MRE11 germline single nucleotide polymorphism (SNP), rs1805363 (G > A), to be associated with poor outcome following radiotherapy (RT) and increased expression of MRE11 isoform 2 in a limited panel of bladder cancer cell lines and tumours.DNA double strand breaks are the cytotoxic lesions produced by ionising radiation. Critical for the repair of these lesions is the DNA damage response protein MRE11 which, in a complex with RAD50 and NBS1, mediates DNA damage signalling and double-strand break repair. We previously found the presence of an MRE11 isoform 2 would be more radio resistant than cells expressing MRE11 isoform 1.To look for further evidence in support of the SNP/isoform association in a larger panel of germline and tumour samples donated by patients diagnosed with invasive bladder cancer, and to test the hypothesis that bladder cancer cells expressing G > A SNP. Loss of heterozygosity was determined by genotyping tumour DNA in 17GA germline patients. The Cancer Genome Atlas was mined to correlate presence of the GA germline genotype with MRE11 isoform expression. We used colony formation assays and γH2AX foci kinetics after ionising radiation in RT112 MRE11 knockdown cells expressing ectopic MRE11 isoform 1 or 2.Germline DNA from 189 patients with invasive bladder cancer  was genotyped for the rs1805363 MRE11 isoform 2 expression was found to be significantly higher (p = 0.007) in patients’s samples containing the A minor allele compared to those with only the G major allele (n = 23). In the TCGA database we found 16% (66 out of 406) of bladder tumours heterozygous for the SNP and only two homozygous, and a significant relative increase of isoform 2 usage (p = 0.017). We identified no significant difference in radio sensitivity between bladder cancer cells expressing either MRE11 isoform.Of the 189 germline DNA samples, 22 contained both the A minor allele and G major allele with the remaining wild type containing only the G major allele. LOH was identified in seven of 17 available tumour samples. Tumour MRE11 isoform 2 was not found to be associated with increased cellular sensitivity to radiation. We conclude that the previously reported association between the germline rs1805363 SNP and poor survival in MIBC patients following RT is unlikely to be related to the DNA damage response function of MRE11 isoform 2.In this study the  Organ-confined muscle-invasive bladder cancer (MIBC) is routinely treated using either radical cystectomy or bladder-sparing approaches including chemoradiotherapy (CRT) or radiotherapy alone, with or without neoadjuvant chemotherapy. Cystectomy and radiotherapy have been shown to result in similar cause-specific survival rates  but no rMRE11 results in two transcript variants encoding MRE11 isoform 1 and MRE11 isoform 2, respectively. The two MRE11 isoforms are similar, with the only difference in the translated protein being the absence of a short 27 amino-acid-long exon, exon 16, in MRE11 isoform 2. At the transcript level MRE11 isoform 2 has a longer 5’ UTR than the MRE11 isoform 1.DNA double strand breaks (DSB) are the cytotoxic lesions caused by ionising radiation and are repaired via two major pathways: non-homologous end joining and homologous recombination . MRE11 iMRE11 (rs1805363) which was associated with worse cancer-specific survival following radiotherapy  was diluted 1:2000 in PBST and applied for 1 hr in room temperature. Cells were washed with PBS as before and mounted by using proLong diamond antifade mountant with DAPI . Cells were visualised for γ-H2AX foci kinetics, P values were calculated using Kruskal Wallis test with Dunn’s multiple comparisons in GraphPad Prism. Counting of γ-H2AX foci was determined using Image J software. For clonogenic assays, surviving fraction was calculated on the basis of the number of colonies on non-irradiated plates. Data are presented as the log of the surviving fraction with error bars representing the SEM.Clonogenic assays, western blots, and immunofluorescence data are representative of three independent experiments. For G > A SNP. Of these patients, 141 were diagnosed with muscularis propria invasion (stage T2+) and 48 with lamina propria invasion (T1). Of the 189 samples, 22 were identified as containing both A and G alleles, with the remainder identified as only containing the major G (WT) allele. Loss of heterozygosity (LOH) within heterozygous tumours was investigated by genotyping extracted tumour DNA, which was available for 17 of the germline genotyped casesidentified as positive for both A and G alleles. LOH was identified in 7 of the 17 samples .We obtained 189 samples of germline DNA from the University of Birmingham’s Bladder Cancer Prognosis Programme which were genotyped for the rs1805363 MRE11 isoform 2 expression was found to be significantly higher (p = 0.007) in patients positive for the rs1805363 A minor allele  compared to patients positive for only the G major allele (mean proportion of MRE11 isoform 2 in the G only germline genotyped patient group: 2.3%). Isoform 2 was also found to be generally increased in non-wild-type samples in TCGA , as well as in direct comparison to isoform 1 (p = 0.017) , the area of tissue being too small for sufficient material to be collected (5/12) or the tissue being identified as highly inflamed (1/12). Of the remaining 24 blocks, 10 were from patients’ germline positive for both A and G alleles and 14 from patients positive for only the G (WT) allele. RNA extracted from patient tumours was then used to quantify the relative expression of isoform 1 and 2 within the samples and grouped according to germline genotype . One sam017) see . Althoug017) see A. Furthe017) see B.MRE11 isoform 2 could influence cellular radiosensitivity and DNA damage response. We used an RT112 bladder cancer cell line knocked down for endogenous MRE11 and re-expressing an MRE11 construct containing either the full length MRE11 coding sequence (MRE11 isoform 1) or the MRE11 coding sequence lacking MRE11 exon 16 (MRE11 isoform 2). As this question referred to the function of the translated protein we did not include the MRE11 isoform 1 or 2 5’UTR sequences in these constructs.We then investigated whether the increased presence of MRE11 isoform 1 or isoform 2 in these cells led to a greater than 2.5-fold increase in full-length MRE11 protein level , which corresponds to approximately 30% of the endogenous full-length MRE11 level in the parental cell line  cells  higher levels in the MRE11 knockdown cells compared to the parental cell line that persisted for 2, 6 and 24 hrs after irradiation. Consistent with the level of expression achieved compared to the parental cells, MRE11-transfected cells showed a partial reduction in γ-H2AX foci at 2 hr after 2 Gy irradiation, at an intermediate level between the parental and MRE11 knockdown cells. The reduction in γ-H2AX after rescue with either the MRE11 full length or Δexon16 constructs was 1.3-fold at 30 minutes after 2 Gy IR, similar to the MRE11 protein levels achieved relative to the parental cells. There was no significant difference in γ-H2AX foci at any of the time points between the MRE11 full length and the Δexon16 cells (γ-H2AX foci level (No IR) for all four cell lines were re-achieved at the 6 hr time point. This suggests that the absence of exon 16 has minimal effects on cell survival and repair of DNA damage after IR.A colony survival assay was performed to determine the radiosensitivity of the MRE11 RT112 cell lines. The MRE11 knockdown cell line displayed a significant increase in radiosensitivity compared to the parental RT112 cell line at doses of 2 to 10 Gy. This was rescued by the re-expression of either the MRE11 full length (isoform 1)  or the MRE11 exon16-negative construct (isoform 2) , while MRE11 knockdown cells had 62.5-fold and 52.6-fold reduction on survival compared to parental cells at 8 and 10 Gy. However, there was no significant difference in radiosensitivity between cells expressing the full-length version MRE11 and exon16-deleted (6) cells B. To det16 cells A and B. MRE11 rs1805363 SNP in patients with invasive bladder cancer, we genotyped the rs1805363 status of germline DNA and in this study provided further evidence for a correlation between the presence of the MRE11 germline rs1805363 A minor allele and an increase in the tumour expression of MRE11 isoform 2 over isoform 1. However, it would be useful to establish if the tumour genotype is a more accurate indicator of the expression of MRE11 isoform 2 in invasive bladder tumours.In line with our previous report on the MRE11 isoforms from RNA-seq data. In addition, it would be of interest to study other isoforms, as for example ENST00000540013.5 coding for a much shorter protein without exon 16, which seems to be supressed in the non-wild-type cases  than reported earlier  or cells expressing MRE11 lacking exon 16 (MRE11 isoform 2). We therefore conclude that the presence or absence of MRE11 exon 16 may not influence cell survival or DSB repair capacity after IR, contrary to our initial hypothesis, although we appreciate that our conclusions are limited by the use of only one cell line system.Given that the presence of the rs1805363 A minor allele in the germline DNA of patients with invasive bladder cancer was associated with worse survival after radiotherapy , we hypoMRE11 isoform 2 we were interested in directly comparing the protein function of our MRE11 constructs. Therefore, we selected cells for our assays that stably re-expressed our constructs. Unfortunately, this strategy did not allow us to address whether there was a difference in the protein stability or turnover between the two isoforms. Ideally, the protein levels of MRE11 isoform 1 and isoform 2 would have been assessed in patient samples where we had quantified the relative expression of mRNA. However, unfortunately with no available antibody distinguishing between the two isoforms and with only a 27aa difference between the two protein variants, it was not possible to reliably distinguish the isoforms using techniques such as western blotting or immunohistochemistry. We did have an antibody raised to exon 16 but this unfortunately proved unsuitable for western blotting, and so we could not verify its specificity for use in determining the isoforms’ presence in the samples. Interestingly, the phosphorylation of MRE11 threonine 597, which resides within exon16, has been found to promote MRE11 degradation in PTEN deficient cells [In our cellular analysis of nt cells , which dMRE11 isoform 1 compared to 2 is that we only re-expressed either MRE11 isoform 1 or 2 in our KD cells. However, in the human tissue we analysed MRE11 isoform 1 was present in conjunction with isoform 2. Therefore this method may be an oversimplification of the underlying biology and expressing different ratios of the two isoforms may be a more representative model in which to study their function. Indeed, in cells, MRE11 exists as a homodimer with dimerisation being critical for the correct assembly of the MRN complex and the binding and aligning of DNA ends [A limitation in the system we used to investigate the efficiency of DSB repair by DNA ends . It is pDNA ends .MRE11 isoform 2 levels to radiotherapy.Aside from its role in DNA damage repair, MRE11 also functions in the repair of stalled and collapsed replication forks . While wThe authors report no funding.The authors have no conflict of interest to report.AW performed experiments, interpreted the data and drafted and revised the manuscriptJN performed experiments, interpreted the data and drafted and revised the manuscriptLB provided histopathological expertise for tumour identification and interpretation, and revised the manuscriptRA performed bioinformatics analysis, interpreted the data and drafted and revised the manuscriptN H-Z performed bioinformatics analysis and interpreted the dataMZ contributed patient samplesKC contributed patient samplesNJ contributed patient samplesRB contributed patient samples, interpreted the data and revised the manuscriptAK conceived the study, supervised the work and revised the manuscript.All authors approved the final version of the manuscript and agreed to be accountable for the accuracy and integrity of the work.Click here for additional data file.


