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          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">DNA fragments on agarose gel electrophoresis after restriction enzyme digestion of exon 1 of the mannose-binding lectin &#40;<span class="elsevierStyleItalic">MBL2</span>&#41; gene codon 54&#46; In all&#44; 349<span class="elsevierStyleHsp" style=""></span>bp PCR product was digested with <span class="elsevierStyleItalic">BanI</span> for codon 54 polymorphism&#46; The normal allele &#40;allele A&#41; is cut into two fragments with <span class="elsevierStyleItalic">BanI</span> &#40;lanes 2 and 5&#41;&#44; 89 and 260<span class="elsevierStyleHsp" style=""></span>bp&#46; The variant allele &#40;allele O&#41; remains uncut &#40;lanes 1 and 6&#41;&#46; Both uncut and digested fragments are seen in AO heterozygote &#40;lanes 3 and 4&#41;&#46; L&#58; 100<span class="elsevierStyleHsp" style=""></span>bp DNA ladder&#46;</p>"
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and mutant <span class="elsevierStyleItalic">MBL2</span> alleles occur as a result of three single-point mutations in this gene &#40;B&#44; C&#44; and D&#41;&#46; Although functional MBL levels are low in heterozygous polymorphisms&#44; MBL levels in homozygous polymorphisms are so low that they may not even be determinable&#46;<a class="elsevierStyleCrossRefs" href="#bib0145"><span class="elsevierStyleSup">4&#44;5</span></a> MBL activates the complement system by binding to mannose or sugar motifs&#44; which are found in many microorganisms&#44; and plays an important role in innate immunity and inflammation&#46;<a class="elsevierStyleCrossRefs" href="#bib0135"><span class="elsevierStyleSup">2&#44;6</span></a> In newborns&#44; an increase in sepsis frequency is observed when MBL levels are low&#46;<a class="elsevierStyleCrossRefs" href="#bib0135"><span class="elsevierStyleSup">2&#44;3&#44;7</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">The mortality and morbidity rates in preterm infants are higher than those in term infants&#46; As gestational week and birth weight decrease&#44; the risk of complications increases&#46; In preterm infants&#44; complications are observed in the early &#40;neonatal period&#41; and late periods &#40;after discharge&#41;&#46; Although the survival rate of most preterm infants has improved because of advances in medical care&#44; the incidence of short-term complications remains relatively stable&#46; Short-term complications increase the risk of long-term sequelae&#46;<a class="elsevierStyleCrossRefs" href="#bib0165"><span class="elsevierStyleSup">8&#44;9</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">In recent years&#44; many studies have been conducted on the importance of MBL during the neonatal period&#44; and most of these are associated with sepsis&#46; In sepsis&#44; proinflammatory and anti-inflammatory cytokine ratio is vital in terms of defense against infectious agents&#46; The imbalance in this ratio is manifested by increased morbidity and mortality during the neonatal period&#46;<a class="elsevierStyleCrossRefs" href="#bib0150"><span class="elsevierStyleSup">5&#44;6</span></a> Increased cytokine levels have a significant role in the pathophysiology of morbidities such as respiratory distress syndrome &#40;RDS&#41;&#44; intraventricular hemorrhage &#40;IVH&#41;&#44; necrotizing enterocolitis &#40;NEC&#41;&#44; bronchopulmonary dysplasia &#40;BPD&#41;&#44; and retinopathy of prematurity &#40;ROP&#41;&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">10</span></a> This prospective study aimed to investigate the association of <span class="elsevierStyleItalic">MBL2</span> polymorphism with short-term outcomes in preterm infants&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Materials and methods</span><p id="par0020" class="elsevierStylePara elsevierViewall">All preterm infants of &#60;37 gestational weeks who were admitted to the neonatal intensive care unit &#40;NICU&#41; of the Uludag University Medical School during a two-year period were enrolled in this prospective study&#46; The neonates