Demographic characteristics in patients with short-gap and long-gap esophageal atresia: a comparative study

Background The knowledge of the size of the gap between esophageal ends in esophageal atresia (EA) before surgery is of clinical importance. The aim of this study was to compare the demographic characteristics between patients with short-gap esophageal atresia (SGEA) and long-gap esophageal atresia (LGEA).Patients and methods Medical records of all patients managed for EA spectrum in our department between 2003 and 2012 were evaluated, retrospectively. Demographic data included the maternal age, the number of parities and deliveries, the presence of polyhydramnios and the prenatal diagnosis, sex, the gestational age and prematurity, the type of delivery and the birth weight, age at the time of diagnosis and treatment, the presence of associated anomalies including VACTERL-type and non- VACTERL-type anomalies, the type of EA according to Gross classification, and discrepancies between the diameters of atretic esophageal ends. SGEA and LGEA were defined as a gap of less than three vertebral bodies or three or more vertebral bodies in length between the atretic esophageal ends, respectively.Results There were 99 patients treated for the diagnosis of EA spectrum during the study period: 81 in the SGEA group and 18 in the LGEA group. Most of the parameters studied did not differ between the two groups. Type-C EA was more prevalent in patients with SGEA (n= 77/81) and type-A was more frequent in children with LGEA (n= 8/18) (P < 0.05). The frequency of prenatal diagnosis (2.5% for SGEA vs. 22.2% for LGEA) was more common in the LGEA group (P < 0.05). Cesarean section compared with normal vaginal delivery was more commonly seen in both groups (56.8% for SGEA vs. 66.7% for LGEA).Conclusion Most of the demographic parameters were similar between the two groups of patients. However, the frequency of prenatal diagnosis was more common in patients with LGEA. Most of the patients in both groups were delivered by a cesarean section. Type-C EA was more prevalent in patients with SGEA and type-A was more frequent in children with LGEA. Further analysis of this topic is warranted and may be beneficial in revealing the true demographic differences between patients with SGEA and LGEA

A pancreatic neuroendocrine tumor diagnosed during the management of acute appendicitis

Pancreatic neuroendocrine tumors (PNET) are increasingly being discovered. A case of PNET diagnosed and treated during the management of acute appendicitis is presented and discussed. The importance of imaging modalities in patients with acute abdominal pain is emphasized. To the best our knowledge, this is the first pediatric report of PNET and acute appendicitis combination.Keywords: acute appendicitis, children, pancreatic neuroendocrine tumo

Evolution of the miR-290–295/miR-371–373 Cluster Family Seed Repertoire

<div><p>Expression of the mouse miR-290–295 cluster and its miR-371–373 homolog in human is restricted to early embryos, primordial germ cells, the germ line stem cell compartment of the adult testis and to stem cell lines derived from the early embryonic lineages. Sequencing data suggest considerable seed diversification between the seven homologous pre-miRNAs of miR-290–295 but it is not clear if all of the implied miR-290–295 seeds are also conserved in the human miR-371–373 cluster, which consists of only three homologous pre-miRNAs. By employing miRNA target reporters we show that most, if not all, seeds in miR-290–295 are represented in miR-371–373. In the mouse, pre-miR-290, pre-miR-292 and pre-miR-293 express subsets of the miRNA isoforms processed from the single human pre-miR-371. Comparison of the possible miR-290–295/miR-371–373 seed repertoires in placental mammals suggests a model for the evolution of this miRNA cluster family, which would be otherwise difficult to deduce based solely on pre-miRNA sequence comparisons. The conservation of co-expressed seeds that is characteristic of miR-290–295/miR-371–373 should be taken into account in models of the corresponding miRNA-target interaction networks.</p></div

Comparisons of the miR-290–295/miR-371–373 clusters in <i>Placentalia</i>.

