To investigate the time-course of changes in knee-extensors muscle mass, architecture and function in response to plyometric training (PLT) performed on a novel training device, the Tramp-Trainer. This machine consists in a trampoline connected to an inclined sledge which allows the performance of repeated jumps while the subject is sitting on a chair.mean, together with vastus lateralis (VL) architecture  were assessed after 2, 4, and 6 weeks of PLT.Eight healthy males  underwent 6 weeks of bilateral PLT on the tramp-trainer machine. Training was performed three times per week (between 120 and 150 bounces per session). Knee-extensor maximum voluntary torque (MVT) and power, quadriceps femoris (QF) volume (VOL), cross-sectional area from the 20% to the 60% of femur length and CSAp < 0.001) and 22.2% , respectively, and declined to 13.3% (p < 0.05) at week 6 of PLT. Power increased by 18.2%  and 19.7% . QF VOL increased by 4.7%  and 5.8% ; VL VOL increased by 5.2%, (p < 0.05), 8.2%, (p < 0.01), and 9.6% (p < 0.05) at weeks 2, 4, and 6, respectively. An increase in Lf was detected already at wk 2 , with further increase at 4 and 6 weeks of PLT . PA increased by 5.8% (p < 0.05) at week 6. Significant positive correlations were found between CSAmean and Power  and between QF VOL and Power .All results are presented as changes versus baseline values. MVT increased by 17.8%  may have contributed to such fucntional changes.PLT induced rapid increases in muscle volume, fascicle length, pennation angle, torque and power in healthy younger adults. Notably, changes in VL VOL and Lf were detectable already after 2 weeks, followed by increases in knee extensors VOL and power from week 4 of PLT. Since the increase in CSA Muscle power is a major contributor of performance both in sports and in daily life activities, and several studies have been carried out in order to understand how power could be improved in young, older, and clinical populations . In thisIncrements in maximum isometric and explosive strength have been reported after plyometric training (PLT) protocols . Over a Although previous studies investigated the changes of skeletal muscle size, strength and power following plyometric training programs, to the present date and to the best of our knowledge, none of these studies focused on the time-course of the muscular adaptations to PLT. Defining when and how these changes occur may be beneficial in order to plan time-efficient protocols in healthy or clinical populations. In fact, when programming a training protocol aiming to elicit specific muscular adaptations  it is of extreme importance to know the timing and amplitude of such adaptations in order to plan personalized and efficient intervetions, with a well-defined progression of the load based on truly achievable goals.However, one important aspect of the plyometric training methods reported in literature is that the majority of the studies exploited free-body plyometric exercise programs using different intensities, workloads, durations, number of sessions per week, and unilateral or bilateral modalities . It follHence, the aim of the present study was to investigate the time-course of knee-extensors changes in muscle size, architecture and function in response to a 6-week plyometric training performed on the tramp-trainer device. Based on previous reports that show that early adaptations in muscle architecture were observed with resistance exercise , our hypFourteen young males were recruited to undergo a 6-weeks PLT program . The pre-to-post adaptations to PLT (week 0 and week 6) are described in a previous study from our laboratory . Among tTraining was performed on the “Trampoline-Trainer” (tramp-trainer) exercise machine  FREI AG, the traVolunteers exercising on the tramp-trainer start from a semi-squat position with their knees flexed to a range between 90 and 80 deg . After a maximal push of the lower limb muscles , the body and chair are displaced along the rail, followed by a landing on the elastic trampoline and an immediate recoil as the subject bounces back off the trampoline, and the entire cycle is then repeated for successive jumps . Thus, tPrior to the first training session, on a separate day, participants were familiarized with the tramp-trainer, being asked to perform a maximum of only 5 submaximal bounces. During such familiarization session, each participant practiced starting and landing from a knee angle of ca. 90° measured with manual goniometer .Training was performed 3 times per week (on the same days and at the same time for each session) for a period of 6 consecutive weeks. At least one resting day between training sessions was programmed. Training volume was based upon the guidelines of Training load was matched across subjects by determining 30RM values before the start of the program. 30RM is the training level at which subjects could perform no more than 30 repetitions without a decrease in bounce performance. To quantify bounce performance, a meter ruler was attached to the carriage of the training device to monitor, via an operator visual inspection, average bounce height during the assessment. Subjects were instructed to bounce at their 30RM, and if they fell significantly below their 30RM height for up to 3 consecutive bounces, they were prompted to increase bounce height in order to maintain a constant training load. A significant decrease was determined as 5 cm or more below the 30RM height.As training progressed, loading increased progressively because the velocity of impact increased due to higher bounce distances. As bounce distance increased due to training adaptations, new 30RM values were identified and implemented to progressively increase the training load. 30RM values were re-checked every 7 days (before the start of the first session each new training week). When volunteers reached the highest value of the meter ruler attached to the carriage  for most (∼90%) of the repetitions in a single training session, a 15 kg weighted vest was provided from the successive session in order to further increase the training load. In addition, training load increase was provided by increasing the number of series from week 4 of the training period.During each training session a red mark was placed on the side of the inclined plane rail-track as a visual target corresponding to a knee flexion of ca. 90°.n = 4) or (ii) power testing first and MVT testing after (n = 4).To assess the time-course of training adaptations, test-sessions were performed before the beginning of the training (baseline), and after 2, 4, and 6 weeks of training. Testing sessions were scheduled right before the first training of the week, every 2 weeks, to allow a time lag of at least 48 h between the last training bout of the previous week and the testing day, in order to minimize exercise-induced edema. During all the test-sessions the following parameters were assessed: knee extensors isometric maximum voluntary contraction torque (MVT) and peak power production; vastus lateralis (VL) and quadriceps femoris muscle (QF) cross-sectional area (CSA) from the 20% to the 60% of the femur length, from which VL and QF volume were then estimated (see specific section below); VL muscle architecture at the 50% of the femur length. During the testing sessions, all participants were firstly scanned for US (VL architecture and QF CSAs) and afterward were randomly assigned to the following sequences (i) MVT testing first and power testing after  calculation.Isometric maximum voluntary torque was measured using an isokinetic dynamometer  at a fixed joint angle of 70°, with full knee extension corresponding to 0°. The dynamometer had been previously calibrated following the manufacturer instructions, and gravity correction had been performed. After a brief warm-up, consisting of 10 short sub-maximal contractions at the 50% of MVT (1 s each), participants performed two maximum voluntary contractions, which lasted for 4 s, with a rest period of 30 s between contractions. Volunteers were provided with both real time visual feedback of torque production and a strong verbal encouragement during isometric contractions. The maximum isometric torque value (MVT peak) was chosen for data analysis.Knee extensors power was assessed using the Nottingham Power Rig . The power rig provides a measure of peak power during a hip and knee extension push againts a pedal . Brieflyin vivo using extended-field-of-view (EFOV) ultrasonography imaging . A 47 mm, 7.5 MHz linear array probe was used to obtain images at different muscle length percentages. Different region of interest were identified at the 20, 30, 40, 50, and 60% of the femur length (measured from greater trochanter to the mid patellar point) and marked on the skin  or estimated  CSAs. In addition, QF CSAin vivo using B-mode ultrasonography . Ultrasound images were obtained on the right leg, while the participant was laying in supine position; the 50% of the femur length (measured from the great throcanther to the mid patellar point) was found and marked with a skin marker, and afterward the probe was orientated transversally and the medial and lateral borders of the VL were identified. The 50% of the distance between the VL borders was chosen for image acquisition. VL fascicle length (Lf) and pennation angle (PA) were measured using a 100 mm, 10–15 MHz, linear-array probe as described by 1. Briefly, fascicles were digitized using the segmented line tool (thus not neglecting any fascicle curvature within their visible portion). Then, fascicle length was determined through manual extrapolation  of fibers and aponeuroses if a portion of the fascicle extended outside of the captured ultrasound image. Usually 2–3 fascicles were eveluated for each scan and their average value was taken as final Lf value. PA was measured at the intersection between the fascicles and the deep aponeurosis. An average of at least 3 PA measures per image was performed  performed throughout the study was calculated as previously described , TT inclination, and average height reached during each bounce performed in each session relative to the maximum height of the track/carriage .The total external mechanical work  = [(BM + VM) × g × (sin22)] × (ex)1.265-h × 30BM (kg) represents the subject body mass, VM (kg) the mass of the weighted vest, g is the gravitational acceleration (9.81 m⋅s–2) multiplied by the sine of the pre-set inclination of the track, 22°(which was kept constant throughout the whole study), hex (m) represents the average height reached by every bounce in the set , and 30 is the number of bounces performed per set.where All data are presented as average values (and standard deviation).