of growth, fertigation was initiated with Peters Professional 15-5-15 Cal-Mag at 150 ppm, and maintained during the entire time of growth. At 4 weeks basil plants were transferred to individual pots.Seeds from basil (−2 s−1 for exclusive B or R and for the sum of B and R (1:1 ratio—50 μmol m−2 s−1 each). When a third wavelength was added , its contribution was 50 μmol m−2 s−1 (ratio 1:1:1), giving a total fluence rate of 150 μmol m−2 s−1. Photoperiod was 12 h light/12 h dark in order to approximate greenhouse photoperiod conditions.Quantum energy distribution of full sunlight in the greenhouse and in LED chambers was determined at 1 pm in July at the University of Florida, Gainesville, FL (29.67°N), using a StellarNet spectroradiometer. Fluence rates were measured with a LI-COR light meter (model LI-250), and far-red fluence rates were determined with an International Light meter (model IL1400A). LED chambers were maintained at 24°C, ventilated and lined with reflective mylar. Light was provided by Plant Whisperer light units . The wavelengths used were 450 (Blue—B), 520 (Green—G), 600 (Yellow/Amber—Y), 600 (Red—R), and 735 (Far-red—Fr) nm. Fluence rates were set at 100 μmol mBasil plants germinated in the greenhouse were allowed to develop for 6 weeks. At week 2 and once every week (5 measurements), plants were imaged from above. These images were used for leaf area measurement using Image Tool 3.0. Top leaves and internodes from individual seedlings and plants were used for volatile collection. At weeks 2 and 3 entire seedlings were used, whereas from weeks 4 to 6, plants were cut 1 cm below the top internode with a razor blade. This approach ensures that quantitative changes detected are not due to the age of leaves, given that the density of leaf trichomes, where volatile production occurs, decreases as leaves age  and connected to a push-pull dynamic headspace collection system. Filtered air was pushed into the headspace of each tube during 2 h so that the emitted volatiles were forced to pass into a glass column with a porous copolymer volatiles adsorbent trap . Methylene chloride was used to elute the volatiles from the trap and nonyl acetate was used as elution standard  ion source was set to 70 eV. The full scan mass range was 40–550 m/z (threshold: 150).After elution, the samples were analyzed on a 7890B/5977A Series Gas Chromatograph/Mass Selective Detector . The volatiles were separated in a DB-5 capillary column  using helium as carrier gas at 11.5 psi, temperature: 220°C. The initial oven temperature was 40°C (hold time: 0.5 min) and increased to 250°C (hold time: 4 min) at 5°C minVolatile compound MS spectra were compared to NIST 11 database search for identification, and authentic standards  for 44 compounds  were used to confirm identity by comparing the retention times. Where standards were not available compounds were identified based on results of previous analyses in different basil cultivars . Seedlings were cut at the basis of the hypocotyl, weight was recorded, and total material was used for antioxidant capacity analysis. In the greenhouse trial performed in June, only the top leaf internode, cut 1 cm below the internode, was used. The oxygen radical absorbance capacity-fluorescein (ORAC-FL) method was followed as previously established .Means of log-transformed weights and heights values were compared by One-way ANOVA and comparisons for each pair were evaluated by Student's q-value  were germinated in a glass-covered greenhouse and plants allowed to grow for 6 weeks. Beginning at 2 weeks after sowing, measures of plant development  were recorded. Every week leaf and stem growth along with the appearance of new leaf-pairs was measured . This trend progressively changed thorough week 5, and by week 6 phenylpropanoids levels were at 15% of the levels measured at week 4 , while monoterpenoids showed a 1.5-fold change . Differences in more than 65% of sesquiterpenoids and monoterpenoids  were observed at week 6. A developmental heat map was constructed showing trends of individual volatile compounds within each class . All compounds were also affected by developmental cues. Monocarboxylic acids and fatty acyl volatiles peaked at week 4 , whereas the remaining classes maintained relatively higher levels after week 3. In terms of individual species, for example, the phenylpropanoid eugenol . Higher amounts were observed at week 6, despite the decreasing levels of phenylpropanoids as plants matured . The monoterpenoid linalool (CAS 78-70-6) was detected at similar amounts for weeks 4 and 5 and peaked at week 6 . Other monoterpenoids such as 1,8-cineole (CAS 470-82-6) and two pinene compounds (CAS 80-56-8 and CAS 18172-67-3) had smaller variations and a significant increase at week 4 .The level of chemicals at the basis of basil flavor vary within plant tissues and developmental stages  while there was not statistical difference under R light. Relative amounts of major classes of volatiles remained consistent with greenhouse conditions, except for fatty acyls and carboxylic acid esters, which were