<p>(<b>A</b>) Multiple sequence alignment of the individual pre-miRNAs from species belonging to 14 distinct placental orders. Sequences are ordered according to the UPMGA tree (shown on the left) and are labeled with the species abbreviation. pre-miRNAs are numbered according to their position with respect to the transcription start site (the most promoter proximal pre-miRNAs are at position 1). The alignment consensus and sequence logo are shown at the top and the pre-miRNA secondary structure elements at the bottom. The 5p0 and 3p0 reference positions discussed in the text are also indicated. Active (light) and inactive (dark) miRNA seed positions within the human and mouse clusters are highlighted. The activities of miR-295-3p+1 and miR-372-3p+1 are unknown (Active?). Species abbreviations are as follows: bos-tau – <i>Bos taurus</i> (domestic cow), can-fam – <i>Canis familiaris</i> (dog), das-nov – <i>Dasypus novemcinctus</i> (armadillo), ech-tel – <i>Echinops telfairi</i> (lesser hedgehog), equ-cab – <i>Equus caballus</i> (horse), eri-eur – <i>Erinaceus europaeus</i> (European hedgehog), hom-sap – <i>Homo sapiens</i> (human), lox-afr – <i>Loxodonta africana</i> (African bush elephant), mus-mus – <i>Mus musculus</i> (house mouse), myo-luc – <i>Myotis lucifidus</i> (little brown bat), och-pri – <i>Ochotona princeps</i> (American pika), pro-cap – <i>Procavia capensis</i> (rock hyrax), ict-tri – <i>Ictidomys tridecemlineatus</i> (thirteen-lined ground squirrel), tur-tru – <i>Tursiops truncatus</i> (bottlenose dolphin) (<b>B</b>) Evolutionary relationships between the species in (A). Species abbreviations are followed by the number of pre-miRNA hairpins in the corresponding cluster. The names of orders and relevant superclades are indicated. The evolutionary tree is according to ref <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519-OLeary1" target="_blank">[29]</a>.</p

Active seeds in miR-290–295 and miR-371–373.

<p>Active seeds in miR-290–295 and miR-371–373.</p

Silencing of seed only miR-290–295/miR-371–373 reporters.

<p>(<b>A</b>) Sequence variation between multiple sequence alignment positions 3p0 and 3p+9 in pre-miR-290–295/pre-miR-371–373. This panel should be used as key to the various reporter designations. 3p0 miRNA positions 1–7 (equivalent to alignment positions 3p0-3p+7) are invariant in all pre-miRNAs as is 3p0 miRNA position 10 (alignment position 3p+9). Mature miRNAs that correspond to the sequence paths in the graph are shown on the right. (<b>B</b>) Silencing of seed only reporters matching all possible miR-290–295/miR-371–373 3p0 miRNA 7mer seeds (2-7C-S, 2-7U-S, 2-7G-S and 2-7A-S) and their position 2 mismatch controls (3-7C-S and 3-7U-S) in wild type (WT) and miR-290–295 knockout (KO) mouse ES cells. (<b>C, D</b>) The miR-292-3p0 7mer seed reporter 2-7C-S and its position 2 mismatch control (3-7C-S) were co-transfected into miR-290–295 null ES cells as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone-0108519-g003" target="_blank">Figure 3C, E</a>. +miR-294 is a rescue construct driving a single pre-miR-294 hairpin. (<b>E</b>) Silencing of reporters with sequences matching the 7mer seeds of miR-292-3p+1 (3-7CG-S), miR-293-3p+2 (4-7CGC-S) and miR-295-3p+1 (3-7UA-S) and their corresponding seed position 2 mismatch controls (4-7CG-S and 5-7CGC-S) in wild type and miR-290-295 knockout ES cells.</p

Silencing of luciferase reporters containing perfectly complementary miR-290–295 and miR-371–373 target sites in mouse ES cells.