were categorized into two groups according to their <span class="elsevierStyleItalic">MBL2</span> genotypes&#46; Normal <span class="elsevierStyleItalic">MBL2</span> genotype was defined as <span class="elsevierStyleItalic">MBL2</span> wild-type &#40;AA genotype&#41;&#44; whereas mutant <span class="elsevierStyleItalic">MBL2</span> genotype was defined as <span class="elsevierStyleItalic">MBL2</span> variant-type &#40;AO or OO genotype&#41;&#46; The exclusion criteria included refusal of parental consent&#44; infants with major congenital abnormalities&#44; and those undergoing a major surgical procedure&#46;</p><p id="par0025" class="elsevierStylePara elsevierViewall">Gestational age&#44; birth weight&#44; gender&#44; mode of delivery&#44; Apgar score at 1 and 5<span class="elsevierStyleHsp" style=""></span>min&#44; prenatal demographics&#44; antenatal steroid administration&#44; premature rupture of membranes&#44; history of chorioamnionitis and durations of invasive mechanical ventilation&#44; total supplemental oxygen&#44; central catheterization&#44; and total parenteral nutrition were recorded&#46; The presence of neonatal morbidities such as RDS&#44; late-onset sepsis &#40;LOS&#41;&#44; IVH&#44; NEC&#44; BPD&#44; ROP&#44; and the mortality data of the preterm infants were recorded&#46;</p><p id="par0030" class="elsevierStylePara elsevierViewall">RDS was diagnosed based on clinical findings &#40;tachypnea&#44; retractions&#44; nasal flaring&#44; and cyanosis&#41; or radiological findings &#40;reticular granular pattern or air bronchograms&#41;&#46; All neonates underwent the same management according to the NICU protocols and as recommended by European consensus guidelines on the management of neonatal respiratory distress syndrome in preterm infants&#46;<a class="elsevierStyleCrossRefs" href="#bib0180"><span class="elsevierStyleSup">11&#44;12</span></a> Neonatal sepsis was defined as the presence of clinical signs of sepsis with a positive blood culture&#46; Blood cultures were analyzed using the fully automated BACTEC method in a BACTEC 9240 device &#40;Becton Dickinson&#44; Heidelberg&#44; Germany&#41;&#46; LOS was determined by the time at which sepsis occurred between 4 and 30 days after birth&#46;<a class="elsevierStyleCrossRef" href="#bib0190"><span class="elsevierStyleSup">13</span></a> IVH was evaluated by cranial ultrasound examinations&#44; which were performed by the same pediatric radiologist and diagnosed using the Papile classification system&#46;<a class="elsevierStyleCrossRef" href="#bib0195"><span class="elsevierStyleSup">14</span></a> NEC was diagnosed according to clinical and radiographic findings and classified according to modified Bell&#39;s criteria&#46;<a class="elsevierStyleCrossRef" href="#bib0200"><span class="elsevierStyleSup">15</span></a> BPD was classified into three groups in terms of BPD severity depending on the duration and level of supplemental oxygen and mechanical ventilatory support at 36 weeks postmenstrual age&#46;<a class="elsevierStyleCrossRef" href="#bib0205"><span class="elsevierStyleSup">16</span></a> ROP was classified according to the International Classification of Retinopathy of Prematurity&#46;<a class="elsevierStyleCrossRef" href="#bib0210"><span class="elsevierStyleSup">17</span></a></p><p id="par0035" class="elsevierStylePara elsevierViewall">The MBL levels and gene polymorphisms were assessed within 3<span class="elsevierStyleHsp" style=""></span>h in most infants and within 24<span class="elsevierStyleHsp" style=""></span>h after birth in all infants&#46; The blood samples for the measurement of MBL levels were collected in a test tube and these blood samples were centrifuged within thirty minutes after obtaining&#46; After the centrifugation process&#44; serum of the samples were immediately stored at &#8722;80<span class="elsevierStyleHsp" style=""></span>&#176;C until analysis&#46;</p><p id="par0040" class="elsevierStylePara elsevierViewall">Blood samples were analyzed using enzyme-linked immunosorbent assay&#46; PCR and restriction fragment length polymorphism were used