PMS2 mutation carriers. We aimed to evaluate MMRpredict and PREMM5 in a clinical cohort and for PMS2 mutation carriers specifically. In a retrospective, clinic-based cohort we calculated predictions for LS according to MMRpredict and PREMM5. The area under the operator receiving characteristic curve (AUC) was compared between MMRpredict and PREMM5 for LS patients in general and for different LS genes specifically. Of 734 index patients, 83 (11%) were diagnosed with LS; 23 MLH1, 17 MSH2, 31 MSH6 and 12 PMS2 mutation carriers. Both prediction models performed well for MLH1 and MSH2 (AUC 0.80 and 0.83 for PREMM5 and 0.79 for MMRpredict) and fair for MSH6 mutation carriers (0.69 for PREMM5 and 0.66 for MMRpredict). MMRpredict performed fair for PMS2 mutation carriers (AUC 0.72), while PREMM5 failed to discriminate PMS2 mutation carriers from non-mutation carriers (AUC 0.51). The only statistically significant difference between PMS2 mutation carriers and non-mutation carriers was proximal location of colorectal cancer . Adding location of colorectal cancer to PREMM5 considerably improved the models performance for PMS2 mutation carriers (AUC 0.77) and overall (AUC 0.81 vs. 0.72). We validated these results in an external cohort of 376 colorectal cancer patients, including 158 LS patients. MMRpredict and PREMM5 cannot adequately identify PMS2 mutation carriers. Adding location of colorectal cancer to PREMM5 may improve the performance of this model, which should be validated in larger cohorts.Until recently, no prediction models for Lynch syndrome (LS) had been validated for The online version of this article (doi:10.1007/s10689-017-0039-1) contains supplementary material, which is available to authorized users. Lynch syndrome (LS) is a hereditary predisposition to colorectal cancer, endometrial cancer and other extra-colonic cancers at a young age , 2. MorbMLH1, MSH2, MSH6 or PMS2, or in the 3′ end of the EPCAM gene and consequent hypermethylation of the MSH2 promoter region  and 0.83 [95% CI 0.73–0.94] for PREMM5 and AUC of 0.79 [95% CI 0.69–0.89 and 0.67–0.91] for MMRpredict. Both models had a fair discriminative power for MSH6 mutation carriers (AUC of 0.69 [95% CI 0.58–0.80] for PREMM5 and AUC of 0.66 [95% CI 0.56–0.76] for MMRpredict). MMRpredict still had fair performance for PMS2 mutation carriers (AUC of 0.72 [95% CI 0.57–0.87]), while PREMM5 failed to discriminate PMS2 mutation carriers from non-mutation carriers at all with an AUC of 0.51 [95% CI 0.35–0.66].Overall, PREMM5 predicted higher probabilities of carrying a LS mutation than MMRpredict . For mutation carriers, risk scores varied from 0.02 to 0.99 for PREMM5 and from 0.002 to 0.99 for MMRpredict. Both prediction models could fairly discriminate between index patients with and without an MMR mutation.. This higher sensitivity came at the expense of a lower specificity (46 vs. 67%). For PREMM5, using a cut-off of 5%, resulted in a sensitivity for MLH1 mutation carriers to 83% for PMS2 mutation carriers . Overall, the revised Bethesda guidelines had a sensitivity of 90% with a specificity of 24%. In order to reach the same sensitivity, PREMM5 and MMRpredict had a similar specificity (25%).Sensitivity of the revised Bethesda guidelines decreased from 96% for PMS2 mutation carriers and non-mutation carriers. Only one significant difference remained; PMS2 mutation carriers more often had proximal CRC than patients without an MMR mutation    and MSH6 (AUC 0.75 [95% CI 0.65–0.84] vs. 0.69 [95% CI 0.58–0.80]) mutation carriers . Overall, the adjusted PREMM5 model performed better than the original PREMM5 model (AUC 0.81 [95% CI 0.76–0.86] vs. 0.72 [95% CI 0.66–0.79]) and MMRpredict (AUC 0.81 vs. 0.73 [95% CI 0.66–0.79]). The adjusted prediction model can be found as supplemental material.Adding tumour location also improved the performance of PREMM5 for identifying At a 5% cut-off, sensitivity of the extended PREMM5 model was higher than the sensitivity of the original PREMM5 model (92 vs. 78%) with similar specificity (45 vs. 46%). Sensitivity and specificity of the extended PREMM5 model at a 5% cut off were both higher than those of the revised Bethesda guidelines (sensitivity 92 vs. 90% and specificity 45 vs. 24%).MLH1, 26 MSH2, 28 MSH6 and 73 PMS2 mutation carriers. Similar to the results in the initial cohort, the extended PREMM5 model had better predictions than the original PREMM5 model for PMS2 mutation carriers (AUC 0.90 [95% CI 0.86–0.94] vs. 0.82 [95% CI 0.76–0.87]) and overall (AUC 0.92 [95% CI 0.89–0.95] vs. 0.87 [95% CI 0.84–0.91]). Performance for MLH1, MSH2 and MSH6 mutation carriers was also slightly better for the extended PREMM5 model than for the original PREMM5 model .In our validation cohort, 60% of the patients were male and median age was 55 years (IQR 45–63 years). The cohort included 31 PMS2 mutation carriers. The performance of the PREMM5 model improved considerably by adding the location of CRC to the model. In our clinical cohort of 734 CRC patients as well as in a validation cohort of 376 CRC patients, this extended PREMM5 model not only identified PMS2 mutation carriers more accurately, its overall performance was also better than the original PREMM5 model and the MMRpredict model.The results of our study indicate that while the models MMRpredict and PREMM5 can adequately predict whether an individual is likely to have Lynch syndrome, they fail to identify 1,2,6 model had a slightly better overall performance than MMRpredict [1,2 also performed better than MMRpredict in several studies [Our results are in line with those of previous studies, where the PREMMRpredict , 32, 33. studies , 24, but studies , 26 or l studies  predicti studies .PMS2 mutation carriers specifically. An explanation for this could be that the location of CRC is incorporated in the MMRpredict model but not in the PREMM5 model. Proximal location of CRC is a known predictor for Lynch syndrome and in our cohort was the only significant difference between PMS2 mutation carriers and non-mutation carriers. After adding this new variable to the existing PREMM55 model, this new model performed better than MMRpredict for PMS2 mutation carriers. The extended PREMM55 model also performed better than the original model for MLH1, MSH2 and MSH6 mutation carriers and had a better overall performance.Although PREMM5 had better overall predictions, MMRpredict had a better performance for PMS2 mutation carriers. Selection of patients for analysis of MSI and IHC may have been less stringent at the Erasmus Medical Center Rotterdam than at the Leiden University Medical Center. Therefore, mutation carriers in our validation cohort, who were all from Leiden University Medical Center, may have had a family history more suspect for Lynch syndrome than family history of the patients in our original cohort. This could explain the higher AUCs in the validation cohort. However, in both cohorts we showed that the extended PREMM5 had better performance.In our validation cohort, all AUCs were much higher than in our original cohort, including those for Prediction models for Lynch syndrome are not yet regularly used in current clinical practice. However, the US Multi-Society Task Force on Colorectal Cancer recommends genetic evaluation if an individual’s risk of carrying an MMR gene mutation is ≥ 5% according to one of the prediction models MMRpro, MMRpredict or PREMM . The Ame1,2,6 [The US Multi-Society Task Force on Colorectal Cancer recommends the use of either PREMM, MMRpredict or MMRpro to assess the probability of an individual carrying an MMR mutation . Since w1,2,6 –27, 32.MLH1 and MSH2 mutation carriers than for LS patients carrying a mutation in MSH6 or PMS2. This finding is in line with a previous study that showed that carriers of mutations in MSH6 or PMS2 had lower risk scores than carriers of a mutation in MLH1 or MSH2 [PMS2 mutation carriers was the least accurate, in line with its more limited penetrance.Both PREMM5 and MMRpredict were far more accurate for  or MSH2 . In our PMS2 mutations [PMS2 mutation carriers. To our knowledge, our study is the first to validate LS prediction models for PMS2 mutation carriers specifically since the development of the PREMM5 model. At a 5% cut-off, our extended PREMM5 model was able to detect 5/6 PMS2 mutation carriers who would have been missed by the original PREMM5 model at the same cut-off. Identification of Lynch syndrome carriers is highly important, since this allows not only them, but also their family members carrying the same mutation, to undergo intensive surveillance in order to prevent the development of cancer. Our new model would also identify more Lynch syndrome patients overall than the original PREMM5 model.Around 15% of all Lynch syndrome cases are estimated to be caused by utations . In our The performance of prediction models can differ between high-risk settings and population-based cohorts. Further validation studies should indicate whether our results can be generalized to settings with patients at low to median risk of having Lynch syndrome. Since patients in our study cohort were all referred for genetic counselling, family histories were obtained in detail and in many cases also verified by medical documents. In other settings where patients are at lower risk of having Lynch syndrome, family history is not verified and might be less reliable. Therefore, prediction models should also be validated in population-based cohorts. However, in a meta-analysis, prediction models performed better in population-based cohorts than in clinic-based cohorts .1,2,6 was more accurate than MMRpro and MMRpredict, but still only reached an AUC of 0.71 [PMS2 was not performed in the Korean study, so there might have been more mutation carriers in their cohort. Another non-Western population has been studied by Khan et al., who analysed the performance of prediction models in 15 African American patients [1,2,6 both had a high AUC of 0.89.It is not known whether the current prediction models for Lynch syndrome are useful in non-Western populations. In a recent study among Korean patients, PREMM of 0.71 . There wpatients . In thesMSH6 and PMS2 mutations. Since 12 patients were identified as a PMS2 mutation carrier, we were able to evaluate the prediction models for each MMR mutation specifically, admittedly with considerable uncertainty [PMS2 mutation carriers.A main strength of our study was the large cohort, which consisted of more than 700 index patient including 83 Lynch syndrome patients. Also, our cohort included patients with ertainty . FurtherA limitation of our study was that germline mutation analysis was not done for all index patients. Patients who had microsatellite stable tumours with normal IHC were assumed to be non-mutation carriers. However, some of these patients might still have an MMR mutation. Also, the sample size per gene was still relatively small and it is unclear whether our results from a high-risk population apply to a population-based setting.PMS2 mutations. Adding the location of CRC to the PREMM5 model improves the performance of the model for PMS2 mutation carriers as well as its overall performance. These findings should be validated in large cohorts from population-based settings.In conclusion, we have shown that although MMRpredict and PREMM5 can accurately predict an individual’s risk of carrying a causative MMR mutation, neither model is able to identify patients with Below is the link to the electronic supplementary material.Supplementary material 1 (DOCX 14 KB)Supplementary material 2 (DOCX 15 KB)Supplementary material 3 (DOCX 19 KB)