The authors would like to provide clarifications and correct errors in Figs PLOS ONE study, as both studies used the same repository of cells and treatments, and therefore some data was re-used. A revised The wildtype flow cytometry DNA panels in PLOS ONE study [The NE study  was carrNE study ; howeverNE study  are novechl1 mutant cells. The incorrect panel is replaced in the revised In scc2-4 cells was incorrectly used in the NZ 1hr panel for wildtype cells. The incorrect panel is replaced in the revised In chl1 mutant cells. The incorrect panel is replaced in the revised In The authors confirm that the above errors occurred in figure assembly and do not affect the analyses or findings of the study. The authors apologize for the errors.The legends for https://doi.org/10.6084/m9.figshare.11412783.v2.The underlying data for the article is provided on Figshare: S1 Figchl1 mutant cells (N = 100 cells for each strain). B) 1N peak quantification for both wildtype and chl1 mutant cells arrested in nocodazole for 3 hours at 23°C. C) Flow cytometer data reveals DNA contents in wildtype and chl1 mutant cells after nocodazole arrest at 23°C for 2.5 hours and 3 hours. D) Flow cytometer data reveals DNA content in wildtype and chl1 mutant cells analyzed in A) Morphology quantification for both nocodazole-arrested wildtype and (TIF)Click here for additional data file.


Motion sensing superpixels (MOSES) is a systematic computational framework to quantify and discover cellular motion phenotypes. Published 26, February 2019During final figure preparation the similar velocity kymographs for 0% serum EPC2:CP-A and EPC2:OE33 in Figure 3A were switched. This mistake was noticed by the authors when rereading the published version. Also in Figure 3A, the red and green font colours used for the cell line names were accidentally switched; these colours should match those in Figure 1 as the same example videos were used for both figures. Figure 3A has been corrected accordingly. These corrections do not affect any of the scientific findings or conclusions of the original paper.The corrected Figure 3 is shown here:The original Figure 3 is shown here for reference:


End-tidal carbon dioxide (EtCO2) monitoring is a noninvasive tool used to estimate Paco2 values.Alterations in the partial pressure of carbon dioxide, arterial  higher than EtCO2. Overall, 187 of all Paco2-EtCO2 pairs (42.0%) agreed. There was larger variation in the Paco2-EtCO2 difference during the first 8 hours compared with 9 to 24 hours after admission to the pediatric intensive care unit. Development of pediatric acute respiratory distress syndrome within 24 hours of admission was associated with a lower likelihood of Paco2-EtCO2 agreement  compared with no development of pediatric acute respiratory distress syndrome. A diagnosis of pediatric acute respiratory distress syndrome 1 to 7 days after admission was associated with a larger first-day Paco2-EtCO2 difference compared with those who never developed pediatric acute respiratory distress syndrome .The analysis included 137 patients  and 445 paired Paco2-EtCO2 agreement was low, especially among patients with pediatric acute respiratory distress syndrome. Low Paco2-EtCO2 agreement early in hospitalization may be associated with future development of pediatric acute respiratory distress syndrome. Data on EtCO2 should not be substituted for data on Paco2 during the first 24 hours.In this study of pediatric traumatic brain injury, Pa co2) and end-tidal carbon dioxide (EtCO2) and associated factors in children with traumatic brain injury (TBI).This secondary analysis of a cohort study examines agreement between partial pressure of carbon dioxide, arterial (Pa However, arterial sampling can be challenging in children, and complications are not uncommon.4 Capnography allows for noninvasive, continuous measurement of end-tidal carbon dioxide (EtCO2) with lower cost and fewer complications compared with arterial cannulation, and it is widely used in clinical pediatric practice as a substitute for arterial cannulation.5 End-tidal CO2 reasonably estimates Paco2 in healthy adults.6 A mean difference of 0 to 5 mm Hg between Paco2 and EtCO2 is generally accepted as good agreement, and good agreement has been demonstrated in patients during general anesthesia who are undergoing prolonged neurosurgical procedures, in clinically stable patients in the pediatric intensive care unit (PICU), and in adult patients with TBI without hypotension, metabolic acidosis, or severe lung injury.10 However, the utility of EtCO2 as a substitute for Paco2 in critically injured children with TBI has not been examined, to our knowledge.Globally, traumatic brain injury (TBI) is a leading cause of pediatric morbidity and mortality.co2 less than 30 mm Hg in the initial 48 hours after admission.11 A recent study showed that adhering to this recommendation improved discharge outcomes.12 The Brain Trauma Foundation does not, however, include a recommendation either for or against the use of EtCO2 measurements because of the lack of data comparing Paco2 with EtCO2 to prevent unwanted hypocarbia and hypercarbia. To evaluate the validity of using EtCO2 as an indicator of Paco2 in children and adolescents hospitalized with TBI, we performed a secondary analysis of a prospective cohort study to describe the agreement between Paco2 and EtCO2 early after TBI and determine the factors associated with Paco2-EtCO2 agreement.Recently, the Brain Trauma Foundation published the third edition of medical management guidelines for severe pediatric TBI, recommending avoidance of prophylactic hyperventilation and Pa12 Waiver of consent was granted by the University of Washington Institutional Review Board for the parent project because this research was a secondary data analysis that was deemed to involve no more than minimal risk to participants and waiver would not adversely affect the rights and welfare of the participants. This study followed the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) reporting guideline.After approval by the University of Washington Human Subjects Division, from December 17, 2018, to January 10, 2019, we conducted a secondary analysis of data from the recently published Pediatric Guideline Adherence and Outcomes program study.Harborview Medical Center is a 450-bed mixed level I adult and pediatric trauma hospital that serves the 5-state  Pacific Northwest region. The 18-bed PICU admits approximately 120 pediatric trauma patients with TBI annually.2 values recorded within 30 minutes of a Paco2 measurement during the first 24 hours after admission and (2) had 1 or more Paco2-EtCO2 pairs with a systolic blood pressure (SBP) measured at any point within the 60 minutes prior to the Paco2 value  had 1 or more EtCOco2, EtCO2, SBP, oxygenation index, oxygenation saturation index, and Glasgow Coma Scale score were abstracted from the electronic health record. Race and ethnicity were self-reported.Data on demographic characteristics, mechanisms of injury, and injury severity measures were obtained from the Harborview Trauma Registry. Ascertainment of diagnosis of TBI via computed tomography, surgical procedures performed, Paco2 measured via arterial blood gas was entered into the electronic health record by laboratory personnel. End-tidal CO2 was measured via Medtronic capnography and manually entered by PICU nurses or respiratory therapists into the electronic health record at a minimum of every hour. Data on SBP associated with EtCO2 and Paco2 levels were recorded with a digital sphygmomanometer or by invasive arterial monitoring and were automatically entered from SpaceLabs monitoring system at least every hour, or more frequently as dictated by clinical care. All paired data points were collected while the patients were intubated and mechanically ventilated.Clinical data collected during the first 24 hours of clinical care after admission to the PICU were examined. Pa13 Patients with at least 3 qualifying values of the oxygenation index or the oxygenation saturation index during a minimum of 6 hours were determined to meet oxygenation criteria for PARDS. Medical record abstraction was performed for all patients meeting oxygenation criteria to evaluate for the presence of a new parenchymal infiltrate detected on a chest radiograph (based on review by the attending radiologist and secondary review by a pediatric intensivist [E.Y.K.]) and for the origin of edema (based on documentation by the attending physician).Patients were determined to have pediatric acute respiratory distress syndrome (PARDS) if they met Pediatric Acute Lung Injury Consensus Conference PARDS criteria within 7 days of their injury.2 value recorded within 30 minutes was used to create 1 Paco2-EtCO2 data pair. The nearest SBP value within 60 minutes prior to the Paco2 value being measured was then added to create a Paco2-EtCO2-SBP trio. Values measured during apnea trials for brain death were excluded because the patients were no longer receiving routine TBI care.For every arterial blood gas measurement obtained during routine medical care, the nearest EtCO2 measured by capnography is a mixture of relatively CO2-rich alveolar gas diffused from capillaries with CO2-poor gas in ventilated lung regions with poor perfusion . Dead space includes the components of the respiratory system that are ventilated but do not participate in gas exchange and alveoli that receive ventilation but are not perfused because of physiologic or pathologic factors, as well as the airways and breathing apparatus. The Paco2 value is typically 2 to 5 mm Hg higher than the EtCO2 value under physiologically stable conditions.14 Therefore, Paco2-EtCO2 pairs were categorized as either in agreement, if the Paco2 value was between 0 and 5 mm Hg greater than its paired EtCO2 value, or not in agreement.End-tidal COco2 measurement and when EtCO2 was recorded.Potential explanatory variables were determined a priori based on literature review and research team discussion, and they included severity of chest injury, severity of nonhead-nonchest injury, presence of PARDS (within 24 hours or 1-7 days after admission), hypotension, patient age, time since admission, and the time difference between PaThe presence of severe chest injury or nonhead-nonchest injury with a region-specific Abbreviated Injury Score greater than 2 was coded as a binary variable. Hypotension was defined as SBP less than 70 mm Hg +2 × (patient age in years) coded as a binary variable. The PARDS status for each patient was categorized as follows: did not develop within the first week of PICU admission, developed within 24 hours of PICU admission, or developed after 24 hours but within 1 week of PICU admission. Time after admission was dichotomized into 0 to 8 hours and 9 to 24 hours, and age was categorized into younger than 1, 1 to 4, 5 to 9, 10 to 14, and 15 to 17 years.co2 and EtCO2 was assessed for the overall study population and stratified by PARDS status at 24 hours using Bland-Altman plots with limits of agreement adjusted for repeated measures.15 We also assessed the proportion of Paco2-EtCO2 pairs with Paco2 values 0 to 5 mm Hg greater than their paired EtCO2 value.We used descriptive statistics to examine clinical injury characteristics. Agreement between Paco2-EtCO2 pairs in agreement with pairs that did not agree. Stratified Bland-Altman plots were created to visually compare trends in agreement across levels of age and whether patients developed PARDS within the first 24 hours and within 1 to 7 days of PICU admission. We assessed constant and