for <span class="elsevierStyleItalic">MBL</span>2 genotyping&#46; Serum MBL levels were measured using an immunoassay kit &#40;Oligomer ELISA kit&#59; Antibody Shop&#44; Copenhagen&#44; Denmark&#41; according to the manufacturer instructions&#46; The lowest detectable MBL concentration was 10<span class="elsevierStyleHsp" style=""></span>ng&#47;mL&#46; For the definition of the functional MBL deficiency&#44; this study used two different cut-off values of MBL concentration&#46; An MBL level<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>700<span class="elsevierStyleHsp" style=""></span>ng&#47;mL was determined as deficiency and &#60;150<span class="elsevierStyleHsp" style=""></span>ng&#47;mL as severe deficiency&#46;<a class="elsevierStyleCrossRefs" href="#bib0130"><span class="elsevierStyleSup">1&#8211;3&#44;18</span></a> DNA was extracted from the blood samples using a commercially available kit &#40;Puregene&#44; Gentra&#44; MN&#44; United States&#41;&#44; and <span class="elsevierStyleItalic">MBL2</span> genotyping was performed using these samples&#46; DNA samples were maintained at &#8722;20<span class="elsevierStyleHsp" style=""></span>&#176;C until use&#46; All genotypes were detected using PCR and restriction enzyme digestion&#46; Exon 1 of <span class="elsevierStyleItalic">MBL2</span> was amplified by PCR&#46; The primer sequences were 5&#8242;-GTA GGA CAG AGG GCA TGC TC-3&#8242; and 5&#8242;-CAG GCA GTT TCC TCT GGA AGG-3&#8242;&#46; In all&#44; a 349-bp PCR product was digested with <span class="elsevierStyleItalic">BanI</span> and <span class="elsevierStyleItalic">MboII</span> for codon 54 and codon 57&#44; respectively&#46; The normal allele &#40;allele A&#41; was cut into two fragments with <span class="elsevierStyleItalic">BanI</span>&#44; 260 and 89<span class="elsevierStyleHsp" style=""></span>bp&#46; The variant allele B &#40;rs1800450&#41; and allele D &#40;rs5030737&#41; remained uncut&#46; <span class="elsevierStyleItalic">MboII</span> cleaved the variant allele C &#40;rs1800451&#41; into 270 and 79<span class="elsevierStyleHsp" style=""></span>bp fragments&#46; The fragments were visualized using electrophoresis on 2&#37; agarose gel&#46; At electrophoresis&#44; the dual band at the restriction site was defined as a heterozygous mutation&#44; whereas the single band was defined as a homozygous mutation&#46; As stated&#44; the normal structural <span class="elsevierStyleItalic">MBL2</span> allele was named A&#44; whereas alleles B&#44; C and D &#40;mutation in codons 54&#44; 57 and 52&#41; were named O&#46; A representative gel electrophoresis of the <span class="elsevierStyleItalic">MBL2</span> exon 1 codon 54 polymorphisms is shown in <a class="elsevierStyleCrossRef" href="#fig0005">Fig&#46; 1</a>&#46;</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia><p id="par0045" class="elsevierStylePara elsevierViewall">This study was approved by the Ethics Committee of Uludag University Medical School and conformed to the standards set by the Declaration of Helsinki &#40;15&#46;01&#46;2013-1&#47;20&#41;&#46; All parents provided informed consents prior to the inclusion of their children in the study&#46;</p></span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Statistical analysis</span><p id="par0050" class="elsevierStylePara elsevierViewall">Statistical analysis was performed using SPSS v&#46; 20&#46;0 software &#40;SPSS Inc&#46;&#44; Chicago&#44; IL&#44; United States&#41;&#46; The results are presented as median &#40;interquartile range&#41; for the variables showing non-Gaussian distribution and mean<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>standard deviation for data showing normal distribution&#46; Student&#39;s <span class="elsevierStyleItalic">t</span>-test was used for group comparisons of normal distributions&#44; and the Mann&#8211;Whitney <span class="elsevierStyleItalic">U</span> test was used for group comparisons of non-normal distributions&#46; The chi-squared test and Fisher&#39;s exact test were used for the comparison of categorical variables&#46; Logistic regression analysis was