Dapsone is recommended as a second line therapy in immune thrombocytopenia (ITP), but is underused because of its potential side effects. The medical charts of 42 ITP patients treated with dapsone (100 mg/day) were retrospectively reviewed in order to assess its efficacy and safety in daily clinical practice. The overall response rate was 54.8%  with a median time to response of 29 days (24–41 days). Patients with complete response had shorter disease duration whereas no difference was observed between responders and non-responders regarding age, sex or previous treatments received. Importantly, after dapsone withdrawal, a sustained response was observed in 5 patients, representing 12% of the whole cohort. Twenty percent of patients (n = 8) relapsed on therapy after 8.1 (6.5–13.6) months. Side effects occurred in 31% (n = 13) of patients, and required dapsone withdrawal in 22% (n = 9) or dosage reduction in 10% (n = 4) of the cases. Side effects resolved in all but one case. Overall, these data support dapsone as an interesting second line therapy in ITP, with a good safety and efficacy profile at a low cost. Steroids are used as first-line therapy, while intravenous immunoglobulins (IVIg) should be restricted to patients with severe bleeding symptoms [Immune thrombocytopenia (ITP) is an autoimmune disorder leading to a low platelet count responsible for bleedings of variable severity. Treatments are recommended in case of bleeding symptoms and/or platelet count below 20x10symptoms . Both stsymptoms –6. Of nosymptoms . Hydroxysymptoms . Becausesymptoms ,12. Of nsymptoms ,14, althsymptoms –18. Immusymptoms .Dapsone efficiency was first reported in the 90’s in ITP ,21. SincWe aimed to assess the efficacy and safety of dapsone in a retrospective monocentric study in adult ITP.Comité de Protection des Personnes Est I), who waived the requirement for informed consent.Medical records of all patients registered in Dijon University Hospital Centre between January 2006 and August 2016 for thrombocytopenia according to the diagnosis-related group (DRG) medical information system (PMSI) were retrospectively reviewed. The study was approved by the institutional review board of the University Hospital of Dijon and the local ethics committee (i.e. newly diagnosed ITP (less than 3 months of evolution), persistent ITP (3–12 months) and chronic ITP (more than 12 months). Differential diagnoses for ITP were ruled out in all patients. Bone marrow examination was performed in patients older than 60, or in case of systemic symptoms, or when anaemia or leukopenia were present, as recommended [Patients with a diagnosis of ITP and treated with dapsone were selected. Demographic data  was considered when platelet count remain below 100x109/L, and complete response (CR) when platelet count was over 100x109/L. Non-response (NR) was considered when none of these criteria were achieved or when treatment was stopped because of side effects within the 4 weeks following its introduction. When another treatment (steroids or TPO-RA) was prescribed concomitantly with dapsone, only patients who had this treatment discontinued and maintained platelet counts over objectives were considered as responders. Patients lost to follow-up were considered as non-responders. Time to response was calculated from the day dapsone was initiated to the one of the first response was achieved. Duration of response was censored at the date of the last platelet count fulfilling response (PR or CR respectively) criteria.Response was defined as an increase in platelet count over 30x10st-3rd interquartile) and qualitative data as percentage. P-values were derived using Chi2 test and Mann-Whitney test for qualitative and quantitative values, respectively. Wilcoxon matched pairs test was used to compare platelet count before and after treatments. P<0.05 was considered significant. Statistical analyses were performed with STATATM Software (Stata Corporation).Quantitative values are reported as median  and the bleeding score was 2 (0–5). Only 2 patients had a history of severe gastro-intestinal bleedings before dapsone initiation. Median follow-up was 67.6 months (49.5–105.2).The lowest median platelet count within the 3 months prior to dapsone initiation was 14x10Dapsone was initiated at 100 mg/day for all but one patient, who had a CR with 100 mg 3 times a week. Dapsone was initiated in association with steroids in 54.8% (n = 23): prednisone was used at a median dose of 1 mg/kg/day and was discontinued after a median duration of 28 days (21–37.5). A TPO-RA was associated with dapsone in 4 cases: 2 did not respond to dapsone, 1 had a rash leading to dapsone withdrawal and the other maintained a CR after TPO-RA discontinuation.9/L (6–25) to 119 (94–141) in responders (p<0.0001), but not in non-responders (14x109/L (5–20) vs. 20 (8–27), p = 0.6; vs. 49.9 (38.5–90.9) months, p = 0.009). Moreover, response duration was longer in case of CR (10.4 (4.2–21.3) vs. 3.9 (1.9–7.1) months; p = 0.05). Patients who received dapsone in association with steroids tended to have more CR than PR . Importantly, all patients who were treated with this combination interrupted steroids after a median duration of 28 days, and previously experienced a relapse after the discontinuation of a first course of steroids. Thus, the fact that the response attributed to dapsone was indeed due to steroids appeared unlikely. However, to rule out this hypothesis, the response rates of dapsone started as monotherapy were also considered: a response was achieved in 47.4% (n = 9/19 patients) with a CR in 21.1% (n = 4/19) and a PR in 26.3% (n = 5/19).Response rates on dapsone are reported in vs.1.8 (0.9–2.5) g/dL, respectively; p = 0.16).Haemoglobin level at dapsone initiation was 13.9 (12.9–14.9) g/dL, with a decrease in 1.5 (0.6–2.1) g/dL at response time. There was no significant difference in the decrease in haemoglobin level between responders and non-responders (1 (0.4–2) Response to dapsone therapy is summarized in Dapsone was definitively interrupted in 7 patients after 7.6 (5.1–15.7) months of response, after a progressive tapering for 3 patients. The reasons for dapsone cessation were side effects (n = 2) or desire of the patient to take off medication (n = 5). Two patients relapsed 3 and 3.5 months after dapsone cessation. Interestingly, a sustained response was observed in 5 patients  after a median follow-up of 21.1 (12.6–29.5) months. A CR was maintained in 3, while 2 switched to a PR. Among these 5 patients with sustained response after dapsone discontinuation, 3 had a newly diagnosed ITP and 2 a chronic ITP.Ten patients were still responders under treatment after 5.9 (2.8–19.5) months. Dapsone dosage was tapered in 7 of them, in order to withdraw dapsone (n = 4) or because of side effects (n = 3). One patient was lost to follow-up after 4.2 months of response.Regarding the 19 non-responders, a response was achieved with rituximab in 8/11, with splenectomy in 3/5, with TPO-RA in 3/4, and with hydroxychloroquine in 1/2, while 3 patients had a platelet count allowing a wait and watch attitude.vs. 26% and TPO-RAs were used in 4% vs.21%, respectively, supporting dapsone as an interesting medication to spare the use of expensive drugs or radical treatment such as splenectomy.As expected, responders to dapsone required less second line treatments  compared to non-responders  after a median follow-up of 67.6 (49.5–105.2) months. Rituximab was used in 17% of responders compared to 58% of non-responders; splenectomy was performed in 17% vs. 51.7% (15/29), p = 0.015).Fifteen side effects were reported in 13 patients (31%), consistent with skin rash (n = 5), including sulfone syndrome (n = 2), methemoglobinemia (n = 4), neuropathy (n = 3), dyspnoea (n = 1), fatigue (n = 1) and diarrhoea (n = 1). Symptomatic anaemia or severe haemolysis was not observed. G6PD deficiency was screened in 36/42 patients (85.7%). There was no difference between patients with or without side effects, regarding age, ITP history, or the combination of steroids at dapsone initiation . HoweverDue to side effects, dapsone was withdrawn in 9 patients and the dosage reduced in 4, leading to the resolution of all the side effects, except for one case (neuropathy), for which dapsone accountability was not formally demonstrated. One patient died of pneumonia, which was not related to dapsone.The characteristics of the population of this study are in accordance with previous reports on adult ITP with a median age of 57.1 years and a female predominance . DapsoneRelapse during treatment or dapsone tapering was observed in 22%, which is in accordance with previous reports ,24,26. OMore surprisingly, a sustained response after dapsone discontinuation was observed in 5 out of 7 responders who interrupted dapsone, representing around 21% of the responders. Interestingly, the response lasted more than 6 months in 5 patients, among which 2 had a chronic ITP. Such a sustained response ranged between 3 and 65.2% of the responders in previous reports ,24,26, wThe median time to response was 29 days, which is in line with previous reports (21–59 days) ,24,26, eIn this study, side effects related to dapsone were observed in 31%. Cutaneous effects were the most frequent, with sulfone syndrome, that is indeed a DRESS (Drug Rash with Eosinophilia and Systemic Symptoms), occurring only in 2 cases. It has been suggested that the concomitant prescription of steroids decreases the intensity of cutaneous side effects during ITP  comparedUnfortunately, no predictive factor of response to dapsone was identified. However, patients with ITP of short evolution had a higher rate of CR, which argues for the use of dapsone early in the course of ITP, notably for newly diagnosed or persistent ITP, which is in line with what was suggested in a previous report . HoweverIn the past decade, new molecules such as rituximab and TPO-RA have improved the management of ITP. However, the cost of the long term use of these therapies can be very high and cheaper treatment options such as dapsone should be considered. The annual cost of dapsone is around 50 Euros, not considering biological exams, which should be compared to the one of TPO-RA that is around 26,000 Euros or to one course of intravenous rituximab  that is approximatively 5,300 Euros, not considering hospitalization costs . Even ifOur results support the valuable place of dapsone as second line therapy in ITP with a response in about one half of the patients. Interestingly, a sustained response after its withdrawal can be observed, representing 5 out of our 23 responder patients. As expected, responders to dapsone will experience fewer treatments for ITP, thus decreasing the need for radical treatment such as splenectomy and for expensive therapies, which is of interest in the current economical context. However, side effects are frequent and reported in one quarter to one third of patients in retrospective studies. They are usually mild and resolved after dapsone tapering or discontinuation. Haemolysis is expected in all patients on treatment, and haemoglobin level should be checked during the first weeks of therapy and during any acute diseases that may precipitate anaemia, such as pulmonary diseases . For thiS1 Table(XLS)Click here for additional data file.