∗Power 3.1.9.4., using muscle power data obtained from a pilot study conducted in our laboratory. Power analysis revealed an actual power of 0.825 when considering a sample size of 8, with effect size being 1.02 and α = 0.05.A priori power analysis was performed using GAll data met the normal distribution criteria, established on quantitative inspection by Shapiro–Wilk test. Repeatability of the measurements was tested for muscle mass (CSA) and architectural  values via calculation of the Intraclass Correlation Coefficient (ICC) of the ultrasound operator performing all the data acquisition and analysis. ICC values were rated good to very good, and resulted as follows: QF CSA = 0.996 (CV = 0.01%); VL CSA = 0.993 (CV = 0.04%); VL PA = 0.962 (CV = 0.05%); VL Lf = 0.966 (CV = 0.09%).2); VL CSA = 1.84% (±0.39 cm2); VL PA = 1.93% (±0.35°); VL Lf = 1.38% (±0.11 cm).In addition, basing on the same data used for the repeatability analysis (ICC), the minimum detectable change was calculated, following the approach suggested by p < 0.05.Differences in the functional, morphological, and architectural muscle components of the study during the time-course  were investigated using a one-way repeated measures analysis of variance (ANOVA). Tukey’s multiple comparisons test was used to identify significance between the different time points. Significance level was set at r). Linear regressions were also calculated and the coefficient of determination (R2) was obtained. The level of significance was set at p < 0.05.Linear relationships between the percentage changes of the morphological and functional parameters during the time-course of PLT were tested using the Pearson’s product-moment correlation coefficient  was used to perform all statistical and The average values for functional and morphological adaptations at each time point are presented in p < 0.001) after only 2 weeks of PLT. After 4 weeks, MVT further increased (n.s. vs. week 2) reaching a peak percentage increment by 22.2 ± 12.2% compared to baseline (p < 0.01) (p < 0.05).Mean MVT values increased by 17.8 ± 7.4% ( < 0.01) . In contp < 0.05) after 4 weeks of PLT, reaching a final pre-to-post increment of 19.7 ± 13.5% (p < 0.05 vs. baseline) (Power increased by 18.2 ± 15.4% (aseline) .The time course of changes in muscle size is reported in p < 0.05). Similarly, VL CSA taken at the 40% and the 60% of the femur length increased, respectively, by 7.8 ± 4.9% and 4.9 ± 2.9% (p < 0.05) after 2 weeks of PLT.Quadriceps femoris CSA showed significant adaptations after 2 weeks of PLT at 40 and 60% of the femur length, increasing, respectively, by 1.5 ± 1.0% and 3.5 ± 2.8% . At 6 weeks, all QF CSAs were statistically larger compared to baseline .From 4 weeks onwards, all the QF CSAs (except the one taken at the 20% of the femur length) showed a significant increment compared to baseline  and 6 weeks (p < 0.05) of PLT compared to baseline. Specifically, at 4 and 6 weeks, the percentage increment vs. baseline were, respectively: 10.9 ± 4.7% (p < 0.01) and 10.7 ± 7.1% (p < 0.05) for the 40%; 6.8 ± 4.0% (p < 0.05) and 7.4 ± 4.2% (p < 0.01) for the 50%; and 7.8 ± 2.6% (p < 0.001) and 11.2 ± 9.3% (p < 0.05) for the 60% of the femur length. Although, at week 6, VL CSA at the 20 and 30% of femur length showed a marked increase, this was not statistically significant .Conversely, VL CSA increased only at the 40, 50, and 60% of the femur length after 4 (mean showed a progressive increment by 4.7 ± 2.8% at 4 weeks (p < 0.05 vs. baseline) and by 5.8 ± 2.7% at 6 weeks (p < 0.01 vs. baseline).Quadriceps femoris CSAp < 0.05) and 5.8 ± 2.6% (p < 0.01) after 4 and 6 weeks of PLT, respectively, compared to baseline. Compared to baseline, VL volume increased significantly by 5.2 ± 2.9% (p < 0.05) after only 2 weeks of PLT, by 8.2 ± 2.9% (p < 0.01) after 4 weeks, reaching an increment by 9.6 ± 5.7% (p < 0.05) at 6 weeks.Quadriceps femoris volume increased by 4.7 ± 2.8% (p < 0.05), thereafter increasing linearly up to 4.0 ± 2.1% at week 4 compared to baseline (p < 0.01 vs. baseline and p < 0.05 vs. week 2) and by a further 0.4 ± 2.3% at week 6 (n.s. vs. week 4), reaching a final pre-to-post training increase by 4.4 ± 2.3% (p < 0.01)  .p < 0.01) by 5.8 ± 3.3% compared to baseline  as well as QF VOL and Power .Significant correlations were found between training induced-changes in CSANone of the training induced-changes in muscle architecutre parameters  were significantly correlated with the increase in Power.P < 0.05) and of the 6th   was significantly different from the first session of the 4th week  . The aveIn the present study we investigated the time-course of changes in muscle mass, architecture and function in response to 6-week PLT performed on the tramp-trainer machine.mean. In addition, VL Lf and PA increased by 4.4 and 5.8%, respectively.At the end of the training period, knee-extensors power was found to be increased by 19.7%, and this was accompanied by increments in both QF and VL volume and CSAR2 = 0.44).Notably, early significant changes in muscle function (MVT), architecture (Lf) and morphology  were detectable after only 2 weeks of PLT. Among these changes, VL CSAs were found to be increased especially in muscle regions closer to the mid-belly, and a similar trend was observed for QF CSA. Furthermore, an increase in knee-extensors power (18.2%) was found just after 4 weeks of PLT, together with significant changes in QF volume (4.7%). Changes in QF volume significantly correlated with the increase in muscle power (The 6-week PLT intervention was highly effective for increasing knee-extensors MVT.mean), while it can be speculated that the main increment may be ascribable to an increased neural drive known to account for the early gain in muscle strength/power with resistance training . In this regard, It is noteworthy that, despite the lack of further gain in muscle torque, muscle power kept increasing till week 6.mean, which showed a progressive increment . Hence, consistent with the notion that volume is the product CSAmean and muscle length  and QF volume and power .Since power is the product of force and velocity, the increment of power may be expected to have been due to an increase in contraction speed. In this regard, a significant and progressive increase in fascicle length was found at all time points . A similar observation applies to CSAe length  both an In addition to these morphological factors, we cannot exclude that neural or tendon adaptations may have contributed to the increased force and power outputs. Previous studies have shown an increase in neural drive of knee-extensors  and planIn addition, tendon properties  are also known to be affected by the training-induced load . NotwithTaken together we cannot exclude that tendon adaptations may have occurred during the training period, but we cannot speculate toward which direction. Further research should aim to clarify this.We have also investigated the distribution of morphological changes of the QF and VL muscles, in terms of muscle volume and CSA values collected at different sites of femur length .Both VL and QF muscle volume increased linearly throughout the training period; however, VL volume increment was significant after only 2 weeks, contributing to QF volume changes which were detectable after 4 weeks of PLT, in accordance with the increases in knee-extensors power.Interestingly, the majority of the works focusing on PLT interventions did not evaluate the possible muscle hypertrophic responses to such training modality. Two studies from Our results also show that, consistently with the findings of The observed distal hypertrophy highlights the possible influence of the eccentric component of PLT. In fact, not only there is previous evidence of regional hypertrophy following different training modalities , but alsRegarding the significant changes in both VL and QF CSA after just 2 weeks of training (which in turn influence the changes in muscle volume), some authors have suggested that such early responses may be influenced by the edema provoked by the eccentric component of exercise in the first training sessions . The exaLastly, in order to ensure our results not to be biased by the repeated measures error, we calculated the minimum detectable change, i.e., the minimum change required to assume that a real adaptation had occurred. For VL CSA, the minimal difference was found to be of 1.84%; within our subjects, only one of them showed increments lower than this at week 2 and week 4, and the average increases were at least 4.6% (week 2). For QF CSA, the minimal difference resulted of 1.38%; at week 2, 4 subjects presented increments higher than this value and 4 presented lower increments. However, the mean increment was higher than 1.38% (=minimum detectable change) in the CSAs taken at almost all the femur lengths, excluding the 50% (+1.06%) which resulted not significant. At weeks 4 and 6, all the subjects but 2 (week 4) and 1 (week 6) showed increments higher than 1.38%, and so did the average value calculated for all volunteers.Thus, it is plausible that the observed adaptations in muscle size could be regarded as true hypertrophic responses.We observed a significant increase in fascicle length after 2 (2.2%), 4 (4%), and 6 (4.4%) weeks of training, while PA showed an increment (5.8%) only at the 6-week time point. Similarly to the approach used to detect whether these small  changes could be regarded as true adaptations  we calculated the minimum detectable change for both Lf and PA. For VL Lf, the minimum detectable change was 1.38%; at week 2, 3 out of 8 subjects showed increments lower than this value, while the average was higher. At weeks 4 and 6 only 1 subject displayed increments lower than 1.38%. VL PA minimum detectable change was 1.93%, and this value was not reached in 6 out of 8 subjects at week 2 , and by 1 subject at weeks 4 and 6. Accordingly, PA was significantly improved after 6 weeks and showed a trend after 4 weeks.To the best of our knowledge, only very few studies have considered muscle architecture with respect of PLT. Similarly to The mechanisms responsible for the increase in fascicle length are likely related to the fascicle stretch caused by the repeated SSC involved in plyometric training. These SSCs are thought to involve consecutive muscle shortening (concentric) and lengthening (eccentric) contractions, but, at a muscle level, the real eccentric nature of the deceleration phase during the SSC has been often debated. Theoretically, the muscle is firstly lengthened before it rapidly contracts concentrically : howeverIt follows that the mechanical stretch applied to the fascicles during training protocols involving a SSC may not be completely comparable to the one observed after pure eccentric contractions . NotwithWe acknowledge that the present study has some limitations.First of all, we did not recruit a control group. However, we recommended our participants to not change their habitual levels of recreational physical activity and to not introduce any new exercise modality in the training period. Second, we acknowledge that the small sample size allow us to draw preliminary conclusions. Furthermore, our participants were not trained individuals and were novice to this training modality: it follows that the responses observed could be influenced by this aspect. Indeed, the relatively small  changes in muscle mass and architecture, and the observed increments in MVT and power could be less pronounced if the present PLT protocol would have been applied to an already trained population.In addition, there are limitations linked to the plyometric training device we used. This device enables to increase the work output both by increasing the slope of the inclined sledge as well as by increasing the distance covered during the push as muscle power increases. To maximize the load, we used the maximum chair slope that could be set in this device. As the subject power increases with the training progression, he eventually reaches the end of the rail track along which the seat travels. To increase load intensity and prevent the subject reaching the end of the rail track we overloaded the subject with a 15-kg weighted vest. However, after 4 weeks of training, most participants were nearing the end run again, so to add extra work-volume we increased the number of series of sets of 30 bounces. This procedure was undoubtedly effective for increasing the total work performed but the overloading was probably insufficient to induce a further gain in muscle force.On the other hand, also the use of the 15-kg weighted jacket could represent a potential limitation. Indeed, this was certainly sufficient for ensuring a gain in muscle strength up to 4 weeks of training but not enough to produce further gains after 4 weeks. However, despite these limitations, power output progressively increased up to the end of training period.Lastly, we did not investigate either neural or tendon adaptations to the training protocol used .The findings of the present study show that plyometric exercise training performed on the tramp-trainer is highly effective for inducing rapid gains in muscle volume, architecture, torque, and power in healthy younger adults.Notably, changes in VL volume and fascicle length were detectable already after 2 weeks of training, followed by increases in knee-extensors volume and power from week 4 of PLT.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation, to any qualified researcher.The studies involving human participants were reviewed and approved by The University of Nottingham Ethics Committee and was compliant with the Declaration of Helsinki. The patients/participants provided their written informed consent to participate in this study.MF, MN, and SL designed the study. EM, FB, JQ, and MF performed the experiments. EM and MF analyzed the data. EM, MF, SL, FB, JQ, and MN discussed the data. EM, MF, SL, and MN wrote the manuscript. All authors contributed to the article and approved the submitted version.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.