performed to investigate the effect of <span class="elsevierStyleItalic">MBL2</span> genotype on RDS&#46; The analysis included factors that were demonstrated in the literature to have an effect on RDS&#58; gestational age&#44; birth weight&#44; sex&#44; antenatal steroid administration&#44; and <span class="elsevierStyleItalic">MBL2</span> genotype were included in the analysis&#46; A <span class="elsevierStyleItalic">p</span>-value of &#60;0&#46;05 was considered statistically significant&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Results</span><p id="par0055" class="elsevierStylePara elsevierViewall">Overall&#44; 131 preterm infants were included in this study&#46; Ten were excluded because of blood sample insufficiency&#44; four because of major congenital abnormalities&#44; and one because of major surgery&#46; In the final analysis&#44; a total of 116 preterm infants were included&#58; 69 with <span class="elsevierStyleItalic">MBL2</span> wild-type &#40;AA genotype&#41; and 47 with <span class="elsevierStyleItalic">MBL2</span> variant-type &#40;AO&#47;OO genotype&#41;&#46; Overall&#44; the rate of <span class="elsevierStyleItalic">MBL2</span> variant-type in preterm infants was 41&#37;&#46; MBL levels were significantly lower and MBL deficiency and severe deficiency were higher in <span class="elsevierStyleItalic">MBL2</span> variant-type than in <span class="elsevierStyleItalic">MBL2</span> wild-type &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#41;&#46; <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a> shows the demographic features of the study population&#46; Codon 57 and 52 polymorphisms were not detected in any of the 116 preterm infants during the genetic evaluation&#46; <span class="elsevierStyleItalic">MBL2</span> codon 54 genotype and allele frequencies were 59&#37; for <span class="elsevierStyleItalic">MBL2</span> wild-type &#40;AA genotype&#41; and 41&#37; for <span class="elsevierStyleItalic">MBL2</span> variant-type &#40;AO and OO genotype&#41;&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0060" class="elsevierStylePara elsevierViewall">Evaluation of short-term morbidity based on <span class="elsevierStyleItalic">MBL2</span> genotype revealed that RDS and mortality rates were significantly higher in the <span class="elsevierStyleItalic">MBL2</span> variant-type group &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;03&#59; respectively&#41;&#46; NEC was found to be more prevalent in the <span class="elsevierStyleItalic">MBL2</span> wild-type group &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;01&#41;&#46; There was no difference between the <span class="elsevierStyleItalic">MBL2</span> wild-type and variant-type groups in terms of IVH&#44; BPD&#44; ROP&#44; and LOS &#40;<a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41;&#46; Consideration of short-term morbidity based on MBL levels revealed that RDS was significantly higher in both the MBL deficient and severely deficient groups &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#41;&#46; NEC was found to be more common with normal levels of MBL &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;002&#41;&#46; There was no significant difference in infants with or without MBL deficiency with respect to IVH&#44; BPD&#44; ROP&#44; LOS&#44; and mortality &#40;<a class="elsevierStyleCrossRef" href="#tbl0015">Table 3</a>&#41;&#46; Further&#44; because univariate analyses revealed that RDS development was more common in the <span class="elsevierStyleItalic">MBL2</span> variant-type&#44; the effect of gestational age&#44; birth weight&#44; gender&#44; antenatal steroid use&#44; and <span class="elsevierStyleItalic">MBL2</span> genetics&#44; which are factors that may affect RDS development&#44; was investigated by logistic regression analysis&#46; <span class="elsevierStyleItalic">MBL2</span> variant-type was found to be an independent factor for the development of RDS &#40;OR&#58; 5&#46;1&#44; 95&#37; CI&#58; 2&#46;2&#8211;11&#46;9&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#41;&#46;</p><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><elsevierMultimedia