more abundant in R, B and BR . Plants grown under BRG and BRY showed higher amounts of monoterpenoids, and decreased carboxylic acid esters and phenylpropanoids compared to BR , and showed the highest volatile levels when compared to greenhouse conditions. Green light treatment resulted in higher levels of fatty acyls. When far-red was added to BR, relative levels of sesquiterpenoids were higher compared to BR , whereas the remaining classes remained unchanged.Total volatiles from 3- and 4-week-old sweet basil grown in the greenhouse and controlled environments were collected and analyzed. Phenylpropanoids were the most abundant species, followed by sesquiterpenoids and monoterpenoids . Controlled environment conditions produced basil plants with overall constant weight during the three seasonal replicates. B and R generated 0.15 g of fresh weight per seedling, which was equal to plants grown under B and R . Higher values were measured under BRG (0.25 g) and BRY (0.30 g).Growth and physiological characteristics of sweet basil in the greenhouse and under the different narrow-bandwidth treatments were measured. Seasonal effects were taken into account by performing experiments during three different seasons of the year: fall, winter, and summer. At 3 weeks, seedling fresh weight, height, and total leaf area were measured Figure . Seasona2), similar in the fall (10 cm2), and lower during the winter , and combination of both resulted in intermediate levels of total leaf area .Similar effects of light conditions and seasons were observed on total leaf area Figure . CompareBasil height was greatest during the summer in the greenhouse (6 cm), comparable to artificial light conditions during the fall (1.5 cm), and slightly lower (0.8 cm) than most controlled conditions in the winter . Fall, winter, and summer grown greenhouse plants displayed similar levels of antioxidant capacity: 60–80 μmol TE per unit seedling weight . B treatments resulted in slight increases, comparable to BR (30 μmol TE g−1). An induction in total antioxidant capacity to 40 μmol TE g−1 occurred when a third waveband was added to BR (p < 0.003), and no statistical differences were found between these conditions and the greenhouse.The reported antioxidant properties of basil prompted a test of total antioxidant activity. Sweet basil seedlings grown in the greenhouse and under narrow-bandwidth conditions were compared. The ORAC-FL assay was used to determine concentrations of Trolox  equivalents (TEs) of 3-week-old seedlings . These tests were performed in October and February and showed major differences.Consistent with the developmental series Figure  volatileAt week 5 plants grown in all conditions produced similar volatile levels and relative profiles Figure . Under tOcimum basilicum) is highly appreciated as a food product, being used fresh, dried, or processed. Yield and aroma composition are fundamental for basil growers, and the basis for its sensory qualities is well established. Ocimum species are polyploid, and basil's delicate flavors result from complex combinations between dozens of chemical compounds -methyl cinnamate, methyl eugenol, and isoeugenol are usually the more abundant. Volatile abundance is also highly dependent on the tissue tested, the age of the plant, season, and growing site , phenylalanine ammonialyase (phenylpropanoids), and terpene synthases. Petunia flowers exposed for 8 h to red and far-red light resulted in altered volatile benzenoid/phenylpropanoid emission, and levels of the key floral volatile 2-phenylethanol were increased when compared to white, blue or dark treatments  in enclosed chambers, as higher weight and leaf areas were consistently seen over the year. It is also likely that supplemental lighting could compensate for the seasonal changes.Greenhouse-grown basil plants were highly affected by seasonal effects which brought changes in photoperiod, spectrum and temperature. Producers that want reliable and regular year-round production cycles may employ a combination of BRG, BRY or BRFr . The greenhouse grown plants experienced relatively high sunshine, and photoperiod ranged from approximately 11–14 h 30 min. The improved relative growth observed in the summer in the greenhouse suggests it may be useful to increase temperature in the controlled environments. These attempts must also consider if beneficial effects seen by specific light signals on volatile profiles are maintained at higher temperatures.Light spectra and intensity, temperature, and photoperiod, are crucial parameters that remained static in the controlled environment experiments. Plants in the greenhouse were quantitatively similar to these artificial conditions during the fall, but had better overall growth characteristics in summer, and were generally worse in winter. Controlled environment chambers were set at constant 24°C. In Gainesville, North Florida, averages daily/night temperatures during those periods are 27°C/13°C, 33°C/22°C, and 21°C/7°C, respectively .The volatile compound retention results also suggest that specific light