proportional differences between Paco2 and EtCO2 within each Paco2-EtCO2 pair, and we estimated Pearson correlation coefficients using Passing and Bablok regression and Deming regression.17The characteristics that may have affected the likelihood of agreement between measures were initially assessed descriptively, comparing Paco2 and EtCO2, we used univariate logistic regression models clustered by patient; to minimize confounding, we used multivariable logistic regression models clustered by patient adjusted for age, PARDS within the first 24 hours of PICU admission, severe TBI, severe chest injury, severe nonhead-nonchest injury, and shock. Anatomic dead space in the pediatric population could have considerable variation and may affect Paco2-EtCO2 agreement. Although we were not able to measure dead space individually, we chose to adjust for age and cluster by patient to address this issue. To account for the nonindependence of data points within patients, we used a fixed-effects model to assess the association between hypotension (the only time-varying a priori selected explanatory variable) and Paco2-EtCO2 agreement within individual patients.To determine the characteristics associated with agreement between Paco2 and EtCO2 difference graphically to assess the association between time after PICU admission and Paco2-EtCO2 agreement. To explore how the median first-day Paco2-EtCO2 difference after PICU admission differed by the timing of PARDS diagnosis, we also constructed side-by-side boxplots of patients who had no PARDS, received a diagnosis within 24 hours of PICU admission, or received a diagnosis 1 to 7 days after PICU admission. Two-sample t tests were used to determine if there were statistically significant differences in the mean Paco2-EtCO2 difference between these groups. The 2 analyses were repeated among patients who survived the first 24 hours of PICU admission to examine the association of early death with our findings.We described the temporal trends of Paco2-EtCO2 differences with subsequent PARDS among patients who had not developed PARDS within the first 24 hours, we performed a receiver operating characteristic curve analysis and examined the sensitivity and specificity of various cutoff values. All statistical analyses were performed using R, version 3.5.1 , and Stata, version 14.2 package (StataCorp),19 and analyses were approved by the study epidemiologist (B.M.).To explore the association of Paco2-EtCO2 pairs were excluded owing to a lack of data within the predefined measurement time window  .Of the 199 potentially eligible patients, 137 (103 boys and 34 girls) met the inclusion criteria; 62 patients and 6 Paa points . The medco2-EtCO2 pairs (15.1%) were recorded for patients who experienced PARDS within 24 hours of PICU admission (co2-EtCO2 pairs (20.7%) were recorded for patients who developed PARDS between 1 and 7 days after PICU admission. Compared with data pairs in agreement, pairs not in agreement were more commonly from patients with PARDS diagnosed within 24 hours of admission (56 of 258 [21.7%] vs 11 of 187 [5.9%]).A total of 67 of the 445 Padmission , and 92 co2-EtCO2 pairs, Paco2 was on average 2.7 mm Hg  higher than its paired EtCO2 value . A total of 43 patients (31.4%) had at least 1 paired data point with disagreement. Among the 378 Paco2-EtCO2 pairs obtained from patients who did not experience PARDS within the first 24 hours of admission, Paco2 was on average 1.4 mm Hg  higher than EtCO2 . Among the 67 Paco2-EtCO2 pairs obtained from patients who developed PARDS within first 24 hours of admission, Paco2 was on average 9.9 mm Hg  higher than its paired EtCO2 value .Among all 445 PaO2 value A. Paco2 an EtCO2 B. Paco2 O2 value C. Paco2 co2 and EtCO2 are most similar are shown by Passing and Bablok regression ; there was no meaningful change in association after multivariable model adjustment.co2-EtCO2 difference during the first 24 hours of admission among those who developed PARDS within 24 hours was 9.41 mm Hg greater than those who never developed PARDS . The mean Paco2-EtCO2 difference during the first 24 hours of admission among patients who developed PARDS 1 to 7 days after admission was 4.02 mm Hg greater than those who never developed PARDS  ; restricting the sample to 24-hour survivors resulted in no meaningful change in findings. In the receiver operating characteristic curve analysis, the Paco2-EtCO2 difference among those who did not develop PARDS within 24 hours was associated with subsequent PARDS development between 1 and 7 days after admission . In addition, a Paco2-EtCO2 difference of 3 mm Hg or greater had a sensitivity and specificity of 0.72 for the onset of PARDS between 1 and 7 days after PICU admission, given that PARDS had not developed within the first 24 hours .The mean Pa1 mm Hg) . The meaco2-EtCO2 pairs were in agreement, and correlation was only moderate; (2) the presence of PARDS in the first 24 hours of PICU admission was associated with a lower likelihood of Paco2-EtCO2 agreement; and (3) even if PARDS was not diagnosed in the first 24 hours, the median Paco2-EtCO2 differences were higher among a subset of patients who developed PARDS later in the first week after injury than among those who never developed PARDS. Together, these findings suggest that, despite the appeal of EtCO2 monitoring, EtCO2 data should not be substituted for Paco2 data early after pediatric TBI.The main findings of this study of hospitalized pediatric patients with TBI are that (1) overall, fewer than half of all Pa2 as a Paco2 surrogate. First, we used Bland-Altman analysis, which revealed less than 50% agreement of EtCO2 with Paco2. Second, we used Passing and Bablok regression, which demonstrated that Paco2 and EtCO2 had positive constant and proportional differences between measurements. Finally, we used the Pearson correlation coefficient and demonstrated that Paco2 and EtCO2 were only moderately correlated. Collectively, these indicators demonstrated a moderate level of agreement and correlation between Paco2 and EtCO2. To our knowledge, this study is the first to use multiple measures of accuracy to address Paco2-EtCO2 agreement, and the results suggest a need to further define the role of capnography and the need to incorporate Paco2 measurements early after pediatric TBI.We used 3 complementary statistical approaches to understand the accuracy of EtCO10 reported that among adults with TBI, 77.3% of Paco2 and EtCO2 values were within 5 mm Hg of each other. In contrast, our pediatric study demonstrated lower agreement and did not identify hypotension or severe chest injury as contributing factors to Paco2-EtCO2 agreement. Differences in the definition of Paco2-EtCO2 agreement and in the study design  may contribute to these differences in results. However, to reflect clinical practice, we defined agreement using a narrower range (Paco2-EtCO2 <5 mm Hg) and paired the data within clinically reasonable intervals, which likely decreased measured levels of agreement. Although the number of infants and young children in our study is too small to make definitive conclusions, the relatively larger amount of dead space in infants and children might further increase the Paco2-EtCO2 difference and lower Paco2-EtCO2 agreement.Lee et alco2 values that are higher than EtCO2 values obtained simultaneously.14 Although there is generally no physiologic basis for why Paco2 would be lower than EtCO2, a negative Paco2-EtCO2 difference was observed among many of our Paco2-EtCO2 pairs. It is possible that this negative difference resulted from the accumulation of exhaled CO2 in ventilation circuitry prior to EtCO2 measurement or poor equipment calibration, but most likely it was due to our method of aligning Paco2-EtCO2 pairs during a period of up to 30 minutes, thus allowing the potential for intervening changes in patients’ clinical conditions to result in recorded peaks in EtCO2 values being paired with recorded dips in Paco2 values.Anatomic and physiologic dead space results in Pa22 It is thus reasonable to expect that many critically injured children with TBI, especially those with PARDS or other pulmonary pathologic characteristics, may have an elevated alveolar dead space fraction and thus lower Paco2-EtCO2 agreement. This study identified PARDS as an independent factor associated with a lower likelihood of Paco2-EtCO2 agreement, which provides new information on the utility of capnography among children with TBI with concurrent lung disease. These data also demonstrated that, for patients who had not received a diagnosis of PARDS during the first 24 hours of PICU admission, Paco2-EtCO2 differences were still greater among those who developed PARDS after the first 24 hours of admission compared with those who never developed PARDS, suggesting that patients may have had a ventilation-perfusion mismatch and worsening dead space fraction before meeting the full criteria for PARDS. Future work is needed to further evaluate Paco2-EtCO2 agreement as a potential early indicator of PARDS development in pediatric TBI.We specifically examined PARDS because children hospitalized with TBI are at risk for PARDS owing to a variety of factors, including intracranial hypertension, severe systemic inflammation, direct traumatic lung injury, and secondary lung injury from aspiration or pneumonia.2 measurements in TBI, this study provides new evidence to fill this knowledge and practice gap. Our data show that, despite the relatively constant mean Paco2-EtCO2 difference and despite the moderate agreement correlation during the first 24 hours of PICU admission, there was even larger variability in the Paco2-EtCO2 difference in the first 8 hours of PICU admission compared with the subsequent 16 hours. Therefore, in addition to the overall observation that Paco2-EtCO2 agreement was low in the first 24 hours after TBI and in the presence of PARDS with large and variable alveolar dead space, EtCO2 may be even more unreliable as a surrogate for Paco2 during the first 8 hours after TBI. Therefore, serial Paco2 measurements during the first 24 hours of admission should still be considered the optimal modality to monitor CO2 levels, especially when ongoing resuscitation may be needed and optimizing cerebral perfusion is critical. Given the focus on prevention of prophylactic hypocarbia in severe pediatric TBI, these findings suggest a need for the Brain Trauma Foundation guidelines to address arterial measurements for Paco2.Because neither the Trauma Quality Improvement Program nor the Brain Trauma Foundation guidelines address the utility of EtCOco2-EtCO2 agreement and hypotension with temporal vicinity, inclusion of PARDS diagnosis as an explanatory variable based on laboratory and clinical information, and a comprehensive description of age subgroups. These methodical strengths enabled us to examine in greater detail the association of Paco2-EtCO2 agreement with hypotension and PARDS. In addition, compared with using severe chest trauma to investigate the association of respiratory disorders with Paco2-EtCO2 agreement, the use of the PARDS diagnosis provided more specific information regarding oxygenation dysfunction and resultant ventilation-perfusion mismatch. To our knowledge, this is the first analysis to effectively obtain timely data on blood pressure and PARDS diagnosis and to use them to examine Paco2-EtCO2 agreement.This study has several strengths, such as a relatively large sample size, a priori analytic framework, use of age-specific definitions of hypotension, examination of the association between Pa2 data points were recorded within 10 minutes of Paco2 measurement, some paired data points had a time gap of up to 30 minutes, which may have reduced the precision of our findings. We did not examine data beyond 24 hours. Despite using fixed-effects multivariable regression models and an a priori analytic framework, we cannot exclude residual confounding that could not be captured by measurable variables.We acknowledge the study limitations. Our findings might not be fully generalizable because the data are from a single level I trauma center and the excluded patients who did not have paired data points differed in several clinical characteristics. Although more than 40% of EtCOco2-EtCO2 agreement is below 50%, especially in the presence of PARDS. Agreement is worst in the first 8 hours after PICU admission, and poor Paco2-EtCO2 agreement early in hospitalization may be associated with future development of PARDS. End-tidal CO2 data should not be a substitute for Paco2 data during the first 24 hours.In children hospitalized with TBI, Pa