Mobile phase consisted of solvent A  and solvent B (acetonitrile). Ions were generated by electrospray (ESI) and multiple reaction monitoring (MRM) was used as basis for quantification. The results revealed a linear calibration curve in the range of 5–400 ngmL-1 (r2 ≥ 0.9998) with lower limit of quantification (LOQ) and lower limit of detection (LOD) of 4.66 and 1.53 ngmL-1 in plasma, 4.19 and 1.38 ngmL-1 in RLMs. The intra- and inter-day precision and accuracy in plasma ranged from1.06 to 2.54% and -1.48 to -0.17, respectively. Whereas in RLMs ranged from 0.97 to 2.31% and -1.65 to -0.3%. The developed procedure was applied for quantification of PNT in human plasma and RLMs for study metabolic stability of PNT. PNT disappeared rapidly in the 1st 10 minutes of RLM incubation and the disappearance plateaued out for the rest of the incubation. In vitro half-life (t1/2) was 6.26 min and intrinsic clearance (CLin) was 15.182± 0.477.In the current work, a rapid, specific, sensitive and validated liquid chromatography tandem mass-spectrometric method was developed for the quantification of ponatinib (PNT) in human plasma and rat liver microsomes (RLMs) with its application to metabolic stability. Chromatographic separation of PNT and vandetanib (IS) were accomplished on Agilent eclipse plus C18 analytical column  maintained at 21±2°C. Flow rate was 0.25 mLmin Calculation of accuracy and precision values were carried out using freshly prepared RLMs and plasma calibration curves. PNT QC samples were stored at room temperature for 8 hours for further assessment of their bench top stability. Numerous freeze—thaw cycles (three cycle) were performed to estimate stability of PNT QC samples after freezing these samples at -80°C and then thawing them at room temperature. Additionally, assessment of PNT stability was done through assaying PNT spiked QC samples after one day of keeping them at 4°C and after one month of storing them at -20°C. Additionally, dilution integrity of PNT samples were done to confirm the integrity of the concentrated PNT samples that need e dilution step. PNT stock solution was prepared and spiked in plasma and RLMs to attain a concentration of 1.8 times of the upper concentration in the calibration range then two or four fold dilution were done. Three replicates of each dilution were evaluated and the integrity of the PNT samples was maintained if % nominal lied in ±15% of nominal values and % CVs ≤ 15%.-1 of microsomal proteins in addition to 1mM NADPH in phosphate buffer (pH 7.4) containing 3.3 mM MgCl2 in a final volume of 1 mL. The incubation is started by the addition of NADPH and terminated with 2mL of ACN at 0, 2.5, 5, 10, 15, 20, 40 and 50 min. Precipitated proteins were removed by centrifugation at 14000 rpm for 12 min at 4°C. Supernatants was removed and filtered through 0.22 μm syringe filter. IS (75 μL) was added to 1mL of the filtered supernatant in addition to 0.5 mL of mobile phase . Five μL of this solution was injected into the LC-MS/MS system. Concentrations of PNT in RLMs matrix were calculated from the regression equation of freshly constructed calibration curve using peak area ratios of PNT and IS.The current metabolic stability study is conducted to trace any observable decrease in the concentration of PNT when incubated with RLMs matrix using LC-MS/MS method. Incubations of 1 μM PNT were carried out (in triplicates) with 1 mgmLpH of the mobile phase (its aqueous portion) can assist the ionization of the analytes and also can adjust its peak shape. Different mobile phase compositions were examined where optimum isocratic mobile phase composition was acetonitrile-water (1:1) containing 10mM ammonium formate and pH ~ 4.1 (adjusted with formic acid). PNT and vandetanib (IS) retention times were 2.5 and 1.3 min, respectively, under the optimum chromatographic conditions. A run time of 4 min. was capable for good separation of the two analytes (PNT and IS) and no carryover was observed in blank matrix sample  or PNT-free standard sample . Several assessments were done to obtain the highest stabilized mass response through optimizing all analytical conditions so as to improve the sensitivity and resolution. The m/z 533 for PNT and at m/z 475 for vandetanib (IS). Product ion scan for PNT positive ion at m/z 533 gave major ions of [M+H]+ at m/z 433 and 260. Correspondingly, IS ion at m/z 475 gave one fragment ion of [M+H]+ at m/z 112. These ions were selected for MRM mode scanning of PNT and vandetanib (IS) in the quantification method  ≥ 0.9998 in the two matrices. The calibration curves of PNT in plasma and RLMs have the regression equations of y = 1.517x + 0.3611 and y = 1.537x + 0.1986 respectively. The LOD and LOQ were found to be 1.53 and 4.66 ngmL-1 in plasma, 1.38 and 4.19 ngmL-1 in RLMS. The high r2 value was indicative for the good linearity, and the low values of standard deviations of the intercept and the slope were indicative for the significant validity of the calibration points used for constructing the calibration curve.The suggested method was sensitive and reliable for assaying PNT in human plasma or metabolic stability method. The linear regression analysis for the results was carried out using the least-square method. The results of the calibration curve showed linearity in the range of 5–400 ngmLThe relative standard deviation values of each concentration point (six replicates) did not exceed 2.93%. Calibration and quality control samples of PNT in plasma and RLMs matrices (eleven points) were back-calculated to confirm best performance of the suggested procedure. The precision and accuracy for PNT in plasma samples were found in the range of 0.82 to 2.93% and -2.45 to 2.17%, respectively. On the other hand, precision and accuracy in RLMs were ranged from 0.36 to 2.18% and -1.95 to 2.70%, respectively . The meaThe developed method was confirmed to be reproducible using intra- and inter-day precision and accuracy at PNT QC samples. Percentages error (% error) and percentages relative standard deviation (% RSD) were used for expressing accuracy and precision, respectively. The values for accuracy and precision lied into the suitable range following ICH guidelines , 10 as sThe results of stability studies and dilution integrity in plasma and RLMs were displayed in Tables For evaluation of dilution integrity of PNT samples, The mean recovery % and CV % for 1/2 and 1/4 dilution samples in plasma and RLMs were within 98–102% and <1.8%, respectively.in vitro t1/2 [in vitro t1/2 = ln2/slope). The slope was 0.1107 and hence in vitro t1/2 was found to be 6.26 min. Consequently, CLint was calculated according to in vitro t1/2 method [After stopping the metabolic reaction at different time intervals, concentrations of PNT in RLMs matrix were calculated. These concentrations equal to the % remaining PNT with respect to zero time (which represents 100%). The ln of the % remaining PNT was drawn versus incubation time as displayed in tro t1/2   with low solvent consumption. Assaying PNT samples in one day and/or in various days demonstrated the accuracy and precision of the proposed procedure. The developed method was applied successfully for the quantification of PNT in human plasma and RLMs with the evaluation of PNT metabolic stability in latter matrix. The metabolic stability was expressed in terms of in vitro t1/2 (6.26 min) and CLin (15.182± 0.477).A simple, sensitive and rapid LC-MS/MS method was developed and validated for assaying PNT in plasma and RLMs matrices. The current method showed a linearity range of 5–400 ngmL


Background: We report the first meta-analysis on the impact of prophylactic use of a specific design of negative pressure wound therapy (NPWT) device on surgical site complications.Methods: Articles were identified in which the specific single-use NPWT device  was compared with standard care for surgical site infection (SSI), dehiscence, or length of stay (LOS). Risk ratio (RR) ±95% confidence interval (CI)  or mean difference in LOS ±95% CI was calculated using RevMan v5.3.Results: There were 1863 patients (2202 incisions) represented by 16 articles. Among 10 randomized studies, there was a significant reduction in SSI rate of 51% from 9.7% to 4.8% with NPWT intervention (RR 0.49 [95% CI 0.34–0.69] p < 0.0001). There were six observational studies assessing reduction in SSI rate of 67% from 22.5% to 7.4% with NPWT (RR 0. 32 [95% CI 0.18–0.55] p < 0.0001). Combining all 16 studies, there was a significant reduction in SSI of 58% from 12.5% to 5.2% with NPWT (RR 0.43 [95% CI 0.32–0.57] p < 0.0001). Similar effects were seen irrespective of the kind of surgery , although the numbers needed to treat (NNT) were lower in operations with higher frequencies of complications. There was a significant reduction in dehiscence from 17.4% to 12.8% with NPWT (RR 0.71 [95% CI 0.54–0.92] p < 0.01). The mean reduction in hospital LOS by NPWT was also significant (−0.47 days [95% CI −0.71 to −0.23] p < 0.0001).Conclusions: The significant reduction in SSI, wound dehiscence, and LOS on the basis of pooled data from 16 studies shows a benefit of the PICO single-use NPWT system compared with standard care in closed surgical incisions. The type of study was noted as either RCT (including interim or pilot) or observational for non-randomized comparative studies.2 statistic, and random effects meta-analyses modeling was also reported  p < 0.00001).On the basis of the 10 RCTs of varying patient numbers, there was a significant reduction in the relative risk of SSI with PICOIn comparison, there were six studies that assessed the SSI rate with non-randomized observational methodology, and these also noted a significant reduction in relative risk of SSI with PICO treatment compared with standard care—22.5% to 7.4%—which is a 15.1% absolute reduction and 67% relative reduction (fixed effects RR 0.32 [95% CI 0.18–055] p < 0.00001).% (54/1037) with PICO single-use NPWT treatment compared with 12.5% (140/1117) in the standard care group equating to an absolute reduction of 7.3% and a relative reduction of 58% (fixed effects RR 0.43 [95% CI 0.32–0.57] p < 0.0001).Overall, combining RCT and observational studies, the rate of SSI was 5.2The details of each individual study and the RCT, observation, and overall data analysis (as a forest plot) are shown in A subgroup analysis was performed on the basis of individual surgical specialities if three or more studies existed . Signifi% (78/611) with PICO single-use NPWT treatment compared with 17.4% (118/680) in the standard care group . Other reviews and meta-analyses have included a diverse mix of traditional NPWT and single-use NPWT devices, varying levels of negative pressure, and varying materials/fillers. There was also calculation of an effect on dehiscence again with a homogenous study population (I2 = 0%). On the basis of a higher level of heterogeneity in the LOS data (I2 = 94% when 8 studies combined), however, the conclusions for this specific analysis need to be downgraded but show it is worth investigating LOS further.This is the first comprehensive meta-analysis that has assessed the impact of a specific design of NPWT system on SSCs. By limiting the meta-analysis to a single brand of NPWT with a fixed level of negative pressure (−80 mm Hg), some heterogeneity was reduced. There still remains a heterogeneous mix of surgical indications and patient profiles but the sub-group analysis confirms that the benefit of PICO on SSI remains with stratification. On the basis of the IDe Vries et al  publishethe use of prophylactic NPWT on primarily closed surgical incisions in high-risk wounds for the purpose of preventing SSI, taking resources into account [The PICO-only data in this present analysis confirm that this specific design operating at −80 mm Hg is proven to reduce the incidence of SSI across several surgical indications. This echoes the W.H.O. conditional recommendations that suggest  account . It also account .To support the widespread adoption of the use of prophylactic NPWT, of course there needs to be supporting health economic evidence to show that investing in prevention delivers a net benefit for patients and healthcare systems. These studies are now emerging ,48. At aclinicaltrials.gov. Evidence is broader for SSI than for dehiscence or LOS, and it has not yet been possible to individually analyze for superficial, deep, or organ space SSI.This is a fast moving field. Even with this specific design of single-use NPWT device, new studies are being published regularly, and there are many studies ongoing as noted by published protocols  or listiThis is the first comprehensive meta-analysis that has assessed the benefit of a specific brand of single-use NPWT on SSCs. It showed a clear and significant reduction in SSIs, wound dehiscence, and LOS by application of the PICO single-use NPWT system used prophylactically on a closed surgical incision when compared with standard care.