treatments during germination and early seedling development may install a particular developmental/metabolic pattern that influences the potential to produce flavor and aroma compounds later. After reaching a certain developmental threshold plants delay adjustment to new conditions. This hypothesis would imply increased difficulty to modify sensory characteristics in mature plants, once they have undergone early developmental programming. This hypothesis was tested by measuring volatile profiles at other stages of development, with no favorable outcomes (data not shown).These trials demonstrate that seedling growth and development may be modulated with changes in the light environment, and suggests that narrow-bandwidth light treatments might be a replacement for, or supplement to, ambient solar radiation. The changes in volatile profiles and morphology suggest that tailoring of light conditions may increase the value and quality of high-value herbs grown for human sensory characteristics.SC and MS performed the trials, propagated plants, performed the measurements. MS and CA performed volatile assessments and statistical treatments therein. SC and CA compiled the data and performed statistical analysis. SC, TC, and KF participated in preparation of the manuscript. SC, TC, and KF provided scientific oversight in experimental design and interpretations. TC and KF obtained funding for the study.www.arabidopsisthaliana.com/funding.html.This work was supported by the United States Department of Agriculture's National Institute of Food and Agriculture , Florida Department of Agriculture and Consumer Services (USDA-FDACS), and the UF/IFAS Plant Innovation Center. All of KF's research and outreach funding, as well as travel reimbursements are available at The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer MF-F and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


The China-Myanmar border region presents a great challenge in malaria elimination in China, and it is essential to understand the relationship between malaria vulnerability and population mobility in this region.A community-based, cross-sectional survey was performed in five villages of Yingjiang county during September 2016. Finger-prick blood samples were obtained to identify asymptomatic infections, and imported cases were identified in each village (between January 2013 and September 2016). A stochastic simulation model (SSM) was used to test the relationship between population mobility and malaria vulnerability, according to the mechanisms of malaria importation.χ2 = 0.487, P = 0.485). The average probability of infection in the members of the population that moved to Myanmar was 0.011 (range: 0.0048–0.1585). The values for simulated vulnerability increased with greater population mobility in each village.Thirty-two imported cases were identified in the five villages, with a 4-year average of 1 case/year (range: 0–5 cases/year). No parasites were detected in the 353 blood samples from 2016. The median density of malaria vulnerability was 0.012 (range: 0.000–0.033). The average proportion of mobile members of the study population was 32.56% (range: 28.38–71.95%). Most mobile individuals lived indoors at night with mosquito protection. The SSM model fit the investigated data  contains supplementary material, which is available to authorized users. Globalization and international population migration have caused imported malaria cases to become the predominant threat to the Chinese malaria elimination program , 2. One In addition to receptivity, malaria vulnerability is considered a major characteristic for risk assessment of malaria re-establishment –8. AccorMany researchers have found that vulnerability is related to population mobility , 3, and A community-based, cross-sectional survey was used to obtain data from five villages  in Yingjiang county of China (western Yunnan Province) Fig. , which iThe epidemiological survey was performed during September 2016, which is the peak month for local malaria transmission. Before the survey, a pre-survey of four households was conducted in Hu Que. Ba to adjust the previously developed questionnaire and to improve the planning of the survey. There are 170 households located in the five villages, and an area sampling method that included all households was adopted to collect the basic information for each village and its residents. The basic information for each village was collected by interviewing the primary public health provider, and included the village name, terrain, average temperature, rainfall, main crops, number of households, and number of permanent residents  into the selected village during the previous year. The mobile population was defined as individuals who had resided in at-risk areas for > 1 night during the previous year, based on the malaria transmission route, although individuals were excluded from the mobile portion of the study population if they performed many daytime border crossings. The proportion of the mobile population was defined as the mobile population divided by the number