KIR genes diversity in recurrent spontaneous abortion (RSA). The present paper is a meta-analysis of previous genetic association studies and our previous original study. The results showed that KIR3DL1 was a significantly protecting factor for RSA . KIR2DS2  and KIR2DS3  were significantly risk factors for RSA. For KIR2DS1 there was a high heterogeneity and publication bias. Briefly, the inhibitory gene KIR3DL1 was a protecting factor, and the activating genes KIR2DS2 and KIR2DS3 were risk factors for RSA. However, the effect sizes were not suitable. We suggest further studies on different causes of pregnancy loss, to find the role of KIR2DS1.Natural killer cells (NKs) are the most important cells in the fetomaternal immune tolerance induced through interaction of maternal killer-cell immunoglobulin-like receptors (KIR) and fetal human leucocyte antigens (HLA). Hence, we intend to perform a meta-analysis on the role of maternal  Recurrent spontaneous abortion (RSA) and pregnancy loss have different pathogeneses, consisting of genetic and chromosomal abnormalities , environKIR and HLA genes in human genome have loci (not locus), inherited as haplotypes. In addition, each gene in their loci is polymorphic. Thus, interaction of different KIR molecules with different HLA molecules results in different outcomes consisting of inhibitory and activating responses. KIR gene cluster is located on chromosome 19. This cluster has two types of genes, including 8 inhibitory and 6 activating genes, and 2 pseudogenes. Some of these genes exist in all individuals, like the KIR2DL4. From the viewpoint of medical anthropology, different people from different ethnicities have different KIR-HLA interactions . Usually CD56tory NKs . 2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 3DL1, 3DL2 and 3DL3) and 6 activating genes . Since the involving NKs in implantation of embryo are maternal, we intend to perform a meta-analysis on the role of maternal KIR genes diversity in RSA. Previously, KIR. Nevertheless, their studies were few and therefore our study can serve as an update for that meta-analysis.As we mentioned above, KIR and HLA have different genes and interactions. KIR has 8 inhibitory , the odds ratios (OR) (effect sizes) achieved from these p values were underestimated. This statistical protocol has been previously published . The analyses were carried out through a 2I scale and 2I<50 was considered as homogeneity. In the cases of heterogeneity, we used the random-effect model. In order to find publication bias, we used funnel plots. If a study were to be find outside the funnel, it meant that its effect size was outside the expected 95% confidence interval (CI). In other words, its difference with other studies is statistically significant at p=0.05. Hence, a publication bias does not have necessarily a negative connotation. In the present study, a funnel plot p value < 0.05 means that the mentioned individual study is outside the funnel of 95% CI. We used the In order to cluster the studies for meta-analysis, we designed a dendrogram using the STATA14 software . This cluster analysis involved the complete linkage of binary variables . p values were analyzed through Yate's correction (or fisher's exact test when necessary). Positive effect directions show each gene as a risk factor and negative effect directions show each gene as a protecting factor. Our cluster analysis showed that the study by Dambaeva et al. (2016) had a different design in comparison to other studies . p value <0.05) (The role of e <0.05)  and 3. KIR2DL2 in RSA was not statistically significant . p value <0.05) . No publication bias was found based on the funnel plot . p value <0.05) (The role of e <0.05)  and 5. Tnel plot  and 7. Te <0.05)  and 9. KIR3DL1 in RSA was statistically significant . p<0.05; based on funnel plot) . Inconclusive publication bias was found for this analysis based on the funnel plot . Faridi et al. (2009) study showed significantly more risk effect of this gene in comparison to other studies  . p value <0.05) (The role of el plot)  and 11. nel plot  and 13. e <0.05)  and 15. e <0.05)  and 17. KIR2DS4 in RSA was not statistically significant . p value <0.05) . p value <0.05) . et al. (2009) showed significantly more protective and risk effect of this gene in comparison to other studies, respectively  (The role of e <0.05)  and 19. e <0.05)  and 21. e <0.05)  and 23. +CD56dim and CD16-CD56bright that are called as cytotoxic and immune-regulatory NKs, respectively. In the implantation site, the NKs are mainly CD56bright. Hence, the immune system has a positive and protecting role in implantation and early pregnancy. Embryo implantation and pregnancy are a type of transplantation called semi-allograft. Thus, we need immune tolerance to have a successful pregnancy. The NKs play their roles with their KIRs interacting with the HLAs expressed on trophoblasts , because its activating role  results in higher cytokine releasing of UNKs (AA genotype was a risk factor for the success of assisted reproduction. AA is the most inhibitory genotype and therefore it supports this hypothesis that NK activation is necessary in early pregnancy. Pregnancy loss has numerous causes, in particular embryo genetic and chromosomal abnormalities. Therefore, the immune system's theoretical role is to reject such malformed embryos. Therefore, this risky role of activating KIRs is in fact a protecting role! Of course, it is remarkable that the lack of genetic evaluation of the lost embryos was a limitation for the studies imported to this meta-analysis. It is suggested that this variable should be adjusted in future studies. The results of Although we found significant associations involving 3 genes in the meta-analysis , but theKIR gene diversity in RSA is still in unclear, although our meta-analysis showed 3 genes as associated factors. KIR3DL1 was a protecting factor, and KIR2DS2 and KIR2DS3, which proved to be risk factors for RSA. For KIR2DS1 there was a high heterogeneity. It seems that its role is different among different causes of pregnancy loss. Our previous case-control original investigation showed a significant relation with maternal KIR2DS1 in combination with paternal HLA-C2 as a risk factor. In order to clarify this role we have some suggestions for future studies, such as investigations of this combination concerning the success rate of assisted reproduction, for early first trimester abortions occurring after implantation and early placentation, for stillbirth groups, for abortions secondary to APS, and for successful and unsuccessful pregnancies of malformed embryos and fetuses. We would also like to suggest adjusting models and cohort studies. The role of maternal


Little is known about Parkinson’s disease (PD) epidemiology, demographics, genetics and clinical presentation in Central Asian countries. Western and East Asian studies have suggested that PD might have variable clinico-demographic characteristics in different ethnic groups . In thisWe conducted a retrospective study reviewing outpatient clinical records of PD patients from the three medical centers across the country with an available movement disorders specialist. Only cases compatible with UK Brain Bank PD diagnosis criteria  were selThe clinical records of 990 patients with PD diagnosis were eligible for the analysis. The mean age of PD onset was 55.01±9.7 years  and the mean age at examination was 62.2±9.0 years . Almost half of the cases (43.5%) were between the ages of 61–70 at the moment of the last examination. Mean disease duration was 7.1±5.2 years . . The mean age at an examination in males was also significantly lower than in females . Mean disease duration did not differ between genders (p = 0.4). An autosomal dominant family history of PD was found in 4% of patients being more prevalent among EOPD.EOPD cases composed 30.5% in our cohort, and 7.6% developed PD before the age of 40 years. The male to female (M: F) ratio among the whole cohort and young-onset PD cases was 1 : 1.3 and 1 : 0.99, respectively. The prevalence of females has been noticed in our PD cohort. The mean age of onset was significantly younger in males compared to females . Only 207 (23.7%) patients had an available off stage MDS UPDRS motor score with a mean 34.9±18.2 . EOPD cases had significantly higher MDS UPDRS motor scores and HYS compared to late-onset PD (LOPD) patients . UPDRS motor score did not differ between genders . Dyskinesias were expressed by 17.1% of patients, and 14.5% had both dyskinesias and motor fluctuations. Female and EOPD subjects expressed slightly more levodopa-induced complications.The most prevalent PD subtype was akinetic-rigid. The Mean Hoehn-Yahr stage (HYS) was 2.3±0.8 and it did not differ between genders . Thirty-four percent of patients were on levodopa monotherapy only. In terms of other anti-parkinsonian medications, 9.8% were taking amantadine, 16.1% were taking dopamine agonists (DA), 8.1% were on rasagiline, and 6% were on trihexyphenidyl.The mean age at onset (AAO) in our study (55.01) was younger than in Caucasian (58.0±12) and slightly older than in East Asian (54.0±12) PD patients . The relPD tends to affect males more than females in Western countries, and the opposite has been observed in East Asian countries. . SimilarMale subjects developed PD younger in comparison to females in our study. Although AAO of PD in female subjects was older, mean MDS UPDRS motor scores did not differ between genders. This, bearing in mind the equal mean disease durations in both genders in our study, could suggest similar rates of the disease progression between genders.Less than one in five PD patients in our cohort expressed levodopa induced-dyskinesias and/or motor fluctuations. Considering the mean disease duration 7.1±5.2) and the daily dose of levodopa of 500 mg and more in 58% of patients, one would expect higher rates of motor complications. Interestingly, low rates of dyskinesias and motor fluctuations have also been reported in Western Pacific countries [ and the The analysis of antiparkinsonian medications showed that the main PD medication in Kazakhstan is levodopa, which was taken as monotherapy in one-third of patients. The low proportion of consumed DA and monoamine oxidase inhibitors could suggest either limited access due to the high cost of these medications in the country or that the importance of adjunct levodopa therapy is under-recognized by local neurologists who follow-up PD patients.The retrospective nature of our study appears to be the major limitation. Data were assembled from clinical records, and therefore not every case had available UPDRS scores, detailed information about family history, non-motor symptoms, and mild levodopa-induced complications.Considering all the findings and limitations of this study together, one might infer that well- designed high-quality epidemiological, genetic, and observational studies are warranted in Kazakhstan and other Central Asian countries to fill the gap in the knowledge of PD in the extensive and multinational region.The authors have no conflict of interest to report.Click here for additional data file.