The present study aimed at evaluating the efficacy of a novel technique, the bone lamina technique, in horizontal ridge augmentation clinically & radiographically using a combination of allogenic cortical shell, particulate xenograft and resorbable collagen membrane.Localized horizontal ridge defects, in ten patients , with bucco-palatal ridge width less than 5 mm were included in this study. Localised ridge augmentation was performed using bone lamina technique with mineralised allogenic shell of 1 mm thickness trimmed to the appropriate size using stereo-lithographic models and fixed to the recipient site with stainless steel micro-screws of 1 mm diameter. The space between the shell & host bone was filled with particulate xenograft followed by placement of collagen membrane and primary closure of the site. Clinical parameters including ridge width before & after flap reflection & radiographic (CBCT) ridge width measurements were recorded pre-operatively,and six months after the augmentation procedure. Results obtained were analysed statistically.p< 0.005), 3.15 ± 0.63 mm (p<0.005) & 3.13 ± 0.70 mm (p< 0.005) respectively.The mean clinical ridge width before flap reflection (BFR), after flap reflection (AFR) & radiographically was 3.7 ± 0.74 mm, 2 ± 0.70 mm & 1.77 ± 0.71 mm respectively at baseline which increased to 6.8 ± 0.95 mm, 5.15 ± 0.98 mm & 4.90 ± 0.90 mm with a mean gain in ridge width of 3.1 ± 0.63 mm (The present study demonstrates that bone lamina technique can be effective means of horizontal ridge augmentation and the use of mineralized allograft in combination with xenograft and collagen membrane leads to good amount of bone regeneration for subsequent implant placement. Key words:Dental implant, guided bone regeneration, horizontal ridge defect, ridge augmentation. With increasing demands of the patient regarding aesthetics and function, the need for implant supported reconstructions has substantially increased . ExtensiAlthough socket grafting with some of the bone substitutes have been able to limit the resorption of post-extraction alveolar ridge up to a certain extent, some amount of bone loss occurs inevitably. The quality of the new tissue in the socket varies broadly . Also, cA plethora of surgical techniques have been described in the last four decades regarding reconstruction of deficient alveolar bone, e.g., use of particulate grafts, block grafts, ridge splitting or ridge expansion, and distraction osteogenesis . The choThe clinician must make the appropriate selection of graft material and technique based on the size, shape, and dimensions of the defect and its location in the mouth . When thAllografts can be used in particulate form or as a block graft, either alone or in combination with autogenous, xenogeneic, or alloplastic materials. Recently, a pilot study successfully described the bone lamina technique to manage horizontal ridge defects. They utilised a porcine cortical bone shield along with collagen membrane to maintain space as well as proper contour of the alveolar ridge which resulted in predictable bone regeneration . HoweverHence, the aim of the present study was to evaluate clinically and radiographically the efficacy of the use of allogenic cortical bone lamina along with collagen membrane & particulate xenograft in the augmentation of localized horizontal ridge defects.Ten patients  for this study were selected from the out-patient Department of Periodontics & Implantology. Each patient enrolled in the study was given a detailed verbal & written description of the risks & benefits of the proposed treatment in their own language & a signed consent was obtained from them before commencement of the study. Ethical approval for the study was obtained from the Institutional Ethical Committee.Patients in the age group of 18-45 years with good general health and oral hygiene were considered for the study. These patients with localised horizontal ridge defects measuring < 4 mm in ridge width at the alveolar crest either in maxilla or mandible were included in the study . PregnanOn the first visit, complete medical and dental histories were recorded and thorough clinical examination was performed. Intra oral peri-apical radiograph of the edentulous site was assessed to exclude patients with vertical ridge defects. Routine blood investigations including complete hemogram, glycated haemoglobin HbA1c) assay, rapid ELISA, HbSAg were performed to evaluate the general well-being of the patient. Oral hygiene instructions were given and oral prophylaxis was performed. In the recall visit 2 weeks after oral prophylaxis, the oral hygiene maintenance was assessed. A cone beam computed tomography (CBCT) scan of the region of interest was recorded. Diagnostic casts were prepared. Three dimensional stereo-lithographic models were obtained from the pre-recorded CBCT  and Ketorol DT 10 mg (Ketorolac) was advised 1 hour prior to the surgery. Pre-procedural mouthrinse in the form of 0.2% Chlorhexidine gluconate mouthwash for 30 seconds was given. This was followed by disinfecting the extra-oral region of the face using 5% Povidone iodine. A cortical allograft shell of 1mm thickness  was customised to the required dimensions based on stereolithographic model . Mid-crestal incision using Bard Parker (BP) knife with 15c blade, was given which was extended as a sulcular incision on the surfaces of adjacent teeth. Vertical releasing incisions were given at the disto-buccal line angles of the adjacent teeth, connecting to the horizontal incision and extending few millimetres (mm) beyond muco-gingival junction. A full-thickness muco-periosteal flap was reflected buccally & palatal flap was reflected to just expose about 3 mm of bone to be able to stabilise the collagen membrane underneath the flap. After flap reflection, the recipient site was debrided & irrigated with sterile saline.The clinical ridge width dimensions were measured before, and after flap reflection using UNC-15 periodontal probe . The ridge width was measured bucco-palatally in mid-buccal region at the level of alveolar crest Fig. a. RadiogThe customized allogenic shell with rounded margins was adapted to the recipient site and fixed in place with 1 or 2 stainless steel mini-screws of 1 mm diameter   and analgesics Maxrel (Diclofenac 50mg + Paracetamol 500mg thrice a day) were prescribed for 5 days. Chlorhexidine 0.2% mouthrinse was also prescribed to be used twice daily for 2 weeks. Patients were recalled on third day post-surgery for re-evaluation. At 14 days post-surgery, suture removal was performed followed by temporization in the same visit using acrylic teeth splinted to the adjacent teeth using stainless steel wire and composite. Following this the patient was kept under regular follow-up once a month for 6 months.At 6 months following augmentation procedure, CBCT was taken and measurements were made on the same sagittal section as that of pre-operative CBCT Fig. a. This wp value of < 0.05 was considered to be statistically significant.All the values recorded were entered in a pro-forma & subjected to statistical analysis. Statistical analysis was done using SPSS software (Version 17.0). Wilcoxon signed rank test was used to analyse the changes in ridge width clinically and radiographically. Repeated measures ANOVA test was used to compare the mean values at different time intervals. For this study, All the patients completed the follow up & there were no drop-outs. Post-operative healing of the surgical site was uneventful except for one patient where membrane exposure was seen 3 days after the procedure. This was managed by betadine (5 % Povidone iodine) irrigation of the site on 3rd, 7th & 14th day post-surgery, antibiotic therapy (Augmentin 625mg thrice a day for 5 days) and 0.2% Chlorhexidine rinses twice daily which led to satisfactory healing of the site.p< 0.005). At baseline, the mean horizontal ridge width after flap reflection (AFR) was 2 ± 0.70 mm which increased to 5.15 ± 0.98mm six months after the augmentation procedure. A mean gain in ridge width of 3.15 ± 0.63 mm was observed and this difference was found to be statistically highly significant (p< 0.005). The mean radiographic ridge width was 1.77 ± 0.71 mm prior to augmentation, which increased to 4.90 ± 0.90 at six months following the surgical procedure which was found to be statistically highly significant with a mean gain of 3.13 ± 0.70mm (p< 0.005).At baseline, the mean clinical ridge width before flap reflection (BFR) was 3.7 ± 0.74 mm. At 6 months, it increased to 6.8 ± 0.95 mm with a mean difference of 3.1 ± 0.63 mm and the values obtained were statistically highly significant  was more than vertical reduction  at 6 months . The mean pre-operative radiographic ridge width was 1.77 ± 0.71 mm which increased to 4.90 ± 0.91 mm at the end of 6 months. This mean gain in ridge width of 3.13 ± 0.70 mm was found to be statistically significant (p < 0.05).The mean increase in ridge width clinically at the end of 6 months was 3.1 ± 0.62 (BFR) and 3.2 ± 0.63 mm (AFR). The mean post-operative bone width at 6 months was 5.15 ± 0.92 mm and the change in ridge width was statistically significant (et al., 72 sites with a mean pre-operative ridge width of 2.4 mm at the crest were treated with a combination of demineralized bone matrix, cortical cancellous chips and biodegradable synthetic barrier membrane. A mean gain in alveolar ridge width at the alveolar crest was 2.8 mm (et al. conducted a study where horizontal ridge augmentation with guided bone regeneration technique was done using a combination of autogenous block graft, particulate xenograft and collagen membrane. A mean gain in ridge width of 4.6 mm was found (Results obtained were comparable to other studies where similar combination of graft materials was used. In a study done by Geurs s 2.8 mm . von Arxas found . As the Implants placed in augmented sites are said to have similar survival rate as that placed in natural bone. In the present study, sufficient bone width was achieved following ridge augmentation and thus an implant of average diameter 3.5 to 4.5 mm and 10 mm in length was placed in each site, at the end of 6 months. Further long-term follow-up of these cases is required to determine the success of implants placed and also the augmentation procedure.Within the limitations of the present study, it can be concluded that the bone lamina technique using an allogenic cortical shell in combination with xenograft and collagen membrane can be used as an effective technique to manage the cases of horizontal ridge defects. The key to managing such defects is proper case selection. Further long term studies with larger sample size and long-term follow-up are necessary to substantiate the results obtained.