of permanent residents.Plasmodium spp. using the finger-prick blood samples  was used with the condition that if f(x) > p × q × (1 - e), the individual would be considered infected. All other individuals were considered uninfected.In the model, Mim, Mem, N, f, d, E, p, q, and e), data regarding Mim, Mem, N, f, d, and E were obtained from the epidemiological survey. Data regarding the other 3 parameters  were obtained using a model fitted with the vulnerability data from the selected villages. In the model fitting, the simulated D values were compared to the malaria vulnerability values (density of imported cases and asymptomatic infections) by calibrating each parameter until the chi-square test revealed no significant difference (P > 0.05).Among the 9 parameters in the SSM model . This approach allowed us to obtain the simulated densities of imported infections based on different M values for each village.To explore the relationship between malaria vulnerability and the proportion of the mobile population, the SSM model was simulated 1000 times using different Microsoft Office Excel 2010  was used to run the SSM model. SPSS 13.0  was used to perform the chi-square test and the Fisher’s exact test.χ2 = 56.667, P < 0.001). The male mobile population was slightly larger than the female mobile population, although this difference was not significant in the five villages . Most mobile individuals were 20–39-years-old, although no significant differences were observed in the age distributions for the five villages .The five study villages were located in warm and rainy areas with variable terrain. The main crops produced by all five villages were banana and rice. The median number of permanent residents was 86 (range: 72–146), and 92.11% of the mobile population were temporary emigrants (Table Plasmodium vivax (93.75%) and generally involved male individuals (59.38%), although the cases were generally distributed equally among the seven age groups. The highest proportion of cases (50.00%) was detected in 2015. Fisher’s exact test revealed no significant differences among the five villages in their distributions of species (P = 1.000), sex (P = 0.651), age (P = 0.571), or temporal distribution (P = 0.233), which are all shown in Table χ2 = 3.580, P = 0.466).During 2013–2016, 32 imported cases were reported in the five villages, with a 4-year average of one imported case per year (range: 0–5 cases). These cases predominantly involved No parasites were detected using PCR in the 353 blood samples from 2016. The median density of malaria vulnerability was 0.012 (range: 0.000–0.033) . All reported temporary immigrants lived indoors at night with protection. Among the reported temporary emigrants, 78.29% lived indoors at night with protection, 1.14% lived indoors at night without protection, and 20.57% lived outdoors at night with protection. Most villages had similar patterns of exposure, except Xin Cun Village, where 78.26% of reported temporary emigrants lived outdoors at night with protection . The median probabilities of infection were 0.011 in area 1 (range: 0.0048–0.1585) and 0.003 in area 3 (range: 0.0021–0.0038). The efficacy of protection was 20% and the protection coefficient (p) for living indoors at night was 0.95  or high (0.3 ≤ M < 0.5) rates of migration from medium p1 areas , we found that malaria vulnerability increased slowly with the mobile population proportion. At exceedingly high (M ≥ 0.7) rates of migration from medium p1 areas , or high (0.3 ≤ M < 0.5) rates of migration from high p1 areas , we found that malaria vulnerability increased with the mobile population proportion. At high (0.3 ≤ M < 0.5) rates of migration from exceedingly high p1 areas , we found that malaria vulnerability increased dramatically with the mobile population proportion , 0.5/1000 population for area 3 (95% CI: 0.1–0.6/1000 population), and a total density of 3.2/1000 population (95% CI: 0.2–10.9/1000 population). The imported cases increased with an increasing proportion of mobile individuals in the study population , 1.9/1000 population for area 3 (95% CI: 0.7–5.7/1000 population), and a total density of 32.4/1000 population (95% CI: 1.6–107.4/1000 population).Based on an average simulated Progress towards eliminating malaria in China has recently reduced the malaria transmission rate in Yunnan Province –27. HoweThe high proportion of mobile individuals in the study population was associated with greater malaria vulnerability, which might increase the likelihood of malaria re-establishment in Yingjiang county. In addition, the model that was used for analysis fit the reported data, which indicates that the SSM model can be used to simulate the relationship between mobile population proportion and malaria vulnerability. Interestingly, we observed different infection probabilities among the mobile populations of the five villages, with the greatest probability observed in area 1. These differences might be related to the heterogeneous distribution of malaria transmission in Myanmar. Thus, the mobile populations of Jing Po Zhai, Ka