Streptomyces albus subsp. chlorinus NRRL B-24108. Here, we report the identification and heterologous expression of the previously uncharacterized albucidin gene cluster. Through a series of gene inactivation experiments, a minimal set of albucidin biosynthetic genes was determined. Based on gene annotation and sequence homology, a model for albucidin biosynthesis was suggested. The presented results enable the construction of producer strains for a sustainable supply of albucidin for biological activity studies.Herbicides with new modes of action and safer toxicological and environmental profiles are needed to manage the evolution of weeds that are resistant to commercial herbicides. The unparalleled structural diversity of natural products makes these compounds a promising source for new herbicides. In 2009, a novel nucleoside phytotoxin, albucidin, with broad activity against grass and broadleaf weeds was isolated from a strain of  Pesticides play an important role in modern agriculture. Among all chemicals being produced, pesticides are in second place after fertilizers in their extent of use. A total of 2.4 billion kilograms of pesticides were applied worldwide in 2007 . NeverthFrom all new pesticide active ingredients registered by the Environmental Protection Agency from 1997 to 2010, almost 70% have origins in natural products. Interestingly, only 8% of conventional herbicides are natural product-derived ,6. The wStreptomyces albus subsp. chlorinus NRRL B-24108 was discovered in 2009 [A novel bleaching herbicide, albucidin, from the strain Escherichia coli strains were cultured in LB medium [Streptomyces strains were grown on soya flour mannitol agar (MS agar) [All strains, plasmids and BACs used in this work are listed in B medium . StreptoMS agar)  and in lMS agar)  was usedStreptomyces albus subsp. chlorinus NRRL B-24108 was used [E. coli transformation and E. coli/Streptomyces intergeneric conjugation were performed according to standard protocols [The previously constructed BAC library of was used . DNA manrotocols ,12,15. BStreptomyces strains were grown in 15 mL of TSB in a 100 mL baffled flask for 1 day, and 1 mL of seed culture was used to inoculate 100 mL of SG production medium in a 500 mL baffled flask. Cultures were grown for 7 days at 28 °C and 180 rpm in an Infors multitron shaker. Albucidin was extracted from the culture supernatant with an equal amount of butanol, evaporated, and dissolved in methanol. Albucidin production was analysed on a Bruker Amazon Speed mass spectrometer coupled to UPLC Thermo Dionex Ultimate 3000 RS. Analytes were separated either on a Waters ACQUITY BEH C18 column  or on a Waters ACQUITY BEH C18 column . Water + 0.1% formic acid and methanol + 0.1% formic acid were used as the mobile phases. For the determination of high-resolution mass, analytes were analysed with a Thermo LTQ Orbitrap XL coupled to UPLC Thermo Dionex Ultimate 3000 RS. Analytes were separated on a Waters ACQUITY BEH C18 column  with water + 0.1% formic acid and methanol + 0.1% formic acid as the mobile phase.For metabolite extraction, Streptomyces albus subsp. chlorinus NRRL B-24108 was inoculated into 100 mL of SG medium in a 500 mL baffled flask and cultivated overnight at 28 °C and 180 rpm. The pH of the culture was adjusted to 8.5 with 1 M NaOH. Ten millilitres of culture was transferred into three 50 mL falcon tubes. Then, 64 mg of wet NTG was dissolved in 16 mL of water. Next, 6.666 mL, 4.285 mL and 1.765 mL of NTG stock solution were added to the tubes containing culture to reach final NTG concentrations of 800 µg/mL, 600 µg/mL and 300 µg/mL, respectively. The samples were incubated at 28 °C for 30 min in the overhead shaker. The mycelium was precipitated by centrifugation, and the supernatant was discarded. The mycelium was washed twice with 5% sodium thiosulfate solution. The treated samples were plated on MS agar plates and cultivated for 14 days at 28 °C. The spores were washed with water and plated in dilutions on MS agar. The plates with spore dilutions were incubated for 10 days at 28 °C. Single colonies were picked on 30 mm plates with SG agar. The plates were incubated for 14 days at 28 °C. Agar blocks were cut out from the plates and transferred into 2 mL tubes. Albucidin was extracted from the agar blocks with 500 µL of butanol for 48 h. The extracts were analysed using HPLC-MS.One millilitre of spore suspension of Streptomyces albus 1K1 was grown in 10 L of SG medium, and albucidin was extracted with butanol. The dry extract was dissolved in 50 mL of water containing 5% acetonitrile and 0.1% formic acid. The extract was loaded onto 3 C18 SPE columns (Discovery DSC-18 SPE 52607-U) equilibrated with 5% acetonitrile in water with 0.1% formic acid. The flowthrough was collected. The columns were washed twice with 12 mL of 5% acetonitrile containing 0.1% formic acid. The flowthrough and the wash fractions were combined and evaporated in a rotary evaporator. The presence of albucidin was detected by HPLC-MS.The dry material after the SPE purification step was dissolved in methanol and used for size-exclusion chromatography. Separation was performed on a glass column (30 mm × 1000 mm) packed with Sephadex LH-20 and methanol as the mobile phase. Fractions containing albucidin were identified by HPLC-MS. Albucidin-containing fractions were combined and evaporated. The dry extract was dissolved in 5 mL of 5% methanol in water containing 10 mM potassium phosphate buffer pH 6.4.HPLC separation was performed on a preparative HPLC Thermo Dionex Ultimate 3000 equipped with a Macherey Nagel Nucleodur HTec C18 column . A 10 mM potassium phosphate buffer (pH 6.4) was used as solvent A, and 50% methanol in 10 mM phosphate buffer  was used as solvent B. The following gradient at a flowrate of 15 mL/min was used for separation: 0 min–13% B, 20 min—25% B, 21 min–25% B, 24 min–100% B, 25 min–100% B, 28 min–13% B, 29 min–13% B. Albucidin eluted at 18 min. The albucidin-containing fractions were pooled and evaporated.For the final purification step, the dry material after HPLC purification was dissolved in 5 mL of water and loaded onto a Sephadex LH-20 column (30 mm × 450 mm) previously equilibrated with water. Water was used as the mobile phase. The fractions containing albucidin were identified by HPLC-MS, pooled and evaporated.1H-NMR spectra were recorded on a Bruker Avance 500 spectrometer  at 300 K equipped with a 5 mm BBO probe using deuterated trifluoroacetic acid  as the solvent containing tetramethylsilane (TMS) as a reference. Albucidin was measured in deuterated water . The chemical shifts are reported in parts per million (ppm) relative to TMS. All spectra were recorded with the standard 1H pulse program using 128 scans. The structure of albucidin was confirmed by comparison of the recorded 1H NMR data  was used. The strain in the exponential growth stage was treated with various NTG concentrations  for 30 min. After mutagenesis, the cells were washed with 5% thiosulfate solution and plated in dilutions on MS-agar medium for segregation of mutations. The spores of the obtained mutant populations were washed and plated on MS-agar plates in dilutions to obtain single colonies. Altogether, 4000 individual mutants were analysed for albucidin production. The mutants were cultivated on individual plates with the production medium SG agar. The metabolites were extracted with butanol, and albucidin production was assayed by HPLC-MS. Eight mutants that lost the ability to produce albucidin were identified in the course of this screening: 6-238, 6-260, 6-389, 6-444, 6-612, 6-892, 8-610 and 8-639. The genomic DNA of the obtained zero mutants was sequenced using Illumina technology. The point mutations in the genomes of the mutants were detected by mapping the sequencing reads to the reference genome of the wild type albucidin producer. Up to 100 transition mutations were identified in the genomes of the mutant strains. By comparing the mutation patterns of the separate mutants, a short genomic region was identified that was affected by point mutations in all analysed zero mutants, implying its potential involvement in albucidin production .  and albB (gene 16) were substituted with the ampicillin resistance gene in BACs 1K1_KO14, 1K1_KO15 and 1K1_KO16, respectively -28 was substituted by the hygromycin resistance marker . b To dete to 29). c. In BAC to 29). c. The geectively c. In thee marker c. The coS. albus 1K1_KO14  and albB (gene 16) completely abolished albucidin production in the strains S. albus 1K1_KO15 and S. albus 1K1_KO16 -28 did not affect albucidin production by the S. albus RS strain  is surprising since it belongs to the same operon as the essential genes albA (gene 15) and albB (gene 16). The deletion of the albC gene (gene 17) might be cross-complemented by an unidentified gene in the genome of the host strain S. albus Del14.Deletion of the genes S strain . It was S. albus 1K1_LS  were substituted with the ampicillin resistance gene and the genes upstream of the operon (genes 1–14) were substituted with the hygromycin resistance gene  Gene cluster for the biosynthesis of albucidin.