Herein a highly active non-precious transition metal catalyst for the homogeneous hydrogenation of carbon dioxide to formate is presented. Detailed ligand optimisation enabled the development of a nickel-based catalytic system with exceptional productivity. ii) salts in combination with tailored multidentate ligands enabled the effective transformation of carbon dioxide with an exceptional TON of up to 4.65 × 106. This unprecedented productivity based on the novel nickel catalyst not only outmatches that of existing systems containing first row transition metals, but also established catalysts based on precious transition metals.Herein a highly active non-precious transition metal catalyst system for homogeneous hydrogenation of carbon dioxide to formate is presented. The application of selected nickel( The first system showed feasibility of CO2 hydrogenation in water with a homogeneous nickel catalyst.2 hydrogenation, again relying on the strong Verkade's base. A bimetallic nickel–gallium catalyst was able to perform the reaction at room temperature with TONs of up to 3150 2 and DBU as the base was performed (In the first set of COerformed .4)2 a low TON of 36 could be obtained methyl)amine) a higher TON of 184 was obtained (3 (tris(2-(diphenylphosphino)ethyl)phosphine) a TON of only 41 could be achieved ethyl)amine) showed a high productivity with a TON of 846 (3Cy (NP3Cy = tris(2-(dicyclohexylphosphino)ethyl)amine) a decreased turnover number of 384 was obtained 2.Consequently, further investigations focused on the application of the versatile NPnformate/nDBU) of approximately 0.33 2, NP3 ligand and DBU as the basic requirement for the exceptionally high activity  to a 1 : 1 mixture of Ni2+/NP3 resulted in a highly decreased TON and a low AAR . This productivity denotes an overall yield of roughly 6.30 g (31.62 mmol) of isolable [DBUH] formate and further shows the stability of the nickel catalyst system in this study.3 system 3) or zinc triflate (Zn(OTf)2) did not enhance the activity of the established system (3) inhibits catalysis and nearly no formate could be observed (4) a clear enhancement in the catalytic activity was seen achieving a TON of up to 5440 and an AAR of 1.09 (4) has a clearly inhibiting effect resulting in only a poor yield of formate in a TON of 400 2. Analysis after the catalytic experiment revealed a reduction of the initially formed NP3Ni(ii) complex, leading to the formation of a major species with a chemical shift of 21 ppm. Confirmation of the defined species NP3Ni(0) could be obtained with a further control experiment. Consequently, a mixture of NP3/Ni2+, DBU and molecular hydrogen (p = 3 bar) resulted in a 31P-NMR spectrum comparable to that of a post-reaction mixture  salts enabled the effective transformation of CO2 in the presence of molecular hydrogen in acetonitrile solution. Under optimized conditions, using 0.002 μmol nickel catalyst, an exceptional TON of approximately 4.65 × 106 was achieved. This unprecedented productivity based on the novel nickel catalyst not only outmatches that of existing systems containing first row transition metals, but also established catalysts based on precious transition metals. Further development and detailed mechanistic investigations on the catalyst system are ongoing in our laboratories.In conclusion, a non-precious transition metal catalyst system could be developed for homogeneous hydrogenation of CO4)2·6H2O and NP3: under an argon atmosphere, a Schlenk tube was charged with a freshly prepared stock solution (c3Ni/NP = 2.5 μmol mL–1) of Ni(BF4)2·6H2O and NP3. In the case of highly diluted reactions the stock solution was further diluted (see the ESI2 (30 bar) until saturation, followed by hydrogen to a total pressure of 90 bar at room temperature. The reaction mixture was stirred and heated to the respective reaction temperature in an aluminium heating cone. After the reaction period, the autoclave was cooled to room temperature and then carefully vented. Mesitylene was added as the internal standard and the resulting solution was analysed by 1H-NMR-spectroscopy. The TONs were found to be reproducible within ΔTON = ±10% in at least two independent runs in selected experiments.The general procedure for homogeneous hydrogenation of carbon dioxide to formate using Ni(BFThere are no conflicts to declare.Supplementary informationClick here for additional data file.


TM6SF2 and MBOAT7 genes as risk factors for alcoholic liver cirrhosis and non-alcoholic fatty liver disease. In this study, we tried to evaluate the association between TM6SF2 variant rs58542926 and MBOAT7 variant rs641738 and the risk of hepatic fibrosis or liver cirrhosis of different etiology. In parallel, we also aimed to evaluate whether these two SNPs modify the effects of the PNPLA3 rs738409 risk variant for the development of hepatic fibrosis and liver cirrhosis. The study was conducted at the Department of Gastroenterology, Lithuanian University of Health Sciences Hospital, and included 334 patients with liver cirrhosis, 128 patients with liver fibrosis, and 550 controls. SNPs were genotyped by quantitative PCR, using TaqMan allelic discrimination assays. Overall, TM6SF2 rs58542926 as well as MBOAT7rs641738 were not linked to hepatic fibrosis, alcohol or hepatitis C virus induced liver cirrhosis in an Eastern European population. These genetic variations also did not mediate the effect of PNPLA3 rs738409 SNP for liver developing liver fibrosis or liver cirrhosis.Previous large-scale genetic studies identified single nucleotide polymorphisms (SNPs) of the  Liver cirrhosis is one of the dominant causes of global disease burden and is associated with life threatening complications . In 2016PNPLA3) gene SNP is linked with liver injury [TM6SF2) and membrane-bound O-acyltransferase domain-containing protein 7 (MBOAT7) gene polymorphisms have been identified as risk factors for alcoholic liver cirrhosis [TM6SF2 variant rs58542926 has also been associated with an increased risk of developing advanced hepatic fibrosis and cirrhosis in patients with non-alcoholic fatty liver disease (NAFLD) [MBOAT7 polymorphism rs641738 was associated with the severity of liver fibrosis in individuals with chronic hepatitis C virus infection [MBOAT7 rs641738 SNP was also reported as an independent predictor of severe liver steatosis in patients with chronic hepatitis C [Genome wide association studies (GWAS) are now widely used to identify associations between single nucleotide polymorphisms (SNPs) and the risk of different diseases employing next-generation sequencing based techniques. Over the last decade a significant number of GWAS have been performed in the field of hepatology , revealir injury ,12. More (NAFLD) . Furthernfection  and NAFLnfection . In a reatitis C .TM6SF2 variant rs58542926 and MBOAT7 variant rs641738 and the risk of hepatic fibrosis or liver cirrhosis of different etiology in an Eastern European patient cohort. Additionally, we also aimed to evaluate whether these two SNPs modify the effects of the PNPLA3 rs738409 risk variant on the development of hepatic fibrosis and liver cirrhosis.In this study we analyzed the association between p < 0.001) than patients with liver fibrosis and controls. Men were predominant in the liver fibrosis group (61.7%. p < 0.05). The major cause of liver cirrhosis was alcohol, the second most common cause was chronic HCV infection. In the liver fibrosis group, the most common cause of liver injury was HCV infection. To eliminate the potential bias of differences in age and sex distribution among the groups, these parameters were included as covariates in logistic regression analysis. The frequencies of alleles and genotypes were in Hardy–Weinberg equilibrium for all SNPs analyzed .TMS6SF2 rs58542926 and MBOAT7 rs641738 polymorphisms in patients with different etiology of liver cirrhosis, hepatic fibrosis and the control groups are presented in TM6SF2 rs58542926 allele frequencies in controls, patients with hepatic fibrosis, liver cirrhosis, alcohol-induced and HCV-induced cirrhosis were: T allele 7.5%, 5.5%, 8.4%, 8.1% and 7.5%; C allele 92.6%, 94.5%, 91.6%, 91.8% and 92.5%, respectively. None of the TM6SF2 rs58542926 alleles or genotypes were linked with the risk of developing liver fibrosis or cirrhosis. MBOAT7 rs641738 SNP alleles and genotypes showed similar distributions across all four groups of the study. In controls, the frequencies of T and C alleles were 43.3% and 56.7%, in the liver fibrosis group 41.4% and 58.6%, in the cirrhosis group 44.6% and 55.4%, in alcohol-induced cirrhosis 42.1% and 57.9%, in HCV-induced cirrhosis 45.8% and 54.2%, and in patients with cirrhosis due to other causes 51.2% and 48.8%, respectively. No significant differences between MBOAT7 rs641738 alleles and genotypes among the different study groups were observed.Allele and genotype distributions for the PNPLA3 rs738409 and liver fibrosis/cirrhosis [PNPLA3 rs738409 and MBOAT7 or TM6SF2 genetic variants. This analysis showed no significant differences between combined MBOAT7 and TM6SF2 SNPs and PNPLA3 risk genotypes.In this part of the study we used genotyping data from our previous study on irrhosis . Table 4TM6SF2 rs58542926 and MBOAT7 rs641738 genes polymorphisms and the risk of hepatic fibrosis and liver cirrhosis. Overall, TM6SF2 and MBOAT7 SNPs were not linked with hepatic fibrosis and liver cirrhosis of different etiology within our study. Additionally, we ran combined analysis to assess whether TM6SF2 and MBOAT7 SNPs mediate the risk of liver fibrosis or liver cirrhosis in the presence of certain PNPLA3 genotypes. The effects of PNPLA3 have been well established in previous research, but we wanted to see if the risk of liver fibrosis is changed by a certain combination of PNPLA3 and TM6SF2 or MBOAT7 genotypes. To the best of our knowledge, this is the first study of an Eastern European population that assessed the impact of TM6SF2 rs58542926 and MBOAT7 rs641738 on developing liver injury.In the current study, we aimed to evaluate the associations between TM6SF2 gene is important for the secretion of very-low density lipoprotein from hepatocytes. Its reduced function leads to increased liver triglyceride and lipid droplet content which contributes to NAFLD [TM6SF2 gene have been linked with a number of liver conditions; however, reported results are varying in between. TM6SF2 rs58542926 was identified as a modifier of hepatic fibrogenesis [TM6SF2 rs58542926 is linked with development of severe liver steatosis in patients with