Ya He, and Xin Cun might have moved to high transmission areas, while the mobile populations of Zhuan Po Zhai and Hu Que. Ba might have moved to low transmission areas in Myanmar.Most mobile individuals lived indoors at night with protection, although the efficacy of protection and rate of sleeping indoors was low during their stays in Myanmar. Furthermore, we detected high values for the mobile population proportion and malaria vulnerability in the China-Myanmar border region. Moreover, the SSM model predicted a linear relationship between mobile population proportion and malaria vulnerability. Therefore, to reduce the risk of malaria re-establishment in the border regions of China, we recommend introducing mobile population-specific measures, such as health education to reduce malarial vector exposure and blood screening with ultrasensitive reverse transcription PCR to identify asymptomatic infections when mobile people return to China .The present study is limited in that it only evaluated PCR-based data regarding asymptomatic infections from a single community-based, cross-sectional survey in 2016, and it is probable that asymptomatic infections were not detected during 2013–2015 when PCR was not used. Therefore, the regional vulnerability might be underestimated by using the 4-year average for imported cases, although we believe that this would only have a minor effect on our findings. It is important to match temporal behaviors with infection data from the same period, and the present cross-sectional study collected mobile population data from the previous year to ensure that it matched the imported case data from 2016. However, we found that no imported cases were reported in three villages  implements an epidemiological investigation to categorize the case as imported or indigenous. The municipal CDC then checks the information that was reported by the county CDC, and each case is finally verified by a provincial reference laboratory after being reported by the local public health institute. The final confirmation of each malaria case is approved by the NHFPC expert group. This process and the step-by-step protocol ensure that each malaria case is diagnosed and categorized correctly, and minimizes the likelihood of diagnostic bias in China.Another limitation is the possibility of bias in linking nfection  can makeThis community-based, cross-sectional study was performed to develop an SSM model that simulates mobile population dynamics and malaria vulnerability in the China-Myanmar border region. High population mobility was observed with different epidemiological characteristics and exposure patterns, which were associated with varying levels of vulnerability in the studied villages. Thus, the SSM model could be used as a tool to quantify the linear relationship between vulnerability and mobile populations, and it may be useful for assessing the risk of malaria re-establishment in China.Additional file 1Registration Form for Village Profile in Yingjiang County. (DOCX 27 kb)Additional file 2Questionnaire for Household Survey in Yingjiang County. (DOCX 29 kb)Additional file 3Parameter definitions and values for the five selected villages. (DOCX 42 kb)


Higher level of nitrogen and boron resulted in increased growth , total microcystin concentrations , and mcyA transcript but its expression was not directly correlated with total microcystin concentrations in the target strains. Interestingly, Durgakund strain had much lower microcystin content and lacked microcystin-YR variant over NIES 843 and KW. It is inferred that microcystin concentration and its variants are strain specific. We have also examined the heterotrophic bacteria associated with cyanobacterial bloom in Durgakund Pond and Wangsong Reservoir which were found to be enriched in Alpha-, Beta-, and Gammaproteobacteria and that could influence the bloom dynamics.Roles of nutrients and other environmental variables in development of cyanobacterial bloom and its toxicity are complex and not well understood. We have monitored the photoautotrophic growth, total microcystin concentration, and microcystins synthetase gene ( Microcystis, Oscillatoria, Anabaena, and Nostoc produce toxins and other bioactive compounds that can poison and kill humans and livestock  containing a linear 30–60% denaturant gradient (100% denaturant corresponded to 7 M urea and 40% (v/v) formamide). DGGE was run for 18 h at 110 V on the DCode system . The gel was stained with ethidium bromide and visualized by UV transillumination. Thereafter, the bands were excised from the DGGE gel and incubated in Milli-Q-water  and plated on Luria-Bertani agar plates in the presence of ampicillin. The white colonies were selected and incubated in Luria-Bertani broth containing ampicillin (50 μg/mL). DNA was purified using the QIAprep® Spin Mini-Prep Kit . The purified products were sequenced by Solgent Inc. , using an ABI 3730XL automatic DNA sequence .The amplified PCR products were purified using the QIAquick® Gel Extraction Kit  and ligated into the pGEM-T Easy Vector  according to the