The purpose of this study was to compare the outcomes of the buccal mucosa and lingual mucosa used in children who received multiple failed hypospadias surgeries.We conducted a retrospective study of 62 children who received buccal or lingual mucosa graft urethroplasty in our hospital between 2012 and 2015. The ages ranged from 3.5–11 y. All cases included multiple failed hypospadias procedures, and the subjects received previous operations 2–3 times. All patients underwent one-stage operations. Thirty-three cases were treated with lingual mucosa grafts. The patient ages ranged from 3.5 to 11 y (median 7.5 y), and they had previous operations 2–3 times (mean 2.8±0.7). Grafts ranged from lengths of 2–6 cm (mean 5.1±0.46 cm) and widths of 0.5–1.5 cm (mean 1.2± 0.16 cm). Our follow-up was 5 to 12 m (mean 8.3±1.2 m). Twenty-nine cases were treated with buccal mucosa grafts. The patient ages ranged from 4 to 9.2 y (median 7.0 y), and they had previous operations 2–3 times (mean 2.5±0.2). Grafts ranged from lengths of 2–5.3 cm (mean 4.9± 0.28 cm) and widths of 0.5–1.5 cm (mean 1.0±0.11 cm). Our follow-up was 5 to 12 m (mean 7.9±0.5 m). The results were tested with SPSS 18.0. The rates of complications were compared by a chi-square test, and pre-operative conditions were compared by t test.For the outcomes of the two groups, there was no significant difference between the groups in terms of age, preoperative surgery time, and the length and width of the grafts (p>0.05). For the lingual mucosa graft group, fistula: 2/33 (6.0%), stricture: 1/33(3.0%), ventral curvature: 2/33(6.0%), complications: 5/33(15.0%), success rate: 28/33(84.8%), Hose score: 14.34±0.95, peak flow: 6.5 ml/s-12.0 ml/s, and mean peak flow: 9.3±0.4 ml/s. For the buccal mucosa graft group, fistula: 2/29(6.8%), stricture: 2/29(6.8%), ventral curvature: 1/29 (3.4%), complication rate: 5/29(17.0%), success rate: 24/29 (83.0%), Hose score: 14.28±1.03, peak flow: 6.5 ml/s-12.0 ml/s, and mean peak flow: 9.2±0.2 ml/s. There were no differences between the two groups for overall success, complication rates, peak flow, and the Hose scores(P>0.05).The lingual mucosal graft and the buccal mucosa graft both achieved good outcomes, and the lingual mucosa graft made up for the shortcomings of the buccal mucosa graft, which provided a reliable way to treat the multiple failed hypospadias surgeries in pre-pubertal boys. The incidence of hypospadias is approximately 1/300, which has rapidly grown in recent years. Because surgeries are the only way to solve hypospadias, many studies of surgeries for the condition have been reported in the literature. We found different types of surgeries with different outcomes . Since HWe recruited sixty-two pre-pubertal boys who received buccal or lingual mucosa graft urethroplasty in our hospital between 2012 and 2015. The patThe preparation of the LMG before the operation was similar to BMG urethroplasty. Because all of the 62 patients could have received either surgery, we assigned the two procedures at different periods. All patients were operated on general anesthesia with nasotracheal intubation. We documented the appearance of the failed hypospadias procedures . For theHarvesting method for the lingual mucosa grafts: We measured the urethral defect , keepingHarvesting method for the buccal mucosa grafts: We measured the defect and fully exposed the lower lip mucosa. Nor-epinephrine  was applied to the area in both groups, and the graft was harvested Figs  and 8.The graft was defatted . The donThe urethral catheter was removed after 3 weeks. For data analysis, our hospital used IBM SPSS version 18.0 for statistical analysis. The comparison of urethral meatus measurements and the outcomes was performed with a chi-square test, and P < 0.05 indicated that the differences were statistically significant. We assessed the outcome of the cosmetic results with the HOSE score table , diverticula (which could affect the urine stream and require surgery), meatal stenosis/stricture (with stricture apparent on a urethrogram and requiring subsequent intervention), persistent ventral curvature, glans dehiscence and/or skin reoperation with some bleeding, and a loss of stent .For the outcomes of the two groups, there was no significant difference between groups in terms of age, previous repairs, and the length and width of the grafts (p>0.05) . For theIn our study, 10 cases were defined as failures based on the definition of success. Three patients were diagnosed with urethra strictures  based on the urethrography. All returned to normal after receiving dilation. Four patients with fistulas had received fistulous neoplasty. Three cases with ventral curvature  were conducted with dorsal tunica albuginea plication based on the methods of Baskin (1994) [Urethroplasty is the key procedure for treating hypospadias and different types of procedures have appeared over the years. Many patients have to face multiple surgeries due to failed procedures, and these patients typically develop problematic conditions, such as stricture, fistula, and ventral curvature , 12. It However, this method is not suitable to patients with longer urethral defects, physiological deficiency of oral grafts or oral mucosa damage due to previous harvestings. Additionally, many scholars have reported that the buccal mucosa graft donor site has some complications, such as persistent perioral numbness, salivary changes, and difficulty in opening the mouth . This diWe could barely find any studies about the comparison between the two grafts used in inlay urethroplasty, especially in the pre-puberty boys from a Chinese population. To evaluate the outcomes, we conducted a retrospective study of sixty-two children who received either buccal (twenty-nine) or lingual (thirty-three) mucosa graft urethroplasty in our hospital between 2012 and 2015. The ages ranged from 3.5-11y. For the outcomes of the two groups, there was no significant difference between groups in terms of age, preoperative surgery time, and the length and width of the grafts (p>0.05) . For theOur results showed that we have achieved satisfactory outcomes for the two groups. The overall successful rates of the BMG and LMG were 83.0% and 85.0%, respectively, which were similar to the 87% success rate reported by Abdelhameed et al  in adultWe know from the literature that the lingual mucosa shares many characteristics with the urethra, such as the same embryologic origin as the urethra as well as the immunologic advantage of resistance to infection and tissue characteristics , which are more obvious prior to puberty , 21. TheWhen multiple hypospadias procedures fail, 1) the failed hypospadias repairs are often associated with penile skin loss and deficient local tissues, 2) BMG are often not possible due to oral complications from a prior procedure, 3) patients undergoing failed hypospadias often have long urethral defects, and 4) the buccal mucosa grafts are not available due to frequent tissue harvesting. If buccal mucosa grafts are confronted with scarred tissues, BMG urethroplasty cannot be chosen again. So if surgeons are confronted with these problems in pre-pubertal boys, there is still a reliable graft to fix the poor repair.It should be noted that compared to the LMG used in adults, the advantages of LMG used in children before puberty are that the tissue material is soft and thick in mucosal tissues with a better blood supply, high cell activity and survival rate. However, there are also disadvantages in that the tongues of children have a smaller volume and are harder to secure during the procedure than tongues in adults.The length of the lingual graft in children is long enough for the defect in general, and if the urethral defect is longer, we could choose the two sides of the ventro-lateral surface of the tongue. The thickness and the articulation are related to the complications based on the outcome and the long time follow up in children. We have the relatively shorter follow up time with a good outcome, and maybe it’s a shortage of our research.The limitations of our study include 1) a short follow-up period. A short follow-up period may have had an effect on the complication rates. We had The lingual mucosal graft and the buccal mucosa graft both achieved good outcomes, and the lingual mucosa graft made up for the shortcomings of the buccal mucosa graft and provided a reliable way to treat multiple failed hypospadias procedures in pre-pubertal boys.S1 File(PDF)Click here for additional data file.