The limit of detection (LOD) and limit of quantitation (LOQ) of the method were 0.02–0.04 mg/kg and 0.05–0.13 mg/kg respectively. A good recovery range between 85.3 – 121.2% were obtained for a spike level of 1.0 mg/kg in real samples. ANOVA analysis at 95% CL showed statistically no significant difference (p > 0.05) in the recoveries between samples and also between the individual compounds. The method experimented and proposed in this study is fast, easy, cheap, robust and ecologically safe, presenting an alternative method for routine analysis for increased rate of surveillance against the illicit use of Sudan dyes as food additives.Illicit use of Sudan dyes, a group of harmful and carcinogenic azo dyes, in the food industry has taken a surge in various parts of the world, especially in Africa. Their use in food as additives pose a dire health risk to consumers and have been banned by various food regulatory bodies worldwide. To help increase surveillance, various methods have been proposed for their analysis in literature. This study also sought to experiment and propose an alternative method for quick, easy, cheap, robust and ecologically safe analysis of Sudan dyes in chilli pepper powder and similar matrices. The optimized method used a 6.0 mL mixture of acetone:acetonitrile (1:5 v/v) solvent in a modified QuEChERs method for extraction of Sudan dyes I-IV. The simultaneous analysis of the dyes were achieved on Shimadzu prominence UFLC 20AD coupled with SPD 20AX UV detector operated at dual wavelength of 500 and 480 nm. A total of twenty four (24) chilli pepper powder samples from eight different vendors on the Ghana market were analysed using the optimized method. Quantitation of analytes were done using the external standard calibration method with determination coefficient, R Food Analysis; Food Safety; Analytical Chemistry; Sudan dyes; Chilli pepper powder; Modified QuEChERs; UFLC-UV They are cheap, highly lipophilic and have attractive intense red colours, hence have illegally been used in some food industry as food additives to enhance the colour of foods such as palm oil, spices, powder Chilli pepper and sauces as well as some meat products . The mosUnfortunately, to date, the menace of using these carcinogenic azo dyes as food additives in the food industry has not stopped. For instance, in late 2006 the presence of Sudan IV dye was reported in some duck and hen eggs in Mainland China . Amoako-Because of prevalence illegal use of Sudan dyes in foods, and to enhance surveillance, several analytical methods have been proposed in literature . Most of22.14 + 1.5 g NaOAc) were purchase from Phenomenex Inc.Single component Sudan dye standards (purity >96.0%) for Sudan I, II, III, and IV (Sigma Aldrich). GC grade acetone (purity >99.0%), and LC grade acetonitrile (purity >99.9%) were purchased from Merck KGaA, Darmstadt, Germany. The glacial acetic acid (purity >99.7%) was from Daejung Chemicals, Siheung, Korea. MilliQ ultrapure deionized water (18.2 MΩ) filtered through MilliQ polisher (DI water), AOAC Method 2007.01 roQ QuEChERS extraction salts  different samples were purchased from eight vendors. Spiked and unspiked samples were prepared for each sample, and extracted for the analysis.2.2.1The samples were extracted using three (3) different types of solvent systems, i.e. 6.0 mL acetonitrile only, 1.0 mL acetone and 5 mL acetonitrile mixture [17:83% (v/v)] and 6.0 mL acetone: acetonitrile [50:50%(v/v)]. This was done to optimize the extraction process with the right solvent combination for efficient and reproducible extraction.In all extractions, 2.0 g of powdered chilli pepper samples were taken. Spiked samples at 1.0 mg/kg spike levels were prepared by the addition of a mix Sudan dye standards. A volume of 2.0 mL DI water was added to the sample in a centrifuge tube. The sample was shaken for 30 s and 6.0 mL of the extraction solvent was added followed by the addition of 2.0 g QuEChERS extraction salt mixture. The resulting mixtures in the tubes were manually shaken for 1.0 min and finally centrifuged at 4000 rpm for 5 min. Prior to UFLC-UV analysis, the supernatants were filtered through a 0.45 micron syringe filter and 1.5 mL was taken for the analysis. All samples both spiked and unspiked were extracted and analysed in replicates (n = 3). A reagent blanks were also extracted in similar manner and analysed.2.3A 1000 mg/kg of a mix standard was prepared by dissolving 0.1 g of each Sudan dye standard in acetonitrile and made up to the mark in a 100 mL volumetric flask. An amount of the stock solutions was taken and diluted with acetonitrile to prepare a working standards of concentrations of 0.1, 0.5, 1.0, 2.0, 5.0, and 10.0 mg/kg. Continuous calibration verification standards of 10.0 mg/kg were also prepared daily. Also a 10.0 mg/kg standard for system suitability was prepared.2.4The analysis was carried out using Shimadzu Prominence UFLC 20AD coupled with UV detector SPD 20AX.2.4.1The solvent systems used for elution were:Mobile phase A: 0.1% (v/v) acetic acid in DI water.Mobile phase B: Acetonitrile.Elution Solvent mixture: A:B = 10:90% (V/V).The LC system was operated in isocratic mode with a flow rate 1.5 mL/min. The column used was 150 mm × 4.6 mm (i.d.) Phoenix C-18 column and it was maintained at a temperature of 40 °C in an oven.The UV-Vis detector SPD-20AX was operated in dual wavelength mode, i.e. 500 nm (channel 1) and 480 nm (channel 2). The flow cell temperature was maintained at 40 °C.The injection volume for standards and samples was 5.0 μL and the total run and detection time was 10 min.2.4.2All the UFLC-UV analytical data were acquired using Labsolution version 3.0 software. Data were obtained by comparing retention time and area of each compound in the standards with that of each real sample. The external calibration method was used in acquiring concentration levels.2.4.3System suitability was conducted for each batch using FDA Criteria. Solvent A, was freshly prepared prior to each daily batch of analysis. Spiked and unspiked samples were analysed in replicates. Reagent blank was prepared and analysed in a similar manner to the samples used. Initial Calibration verification (ICV) standards of 0.4 and 2.0 mg/kg concentrations were ran to initially validate the calibration curve. Continuing calibration verification standard at 10.0 mg/kg concentration were also ran in triplicates (n = 3) after each ten (10) sample analysis to validate the calibration curve.2.5Analysis of variance (ANOVA) and other statistics on the data were conducted with excel toolpak. The Shimadzu labsolution browser was also employed in the calculations of LOD, LOQ and the statistics of the other analytical merits.33.12 were greater than 0.9999  real chilli pepper samples from eight different major vendors on the Ghanaian market. The mean percent recoveries (n = 3) of the spiked samples ranged between 85.3 – 121.2% . Sudan IIt is also worth noting that the use of a maximum total organic solvent volume of 6.0 mL for extraction, may pose a minimal risk to laboratory personnel and the environment. The method may thus be said to be ecologically safe for use in all settings.Two-way ANOVA conducted at the 95% CL showed no statistical significant difference (p > 0.05) between samples with respect to the total Sudan dyes recovery and also between the recoveries of each Sudan dye analytes in each sample. These suggested, the method employed in this study had good reproducibility and repeatability and may produce consistent recoveries for Sudan dyes in powdered chilli pepper samples and similar matrices.4This study employed a modified QuEChERs extraction protocol combined with UFLC-UV simultaneous analysis of the four Sudan dyes. The method, both extraction and instrumental analysis, was quick using a maximum total time of twenty minutes (20 min). The method was also robust and cheap when compared with other traditional and recent methods published in literature. The use of only 6.0 mL of organic solvent for extraction, poses a minimal risk to laboratory personnel and the environment. This method may help to increase productivity in the routine analysis of Sudan dyes in real samples, and help regulatory bodies in quick and accurate analysis of more samples per day to help with surveillance in the food industries to help curb the illicit use of these dyes.Joseph Kweku Adjei: Conceived and designed the experiments; Performed the experiments; Analyzed and interpreted the data; Contributed reagents, materials, analysis tools or data; Wrote the paper.Vigil Ahormegah, Alex Kissi Boateng, Harry Kwaku Megbenu, Samuel Owusu: Performed the experiments; Contributed reagents, materials, analysis tools or data.This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.The authors declare no conflict of interest.No additional information is available for this paper.


Aedes aegypti populations.Owing to the increased reports in Aedes-borne diseases in the Caribbean and Latin America, the United States Agency for International Development assisted the Jamaican Ministry of Health and Wellness in conducting insecticide susceptibility tests on Aedes aegypti eggs were collected, reared to adults per collected population and their susceptibility to varying pyrethroids and organophosphates were tested using the World Health Organization paper bioassays for these insecticides. The Centers for Disease Control and Prevention bottle bioassay was used to assess susceptibility to the carbamate, bendiocarb. The voltage gated sodium channel gene mutations V1016I and I1011V, normally associated with pyrethroid resistance, were also analysed.Sentinel sites were established in seven parishes of Jamaica  and Aedes aegypti collected from all parishes exhibited resistance to pyrethroids at the following concentrations, permethrin 0.25–2.5%; deltamethrin 0.03–0.15%; lambda-cyhalothrin 0.03–0.3%; and etofenprox 0.5–2.5%. The insecticide deltamethrin at concentration 0.3% was the only pyrethroid tested that resulted in high mortality, 94.9 ± 0.34% knockdown within 1 hour of exposure and 98.95 ± 0.01% mortality (p <0.01) at 24 hours post exposure. The frequency of the voltage gated sodium channel gene mutation V1016I was high in the tested population, possibly accounting for the reduced sensitivity to pyrethroids. Organophosphate resistance was also observed in all populations tested. Mortality rates for 0.8% Malathion was 0.8 ± 0.70–60.68 ± 0.38% after 24 hour and 0.00–47.10 ± 3.02%, for pirimiphos-methyl 0.21%. Bendiocarb applied as 12.5 μg/ bottle resulted in mortality rates of 76.25 ± 4.30–100 ± 0.00% after 30 minutes of exposure.The results showed that Ae. aegypti from the seven parishes analysed demonstrated resistance to the insecticides tested. Deltamethrin and bendiocarb at concentrations 0.3% and 12.5μg respectively, were considered most effective, causing high mortality in the local populations. Routine monitoring and evaluations of Ae. aegypti populations from the included parishes are recommended. Additionally, the study results represent the most comprehensive testing to date with local Aedes aegypti populations distributed across different parishes of Jamaica and should be useful to guide national and sub national strategies for vector control and surveillance.The results showed that  Aedes aegypti populations, and to determine which insecticide is more effective to reduce vector populations of Jamaica, we conducted an array of bioassays with three different types of insecticides  to use local evidence and inform public health authorities on vector control status and alternatives.The recent outbreaks of chikungunya, Zika and dengue viruses in the Caribbean and Latin America during 2013–2018, have motivated local health authorities in individual countries to seek international assistance from multiple agencies and adjust their vector management strategies in order to contain the transmission of Aedes-borne diseases. Countries within the region rely mainly on the use of commercially available insecticides to manage the mosquito populations and by extension, reduce the transmission of vector-borne diseases. However, this practice is stymied by the ability of these insects to develop resistance mechanisms to insecticides of frequent use. Many countries in the Caribbean and Latin America regions lack the facilities and supplies to conduct routine insecticide susceptibility testing. In an effort to ascertain knowledge on the presence of insecticide resistance in  Aedes aegypti, found ubiquitously throughout Jamaica and the wider Caribbean [Aedes-borne diseases, such as chikungunya and Zika viruses [Ae. aegypti. A key element of this collaboration was the assessment of the susceptibility status of this mosquito vector to commonly