chronic hepatitis C (CHC), although no significant association between TM6SF2 and severe liver necro-inflammation or fibrosis was found [TM6SF2 rs58542926 not only had an impact on liver steatosis, but was also relevant in the development of severe fibrosis in individuals with CHC [rs58542926 has a modest effect on NAFLD [TM6SF2 genotype did not impact histological features with biopsy-proven NAFLD in Japanese patients [TM6SF2 on fibrosis in NAFLD, CHC and CHB patients, Eslam et al. used a large cohort of Caucasians and found that rs58542926 has more influence on the serum metabolic profile and predisposed hepatic steatosis, rather than acting directly on liver inflammation and fibrosis [TM6SF2 variant and hepatic fibrosis [TM6SF2 rs58542926 between fibrosis and cirrhosis groups was different, but the significance level was not reached and this genetic variant alone or combined with PNPLA3 risk genotype was not linked with liver fibrosis or cirrhosis.to NAFLD ,18,19. Gogenesis  and was ogenesis ,21. Coppas found . Furtherwith CHC . Meanwhion NAFLD . Moreovepatients . To conffibrosis . AnotherMBOAT7 rs641738 SNP with the entire spectrum of NAFLD in individuals of European descent was first reported by Mancina et al. [MBOAT7 genetic variant was not associated with hepatic steatosis, but significantly linked to hepatic fibrosis by Krawczyk M. and colleagues [MBOAT7 rs641738 was not linked with risk of developing NAFLD [rs641738 and liver steatosis, but an association with hepatic inflammation and fibrosis in CHC was detected [MBOAT7 locus rs641738 was not linked with liver fibrosis or cirrhosis. When combining different genotypes of this SNP with a well know PNPLA3 rs738409 variant no significant differences were observed.The association of a et al. . The samlleagues . In an Ang NAFLD . Yet anodetected . NeverthPNPLA3 SNP sub-analysis needs further re-evaluation in larger well-defined cohorts in order to evaluate observed synergistic effects of different genetic variations.There are certain limitations associated with the design of our study that need to be acknowledged. The major limitation of our study is related to etiological heterogeneity of liver cirrhosis cohort and not a large number of patients within the subgroup analysis. Most of our patients in the liver fibrosis group had HCV induced fibrosis. Moreover, relatively small sample sizes within combined TM6SF2 rs58542926 and MBOAT7 rs641738 genotyping study included 1012 individuals . For combined analyses of carriers of PNPLA3 rs738409 risk genotype, we used data from our previous study with the same cohort [The e cohort  and inclTM6SF2 rs58542926 and MBOAT7 rs641738 SNPs were genotyped by real-time PCR (RT-PCR), using TaqMan® allelic discrimination assays with a 7500TM Fast real-time PCR system .Genomic DNA from samples was isolated from whole blood mononuclear cells using a salting-out method and stored at −20 °C until analysis, as described previously . TM6SF2 TM6SF2 and MBOAT7 alleles and genotypes frequencies between cases and controls were compared by Pearson’s goodness-of-fit χ2 and Fisher exact tests. Associations between control and cases groups with SNP alleles and genotypes were calculated using logistic regression analysis with adjustment for age and sex. 2 tests. For additional analysis, we divided the cohort in two groups with respect to PNPLA3 rs738409 genotypes. p-value was adjusted for multiple comparisons and the value of <0.025 (0.05/2) was considered to be statistically significant. Statistical analysis of the genotyping data was performed using PLINK software version 1.07 [ion 1.07 .TM6SF2 rs58542926 as well as MBOAT7 rs641738 were not linked to hepatic fibrosis, alcohol or hepatitis C virus induced liver cirrhosis in an Eastern European population. These genetic variations also did not mediate the effect of PNPLA3 rs738409 SNP on the development of liver fibrosis or liver cirrhosis.In conclusion,


The CCTG repeat tract is part of a complex repeat structure comprising not only CCTG tetraplets but also repeated TG dinucleotides and TCTG tetraplet elements as well as NCTG interruptions. Here, we provide the distribution of normal sized alleles in the German population, which was found to be highly similar to the Slovak population. Sequencing of 34 unexpanded healthy range alleles in DM2 positive patients (heterozygous for a full expansion) revealed that the CCTG repeat tract is usually interrupted by at least three tetraplets which according to current opinion is supposed to render it stable against expansion. Interestingly, only the largest analyzed normal allele had 23 uninterrupted CCTGs and consequently could represent an instable early premutation allele. In our diagnostic history of DM2 cases, a total of 18 premutations were detected in 16 independent cases. Here, we describe two premutation families, one with an expansion from a premutation allele and the other with a contraction of a full expansion down to a premutation allele. Our diagnostic results support the general assumption that the premutation range of unstable CCTG stretches lies obviously between 25 and 75 CCTGs. However, the clinical significance of premutation alleles is still unclear. In the light of the two described families we suggest incomplete penetrance. Thus, as it was proposed for other repeat expansion diseases , a fluid transition of penetrance is more likely rather than a clear cut CCTG number threshold.Autosomal dominant inherited Myotonic dystrophy type 1 and 2 (DM1 and DM2) are the most frequent muscle dystrophies in the European population and are caused by repeat expansion mutations. For Germany cumulative empiric evidence suggests an estimated prevalence of DM2 of roughly 9 in 100,000, therefore being as prevalent as DM1. In DM2, a (CCTG) Myotonic dystrophy type 2  is an autosomal dominant multisystemic neuromuscular disorder that occurs in adults , 2. Althn repeat located in the first intron of the CNBP  gene in the 3q21.3 chromosomal region (DM2 is caused by the expansion of an unstable (CCTG)l region . Like inl region . It disrCNBP repeat tract is generally described as (TG)n(TCTG)n(CCTG)n. Normal alleles as a rule have <25 copies of the decisive CCTG repeat, whereas expanded alleles have as many as 75–11,000 copies w(CCTG)x(NCTG)y(CCTG)z. In contrast, unstable expanded alleles obviously have a pure CCTG stretch without the NCTG interruption making them more susceptible to strand slippage and unequal crossing over processes in cell division tability , 6, 7. Cdivision .CNBP repeat motif has been conducted for the German population. In the present report, an estimate of the DM2 prevalence in Germany is given and a statistical analysis of the normal allele length frequency and distribution was performed. Alleles of the most frequent repeat lengths (n = 31) as well as of the longest normal alleles (n = 3) were sequenced to determine the detailed motif composition and its variance within the German population. Furthermore, we present two small families, one with a transgenerational contraction into the premutational range and another with a transgenerational expansion of a premutation allele to a full expansion.The composition of the complex repeat motif and allele frequency of normal and expanded alleles has already been studied in the Slovak and American population , 7. So fCNBP repeat sequence, using a fluorescently marked 6-F-primer (DM2-Cl3N58-F) 5′-GGC CTT ATA ACC ATG CAA ATG-3′, binding 47 bp upstream of the TG repeat sequence. The reverse primer (DM2-Cl3N58-R) 5′ GCC TAG GGG ACA AAG TGA GA 3′ binds directly after the last single TCTG tetraplet following the repetitive sequence  on an Applied Biosystems 3130 Genetic Analyzer. Sequencing conditions were the following: Step 1: 96°C (2 min), step 2: 96°C (20 s), step 3: 56°C (20 s), step 4: 60°C (3 min), and step 5: 60°C (5 min). Steps 2–4 were repeated in 26 rounds. Data were analyzed with the software Chromas lite (Technelysium). All sequenced patients had given informed consent for research purposes in the DM2 field, as required by the declaration of Helsinki 2013.There are no well-documented prevalence data for DM2 except of the reports from Finland  and RomeFor this study, fragment analysis data were collected from 739 probands of presumed German origin in order to define the polymorphic spectrum of DM2 repeat tract lengths in the German population. Figure ~55 allele to a full expansion (family 1) and the contraction of a full expansion to a (CCTG)~30 allele The composition of the complex repeat motive of normal length alleles in the German population was analyzed by Sanger sequencing of DNAs from 34 DM2 patients with repeat expansion determined by routine diagnostic fragment analysis. Thirty one of the 34 DM2 positive patients were specifically selected because they possessed one of the three most frequent repeat lengths . The remaining three DNAs were derived from those three DM2 patients with a full expansion that revealed the three longest healthy range alleles found in our patient cohort . Table CNBP repeat the CCTG tract was interrupted by three tetraplets (NCTG)3: GCTG CCTG TCTG. The last CCTG repeat unit always comprised 7 CCTGs whereas the first CCTG unit before the interrupting (NCTG)3 was found to vary between 5 and 6 CCTG tetraplets. Of the 31 sequenced alleles 26 revealed (CCTG)5(NCTG)3(CCTG)7 (84%) while only 5 alleles (16%) had 6 leading CCTG tetraplets [(CCTG)6(NCTG)3(CCTG)7]. In contrast to the highly stable CCTG tract, the (TG)v and (TCTG)w motifs at the beginning of the CNBP repeat stretch vary widely within the range of v = 17–25 (TG)v and w = 7–10 (TCTG)w. The most frequent normal allele was identified to contain 9 TCTG tetraplets [(TG)v(TCTG)9(CCTG)5(NCTG)3(CCTG)7], with a frequency of 42% of the 31 analyzed alleles and a range of (TG)v of v = 19–23.In case of the 31 patients with the high abundance alleles , in all of the analyzed healthy range alleles of the 22 (TCTG)w(CCTG)8(NCTG)5(CCTG)6. Specifically, in both alleles the interruption comprised five tetraplets: GCTG CCTG TCTG CCTG TCTG. The allele of 168 bp was found to be an exception with 23 uninterrupted CCTG tetraplets  was similar to each other: (TG)21(TCTG)9(CCTG)5(NCTG)3(CCTG)7 (138bp) while the other for (TG)21 (TCTG)10(CCTG)5(NCTG)3(CCTG)7 (142bp). Another patient with homozygous allele lengths but heterozygous motif composition revealed a rare allele (TG)21 (TCTG)7(CCTG)6(NCTG)3(CCTG)9 where the second CCTG motif had 9 instead of the constant number of 7 CCTGs (data not shown).Interestingly, only 2 of the 11 DM2-negative probands that appeared to be homozygous in the fragment analysis were indeed homozygous while the other 9 probands had repeat tracts with an identical length but a different motif composition where particularly TG and TCTG repeats varied widely. One of these true homozygous patients was homozygous for the allele (TG)The estimation of DM2 prevalence in Germany of about 9 in 100,000 people is equivalent to findings in the Italian population . For theCNBP repeat tract seem to be essentially similar among European populations. Comparing the German population's combined tract lengths [(TG)v(TCTG)w(CCTG)x(NCTG)y(CCTG)z] with those of the Slovak population underscores this hypothesis. The distribution of most healthy range alleles was found to be almost identical to the statistical results of the Slovak population . Radvanszky et al. identified 138 bp as the most common tract length, which was found to be the most frequent in the German population, too. The second-most frequent allele length of 134 bp among