Mussismilia braziliensis during a diurnal cycle, and compared them to the water column. We determined microbial abundance and nutrient concentration in samples taken within a few centimeters of the coral's surface every 6 h for 48 h, and sequenced microbial metagenomes from a subset of the samples. We found that dominant taxa and functions in the coral MBL community were stable over the time scale of our sampling, with no significant shifts between night and day samples. Interestingly, the two water column metagenomes sampled 1 m above the corals were also very similar to the MBL metagenomes. When all samples were analyzed together, nutrient concentration significantly explained 40% of the taxonomic dissimilarity among dominant genera in the community. Functional profiles were highly homogenous and not significantly predicted by any environmental variables measured. Our data indicated that water flow may overrule the effects of coral physiology in the MBL bacterial community, at the scale of centimeters, and suggested that sampling resolution at the scale of millimeters may be necessary to address diurnal variation in community composition.Corals display circadian physiological cycles, changing from autotrophy during the day to heterotrophy during the night. Such physiological transition offers distinct environments to the microbial community associated with corals: an oxygen-rich environment during daylight hours and an oxygen-depleted environment during the night. Most studies of coral reef microbes have been performed on samples taken during the day, representing a bias in the understanding of the composition and function of these communities. We hypothesized that coral circadian physiology alters the composition and function of microbial communities in reef boundary layers. Here, we analyzed microbial communities associated with the momentum boundary layer (MBL) of the Brazilian endemic reef coral  Coelastrea aspera increases in a step-wise pattern from juveniles to medium colonies, decreasing again in older, larger colonies  and the filters were frozen. Three 2 l subsamples were filtered through 0.7 μm pore GF/F glass fiber filter and aliquots of 50 ml of the filtrate were conserved in amber bottles with 1 ml of phosphoric acid for Dissolved Organic Carbon (DOC) determination , inside the Abrolhos Marine National Park (SISBIO permit 27147-2) in August 14th to 16th, 2011, during the austral winter. The main organisms contributing to benthic cover in Santa Bárbara are: coral 7.7%, crustose coralline algae (CCA) 6.6%, fleshy algae 22.4%, and turf algae 66.4%. Our sampling strategy targeted the MBL above visually healthy Inorganic nutrients concentrations were determined using standard oceanographic methods . Three “day” time points were sequenced: 18:00 on August 14th (the first sampling day), 18:00 on August 15th, and 12:00 on August 16th, the last sampling day. We also sequenced one “night” water column sample from 00:00 on August 15th and one “day” water column sample from 12:00 on August 15th. Samples from 18:00 were taken before sunset, at the end of 12 h of daylight exposure, and therefore were grouped as “day.” The same criteria was applied to 06:00 samples grouped as “night.”To investigate the composition of microbial assemblages associated to https://doi.org/10.6084/m9.figshare.4614745.v1) and MG-RAST under the project Diel Mussismilia . Reads shorter than 100 bp and with mean quality scores lower than 25 were removed using PrinSeq , while orthophosphate and nitrate were positively correlated . Nitrate contributed to most of the inorganic nitrogen pool both in the coral MBL and the water column . Concentration of DOC and Chlorophyll a are shown in Table r = 0.79, p < 0.001). Chlorophyll a, DOC, and nutrient concentrations were not significantly different between day and night samples . Therefore, we were not able to identify diurnal patterns in nutrient concentration in the MBL above M. braziliensis or the water column at this time scale.Variation in organic and inorganic forms of nitrogen and phosphorous over time are shown in Table 5 to 7.0 × 105 ml−1 in the coral MBL, and from 4.4 × 105 to 6.6 × 105 ml−1 in the water column. Viral abundances ranged from 4.8 × 106 to 10.6 × 106 ml−1 on the coral and 5.1 × 106 to 7.3 × 106 ml−1 in the water column. When analyzing MBL and water column samples together, viruses and prokaryotes showed a significant negative correlation to each other . Prokaryotic abundance also correlated negatively to total phosphorous concentration , while viral abundance correlated positively to total phosphorous . The abundance of autotrophic pico- and nanoeukayotes ranged from 4.2 × 103 to 10 × 103 ml−1 and from 3.6 × 102 to 10.8 × 102 ml−1, respectively. Pico and nanoeukaryote abundances were positively correlated to each other  and were both negatively correlated to viral abundance . Prokaryotic and picoeukaryote abundances correlated positively . No diurnal patterns were observed in the microbial and viral abundances on the coral MBL or in the water column . However, over the course of the diurnal monitoring we detected a viral burst event that occurred at 18:00 of the first night. At this time, prokaryotic abundances in the MBL dropped to the lowest values recorded during the cycles, while viral abundances peaked , Firmicutes (21.5%), Thaumarchaeota (9.6%), Euryarchaeota (8.4%), Cyanobacteria (5.9%), and Bacteroidetes (4.4%). During the day, the same phyla were dominant, except for Euryarchaeota: Proteobacteria (41.6%), Bacteroidetes (13.79%), Cyanobacteria (8.3%), Firmicutes (8.2%), Thaumarchaeota (4.3%), and Tenericutes (4.2%). The main phyla in the water column at night were Proteobacteria (39.4%), Cyanobacteria (17.2%), Firmicutes (9.4%), Thaumarchaeota (7.4%), and Euryarchaota (5.7%). Overall, the same phyla dominated all three groups of samples.The microbial community associated with M. braziliensis MBL at night was dominated by members of the genera Anaplasma , Coprococcus (6.05%), Staphylococcus (5.6%), unclassified Alphaproteobacteria (5.0%), Coprothermobacter (5.0%), and the archaeal genus Nitrosopumilus (5.0%) , Fluviicola (4.8%), Vibrio (4.6%), Synechococcus (4.6%), Glaciecola (3.8%), and Alteromonas (3.5%). The most abundant genera in the water column were Synechococcus (12.5%), unclassified Alphaproteobacteria (8.3%), Hirschia (5.3%), Anaplasma (4.9%), Prochorococcus (4.4%), and Nitrosopumilus (3.0%). NMDS analysis based on relative abundances of genera showed no significant clusters among the three tested groups , and Pielou's evenness, respectively).At the genus level, the microbial community associated with ) Figure . During aciecola .8%, and SD); amino acids and derivatives (11.9 ± 1.4%); protein metabolism (9.6 ± 1.2%); cofactors, vitamins, prosthetic groups, pigments (8.7 ± 0.5%), and RNA metabolism (6.3 ± 1.1%). At the level 2, most of the predicted proteins were unclassified , followed by protein biosynthesis (6.3 ± 0.5%), central carbohydrate metabolism (6.3 ± 0.3%), lysine, threonine, methionine, and cysteine (3.7 ± 1.4%), RNA processing and modification (3.4 ± 0.7%), and transcription (2.9 ± 0.6%). NMDS analysis was performed based on the abundance of functions classified at the level 2 subsystems (as opposed to level 1 described above). PERMANOVA-tests showed no significant difference in functional profiles between MBL and water column or between daytime and nighttime MBL . Following, DOC contributed to 27%, prokaryotic abundance and source (MBL or water column) each contributed with 17.8%, time of collection with 14%, and total phosphorus with 13.9%. All the other variables contributed to 10% or less the community dissimilarity. We tested significant correlations between nitrate, which had the highest explanatory proportion, and each taxon. Coprothermobacter was negatively correlated with nitrate . We then performed a reverse DistLM-test where taxonomic dissimilarity was tested as predictor for environmental variable dissimilarity. Synechococcus was the genus explaining the highest proportion of environmental variable dissimilarity .We investigated if taxonomic profiles in both MBL and water column samples were determined by nutrient concentrations. We performed these analyses with 26 dominant genera, defined as those with at least 1% relative abundance, which contributed to a total of 70.6% of the community composition. The relationship between environmental variables and dominant taxa can be visualized in the Canonical Correspondence Analysis (CCA) plot Figure . DistLM p-values were not significant . Accordingly, the reverse test where functional profiles were tested as predictors of environmental variables showed no significant functions.Analyses of relationships between environmental variables and dominant functional profiles were performed on 25 SEED level 2 subsystems, each one with abundances of at least 1%, contributing to a total of 51.9% of all functionally annotated reads. Correlations between dominant functions and environmental variables can be visualized in the CCA Figure . DistLM M. braziliensis at the diel scale. We found no evidence for a diel pattern in bacterial taxonomic and functional profiles at the time scale sampled, and observed that the variability of taxonomic profiles correlated with nutrient concentrations, suggesting that the coral MBL was more influenced by water column dynamics than by coral physiology. This result is consistent with previous findings in water column bacterial communities in the Great Barrier Reef, and supports the hypothesis that diurnal patterns in community composition are related to planktonic processes rather than to benthic–pelagic coupling . This suggests that a significant fraction of the active community might be comprised of eukaryotic viruses, despite the fact that these viruses comprise only about 10% of total the viral community (Brussaard et al., Despite the lack of a diel pattern in nutrients and microbial abundance during the sampled period, we identified a synchronized change in total prokaryotic and viral abundance that indicated a lytic event. Studies of short time scale variation in viral abundances in marine environments have conflicting results, and most of them did not display diel cycles (Jiang and Paul, S. pistillata is significantly thinner when a 5 cm/s current is applied (Shashar et al., The lack of diel patterns and the correlations between taxonomic dissimilarities and nutrients suggest that water flow over corals determines microbial community composition in the MBL, rather than diel switches in coral physiology. Community assembly patterns observed here in the MBL, and before in the coral holobiont, indicate that stochastic processes such as migration or nutrient fluctuations due to hydrodynamics are important forces shaping the short-term fluctuations in the microbiome, as opposed to deterministic processes (Sloan et al., M. braziliensis do not display significant diurnal patterns. Instead, this community is similar to the water column 1 m above the corals and responds to environmental fluctuations in nutrient concentrations. Our results suggest that despite coral mucus contribution to the MBL community composition, the overlying water currents contribute more to short term variability in community assembly than diurnal changes in coral physiology. These results expand the view of short-term temporal dynamics of coral microbiome, often bypassed in studies on coral health and reef ecology.In conclusion, we show that the microbial community in the MBL of the coral CS, GG, and FT designed the study; CS, GG, FC, GS, JH, LO, AC, CR, CT, RF, RE, and ED generated and analyzed the data; CS and FT wrote the manuscript and all authors contributed to revisions.