used insecticides.Each year, more than 284 million people are affected by dengue . The maiaribbean . The ciraribbean , reportsaribbean , 7, the  viruses  has beenAe. aegypti populations. However, this method is limited by the ability of insects to develop resistance to the chemicals of frequent use [Ae. aegypti densities. Organophosphates and carbamates are acetylcholinesterase inhibitors, toxins that trigger prolonged neurological synapses in mosquitoes, eventually resulting in paralysis, while pyrethroids on the other hand, are fast-acting neurotoxins. Unlike organophosphates and carbamates, their mode of action prevents the closure of protein-voltage-gated sodium channels (vgsc) on neurons, causing excess excitation, and eventually paralysis. Alterations in the specific binding site of a neurotoxin in mosquitoes can confer resistance to the insecticide [The use of chemical insecticides for community spraying is the main method to control the adult uent use . Many couent use –13. The uent use  and mostuent use  to reducecticide –16. Mosqecticide . The levecticide . Dependiecticide , which mAe. aegypti populations from one or two parishes of Jamaica are available [Resistance may vary across different geolocations despite proximity . Effectivailable , 14, 25 vailable , 26–29.Ae. aegypti populations from Jamaica to various adulticides and provide evidence on the status of mosquito vector susceptibility to the three commercially available insecticides . The overall goal was to support the local health authorities and foster evidence-based decisions for Aedes management strategies.The aims of this study were to determine the susceptibility of This study was developed in Jamaica, in the eastern parishes of the island: St. Catherine, Kingston & St. Andrew (KSA), St. Thomas, Portland, St. Mary, and St. Ann. The specific locations were selected in agreement with the MOHW mainly based on the epidemiological records of 2015 and 2016. Parishes with the highest incidence of Zika cases and classified as the most populated and accessible in the eastern territory were included in the study . SpecifiWHO susceptibility test kit tubes, with impregnated papers, pyrethroid (PY) control in Silicone oil; permethrin 0.25%, 1.25%, 2.5%; deltamethrin 0.03%, 0.15%, 0.3%; lambda-cyhalothrin 0.03%, 0.15%, 0.3%; etofenprox 0.5%, 2.5%; organophosphates (OP)/ carbamate (Carb) control in olive oil; malathion 0.8%; pirimiphos-methyl 0.21% were purchased from Universiti Sains Malaysia (USM), Vector Control Research Unit, Infotech , which is the only entity approved by the World Health Organization (WHO) to supply insecticide kits (mosquito diagnostic test kit WHO/VBC/81.806) and materials for regular entomological surveillance in public health. Analytical grade bendiocarb , and  acetone were also employed. Black plastic cups and pellon paper, which were used as ovitraps (described below) and egg papers respectively, were locally sourced in Kingston, Jamaica.Aedes aegypti populations. The surveillance system established by ZAP selected a number of sentinel sites for frequent visits and deployment of ovitraps to collect Ae. aegypti eggs. Sentinel sites were established in 100 homes in each of the parishes on the eastern half of the island: St. Catherine, Kingston & St. Andrew (KSA), St. Thomas, Portland, St. Mary, and St. Ann. Field activities in the parishes of Kingston and St. Andrew were regularly managed under one umbrella within the MOHW vector program in Jamaica. Therefore, to maintain the zoning, monitoring and evaluation surveillance of the MOHW, ovitraps set throughout the parishes of Kingston and St. Andrew were described as being derived from KSA. The ovitrap surveillance was conducted weekly, and once the field teams visited the homes selected for this process, all pellon papers containing mosquito eggs were transported to a laboratory for analysis. All laboratory activities were conducted at the Mosquito Control and Research Unit (MCRU) Insectary, funded by ZAP and The University of the West Indies (UWI).The biological material employed in this study was obtained through a continuous entomological monitoring system, deployed by ZAP in collaboration with the MOHW of Jamaica. ZAP’s support to the MOHW included teams of field workers distributed in different parishes of Jamaica, consistently implementing households visits and data collection on Prior to the operational activities, local communities within each parish were engaged by ZAP and the local health authorities. The community members were informed about the intended activities planned by the project. All householders who participated in the study approved the operational activities of ZAP and granted permission to their property on a weekly basis between September 17 to December 14, 2018, to collect samples and record information.th 2018 to December 7th 2018. Yeast was used as a mosquito attractant. The traps were replaced weekly and papers were collected from each parish and brought to the MCRU insectary. Once egg papers received at the insectary, these were air dried and assessed for the presence of Ae. aegypti eggs; a sample of papers was periodically set to hatch for susceptibility bioassays.Black oviposition cups  lined with pellon paper (7 cm wide) and filled with 50 ml water containing 5 g of instant yeast  were set up through the period of September 170C temperature, 75 ± 5% relative humidity and 12:12 h light: dark photoperiod. The larvae were fed on a diet of ground cat food (Friskies). Pupae were placed in emerging chambers  until the eclosion to adults and then transferred to 30 x 30 x 30 cm acrylic cages .The egg papers (25 per parish) were submerged in acrylic containers (37.5 × 25 × 15 cm) with 3 L of water with 0.1 g of instant yeast . To increase egg hatching, we replicated the media and methods described by Zheng et al . Tap watAe. aegypti) eggs were initially donated from the ZAP Dominican Republic project , Atlanta). This strain was used as the insecticide-susceptible reference strain for mortality comparison with the Jamaican wild populations. The strain was reared in isolation at the MCRU insectary, Jamaica, under the local insectary conditions described above.Rockefeller laboratory strain  collected from St. Catherine, Jamaica. Mosquitoes from the tests were placed in 15 ml Eppendorf tubes per test (n = 20–25 Ae. aegypti), tubes were labelled with the test compound as well as either dead or alive to denote the status of the mosquitoes after the completion of the bioassays. Each tube was previously packed with silica gel beads and cotton prior to adding mosquitoes. The samples were shipped to the Lobo Lab, University of Notre Dame, Notre Dame, IN 46556 for genetic analysis.The pyrethroid WHO susceptibility assays with permethrin 0.25%; deltamethrin 0.03%; and lambda-cyhalothrin 0.03% were repeated on Val1016, and Ile1011, from here on referred to as V1016I and I1011V respectively) [Ae. aegypti were assessed. Control mosquitoes with and without insecticide resistant alleles, used for V1016I diagnosis were the New Orleans and the Liverpool strains of Ae. aegypti respectively. Amplification was performed in 25 μl volumes in 96-well PCR plates (Dot Scientific) in a Mastercycler Gradient thermocycler (Eppendorf). Each reaction contained 1X Taq buffer , 1.5 mM MgCl2, 200 μM dNTPs, 5 pmoles of each primer   in Ae. ah primer , 1 unit 95% CI) per population. Abbott’s formula [The data are presented as mean ± 95% confidence interval / (100 –C)) x 100; whereby T is equal to the total percent mortality in the treated group, and C is equal to the percent mortality in the control group, providing that the control mortality was ≤ 20%. The mortality in the control group for all bioassays per parish was less than 20%.Posthoc Tukey test were used to analyze significant differences (p<0.05) in susceptibility to insecticide between the Rockefeller strain and tested Jamaican populations, where possible. R (version 3.6.2) was used to calculate confidence intervals.All statistical analyses were completed using SPSS for Windows (version 17.0). One-way analysis of variance (ANOVA) and Ae. aegypti mosquito populations in seven parishes of Jamaica. The results showed evidence of high levels of resistance against six commonly used insecticides. In comparison to the Rockefeller strain, all populations from Jamaica sampled in this study demonstrated resistance to the pyrethroids, permethrin, deltamethrin, lambda-cyhalothrin and etofenprox, as well as the organophosphates, malathion and pirimiphos-methyl with the highest mortality displayed at 68.17 ± 3.26% over 24-hours exposure. However, the tested populations appeared responsive to the carbamate, bendiocarb. The differences in the levels of resistance for each insecticide tested were further compared between the populations.This study was undertaken with the aim of examining the susceptibility status of Ae. aegypti mosquitoes from the seven parishes under study. As can be seen in Ae. aegypti F0 females for all populations, where the St. Ann population (p < 0.01) was the least affected by the pesticide. However, 24 hours post-exposure to 0.03% deltamethrin resulted in mortality ranging from 1.63 ± 1.14–68.17 ± 3.26%, with the St. Ann population being the most affected (p < 0.01). The results at 24 hours post-exposure to 0.03% lambda-cyhalothrin were not effective either, with mortality ranging from 15.43 ± 3.68–38.61 ± 1.32%. Similarly, post-exposure to 0.5% etofenprox caused 0.00 ± 0.89–42.14 ± 2.10% mortality with the insecticide having negligible effect on St. Catherine (0.98 ± 4.4%), KSA (0.00 ± 0.89%), St. Thomas (5.5 ± 1.81%) and Portland (0.00 ± 4.34%) populations (p < 0.01).The percent mortality after 24 hour post-exposure to 0.25% permethrin ranged between 5.24 ± 0.59–36.19 ± 3.51% in kdr mutation assays. The results of the WHO resistance intensity assays on the St. Catherine population are summarized in There were no significant differences between the pyrethroids tested and mortality rates amongst the populations, with the exception of St. Ann. As such, one population, St. Catherine, was selected for the WHO resistance intensity assays as well as for Ae. aegypti.The effects of the pyrethroids, permethrin, deltamethrin, lambda-cyhalothrin and etofenprox at 5x or 10x concentration were assessed on samples collected from St. Catherine and the results compared to that of their 1x concentration. Exposure to 0.3% deltamethrin, appeared most effective, resulting in mortality greater than 98% at 24 hours post exposure. Exposure to either 1.25% or 2.5% permethrin resulted in 4.14 ± 0.03 and 65.93 ± 0.05% mortality, respectively after 24 hours post exposure. Exposure to lambda-cyhalothrin caused less than 85% mortality, even at its 10x (0.3%) concentration after 24 hours post exposure. Only the 5x (0.5%) etofenprox was used in this study, 24 hours post exposure resulted in only 65.93 ± 0.05% mortality of p < 0.01). Similarly, 0.21% pirimiphos-methyl at 24 hours post-exposure was negligible, failing to cause mortality greater than 50% in the test populations, in comparison to the Rockefeller control, where Malathion or pirimiphos-methyl at the given concentrations, resulted in 95–100% mortality within 1 hr and 100% mortality at 24 hrs.The effects of the organophosphates 0.8% Malathion or 0.21% pirimiphos-methyl after 1-hour exposure were considered negligible See . The morp = 0.10  were assessed for V1016I and I1011V. All samples tested had the wild-type allele for I1011 . DNA was extracted from a total, N = 69 mosquitoes , St. Thomas, Portland, St. Mary, and St. Ann).The current study presents an updated and comprehensive profile on the state of insecticide susceptibility in Jamaican p <0.01) range for all pyrethroids tested at the 1x concentrations . As such, Ae. aegypti resistance to pyrethroids in eastern Jamaica can possibly be attributed to the unregulated overuse of household or animal-care products considering that these mosquitoes are usually found in close proximity to human dwellings or possibly from agricultural spraying.From the study, the Jamaican populations tested demonstrated resistance to pyrethroids and organophosphates. Deltamethrin was the only pyrethroid that caused high mean mortality in one of the six established populations. The highest mortality observed for deltamethrin 0.03% was 68.17 ± 3.26% in the St. Ann population. Surprisingly, this was the highest mortality  concentrations, deltamethrin at 10x concentration (0.3%) was toxic to the St. Catherine population that resulted in > 90% (94.90 ± 0.34%) knockdown within 1 hour of exposure and 98.95 ± 0.01% mortality (p <0.01) at 24 hours post exposure Click here for additional data file.S2 Table(XLSX)Click here for additional data file.