ident="tbl0015"></elsevierMultimedia></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Discussion</span><p id="par0065" class="elsevierStylePara elsevierViewall">It was observed that MBL levels were lower in preterm infants with <span class="elsevierStyleItalic">MBL2</span> variant-type than in those with <span class="elsevierStyleItalic">MBL2</span> wild-type&#46; RDS was significantly more common in the <span class="elsevierStyleItalic">MBL2</span> variant-type group and also in the MBL deficient group&#46; Additionally&#44; the mortality rates were higher in preterm infants with <span class="elsevierStyleItalic">MBL2</span> variant-type&#46; In the study model&#44; <span class="elsevierStyleItalic">MBL2</span> variant-type was a significant independent factor for RDS after adjusting for the effects of other factors&#46; Besides&#44; the prevalence of NEC was higher in the <span class="elsevierStyleItalic">MBL2</span> wild-type group and with normal MBL levels&#46; It is believed that these findings will contribute toward accumulating evidence on the effect of MBL in preterm morbidities&#46;</p><p id="par0070" class="elsevierStylePara elsevierViewall">The collectin family and MBL play an important role in the primary immune elimination of invasive microorganisms in the innate immune response as well as in the regulation of ongoing immune responses against microbial invasion&#46; Studies have reported an association between MBL deficiency or variant genotype as well as infection and pulmonary pathologies&#46;<a class="elsevierStyleCrossRef" href="#bib0220"><span class="elsevierStyleSup">19</span></a> Pulmonary function impairment has been reported in patients with MBL deficiency and cystic fibrosis&#46; Patients with bronchiectasis with MBL deficiency or variant-type have a higher rate of chronic microbial colonization and frequent recurrence of pulmonary problems&#46;<a class="elsevierStyleCrossRefs" href="#bib0220"><span class="elsevierStyleSup">19&#44;20</span></a> In some studies&#44; similar to the results obtained in the present study&#44; it has been shown that MBL deficiency or variant-type cause respiratory morbidity independent of infection&#46;<a class="elsevierStyleCrossRefs" href="#bib0130"><span class="elsevierStyleSup">1&#44;21</span></a> There is high sequence homology between MBL and SP-A and SP-D&#46; The genes encoding these proteins are located on the long arm of chromosome 10 and belong to a similar lineage&#46;<a class="elsevierStyleCrossRef" href="#bib0235"><span class="elsevierStyleSup">22</span></a> SP-A and SP-D are involved in the removal of many pathogens in the lungs&#44; and although SP-A is particularly known for its immune functions&#44; RDS is associated with decreased SP-A levels&#46;<a class="elsevierStyleCrossRef" href="#bib0240"><span class="elsevierStyleSup">23</span></a> Mutant <span class="elsevierStyleItalic">MBL2</span> genetics are associated with insufficient surfactant protein-A production&#44; which may facilitate the development of RDS&#46; Similarly&#44; the present study also found a significant increase in RDS prevalence and mortality in patients with mutant <span class="elsevierStyleItalic">MBL2</span> genetics&#46; Early selective surfactant therapy in RDS has been reported to reduce pulmonary injury and mortality&#46;<a class="elsevierStyleCrossRef" href="#bib0245"><span class="elsevierStyleSup">24</span></a> It is believed that during the evaluation of <span class="elsevierStyleItalic">MBL2</span> genotype at the time of delivery in premature infants with high RDS risk and &#8804;32 gestational weeks and in borderline cases with an indication for surfactant&#44; the early administration of surfactants to patients with mutant <span class="elsevierStyleItalic">MBL2</span> genetics will reduce mortality and pulmonary morbidities&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">In recent years&#44; there has been an increased interest in the association between MBL and inflammatory morbidities&#46; It has been reported that