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 satisfactory outcome in the surgical treatment of obesity must include, in addition to weight loss, a significant change in the pre-existing comorbidities and in the quality of life.  To evaluate the quality of life in the late postoperative period in patients that underwent videolaparoscopic sleeve gastrectomy.  It was applied the questionnaire "Bariatric Analysis and Reporting Outcome System" (BAROS) in patients that underwent videolaparoscopic sleeve gastrectomy.  A total of 47 patients between 21-60 years old were evaluated. The total mean of BMI before surgery was 43.06±5.87 kg/m². The average percentage of the reduction of excess weight after surgery was 85.46±23.6%. The score obtained by patients in the questionnaire about the improvement in the quality of life showed excellent (36.17%), very good (40.43%), good (21.28%) and reasonable (2.13%) results. There was clinical improvement after surgery in all comorbidities investigated.  The weight loss was critical to improve the quality of life and offered the resolution or clinical improvement in all of the investigated comorbidities in patients submitted to sleeve gastrectomy. Overweight and obesity are defined as an excessive accumulation of body fat that can affect health. Obesity is considered a chronic and multifactorial disease, and it is associated with several comorbidities and severe losses in quality of life,,The surgical approach for severe obesity is called bariatric surgery and appeared around 1950. Over the following years, several techniques and analysis were developed and they recognized this procedure as effective both in weight loss and in reduction of comorbidities, providing increased survival compared to clinical approaches only2 when holding comorbid conditions that threaten life, as type 2 diabetes mellitus, hypertension, dyslipidemia, arthropathy, and others. This approach is also possible for patients with a poor clinical treatment of at least two yearsSurgery is indicated for patients with Body Mass Index (BMI) greater than 40 kg/m² or greater than 35 kg/m,,The procedure that patients underwent in this article is called "vertical gastrectomy" or "gastric sleeve". It is a restrictive procedure that involves the removal of the greater curvature of the stomach, starting from 4 to 6 cm from the pylorus to the esophagogastric angle, leaving the new shell with a tubular and elongated shapeHowever, as many factors other than weight are modified after this procedure, it is essential to evaluate the quality of life of these patients. Thus, in order to reinforce the importance of continuity of care in the postoperative period it was developed in 1998 the "Bariatric Analysis and Reporting Outcome System Protocol" (BAROS). This instrument has emerged as a simple, cheap and reliable evaluation of self-perceived quality of life in patients in the postoperative period of weight reduction surgeryThe BAROS protocol consists of three major areas of analysis , which obtains a maximum score of three points for each category, totaling a maximum of nine points. The score of the category "weight loss" is given by the percentage of excess weight loss (% EWL). In "medical conditions", the individual reaches a higher score as there is clinical improvement or cure of one, several or all preexisting comorbidities before surgery. Finally, the "questionnaire of quality of life" includes questions about physical activity, social interaction, working performance, sexual interest and improvement in the general conditionThus, the purpose of this study was to apply this protocol in patients who underwent stomach reduction surgery through laparoscopic vertical gastrectomy.This article is the result of a cross-sectional study conducted between the months of April and May of 2016. The sample was selected from a clinical database of patients undergoing laparoscopic vertical gastrectomy surgery by the staff of the Clinic of Surgery of Digestive System - VIDAR, Blumenau, SC, Brazil between 2012 and 2015. The project was approved by the Ethics Committee of the Regional University of Blumenau number 1519860.Patients were contacted and invited to participate in person filling the printed BAROS questionnaire. The same questionnaire adapted in "Google Forms" platform was sent by e-mail if impossibility of displacement to the clinic. Exclusion criteria were: a) Patients who had any cognitive limitation to compromise responses to data collection instrument; b) who refused to answer the questionnaire; c) who did not answered a question of the form. The study included only patients who were in the late postoperative period. The data were divided into three categories, according to the period elapsed after surgery: a) up to six months; b) seven to 12 months; c) more than 12 months. Was performed using Epi Info software, version 7. The data level of significance was p<0.05.2. The average overweight of the patients was equal to 51.01±18.48 kg, an equivalent to 40.93±7.65% excess of average body weight.A total of 47 patients were evaluated, of whom 76.6% (n=36) were female and 23.4% (n=11) male. The average age was 37,3±10,75 years. The average weight and BMI before surgery were, respectively, 121.05±22.56 kg and 43,06±5.87 kg/mAll 47 patients completed the questionnaire BAROS, generating the following results: 36.17% (n=17) were classified as "excellent"; 40.43% (n=19) as "great"; 21.28% (n=10) as "good"; and 2.13% (n=1) as "reasonable" improvement of quality of life. No patient had a score in the "bad" category .There was no statistically significant difference between the score of men and women (p>0.05). The time of postoperative period was also not significant in influencing the BAROS score (p>0.05). In patients with more than six months after the surgery, it was observed predominance of excellent (71.58%) and great (76.84%) results. It was also found that the highest scores were obtained for those patients who had lost more than 75% of overweight (p<0.05).For the five domains assessed by questionnaire, the results were as follows: a) General condition: all patients responded "very good" (n=40 or 85.11%) or "good" (n=7 or 14.89%); b) Social or family activities: 21 of 47 patients (44.68%) claimed to have been no change in this area, and one patient (2.13%) reported worsening of social relationships; "improved a lot" got 29.79% (n=14) and the "improved" 23.40% (n=11); c) Physical activities: "increased a lot" 40.43% (n=19), "increased" 34.04% (n=16), "unchanged" 23.40% (n=11) and "decreased" 2.13% (n=1); d) Sexual interest: "greatly increased" 14.89% (n=7), "increased" 29.79% (n=14), "unchanged" 44.68% (n=21), "decreased" 6 38% (n=3) and "greatly diminished" 4.26% (n=2); e) Working performance: "increased a lot" 51.06% (n=24), "increased" 38.30% (n=18), "unchanged" 4.26% (n=2) and "decreased" 6.38% (n=3) .The gender of the patient was not a significant factor in EWL% (p>0.05). A total of 69.4% of women lost more than 75% overweight vs. 45.4% for men. However, men lost more total weight in kilograms (72.7% lost between 40-80 kg against 38.9% of women). Patients with grade 3 obesity  statistically (p<0.05) obtained better results in the loss of excess weight: 65.9% of patients lost more than 50% of the excess of weight.The time of postoperative period exerted a significant influence (p<0.01) in reducing the BMI of patients: limited reductions, of less than 10 kg/m², were found in individuals living a postoperative period of less than six months; in contrast, 52.63% with more than 12 months lost between 15-30 kg/m².Regarding comorbidities among patients, 55.3% (n=26) had hypertension, 15% (n=7) had type 2 diabetes mellitus, 27.6% (n=13) had dyslipidemia, 44.7% (n=21) had obstructive sleep apnea, 49% (n=23) suffered from some form of joint problem and 49% (n=23) had depressive disorder before performing the surgical procedure to reduce weight. These comorbidities were not diagnosed or classified by the authors, but self-reported by patients included in this study.It was observed in the postoperative period that 77% (n=20) of patients who had high blood pressure achieved complete resolution of this disease and 38.4% were able to maintain control of hypertension with fewer antihypertensive medications. Among patients with type 2 diabetes mellitus, 71.4% (n=5) had complete resolution of disease and 28.5% (n=2) were able to reduce medication for glycemic control. Those who had dyslipidemia, 92.3% (n=12) showed no more such comorbidity and among those suffering from obstructive sleep apnea syndrome, 57.1% (n=12) did not report symptoms. Of individuals who had some kind of joint problem, 61% (n=14) extinguished this disease and did not use medications anymore; 34.7% (n=8) were able to reduce the number of painkillers and other medicines. Finally, of patients who had depression in preoperative period, 39.1% (n=9) reduced the number and/or concentration of antidepressant medications and 48% (n=11) did not have symptoms anymore , 80.82±20.43% (n=25) and 93.13±27.42% (n=19), respectively.In relation to the average weight loss, patients who had undergone surgery within six months (n=3) lost 31.67±16.77 kg; between seven and 12 months (n=25), 38.36±8.94 kg; more than 12 months (n=19), 45.83±12.12 kg. The average reduction of BMI was 10.53±3.83 kg/m2; overweight, 10.01±14.08 kg equivalent to 9.72±15.25%; the EWL% of all patients was 85.46±23.6% , dizziness (n=2), gastroesophageal reflux disease (n=1), hypotension (n=1), weakness (n=1), epigastric pain (n=1), and tremor (n=1). No patient reported more than one complication, and only one required reoperation, but did not justify the reason.,,,,,Several studies document the influence of weight loss in improving the quality of life The average score of patients in the questionnaire of quality of life was 1.85±0.64, a maximum of three points. Similar data were obtained by Janik et al.,The BAROS' domains most highly evaluated by patients were "general condition" (average score of 0.92 up to 1.0), "working performance" (0.33/0.5) and "physical activity" (0.28/0.5). The areas "social activities" (0.2/0.5) and "sexual interest" (0.11/0.5) had the worst average. This happened because most patients have answered that there was no change in these two domains generating zero score. Other studiesLike other studiesA retrospective study of 407 patients published by Ortega et al.,Several studies show that the sleeve gastrectomy reduces mortality and the development of new comorbid conditions and worsening of already present diseases in obese individuals,The prevalence of diabetes among the obese patients in the preoperative period was similar to the study of Blume et al.In addition to the above comorbidities, the high frequency of psychological diseases of obesityLimitations of this study include the subjectivity of BAROS questionnaire, the non-equivalence of the samples in the postoperative period of time, not representativeness of the sample compared to the prevalence in the population. In addition, analysis of the quality of life would be more accurate if the assessment was carried out in different postoperative times of each patient, obtaining thus an evolutionary clinical parameter.Quality of life was affected by the reduction of excess weight and the metabolic changes with resolution of most of the evaluated comorbidities. The change of patient dissatisfaction regarding the ability to work, physical activity and general health were also considerable. The least significant results with respect to sexual and social activities interest are possibly related to self-assessment capacity.