Lateral lymph node metastasis (LLNM) is very common in papillary thyroid carcinoma (PTC). The influence of tumour location on LLNM remains controversial. The purpose of this study was to reveal the association between PTC tumours located in the upper pole and LLNM.We reviewed a total of 1773 PTC patients who underwent total thyroidectomy with central and lateral lymph node dissection between 2013 and 2018. Patients were divided into two groups according to tumour location. Univariate and multivariate analyses were performed to identify risk factors associated with LLNM and “skip metastasis”.In the upper pole group, LLNM and skip metastasis were significantly likely to occur. Multivariate analysis showed that tumours located in the upper pole, male sex, extrathyroidal extension (ETE), central lymph node metastasis (CLNM) and tumour size were independent risk factors for LLNM, with odds ratios  of 2.136 (1.707–2.672), 1.486 (1.184–1.867), 1.332 (1.031–1.72), 4.172 (3.279–5.308) and 2.496 (1.844–3.380), respectively. Skip metastasis was significantly associated with the primary tumour location in the upper pole and age > 55 years, with ORs of 4.295 (2.885–6.395) and 2.354 (1.522–3.640), respectively.In our opinion, papillary thyroid tumours located in the upper pole may have an exclusive drainage pathway to the lateral lymph nodes. When the tumour is located in the upper pole, lateral neck dissection should be evaluated meticulously. P < 0.001) and were significantly associated with lateral lymph node metastasis (LLNM). Moreover, skip metastasis  was also significantly associated with the primary tumour location in the upper pole, as reported by several study [The morbidity of thyroid carcinoma is increasing worldwide. From 2011 to 2015, 63,324 patients were diagnosed with thyroid carcinoma in America . Many pral study  showed tA retrospective analysis of consecutive patients with PTC who underwent surgery at the Department of Endocrine and Breast Surgery of the First Affiliated Hospital of Chongqing Medical University between January 2013 and December 2018 was performed. The study was approved by the Medical Ethics Committee of The First Affiliated Hospital of Chongqing Medical University (2020-218). Consent has been obtained from each patient or subject after full explanation of the purpose and nature of all procedures used. Patients who met the following criteria were excluded: incomplete clinicopathologic data, other types of thyroid carcinoma, underwent a reoperation, tumour occupying the whole lobe, a bilateral tumour and an isthmus tumour and distant metastasis. Based on these criteria, 1773 patients with PTC were included.All patients were diagnosed through fine-needle aspiration biopsy (FNAB) before surgery, and the diagnosis was confirmed by pathological biopsy as PTC. Physical examinations, ultrasonography and fibrolaryngoscopy were routinely performed on each patient. All patients underwent total thyroidectomy (TT) with central compartment lymph node dissection (LND) and ipsilateral therapeutic or prophylactic lateral neck LND, of which most were therapeutic dissection due to clinical lymph nodes positive(cN1). Prophylactic dissection was performed in few patients with risk factors for lateral lymph node metastasis . The central lymph nodes and lateral lymph nodes  were divided into different regions and examined pathologically by three pathologists. Tumour location was categorized as upper pole, middle pole, and lower pole based on the preoperative ultrasound report from two experienced ultrasound doctors and the findings obtained during the operation. Since there is no clear anatomical division or guideline, the thyroid glands are bisected into three equal volumes  according to the consensus of most medical center. Tumours occupying more than one single pole were included in the upper pole group when the upper pole was involved.t test, the chi-squared test or Fisher’s exact test was used to compare differences in demographic and pathologic data on sex , age , tumour size , tumour location (upper/middle/lower pole), HT , multifocality  and ETE  between the patients with or without LLNM. Logistic regression was used for multivariate analysis to estimate the statistical significance. All variables with significant differences in the univariate analysis were included in the multivariate analysis. Statistical significance was defined as P < 0.05, and the analyses were performed using SPSS version 25.0  and GraphPad 8.0.Tumour multifocality, Hashimoto’s thyroiditis (HT) and extrathyroidal extension (ETE) were confirmed by a pathological examination. The Of the 1773 PTC patients included, 513 (28.9%) were male, and 1260 (71.1%) were female. The male-to-female ratio was 1:2.46. The median age of the patients was 42.1 ± 12.4 years. The mean size of the largest primary thyroid tumour was 1.5 ± 1.0 cm. Primary tumours located in the upper pole were found in 624 (35.2%) patients. A total of 862 (48.6%) patients had a tumour smaller than 1 cm, 624 (35.2%) had a tumour between 1 and 2 cm and 287 (16.2%) had a tumour larger than 2 cm. Gross ETE was observed in 375 (21.2%) patients. HT was present in 247 (13.9%) patients, and multifocal tumours were present in 312 (17.6%) patients. The analysis indicated that 1197 (67.5%) patients had CLNM, with an average number of 2.6 ± 3.3 metastatic lymph nodes. A total of 825 (46.5%) patients had LLNM, with an average number of 1.7 ± 2.9 metastatic lymph nodes. Among these patients, 684 (38.6%) had level II/III LNM, and 374 (21.1%) had level IV LNM. “Skip metastasis” was observed in 139 (7.8%) patients (Table P < 0.05)  patients and 277 (24.1%) patients in each group. A total of 384 (33.4%) patients in the non-upper pole group experienced metastasis to level VI without lateral neck lymph node involvement , which was more than those in the upper pole group (20.7%). In the upper pole group, 53.68% of patients had LLNM. In contrast, 42.65% of patients in the non-upper pole group had LLNM. Skip metastasis was observed in 97 patients (15.54%) in the upper pole group and 42(3.66%) in the non-upper pole group, with a statistically significant difference (5.6% patiP < 0.001), tumour size (P < 0.001), tumour location (P < 0.001), ETE (P < 0.001) and CLNM (P < 0.001). Multivariate analysis (Table P < 0.05).Clinicopathological differences between LLNM and clinicopathological variables are shown in Table is Table  revealedP < 0.001) and in tumours located in the upper pole (P < 0.001). In the multivariate analysis, these factors remained independently associated with skip metastasis; the adjusted ORs with 95% CIs were 2.354 (1.522–3.640) and 4.295 (2.885–6.395), respectively.Table Lymph node metastasis is very common in PTC, with an occurrence rate of 30–80% , 14. EveTumours located in the upper pole were identified as an independent risk factor for skip metastasis in our study. Luo et al.  reviewedOur study also demonstrated that male sex, a large tumour size, ETE and CLNM were significantly correlated with LLNM by multivariate analysis. A random-effects model with 16 studies  showed tThe most common metastatic site of PTC is the central compartment, which has been reported to be an independent contributor to LLNM , 30. ThiThere is no official standard or acknowledged anatomical mark for the location of thyroid tumours. In our study, tumour location was determined with ultrasound and findings during the operation. There should be a concise standard based on imaging  and pathology in future recommendations.There were several potential limitations to this study. Several clinicopathological features, such as histological subtype, Braf mutation and lymphovascular invasion, were not provided. Furthermore, we are examining more detailed pathological reports and long-term follow-up data.The lymphatic drainage pathway of PTC is complex. The extent of LND in each patient should be individualized. When the tumour is located in the upper pole of the thyroid, it is more likely to have LLNM than when a tumour is not located in the upper pole. The risks and benefits of preventive lateral LND should be discussed with the patient.


The ones who were vaccinated against influenza, 91.9% (n = 387 [95% CI: 89.2–94.6]) (p < 0.001) manifested they would do so in future pregnancies. The probability of future non-vaccination was modeled, which was related to an unfavorable opinion towards vaccines (OR = 4.07 [95% CI: 2.01–8.24]) (p < 0.001), having suffered from influenza during pregnancy (OR = 3.84 [95% CI: 1.41–10.42]) (p < 0.05), and not having been vaccinated during previous pregnancies (OR = 38.47 [95% CI: 23.58–62.76]) (p < 0.001). Vaccination during pregnancy increases the intent of vaccination in the future.A good perception of the vaccines administered during pregnancy favors immunization coverage, which is still not optimal for the influenza vaccine. To understand the predisposition towards vaccination in future pregnancies, a study was performed that evaluated the experiences of women with the vaccine or influenza. A cross-sectional study was conducted through telephone interviews given to a total of 683 postpartum women in two health departments from the Valencia Community (Spain). This interview asked about their intention of becoming vaccinated in future pregnancies and whether they were favor or against vaccination. Most of them, 98.5% ( Vaccination of pregnant women , has greatly increased in the 21st century. The protection received by the mother and fetus in a single vaccination implies benefits both for the mother, who is protected from infection and its complications, as well as the fetus and newborn, due to the transfer of antibodies ). (The “previous vaccination during pregnancy” statement yielded a 61.6% flu vaccination coverage (–66.2]). .n = 655 [95% CI: 98.0–99.6]) of women, the statement coincided with the data available in the NVR records, indicating a high agreement and a high degree of sincerity in their responses.The answers provided by the women in their postpartum period about their flu vaccination or not was compared with the existing data found in the Vaccination Registry of the Valencian Community (NVR) to verify the veracity of their responses. The results showed that for 98.8% (n = 673 [95% CI: 97.6–99.4]) confirmed having received the systematic vaccines scheduled in the children’s calendar.As observed in n = 451 [95% CI: 61.6–70.4]) declared that they would be vaccinated again.Women who were vaccinated or unvaccinated during their recent pregnancy were asked if they would be vaccinated again against influenza in a future pregnancy, if it coincided with the period of the seasonal influenza campaign. Of the total women interviewed, 66.0% (n = 387 [95% CI: 89.2–94.6]) as compared to those who had not been vaccinated, 24.4% (n = 64 [95% CI: 13.9–34.9]). . .The reasons provided for not wanting to become vaccinated in a future pregnancy can be observed in n = 421 [95% CI: 56.9–66.2]) of pregnant women who received the vaccine against influenza, 8.6% (n = 36 [95% CI: 0.0–17.7]) manifested suffering secondary effects after the vaccination.Of the 61.6% (n = 17 [95% CI: 23.5–70.9]), pain and inflammation on the arm where they received the injection, 33.3% (n = 12 [95% CI: 6.6–59.9]) and “becoming sick” with influenza after immunization, 16.7% (n = 6 [95% CI: 0.0–46.5]). There was a case of hospital admittance due to the risk of premature birth post-vaccine.The secondary effects experienced were: symptoms of a cold without fever, 47.2% (n = 15 [95% CI: 19.0–69.2]) of the vaccinated pregnant women, suffering a “cold” after vaccination was the main reason for not wanting the vaccine in later pregnancies. However, in the unvaccinated group, the belief that the vaccine was not “necessary or effective” was the main reason for rejection, 53.0% n = 105 [95% CI: 43.4–62.5]) (For 44.1% (4–62.5]) .n = 502 [95% CI: 69.6–77.4]) of the total. Among the unvaccinated group, this ascended to 79.3% n = 208 [95% CI: 73.8–84.8])  .p < 0.05). Only 5.3% affirmed having suffered influenza during another pregnancy. This subgroup was divided similarly between vaccinated and unvaccinated women.Having been sick from influenza in the past, while not pregnant, was a predisposing factor for becoming vaccinated during the present pregnancy (The results obtained after adjusting the multivariate logistic regression models to explain the future intent of non-vaccination against influenza can be observed in p < 0.001), (OR = 4.07 [95% CI: 2.01–8.24]), having experienced influenza during pregnancy (p < 0.05), (OR = 3.84 [95% CI: 1.41–10.42]), to not having been previously vaccinated against influenza during pregnancy (p < 0.001), (OR = 38.47 [95% CI:23.58–62.76]), and lastly, that the pregnant woman was of Spanish origin (p = 0.082), (OR = 1.65 [95% CI: 0.93–2.91]).The probability of non-vaccination was modeled, resulting in the significant association with having an unfavorable opinion about vaccines in general  had been sick with influenza while pregnant, this factor was significantly associated with the future intent of non-vaccination. This data should be clarified, as many pregnant women defined influenza as having had a cold or another respiratory infection, without having a serological diagnosis that could confirm its presence. In this sense, although the secondary effects described by the postpartum women after the vaccination were few (9.1%) and without severity , almost In Europe, to achieve a vaccination coverage against the flu higher than 75% , it is iThe pregnant women who were not native to Spain were vaccinated against the flu more often than Spanish women, as opposed to a study conducted in France, where immigrant women were considered a risk factor for non-vaccination . Our womThe flu vaccination recommendation was directly provided by the midwife during the consultation ,32. We bIf we combine the good and majority opinion of the pregnant women towards vaccines in general with the favorable attitude of the health professionals, then expectations for improvement of the forecasted future vaccination coverage for the child calendar are high . The retAs for the flu vaccine, we received responses such as “it is not an effective or necessary vaccine” and “I have doubts or mistrust about its safety”, which represented more than half of the reasons provided for rejecting it in future occasions ,32,42. PIt would be advisable to create trust in vaccines during pregnancy. The midwives, together with the obstetricians and pediatricians, could collaborate in the creation of training programs related to maternal and child vaccination before the birth .More research studies on this subject could result in the development of future recommendations about the optimal period of time for immunization,  and the most cost-effective professionals, not only in the vaccination, but also in the monitoring and advice phases ,45.As a limitation of the study, it should remembered that the women indicated having fallen ill with influenza in the past or had this disease as a side effect after vaccination, according to her own experience. The vaccination status was also self-reported, and could therefore be subject to recall biases. However, this statement obtained a high level of agreement with the computerized nominal registry of vaccines from the VC (NVR).The experience of influenza vaccination of pregnant women tends to condition future vaccination events. Given the vaccine is usually administered on various occasions, if some reason existed related to the secondary effects that the pregnant women attributed to the vaccine, it could lead to its rejection. The collaborative work by the health professionals involved in pregnancy care should take advantage of the good perception the women have about vaccines to provide efficient advice that will increase vaccination coverage.