the German population was in third place in the Slovak population, whereas the third-most frequent tract length of 142 bp in the German population was at second place among the Slovak allele lengths . Notablyv(TCTG)w stretch. Accordingly, sequencing analysis of probands with only one apparently homozygous allele in the fragment analysis showed that fragment length alone does not predict homozygosity per se due to structural differences of the complex repeat. Indeed, a monomorphic structure was rarely observed (<20%). This is in accordance with other studies where heterozygosity was found to be roughly 90% in different European populations (y(CCTG)z part in contrast was fully stable in all 31 probands. The most common (CCTG)5(NCTG)3(CCTG)7 allele accounted for 26 of the 31 alleles (84%) while only 5 (16%) belonged to the (CCTG)6(NCTG)3(CCTG)7 allele. However, other rare alleles are present in the German population as observed in a “homozygous” but actually heterozygous DM2 negative proband, where a (CCTG)6(NCTG)3(CCTG)9 allele with z = 9 instead of z = 7 was present. Our results correlate well with the observations in the Slovak population where 75.3% of probands contained the (CCTG)5(NCTG)3(CCTG)7 and 17.4% the (CCTG)6(NCTG)3(CCTG)7 allele (7) while only few other alleles could be detected. The interrupting motif (NCTG)y was always found to consist of the three consecutive tetraplets (CGTG)(CCTG)(TCTG) for the three most frequent alleles which has also been previously reported by Radvanszky et al. ulations , 6, 7. Sbserved <0%. This 5 consisted of an identical GCTG CCTG TCTG CCTG TCTG sequence in both alleles.Notably, the structural composition of 2 of the 3 longest analyzed normal alleles (152 and 156 bp) varied only in the amount of TCTG tetraplets. The CCTG interruption (NCTG)1(CCTG)1(TCTG)1 or five (GCTG)1(CCTG)1(TCTG)1(CCTG)1(TCTG)1 tetraplets, respectively. Thus, the high polymorphic structure of the CNBP repeat is also recognizable from its interrupting tetraplets. However, uninterrupted alleles have previously been identified as being distributed over the whole size spectrum of 190 normal sized repeats (118–184 bp) and constituted 2.6% in the randomly selected Slovak individuals  shows incomplete penetrance which might depend on different parameters not known to date but could encompass specific changes in the repeat structure to patients' physical conditions or even environmental impacts. Collecting data from more patients with “small” DM2 expansions and known repeat tract structure as well as a follow up over time is necessary to estimate penetrance and should be a goal of future research to provide better genetic counseling for DM2 patients.Our observations in the German population lead to the assumption that alleles with a combined All subjects gave written informed consent in accordance with the Declaration of Helsinki.AM, TS, and WK data collection, data analysis, and data interpretation. AM, TS, WK, and SR drafting and critically revising the manuscript. BH-K, E-MH, and KS genetic testing.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer PM and handling Editor declared their shared affiliation.


Exoc3l2, which we term Exoc3l2a. We discuss possible functional implications of the resulting domain excision from this isoform of EXOC3L2 based on structural similarities with its paralog M-Sec (EXOC3L3), which is implicated in tunneling nanotube formation. The identification of this Exoc3l2 splice variant expands the potential for subunit diversity within the exocyst and for alternative functionality of this component independently of the exocyst.The exocyst is a molecular tether that retains secretory vesicles at the plasma membrane prior to SNARE-mediated docking and fusion. However, individual exocyst complex components (EXOCs) may also function independently of exocyst assembly. Alternative splice variants of EXOC mRNA and paralogs of EXOC genes have been described and several have been attributed functions that may be independent of the exocyst complex. Here we describe a novel splice variant of murine  EXOC7 permits isoform switching during epithelial to mesenchymal transition (EMT) in breast cancer cells, such that the mesenchymal EXOC7 isoform promotes a migratory phenotype through its ability to recruit remodelers of the actin cytoskeleton [The exocyst complex is composed of eight distinct components EXOC1-8) and tethers secretory vesicles to the plasma membrane prior to SNARE-mediated fusion and exocytosis. Isoform variants exist for several of the EXOCs and these are derived from alternative splicing of the canonical EXOC transcripts or paralogous gene expression. We have previously reported on the EXOC3 paralog EXOC3L2 and established that it is required for directional migration of endothelial cells and can associate with other EXOCs [-8 and teExoc3 and is also known as tumor necrosis factor α-inducible protein 2 (TNFαIP2) and EXOC3L3. M-Sec plays a key role in the formation of tunneling nanotubes (TNTs)[M-Sec is another paralog of es (TNTs), 5. A cres (TNTs). Importaes (TNTs), which ies (TNTs). TogetheExoc3l2 and using a structural homology model based on M-Sec we compare the properties of the canonical isoform of EXOC3L2 with this alternative isoform. The existence of this splice variant of Exoc3l2 expands the potential for EXOC3L2 functional diversity, as an integrated component of the exocyst or as an independent agent, and will need to be accounted for in future studies of EXOC3L2.Here we report an alternative splice variant of murine C57BL6 adult mice were acquired from Charles River Laboratories and housed at the National Veterinary Institute, Uppsala, Sweden. The animal experiments described in this study were conducted under the ethical permit C222/11, which was granted and approved by the Uppsala Animal Experiments Ethics Board , Uppsala District Court, Uppsala, Sweden.RNA was isolated from mouse tissues using the E.Z.N.A. total RNA isolation kit  according to the manufacturers protocol for animal tissues homogenized by needle and syringe. The purity and concentration of the RNA isolates were analyzed using a NanoDrop 2000 . A cDNA library of the RNA isolates was synthesized using the iScript cDNA synthesis kit  according to the manufacturers instructions.Exoc3l2 splice junction were manually designed. Primers were obtained using the Invitrogen Custom DNA Oligos service (Thermo Fisher Scientific). The following primers were used in the study: positive strand primers: 4:5, GTCACAGACGTGAAGGCTCA; P9, GAAGCTCTGGATGGCATCGT; P10:11, GTTTCGGCGGCTGGAGTC; Psj1, GGAGAGAATGGCTTATTGGCTTG,P and negative strand primers: 11, CTCGGAGTGTCCTCCAACTG; Psj2, CAAGCCAATAAGCCATTCTCTC.PPrimers were designed and confirmed to target the templates of interest using the Primer-BLAST tool hosted at the National Center for Biotechnology Information (NCBI) website. Primers that spanned the novel ® DNA polymerase with Buffer II and MgCl2 (Thermo Fisher Scientific). Taq polymerase and Buffer II were added as per the manufacturers instructions; additionally, one PCR volume contained 0.5 mM MgCl2, 5% dimethyl sulfoxide (DMSO), 0.1 μM dNTPs, 0.5 μM forward primer, 0.5 μM reverse primer, and 1.5 μl of sample DNA (1 μg/μl). The PCR was performed on a MiniOpticon PCR System (BioRad Laboratories) or a 2720 Thermal Cycler  with the following protocol: one cycle of 95°C for 3 min; 39 cycles of 95°C for 30 sec; 60°C for 30 sec; 72°C for 1 min; and one cycle of 72°C for 7 min. PCR products and the GeneRuler 100 bp or 1 kb DNA ladder (Thermo Fisher Scientific) were diluted in DNA Gel Loading Dye  and separated by electrophoresis on 2% agarose gels, prepared in 40 mM Tris, 20 mM acetate and 1 mM EDTA (TAE buffer) with 0.01% GelRed fluorescent DNA stain . Gels were scanned using a Gel Do EZ imaging system (BioRad Laboratories) or an Odyssey Fc Imaging System  and images were collected using the ImageLab Software (BioRad Laboratories) or Image Studio™ Software (LI-COR Biosciences).PCR was carried out with AmpliTaq Goldnucleobytes.com) and Adobe Illustrator software.cDNA from bands of interest was extracted from the agarose gel using a QIAquick Gel Extraction Kit  according to the manufacturers protocol for spin-columns. Extracted cDNA concentrations were determined using a NanoDrop 2000 . Sanger sequencing was conducted by the Uppsala Genome Center, Science For Life Laboratory, Uppsala, Sweden. Sequence data was viewed and presented using 4Peaks , according to the manufacturers instructions and performed on an iCycler iQ real-time PCR detection system (BioRad). Melt curves were analyzed using iCycler iQ associated software.Exoc3l2 Sanger sequencing results was performed using the online version of the MAFFT multiple sequence alignment program [Multiple sequence alignment for the  program , and the program . Alignme program , 12. The program . The graExoc3l2 gene is annotated at the Ensemble database as ENSMUSG00000011263 and at the NCBI database as XM_006540475.1. Two transcript variants of Exoc3l2 are currently proposed, the longer of the two would encode a 789 amino acid protein (Uniprot D3YUP5) and consists of twelve exons, while the short transcript is derived from the last four exons of the long transcript and would encode a 242 amino acid protein  or common to both long and short transcripts (P9 and P11). PCR of mouse liver cDNA with primers P9 and P11 produced an amplicon that migrated on agarose gel close to the predicted Exoc3l2 product length of 308 bp, while primers P4:5 and P11 produced an amplicon that migrated close to the predicted Exoc3l2 product length of 818 bp  and reverse (Psj2) primer that targeted the alternative splice junction between exon 6 and 11  [Exoc3l2 is important when designing primers and probes for studying transcript expression, and also when attempting to target specific epitopes of the translated protein with antibody based techniques.A polymorphism (rs597668) near the ase (AD) , and whiExoc3l2, and confirmed its expression in a number of major organs. Based on the recently published structure of M-Sec we built a structural homology model of EXOC3L2 and observed that the C-terminal region lost from this alternatively spliced isoform is similar to that lost from a proposed splice variant of M-Sec (XM_006515792.3). This region contains key residues implicated in M-Sec’s ability to recruit RalA and the exocyst and in doing so facilitate TNT formation. Residues with similar properties are found at comparable locations within the region lost from this alternatively spliced isoform of EXOC3L2. Identification of this splice variant of murine Exoc3l2 expands the family of exocyst-like proteins and future studies are warranted to determine the functional relevance of this alternatively spliced isoform of EXOC3L2.Using a strategy of reverse transcription PCR, exon junction-specific primers and Sanger sequencing we have identified a novel splice variant of murine S1 FigEqual concentrations of mouse heart (adult) and brain cDNA were amplified using primers P4:5+11 and P10:11+11 for Exoc3l2 and the β-actin house keeping gene ACTB. The similar strength of ACTB bands in heart and brain samples indicates that equal concentrations of cDNA (determined prior to PCR by nanodrop) were used from both tissues; therefore, the relatively weaker 818 bp bands for Exoc3l2 in brain samples suggests lower levels of Exoc3l2 transcription in this tissue. Bands at 250 bp representing the splice variant Exoc3l2a were detected in 2/3 mouse heart samples, but in none of the three brain samples.(EPS)Click here for additional data file.