Introduction: The Mediterranean corn borer (MCB), Sesamia nonagrioides, is a major pest of maize, Zea mays, in Mediterranean countries, inflicting significant kernel yield losses. For that reason, it necessary to know the genetic mechanisms that regulate the agronomic and resistance traits. A quantitative trait loci (QTL) mapping study for yield, resistance against MCB attack, and other relevant agronomic traits was performed using a recombinant inbred line (RIL) population derived from the cross A637 × A509 that is expected to segregate for yield, and ear, and stalk resistance to MCB. 171 RILs were evaluated in 2014 and 2015 at Pontevedra, Spain, along with the two parental inbreds A637 and A509 using a 13 × 14 single lattice design with two replications. A genetic map with 285 SNP markers was used for QTL analysis. Our objectives were to detect QTL for resistance to MCB and tolerance-related agronomic traits, to gain insights on the genetic relationship between resistance to MCB attack and yield, and to establish the best way for simultaneously improving yield and resistance to MCB.Results: Twelve significant QTL were detected for agronomic and resistance traits. QTL at bins 1.10 and 5.04 were especially interesting because the same allelic variant at these QTL simultaneously improved yield and insect resistance. In contrast, in the region 8.04–8.05, QTL showed opposite effects for yield and resistance. Several QTL for indexes which combine yield and resistance traits were found especially in the region 10.02–10.03.Conclusions: Selecting genotypes with the favorable allele of QTL on chromosome 5 (bin 5.01) will decrease tunnel length without affect yield, silking and plant height and QTL on the region 5.04 could be used to improve stalk resistance and yield simultaneously. An allele of QTL on bin 9.07 will increase ear resistance to MCB attack but it could produce later varieties while favorable allele in region 1.10 could improve ear and stalk resistance and yield without secondary negative effects. The region 8.03–8.05 mainly but also the region 10.02–10.03 and 5.04 may play an important role to elucidate the association between yield, other agronomic traits and MCB resistance. Ostrinia nubilalis Hübner in North America, Europe, and North Africa; Ostrinia furnacalis Moth in Asia; Diatraea saccharalis Fabricius from the United States to Argentina; Chilo partellus Swinhoe in the southern United States, Central America, and the Caribbean; Diatraea lineolata Walker in Central America, the Caribbean, and South America; and Diatraea grandiosella Dyar in North and Central America , which produces Bt toxins, is a good method for controlling the MCB, but transgenic crops are not authorized in most European countries and are banned in organic agriculture  crops were planted globally  have been successful in developing materials with partial resistance to the MCB F2 population, A637 is an American tolerant line while EP42 is a Spanish sensible line, both are susceptible to corn borer attack. In this study were used two American lines: A637 is tolerant to yield losses and stalk susceptible to corn borer attack while A509 has high yield losses but is resistant to larvae attack. Therefore, the bi-parental population used in the current study offers an additional advantage over the other previously used for revealing the genetic factors behind this unfavorable genetic correlation. The present population is derived from a cross between two lines with contrasting values for yield and resistance to stem tunneling by MCB  resistance, measured as shorter stem tunnels lengths made by MCB larvae, and yield have complex genetic architectures with many minor genes contributing to each trait , to explore the genetic relationship between yield and resistance in a bi-parental population derived from the Corn Belt Dent heterotic group, and to establish the best way for simultaneously improving yield and resistance to MCB.6) derived from the cross A637 × A509 were used in this study. The inbred line A509 was sensitive (higher yield loss than the average) and resistant (shorter stem tunnel length and higher ear resistance than the average) to MCB attack, while the A637 line was tolerant (lower yield loss than the average) but susceptible (larger stem tunnel lengths and higher ear resistance than the average) to the MCB  using a 13 × 14 single lattice design with two replications. Trials were grown at Pontevedra, a location in northwestern Spain . They were hand planted, and each experimental plot consisted of one row of 2.16 m length and 0.80 m between rows, with 13 two-kernel hills spaced 0.18 m apart. Plots were overplanted and thinned, obtaining a final density of ~70,000 plant ha Albajes , and Kha Albajes . Before  Albajes ; tunnel 2) across environments were estimated for each trait on a family-mean basis as described by Holland et al. (rg) and phenotypic (rp) correlations between traits were computed following Holland /EffectTS.QTL analysis was performed using the software package PlabMQTL Utz, . The comx is the observed data, μ is the weighted mean of the trait, and σ is the standard deviation. From the standardized data, four indexes were calculated using standardized data: (1) Y+kr−tl (yield plus kernel resistance minus tunnel length); (2) Y−tl (yield minus tunnel length); (3) Y+kr (yield plus kernel resistance); and (4) Kr−tl (kernel resistance minus tunnel length) and QTL analysis for these indexes were carried out. Two or three traits were combined with the same weight to value together (1) yield and both resistances, (2) yield and stalk resistance, (3) yield and ear resistance, (4) stalk and ear resistance.The data set for each trait was standardized using the formula Z = A QTL joint analysis to co-localizing QTL with additive effects on agronomic and resistance related traits was made using JointQTL option from PlabMQTL Utz, . A LOD trg = 0.65). The genetic correlation coefficients between tunnel length and yield (rg = 0.59) and between tunnel length and plant height (rg = 0.63) were moderate. The highest genetic correlation coefficient was between kernel resistance and silking (rg = 0.86). On the other hand, phenotypic correlations were low; except the correlation between plant height and yield that exceeded 0.50 , 8 (8.04–8.05), and 9 (9.07). These QTL accounted for 50.54% of the total genotypic and 44.50% of the total phenotypic variance. The absolute values of additive effects were 0.71, 1.38, and 0.83 days for the QTL on chromosomes 1 (1.01), 8 (8.04–8.05), and 9 (9.07), respectively. The percentage of bias estimation for additive effects was between 1.1 and 12.7%. The detection frequency in the cross validation of the QTL on chromosome 9 (9.07) was higher (0.90) than that observed for QTL on chromosomes 8 (8.04–8.05) and 1 (1.01). Three QTL for plant height were located on chromosomes 2 (bin 2.04), 4 (4.02), and 8 (8.04–8.05). These QTL accounted for 33.0% of the total genotypic and 21.1% of the total phenotypic variance. The absolute values of additive effects estimated were 4.03, 4.89, and 5.48 cm for the QTL on chromosomes 2 (bin 2.04), 4 (4.02), and 8 (8.04–8.05), respectively. The detection frequency in the cross validation of the QTL on chromosome 8 (8.04–8.05) was higher (0.67) than that observed for QTL on chromosomes 2 (bin 2.04) (0.32) and 4 (0.34).For yield, only one QTL located on chromosome 8 (bin 8.04) was detected. The proportion of genotypic and phenotypic variance explained by the QTL was 23.01 and 15.90%, respectively. The absolute value of additive effect was 0.49 Mg haTwo QTL for the Y+kr−tl index were located on chromosomes 1 (bin 1.10) and 10 (10.02–10.03), and explained more than 37% of the genotypic and 17% of the phenotypic variance. The absolute values for additive effects for the QTL were 0.46 and 0.61 with bias estimations of 54 and 63%. Two QTL for the Y−tl index were located on chromosomes 5 (bin 5.04) and 10 (10.03) and explained more than 31% of the genotypic and 15% of the phenotypic variance. The absolute values of additive effects for the QTL in chromosomes 5 (bin 5.04) and 10 (10.03) were 0.30 and 0.41 with estimation biases of 21.2 and 58.7%, respectively. A QTL for Y+kr index was located in chromosome 8 (bin 8.03–8.04) and accounted for 8.87% of the genotypic and 5.0% of the phenotypic variances, respectively. The absolute value of the additive effect estimated TS for this QTL was 0.31. Another QTL located on chromosome 10 (bin 10.02–10.03) for Kr-tl index accounted for 47.6% of the genotypic and 15.7% of the phenotypic variances. The absolute value of the additive effect estimated for this QTL was 0.58.Analyzing the joint effect of two traits confirmed that the QTL on bins 8.04–8.05 significantly increased yield and tunnel length  were similar to those obtained by other authors using diverse RIL populations under infestation with corn borers , was lower than the heritabilities for agronomic traits but enough high to wait positive results in selection programs to reduce larvae damage. Tunnel length trait has associated a high experimental error because it is affected by plant genotype, larvae pressure and environment and it is more difficult to measure than some agronomic traits as plant height. So, resistance traits show generally low heritabilities.In this study, an intermediate value of The genetic correlation coefficients between tunnel length and agronomic traits in this study indicates that susceptible genotypes would have greater yield, taller plants, and a delayed maturity than those resistant to the MCB as it has been previously suggested by Butrón et al. . This agQTL of MCB resistance appeared in regions where none had been found previously, as is usual when we look for QTL for resistance in biparental populations, it is reinforcing the hypothesis of the polygenic character of this trait. However, the QTL for MCB tunnel length located at bins 8.04–8.05 was already reported in a previous study with RILs derived from a cross between Spanish inbred lines  locus on chromosome 8 at 132 Mb, a major QTL for flowering time, which has been cloned in maize . The favorable allele at bin 5.04 and at bin 10.03 came from A509 and they will be useful for increasing yield and reducing tunnel length in a MAS program, these QTL did not affect kernel resistance. Samayoa et al.  found a The joint analysis corroborated the results of the correlation analysis. Associations between yield and tunnel length were found, as well as QTL for silking and plant height. For all joint QTL, A637 allele produced later maturing and taller plants with greater yield at harvest and longer tunnels than those produced by A509 allele. Therefore, other characteristics, besides yield, such as plant height should be examined to clarify the negative relationship between yield and resistance. It must be emphasized that similar associations between resistance and agronomic traits have been found at bins 8.04–8.05 in lines with different genetic backgrounds, which makes that region especially interesting for further studies about relationship between MCB resistance and yield.Three QTL for tunnel length were found on chromosomes 5, 8, and 10 , one for kernel resistance on chromosome 9 (bin 9.07), one for stalk damage on chromosome 10 (bin 10.02–10.03), one for yield on chromosome 8 (bin 8.04), three for silking on chromosomes 1, 8, and 9 , three for plant height on chromosomes 2, 4, and 8 . At the same time, two QTL for Y+kr−tl index were found on chromosomes 1 and 10 , two QTL for Y−tl on chromosomes 5 and 10 , one QTL for Y+kr on chromosome 8 (bin 8.03–8.04), and finally one QTL for Kr-tl on chromosome 10 (bin 10.02–10.03).The region on chromosome 8 (bin 8.03–8.05) mainly but also the region on chromosome 10 (bin 10.02–10.03) and chromosome 5 (bin 5.04) may be the keys to elucidate the association between yield, other agronomic traits and MCB resistance. So, we recommend fine mapping in these regions for looking for resistance to corn borers and yield genes.Selecting genotypes with the favorable allele of QTL on chromosome 5 (bin 5.01) decreased tunnel length. This same QTL did not affect yield, silking and plant height. The same way, the QTL on chromosome 9 (bin 9.07) could be used to improve ear resistance to MCB attack but it could produce later varieties.The calculation of indexes from the original data set could help plant geneticists and plant breeders to identify QTL which could be used in MAS programs to improve yield and kernel resistance and decrease tunnel length. The region in chromosome 5 (bin 5.04) could be used to improve stalk resistance and yield while regions in chromosome 1 (bin 1.10) and 10 (bin 10.02–10.03) could improve also ear resistance.JCJG carried out field experiments, performed statistical analysis of the data, and drafted the initial manuscript. AB and RAM created and generated the vegetal materials. RAM and BO, advisors, assisted JCJG in the field experiment design, data collection, and statistical analysis. AB and LFS participated in the data collection and statistical analysis. All authors have read and approved the final version of the manuscript.This research was funded by the Plan Estatal de Ciencia y Tecnologia de España within the projects AGL2012-33415 and AGL2015-67313-C2-1-R, both of which were co-financed with European Union funds under the FEDER program, and the project: IN607A/013 funded by Autonomous Government of Galicia, Spain.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.