<p>Reporter activities are expressed as firefly/<i>Renilla</i> luciferase activity ratios normalized to the control transfection of a firefly luciferase construct that does not contain miRNA target sites (No site). In panels that combine multiple experiments with separate no site controls, normalization is to the average of these controls (No site average). (<b>A</b>) Silencing of reporters containing sites perfectly complementary to the putative mature miR-290–295 in wild type mouse ES cells (WT ES Cells), in miR-290–295 knockout ES cells (KO ES cells) and miR-290–295 knockout ES cells co-transfected with a miR-290–295 expression construct (KO ES cells + Rescue). The target site sequences are summarized in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone-0108519-t001" target="_blank">Table 1</a>. (<b>B</b>) The mixtures of firefly and <i>Renilla</i> luciferase constructs shown in A were serially diluted (ND  =  non-diluted, 1∶100, 1∶10000) with a plasmid expressing EGFP so that the total DNA concentration remains constant (to maintain the same transfection efficiency) and were transfected into wild type mouse ES cells. The results obtained from further dilution of the reporters are shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519.s003" target="_blank">Figure S3</a>. (<b>C</b>) Reporter activities in miR-290–295 knockout ES cells co-transfected with the following rescue constructs: full-length miR-290–295 (+miR-290–295), pre-miR-292 deletion mutant (+Δ292), pre-miR-293 deletion mutant (+Δ293), single pre-miR-292 expression construct (+miR-292), single pre-miR-293 expression construct (+miR-293) and control expression vector backbone that does not express any miRNAs (+Empty vector). (<b>D</b>) Serial dilutions (ND  =  non-diluted, 1∶100, 1∶10000) of reporters containing target sites perfectly complementary to miR-371–373 were performed as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone-0108519-g003" target="_blank">Figure 3C</a> and were co-transfected with a miR-371–373 expression construct into miR-290–295 null mouse ES cells. (<b>E</b>) The miR-371–373 luciferase reporters were co-transfected with the following expression constructs: full-length miR-371–373 expression construct (+miR-371–373), pre-miR-371 deletion mutant (+Δ371), single pre-miR-371 expression construct (+miR-371) or an empty expression vector control (+Empty vector).</p

Silencing of bulge-mismatch reporters specific for the 3p+1 and 3p+2 seeds in miR-290–295/miR-371–373.

<p>(<b>A</b>) Predicted secondary structures of the duplexes formed between miR-292-3p+1 and miR-371-3p+1 with the 3-7CG-B target and of miR-293-3p+2 and miR-371-3p+2 with the 4-7CGC-B target. The miRNA sequences are highlighted. (<b>B</b>) Silencing of the miR-292-3p+1 bulge reporter (3-7CG-B) and its position 2 mismatch control (4-7CG-B) and the miR-293-3p+2 bulge reporter (4-7CGC-B) and its wild type mismatch control (5-7CGC-B) in wild type and miR-290-295 knockout ES cells (WT ES cells, KO ES cells). The miR-292-3p+1/miR-371-3p+1 bulge reporter 3-7CG-B and its corresponding position 2 mismatch control 4-7CG-B (<b>C, E</b>) or the miR-293-3p+2/miR-371-3p+2 bulge reporter 4-7CGC-B and its position 2 mismatch control 5-7CGC-B (<b>D, F</b>) were co-transfected as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone-0108519-g003" target="_blank">Figure 3C, E</a>.</p

Short RNA 5′-end distributions in pre-miR-290–295 and pre-miR-371–373 sequencing data.

<p>The frequencies of observed 5′-ends of RNA reads in various short RNA sequencing datasets are plotted as a function of pre-miRNA sequence position. pre-miRNA sequence co-ordinates are given with respect to the 5p0 and 3p0 positions in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone-0108519-g001" target="_blank">Figure 1A</a> and the sum of 5′-end frequencies is normalized to 1 for each individual pre-miRNA. For pre-miR-290-295 the top panels show total ES cell RNA and HEK-293 ectopic overexpression RNA sequencing data (Total1-3, Ectopic) and the bottom panels show HITS-CLIP data. Dataset Total1 is the total RNA dataset from ref. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519-Leung1" target="_blank">[27]</a>, Total 2 and Total 3 are respectively the J1 and Dcr^+/+ total RNA datasets from ref. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519-Calabrese1" target="_blank">[16]</a> and Total 4 is from ref. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519-Chiang1" target="_blank">[6]</a>. CLIP1-3 correspond to datasets WT1A, WT1B and WT2 from ref. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519-Leung1" target="_blank">[27]</a>. The panels corresponding to pre-miR-371-373 show total RNA sequencing data from undifferentiated human ES cells (Undifferentiated) and human ES cells that have been differentiated into embryoid bodies (Differentiated) according to ref <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519-Morin1" target="_blank">[12]</a>. The data for pre-miR-291b, which yields very few reads in all datasets and is, thus, noisy is given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108519#pone.0108519.s001" target="_blank">Figure S1</a>.</p