manufacturer's protocols. They were transformed into HIT™ Origin® Version 6.0. Effects of N, P, and B supplementation to the growth medium on total MCs production, Chl a, and MC content were statistically analyzed using multivariate analysis of variance (ANOVA). Data were explored using SPSS 16.0 software and log-transformed to give these an approximate normal distribution.All experiments were carried out in triplicate with standard deviation (SD) represented as bars wherever necessary using MicrocalμM). These strains were transferred separately in medium having selected higher nutrient concentrations and growth behavior was monitored up to 15th d by measuring Chl a at periodic intervals , P , and B  in all three strains. Increased level of the N in the medium resulted in the higher Chl a yield  in the target strains. The yield was approximately 1.1 times higher in the cultures of all strains with 17.6 mM N. It was interesting that 100 times more concentration of N (1.5 mM) did not affect the Chl a yield (avg. 4.1 mg/L) in the target strains as compared to the growth in the lowest N concentration. The growth of Durgakund strain was lower as compared to NIES 843 and KW strains.Photoautotrophic growth of target strains was monitored in low levels of nutrients  was 1.67 times lower on an average basis as compared to the cultures with low level of N. Increase in the P concentration (100 times) in the growth medium increased the growth of the target strains by 1.21 times (avg. 3.4 mg/L) (a yield (1.1 times) of the target strains at 0.23 mM P indicated that there was no increase in the growth beyond a critical threshold level of P (0.1 mM). This was in contrast to the level of N in the cultures favoring the growth of the target strains. Interestingly, photoautotrophic growth of the strains was favored with increase in B levels  and higher concentrations (23 and 46 μM), respectively.Likewise, the strains grown in varying levels of P behaved differently. The Chl .4 mg/L) . The groB levels . AverageF1,12 = 653, p < 0.001), P , and B  in the target strains. MC content was also significantly influenced by N , P , and B . The highest levels of MC-LR (0.77 mg/L), -RR (8.2 mg/L), and -YR (1.4 mg/L) were recorded at 0.015 mM N in NIES 843. However, at higher concentrations of N (1.5 and 17.6 mM), the same levels of MC variants were recorded in both NIES 843 and KW strains  were detected in both NIES 843 and KW strains at different N concentrations with predominance of MC-RR . MC-YR w strains . In compLikewise MC-YR was not detected in Durgakund strain growing in any of the P concentrations while higher level of MC-YR was recorded in KW strain at 0.23 mM P. Highest level of MC-LR and -RR was recorded in NIES 843 strain followed by KW strain and Durgakund strain . Higher a concentration  in the target strains ( mcyA transcript increased at 1.5 and 17.6 mM N relative to the control (0.015 mM N). Maximum increase (12.4-fold) in mcyA transcript was recorded in Durgakund strain and only 2.3-fold increase in NIES 843. mcyA transcript in KW strain decreased (0.19-fold) relative to the control at 17.6 mM N. However, mcyA transcript increased in the target strains at 1.5 μM N indicating a threshold level of N for optimum expression of mcyA. Similarly, increase in P levels (0.1 and 0.23 mM) increased the mcyA transcript in NIES 843 and Durgakund strains while only 0.23 mM P slightly increased the expression of mcyA. The maximum increase of mcyA transcript (10-fold) was recorded in NIES 843 at 0.1 mM P. In B-containing cultures, concentration-dependent increase in mcyA transcript was recorded in the target strains. The maximum increase (19-fold) in mcyA transcript was recorded in NIES 843 at 46 μM B followed by Durgakund (2.8-fold) and KW strain (1.4-fold).Real-time RT-PCR was used to measure the expression of strains . mcyA tr mcyA expression for the target strains is represented in p < 0.0001, R2 = 0.769). Chl a concentration and MC production were closely related in different concentrations of P in NIES 843  and KW strains  and of B in NIES 843  and KW strains . Such relationship was rather weak in different concentrations of N in NIES 843  and KW strains . In different nutrient regimes the expression of mcyA was not correlated with MC concentration  and content  in NIES 843. Similar trend was evident in Durgakund strain with MC concentration  and content . Contrary to this, mcyA expression pattern is directly reflected on MC concentration  and content  in KW strain.The correlation among growth, MC concentrations, MC content, and M. aeruginosa and were present in all the samples of Durgakund and Wangsong Reservoir. Interestingly, sequence (band 6) related to M. wesenbergii was more intense in August and September samples of Durgakund Pond. Sequence (band 2) related to Merismopedia glauca belonging to order Chroococcales was also seen in all the samples of Durgakund. In general, the banding pattern of DGGE in August and September samples differed from the samples of other months in Durgakund . There are some reports where P limitation slightly