ObjectivesThe main aim is to find out the clinical feature and outcome of status epilepticus (SE) in children managed in a teaching hospital. The secondary aim is to identify the risk factors influencing the adverse outcomes.MethodsIn this prospective cohort, children aged 1 month to 14 years with SE as per the International League Against Epilepsy’s new guideline (2016) who presented to the emergency department during the period of November 2017 to October 2019 were enrolled. Clinical profile, treatment, and outcome of cases (n = 94) were noted.ResultsThe majority of children, 60 (63.82%), were less than five years of age. Prior history of seizures was present in 33 (35.1%) cases, whereas 61 (64.9%) cases presented with SE as the first episode of seizure. In 14 (42.4%) previous seizure cases, SE was due to drug default. No response to first-line antiepileptic drug (AED) was seen in 84 (89.37%) cases. Acute symptomatic etiology was the commonest etiology of SE in 64 (68%) cases, of which neuro-infections accounted for 44 (46.80%) cases. Longer duration (>60 minutes) of status (p < 0.01), ventilator support (p < 0.0001), and circulatory impairment (p < 0.0001) were attributable risk factors for mortality. A total of 28 children died , and 11 showed the persistence of their neuro-deficit.ConclusionsNeuro-infection is the most common etiology of SE in children. Longer duration of SE, more lag time for receiving the first AED, respiratory failure, and presence of shock are independent predictors for poor outcome. Hence, cessation of convulsion at the earliest leads to improved outcomes. Status epilepticus (SE) is a medical emergency, and its neurological outcome is a concern to every pediatrician in developing countries. The estimated convulsive status epilepticus (CSE) prevalence is 14.5 per 100,000 per year in developed countries ,3. It isPrevious studies have described the clinical manifestation and outcomes of SE taking 30 minutes as a cut-off seizure duration ,7. PreseThe objective of the study is to determine the clinical features, etiology, response to treatment with BZD, and immediate outcome, and to identify the risk factors influencing adverse outcomes in children with SE admitted to the pediatric ward/ intensive care unit in a referral teaching hospital.This prospective observational study included children between 1 month and 14 years of age admitted in the pediatrics ward of S. C. B. Medical College situated in a tertiary care teaching hospital from Eastern India over a two-year period (November 2017 to October 2019). Approval of the Institutional Ethical Committee was obtained before initiation of the study. During this period, 94 children with SE as per ILAE’s new guideline (2016)  were admType and etiology of SE were classified based on the SE classification report of the ILAE Task Force . RefractData were analyzed using SPSS Version 21 . A p-value of less than 0.05 was considered statistically significant. Furthermore, univariate analysis was used to find out the significance of various parameters in the outcome of the present study.Of the total 94 children studied, the median (IQR) age was 3 1-7) years. The majority of children 60 (63.82%) were less than five years of age. There was a male predominance (61 [64.9%]). Out of hospital seizure onset was present in 82 (87.2%) cases. A total of 49 (59.7%) patients received pre-hospital treatment with diazepam (30) and phenytoin/phenobarbitone (19). Fever was present in 63 (67%) cases before the onset of seizures. Prior history of seizures was present in 33 (35.1%) cases, whereas the first episode of seizure with 61 (64.9%) cases. Most of the children, 46 (48.9%), had an SE duration of <30 minutes. The seizure was terminated after administration of BZD in 10 (10.63%) cases, but second- and/or third-line drugs were needed in 84 (89.37%) cases. However, 27 (28.72%) patients progressed to RSE. Generalized seizures, mostly generalized tonic-clonic (GTC) seizures, were observed in 87 (92.6%) children. Acute symptomatic etiology was the most common cause of SE in 64 (68%) cases. Neuro-infections accounted for 44 (46.80%) cases and febrile seizures for 5 (5.3%) cases. Again, acute symptomatic etiologies were the major causes of SE in children less than five years of age, whereas remote symptomatic causes were more in children aged 3-10 years  compared to CSE <30 minutes (n = 3 [5.1%]) (p < 0.01).The fatality in children with RSE was 85.2% compared to non-RSE 7.5% (p < 0.0001). Ventilator support (p = 0.0001) and circulatory impairment (p = 0.0001) were the important attributable risk factors for mortality . Other studies have also found a higher prevalence in the younger age group ,13. As tA significantly high incidence of RSE with acute symptomatic etiology was found in the present study. Risk factors for RSE are sepsis, shock at presentation, pre-existing neurological abnormalities, and lag time for receiving the first AED. Children with RSE had a significantly prolonged length of stay in hospital. In this study, a less number (n = 11 [11.70%]) of patients had residual morbidity such as focal neurological deficits and neurocognitive impairment compared to another study ,18.A requirement of mechanical ventilation was relatively higher in the present study because most patients referred to our hospital were in critical condition and refractory status. A similar finding was reported in a study from south India . The comThe mortality rate in the present study was similar to other Indian studies ,22, wherLimitationsFew causes, such as the inborn error of metabolism, autoimmune encephalitis, specific and viral encephalitis, could not be specified due to lack of investigations.Acute symptomatic etiology is the commonest cause of SE, predominantly with neuro-infections. Longer duration of SE, more lag time for receiving first AED, RSE, ventilator requirement, and presence of shock are independent predictors for poor outcomes. Upgradation of intensive care facilities even in district headquarter hospitals might improve outcomes of SE.


In the Surveillance Summary “Abortion Surveillance — United States, 2015,” data on the number of known previous induced abortions for women having abortions in this reporting year were erroneously included for New York City. These estimates did not meet reporting standards and should have been excluded from this report. When corrected, among women with abortions in the reporting year, the proportion with no previous induced abortions increased, and the proportion with one or more previous induced abortions decreased.41.0% of abortions, and women with no previous abortion accounted for 59.0%. Women with three or more previous births accounted for 14.2% of abortions, and women with three or more previous abortions accounted for 6.5% of abortions.”On page 2, the second and third sentences of the second paragraph should have read “Women with one or more previous induced abortions accounted for 38 areas that reported the number of previous abortions for women who obtained abortions in 2015 indicate that the majority (59.0%) had no previous abortions, 34.5% had one or two previous abortions, and 6.5% had three or more previous abortions (Table 17). Among the 34 reporting areas††††† that provided data for the relevant years of comparison , the percentage of women who had no previous abortions increased 2% (from 57.8% to 59.1%), whereas there was a 4% decrease for women who had one or two previous abortions, and the percentage of women who had three or more previous abortions was unchanged (6.4%) from 2006 to 2015. However, the percentage of women who had no previous abortions decreased 1% from 2006 to 2010 (from 57.8% to 57.4%) and then increased 3% from 2011 to 2015 (from 57.2% to 59.1%). By contrast, the percentage of women who had three or more previous abortions increased 11.0% from 2006 to 2010 (from 6.4% to 7.1%) then decreased 9% from 2011 to 2015 (from 7.0% to 6.4%). The percentage of women who had one or two previous abortions remained stable from 2006 to 2010 (35.7% to 35.6%) and then decreased 4% from 2011 to 2015 (from 35.8% to 34.4%).” New York City should have been included in the ††††† footnote, which lists reporting areas that were not included in these estimates.On page 8, the last paragraph of the second column should have read “Data from the 224,163 (59.0), 93,025 (24.5), 37,923 (10.0), 24,519 (6.5), 379,630 (99.4). The * footnote should have read “Data from 38 reporting areas; excludes 14 areas  that did not report, did not report by the number of previous induced abortions, or did not meet reporting standards.” The total in the ** footnote should have been 382,003.In Table 17, the line for New York City should be deleted. For the total line, the numbers and percentages should have read