For this article, the survey data on the effect of perceived excessive workload on faculty members’ commitment was presented. This data was gathered from an academic environment using the full time faculty members of Covenant University. The descriptive research design method was employed. The initial sample size used for the analysis was 228 faculty members but only 189 copies of questionnaire were returned. Statistical package for social sciences (SPSS) was used for the coding of the data. The validity and reliability of the research instrument were carried out using Cronbach Alpha. Descriptive analysis was used for the presentation of the data. This data is made publicly available to assist further study in the area of workload and employees commitment. Specifications TableValue of the data•The data provided in this article shows the basic criteria for measuring excessive workload in organizations and could be adopted for further research work.•This dataset indicates that there is a link between excessive workload and commitment of employees, thus, its availability will foster the need to emphasize on the subject matter.•Improved research work on this survey data would enhance faculty members’ commitment by increased morale, motivation and reduced absenteeism.•Additional investigation of the survey data by the universal professional research bodies will increase the understanding of the effect of work overload on faculty commitment across countries.•Data shared in this data article will open up doors for new research collaborations.1sine qua non for a university׳s survival Number 1 shows the response to the statement “Effective work life balance initiative helps to reduce excessive workload”. 5 (2.6%) respondents were neutral about the statement. 85 (45.0%) respondents agreed that effective work life balance initiative helps to reduce excessive workload while 99 (52.4%) of the respondents strongly agreed that effective work life balance initiative helps to reduce excessive workload.Number 2 presents the responses to the research question are the work life balance initiatives in your organization suitable? Only 1 respondent strongly disagreed with the question representing 0.5%, 3 respondents disagreed representing 1.6%, 14 (7.4%) respondents were neutral about the question. 125 (66.1%) respondents agreed while 46 (24.3%) respondents strongly agreed to the question.Number 3 presents the summary of the data on the responses to the research statement “I often feel that the work life balance initiatives are not effective.”18 (9.5%) respondents strongly disagreed with the statement, 28 (14.8%) disagreed with the statement, 22 (11.6%) respondents were neutral on the statement, 64 (33.9%) respondents agreed with the statement and 57 (30.2%) respondents strongly agreed to the statement.Number 4 provides responses to the statement “I sometimes stay back to finish my work for the day”. 5(2.6%) of the respondents strongly disagreed, 4 (2.1%) of the respondents disagreed while 8 (4.2%) respondents were neutral. 94 (49.7%) of the respondents agreed and 78 (41.3%) strongly agreed that they often stayed back to finish the work for the day.Number 5 shows responses to the statement on I always make sure I go early to work or other related activities. 10 (5.3%) respondents strongly disagreed with the statement, 1 (0.5%) respondent disagreed, 9 (4.8%) respondents were neutral on the statement, 139 (73.5%) respondents agreed and 30 (15.9%) respondents strongly agreed.Number 6 Presents responses to whether the respondents were interested in everything that goes on in the organization. 9 (4.8%) of the respondents strongly disagreed, 18 (9.5%) of the respondents disagreed while 19 (10.1%) of the respondents were neutral as regards the notion. 76 (40.2%) agreed and 67 (35.4%) strongly agreed that they were interested in everything that goes on in the organization.The evolving competition in the university system has placed pressure on employees’ commitment particularly in a recessed economy where organisations are reducing the overhead cost by reducing the workforce and increasing the workload of the surviving employees. Active engagement of faculty members in research, teaching and community impact are 2Data were collected from the sample of two hundred and twenty eight (228) faculty members of Covenant University with the aid of structured questionnaire designed by the researcher based on the similar studies. Stratified and simple random sampling techniques were used. Copies of questionnaire were administered directly to all categories of faculty members ranging from Assistant Lecture to Professor. What informed the choice of the selected Covenant University was because of the fact that Covenant University has been best private university in Nigeria for over four years consecutively. Faculty members were also purposefully selected because of the university drive for quality research outputs and excellent teaching delivery toward achieving their vision of becoming one of the best universities in the world. Two hundred and twenty eight (228) copies of questionnaire were administered; one hundred and eighty nine (189) copies of the questionnaire were retrieved from the field giving the response rate of (82.9%). Meanwhile, the researchers also sought for the permission of the management of the selected institution before the questionnaire were administered to them. In addition, every participant was adequately informed about the objective of the study. In addition, all respondents were given opportunity to stay anonymous and their responses were treated with upmost confidentiality.


This increase in FIO2, however, often leads to errors in the calculation of oxygen consumption (2 concentrations. A possible solution to this problem could be the use of the ‘Eschenbacher transformation’ (ET) as an alternative correction factor. This study examines the concentration of FIO2 at which the HT and the ET are valid, providing plausible data of oxygen consumption corresponding to the wattage achieved during cycle ergometry. Ten healthy volunteers underwent spiroergometric testing under standard conditions (FIO2 = 20.9%), as well as at FIO2 = 40% and 80%. When compared with the predicted values of 2 of 40%. At FIO2 concentrations of 80%, however, the 2 allows a reliable evaluation of stress tests in patients requiring high doses of supplemental oxygen.Spiroergometric measurements of persons who require oxygen insufflation due to illness can be performed under conditions of increased inspiratory oxygen concentration (FIO With this method, patients inhale oxygen-enriched air from a gas reservoir. The performance of the patient can thus be determined by means of spiroergometry, without causing unnecessary physical and psychological stress due to breathing difficulties [2 lead to errors in the calculation of the oxygen uptake ?" .2 leads to an overestimation of the measured 2 of 40% or greater, the error rate in the calculation of 2 = 100%, no valid results can be obtained. Similar findings on the inaccuracy of this measurement method are also provided in studies done by Wilmore et al. (1973) [2 ≥ 40%, 2 concentrations, with an ever-increasing 2 values. The authors also showed a parallel continuous decrease in RER, eventually reaching zero at high FIO2 concentrations. Walsh and Banister (1995) [2 concentrations. They found an increased 2 of 40%. However, higher FIO2 concentrations were not tested in this study. Yet further investigations confirm the effects of increased FIO2 on 2 concentrations [There is consensus in the literature that spiroergometry performed under conditions of increased FIO" 2012) . At an F. 1973) . A validr et al.  and Stan3 . A valthe V˙O2 . This he . At an 2, so the RER (presented as a ratio of 2) takes place, John Scott Haldane (1860–1936) defined a correction factor for the exact calculation of the inspiratory volume (VI). VI is elementary for the calculation of E) [2 = 20.9%), plausible 2 levels lead to different, nonlinear results: (1) The measurement of 2 of 40%. (2) At an FIO2 between 40–60%, O2 concentration measurements must be repeatedly calibrated. (3) At an FIO2 between 60–80%, inaccurate values are to be expected. (4) At an FIO2 greater than 80%, implausible data are provided. (5) At an FIO2 of 100%, calculation of the 2 levels greater than 40%.Ulrich et al.  explain n uptake . On the ume (VE) , 16. Forr (1986)  showed t Haldane 60–1936 d2, Hermann Eschenbacher developed the so-called "Eschenbacher Transformation" (ET) in 1987 [2 levels.To achieve a greater level of accuracy regardless of the selected FIO in 1987 , it seem2. The following hypotheses will be discussed:Following the description of the two computational transformations in terms of their basic theories and implementations , the pre2 = 20.9%) and FIO2 = 40%.Both transformations yield 2 > 40%), the HT produces invalid results in the calculation of At higher concentrations . Subsequent measurements were performed under conditions of increased oxygen concentration (FIO2 = 40% and FIO2 = 80%), each preceded by a 30-minute rest period. All stress tests were carried out with a stepwise increasing workload protocol on a cycle ergometer  and evaluated using the HT. After a two-minute reference phase at a load of 20 watts, the applied step protocol provides a ten-minute test phase with five two-minute stages at increasing loads (50-80-110-140-170 watts). The examination terminated with a two-minute recovery phase (20 watts). The so obtained raw data were also used to calculate the To test these hypotheses, 10 healthy, middle-aged men underwent spiroergometric examination with three different FIOrgometry  were car2 = 0; (2) it takes into account that for RER fraction  VI is different to VE; (3) the transformation must calculate the values corresponding to the load even with increased FIO2; and (4) it must be possible to calculate 2 = 100%. A transformation that fulfills these conditions is a further development of the Haldane Transformation, at least for measurements under elevated FIO2. So far there are no verified and validated studies with this transformation.The new approach of the ET is based on the complete spectrum of all oxygen concentrations. For this, the ET has to meet four basic conditions: (1) the formula is not based on the assumption that VNThe prediction values for ceptable , 14. Was2 on the ventilatory threshold was made to investigate further findings.From the obtained data, a first view at a potential influence of higher FIOSpirometry