reduced or did not influence the toxicity of M. aeruginosa [ Microcystis was also reported with decreasing concentrations of P in culture conditions [ Microcystis strains to N and P [a content at total N : P atomic ratios of 16 : 1 and 50 : 1. Furthermore, it was suggested that cellular MC content is a function of cellular nitrogen status and found to be strongly correlated with cellular protein content at N : P ratios between 18 and 51 in M. aeruginosa [The target strains showed better growth at 0.1 mM P, indicating it is a threshold limit.ruginosa , 37, whinditions , 41. In nditions , 43. Sev N and P , 44, 45.ruginosa .luxS), has been reported by Chen et al. [ Gloeothece sp. [ M. aeruginosa [ Microcystis strains in the current study. It was interesting that B favored growth of all the target strains along with MC concentrations  binding sites were identified in the mcyA/D promoter region, suggesting the role of nitrogen in controlling MC biosynthesis [ mcyA expression were somewhat consistent with MC levels measured under different N concentrations in all the three strains but not directly correlated with the total MC concentrations . This finding is consistent with observation where low N resulted in decrease in transcript levels of MC synthesizing genes [ mcyD expression but the transcript level remained constant and was independent of nitrate availability. Recently, Horst et al. [ mcyB gene but mediated by phenotypic responses of individual cells to N availability. As MC is made up of different modules, each having specific enzymatic function [Transcription ofynthesis . In thisng genes . Howeverng genes  reportedt et al.  suggestefunction , N might mcyA transcript increased under higher P levels in the target strains. In another study, insignificant changes were observed in the relative quantification of mcyD in M. aeruginosa PCC 7806 under excess phosphate, while P-deficiency (N/P = 40 : 1) led to an increase in mcyD transcript and MC-LR per cell [ mcyE expression were also reported in Lake Rotorua with the increase in Microcystis cell concentrations [ mcyE expression occurred over smaller time frames.In this study,per cell . Increastrations . They de mcyA expression in the target strains was consistent but not correlated with MCs levels under selected concentrations of B. Correlation analysis revealed that mcyA expression in only KW strain was directly linked with MC content  and concentrations  under different levels of N, P, and B. Increase in photoautotrophic growth and MC concentrations in the target strains suggested an indirect role of B but its role in nonheterocystous cyanobacteria is still not well documented. Furthermore, it was suggested that accumulation of mcyB in the lysing Microcystis cells stimulated the production of MCs in the remaining intact Microcystis cells [ Microcystis. Various field and laboratory studies failed to assign a single major factor responsible for promoting cyanobacterial bloom and its toxicity. This indicated a complex regulation of bloom development and its toxicity by biological and environmental variables. Induction of the growth of toxic Microcystis bloom is not yet known in spite of the fact that cyanobacterial blooms are regulated by various environmental factors. This study focused on response of Microcystis to differing nutrient concentrations on mcy transcription and MCs production which could help to decipher the possible regulation of the bloom growth and its toxicity and in timely management of the water bodies.Increase in Microcystis. Earlier we have reported the dominance of Microcystis sp. in Durgakund Pond with the highest proportion of potentially toxigenic Microcystis sp. (14%) and MCs concentration in September [ Microcystis bloom changed the microhabitat and quantity and quality of exudates and also microbes have the potential to control bloom development of potentially toxic cyanobacteria [The interaction between bacteria and cyanobacteria is one of the biotic factors that influence the bloom dynamics . Therefoeptember . Our obsbacteria , 57. How mcyA gene expression of three lab-grown Microcystis strains  in different nutrient regimes revealed that MC content per biomass and total MC concentration were highly correlated. MC production increased with increase in growth and this could have important implications for the management of freshwater bodies during Microcystis bloom upsurge. Lack of MC-YR in Durgakund strain indicated that MC concentration and variants were strain specific. Expression of mcyA showed nutrient concentration-dependent tendency but was not correlated with total MC concentration and therefore necessitates a time dependent real-time analysis for understanding the role of nutrients and other ecological variables in MC production. Natural associations of heterotrophic bacteria and cyanobacterial bloom were somewhat similar in freshwater bodies; however bacterial community showed variation during the different phases of the cyanobacterial bloom.In summary, a comparison of photoautotrophic growth, MC content and concentration, and