and diffusing capacity studies were performed with the MasterScreen Body/Diffusion and software JLab 5.72, the spiroergometric examinations with the Vyntus CPX and software SentrySuite 2.17, all provided by CareFusion Germany 234 GmbH . Masks and equipment were supplied by HansRudolph, Inc. . Statistical evaluation of the data sets  was done using the statistics software R . Images were created using Microsoft Excel . Differences were considered significant at p ≤ 0.05. To correct for multiple comparisons, p-values were adjusted with the Benjamini-Hochberg procedure. Using the Bland-Altman limit analysis , the calAll subjects participated voluntarily and received a small compensation. The participants provide their written informed consent to participate in this study. The Declaration of Helsinki was adequately addressed, and the study was approved by the ethics committee of the Hamburg Medical Association (register number PV5327).2 differs from classical spiroergometry only in that the subject breathes a gas mixture with a higher proportion of oxygen compared to indoor climate, from a reservoir equipped with a special valve system. The spiroergometry unit includes ECG leads, blood pressure and pulse oximetry, as well as a respiratory mask. The mask is directly connected to the Digital Volume Transducer and the collection hose for gas measurement. A two-way valve for inhalation and exhalation is connected and is coupled via an adapter to the gas bag into which the subject breathes. The O2 filling hose is joined directly to the O2 source and the gas bag is filled with the predetermined FIO2 concentration through a valve  were included in the study. The medical examination, spirometry, diffusing capacity measurements and the three spiroergometric measurements were carried out from September to December 2016, always over the course of a day. The ambient conditions could be kept almost constant despite the different measuring times . All participants fulfilled the inclusion criteria. 2 under standard conditions was 20.78 ± 0.06%. A mean of 39.85 ± 0.18% and 78.57 ± 0.56% FIO2 were obtained for the measurements with increased FIO2 . 2 concentrations.The mean FIO2-level. At FIO2 of 20.78% a similar behavior of both examined transformations is shown. At FIO2 = 39.85% and especially at 78.57%, the HT has a much higher fluctuation range than the ET. The ET values show significantly less deviation, but are generally a little lower than the HT values. With increasing FIO2-concentration, the ET shows more accurate results , while the HT shows a constant large dispersion and deviation.2 = 20.78%), there were only slight differences in the results calculated with the HT and ET. Both transformations provide plausible results for Under normal ambient air conditions  did not occur until 140 watts (compared to 125 watts at FIO2 = 20.78%). In tests with an FIO2 = 78.57%, the first ventilatory threshold (VT1) was reached 136 ± 63 seconds later than that achieved under normal conditions, at a workload of 158 watts. The second ventilatory threshold (VT2) also occurred later  in the protocol; in the tests with an FIO2 of 39.85%, the VT2 was not observed in seven subjects. The VT2 at an FIO2 = 78.57% was not achieved by any of the participants within the test protocol (2 = 20.78%), the time of VT2 could be determined in six of the ten subjects.Regardless of the transformation method, a shift of the ventilatory threshold in the transition from aerobic to anaerobic cellular respiration was observed with increasing workload in all ten subjects at higher FIOprotocol . Convers2 conditions, pointing to an improved load adaptation , ventilation (VE) and maximum oxygen uptake  and with an FIO2 of 39.85%, we could show that both transformations yield plausible 2 = 20.78%, this phenomenon is observed for both FIO2 concentrations. Despite this, both transformation equations can be used to generate plausible data for the FIO2 concentrations of 20.78% and 39.85%.On the basis of the 10 measurements taken under normal conditions (FIO2 > 40%)—can only partially be confirmed and requires further exploration. Investigation into the first hypothesis shows that the use of the HT for 2 values ≤ 40%; oxygen insufflation at higher concentrations result in data inconsistent with predicted values. Indeed, our test results show significant deviation from predicted 2 > 80%. Determination of exhaled carbon dioxide (2 = 80%). The exact concentration of FIO2 for which the HT can still validly be employed cannot be definitively deduced from this study. The results obtained show a broad scattering of 2 = 40%.The second hypothesis—that the HT does not produce valid 2 concentrations of 40% and 80%, the 2 –can thus be confirmed. Furthermore, the scattering of results does not differ significantly for the varying oxygen concentrations. It can therefore be concluded that the ET is a suitable alternative to the HT for measurements at elevated oxygen concentrations.When the ET is applied at increased FIO2. Based on the power calculation as suggested by Kuzma (1998) [This is a pilot study with a small number of subjects (n = 10). In future studies this field should be further investigated with patient groups and other forms of increased FIOa (1998) , the sama (1998) . Another2 (here 80%). The general application of this method cannot therefore be recommended. The use of the alternative ET at higher FIO2 concentrations, however, produces valid results for 2 of 40% and 80%. In our opinion, the results of the study presented here allow us to extend the range of applications of spiroergometry. Through the implementation of the ET, spiroergometry at elevated FIO2 could provide significant added value for the stress testing of patients with oxygen insufflation.As with the classic spiroergometry the spiroergometry under increased oxygen concentration has to generate valid data. However, results of the HT used so far for the calculation of 2, the subjects also showed a shift of the ventilatory thresholds into higher load ranges. The next step, therefore, would be to confirm the findings obtained here in healthy subjects with those of patients suffering from lung disease. The applicability of "spiroergometry under increased oxygen concentration" could be extended and thus also be of benefit to seriously ill patients.In measurements with increased FIOS1 Appendix(PDF)Click here for additional data file.S1 FigThis picture is 2018 Vyaire Medical, Inc.; Used with permission (RD_0816-0130).(TIF)Click here for additional data file.


We report a preterm female infant with intrauterine growth retardation, dysmorphic facies, missing rib, small hands and feet, and hemihypertrophy. The results of whole genome SNP microarray analysis showed approximately 77 Kb interstitial deletion of the short arm of chromosome 11 (11p15.4). We report novel clinical findings of this rare genetic condition. The deletion in this region in relation to hemihypertrophy (overgrowth syndrome) is novel. We report a novel case of 11p15.4 deletion and a review of the literature.Microdeletion of the short arm of chromosome 11 is a rare chromosomal anomaly. The “pure” deletion of 11p15 region with no other chromosomal imbalance is extremely uncommon. Beckwith-Wiedemann syndrome (BWS)/overgrowth syndrome is known to be associated with genetic and/or epigenetic alterations that modify imprinted gene expression on chromosome 11p15.5  11p15.4 x 1. The delThe clinical features of Beckwith-Wiedemann syndrome (BWS) include hemihypertrophy and/or macroglossia. Hypoglycemia is reported in 30-50% of the babies with BWS .  may play a role in the development of BWS. The ZNF214 and ZNF215 genes are located in one of the three regions on chromosome 11p15, called the Beckwith-Wiedemann syndrome chromosome region-2 (BWSCR2). The H19 (103280), ICR1 (616186), and KCNQ1OT1 (604115) are located at 11p15.5; and the CDKN1C (60856) is located at 11p54 [The genetics of BWS is complex and involves multiple genes on chromosome 11p15 –6. Threeat 11p54 , 8. Paternally imprinted gene would deactivate the activity of CDKN1C gene .The cyclin-dependent kinase inhibitor 1C (CDKN1C) gene encodes for making a protein that helps regulate growth. This protein acts as a tumor suppressor. It is also involved in controlling fetal growth and stops the developing fetus from becoming too large. The activity of the CDKN1C gene depends on which parent it was inherited from. the regulation of several genes that are normally controlled by IC2, including CDKN1C. Reduction in the activity this gene, which normally controls cell growth and division, leads to overgrowth and the other features of BWS. Rarely, BWS has been caused by the deletion of a small amount of DNA from the maternally inherited copy of the IC2 region.BWS is a condition that causes overgrowth and other signs and symptoms that affect various parts of the body. More than half of all BWS cases result from alterations in methylation of the imprinting center 2 region (IC2). The maternally inherited copy of the IC2 region shows a decrease in the methylation, and this abnormality adversely affectsIntrauterine growth restriction has been reported to be associated with both paternal and maternal 11p15 abnormalities , 10. At Deletion of the 11p15.4 region is associated with autosomal dominant hereditary persistence thalassemia  and autosomal recessive beta-thalassemia (OMIM: 141900) . It remains unclear what exactly is the relation between this deletion and the BWS/hemihypertrophy associated genes.We report on a preterm infant who had intrauterine growth retardation and developed hemihypertrophy during the postnatal stay. The microdeletion of 11p15.4 that was found in our case is likely to be paternally derived, since the patient's half-sister had an inherited anemia (family history). The additional features  in our patient have not been previously reported with deletion of the 11p15.4 region. Microdeletions of 11p15 are rare. There have been limited reports of patients with interstitial deletions involving band 11p15.4. To our knowledge none have reported clinical features in a neonate with the same deletion as in our case. Here, we provide additional human genetic evidence that the 11p15.4 deletion contains regulatory elements that play a mechanistic role in the hemihypertrophy BWS phenotype and intrauterine growth retardation (IUGR). microdeletion 11p15.4. These findings may help identify the gene implicated in BWS and IUGR.In summary, we report a novel case of a dysmorphic preterm neonate with