MBL activates the lectin pathway of complement&#44; resulting in ischemia-perfusion damage&#46; In patients with <span class="elsevierStyleItalic">MBL2</span> wild-type&#44; higher MBL levels have been reported and associated with NEC&#44; resulting in reperfusion injury after intestinal ischemia&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">10</span></a> In agreement with these findings&#44; in the present study the prevalence of NEC was higher in preterm infants with <span class="elsevierStyleItalic">MBL2</span> wild-type and normal MBL levels&#46; However&#44; some studies have reported that there is no association between <span class="elsevierStyleItalic">MBL2</span> genotype and NEC&#46;<a class="elsevierStyleCrossRefs" href="#bib0150"><span class="elsevierStyleSup">5&#44;25</span></a> In the present work&#44; the development of NEC may have been relatively more common because of significantly higher mortality rates in patients with <span class="elsevierStyleItalic">MBL2</span> wild-type&#46; Because there are debatable opinions in the literature on <span class="elsevierStyleItalic">MBL2</span> genotype and NEC development&#44; additional studies are needed to clarify this issue&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">In this study&#44; in agreement with previous data&#44; no correlation was found between <span class="elsevierStyleItalic">MBL2</span> genotype and MBL levels with inflammation-associated pathologies&#44; BPD&#44; IVH&#44; and ROP&#46;<a class="elsevierStyleCrossRefs" href="#bib0150"><span class="elsevierStyleSup">5&#44;25</span></a> The evaluation of the association between <span class="elsevierStyleItalic">MBL2</span> genotype and morbidity was the common aspect of these studies&#46; It would be misleading to evaluate the association of <span class="elsevierStyleItalic">MBL2</span> genotype and MBL value obtained at the time of delivery with morbidities alone&#46; Because MBL levels increases as the gestational week increases in the <span class="elsevierStyleItalic">MBL2</span> wild-type group&#44; the evaluation of morbidity development with MBL levels obtained at different postnatal weeks may provide more accurate results to demonstrate the association between lectin pathway and inflammatory morbidities&#46;<a class="elsevierStyleCrossRef" href="#bib0135"><span class="elsevierStyleSup">2</span></a> There is a clear need for extensive studies to investigate the association between MBL and inflammatory morbidities in premature infants&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">Although the association of <span class="elsevierStyleItalic">MBL2</span> genotype with culture-proven sepsis has not been reported in the literature&#44; the association between <span class="elsevierStyleItalic">MBL2</span> genotype and early clinical sepsis has been reported&#46;<a class="elsevierStyleCrossRefs" href="#bib0145"><span class="elsevierStyleSup">4&#44;5&#44;25</span></a> In contrast&#44; the association between MBL deficiency and sepsis has been reported in many studies&#46;<a class="elsevierStyleCrossRefs" href="#bib0140"><span class="elsevierStyleSup">3&#44;18</span></a> In the present study&#44; no association was found between <span class="elsevierStyleItalic">MBL2</span> genotype and MBL levels with LOS&#46; The authors believe that inadequate immune response to infection is observed because of low MBL levels in the early postnatal weeks in preterm infants even if the <span class="elsevierStyleItalic">MBL2</span> genotype is wild-type&#46; Therefore&#44; future studies should evaluate MBL values at the time of sepsis together with genotype&#46;</p><p id="par0090" class="elsevierStylePara elsevierViewall">This study had some limitations&#46; Morbidity was evaluated based on only <span class="elsevierStyleItalic">MBL2</span> genotype and with the levels of MBL within 24<span class="elsevierStyleHsp" style=""></span>h after birth because the MBL levels of the infants were not reassessed during the subsequent postnatal days&#46; Additionally&#44; the results obtained with a limited number of cases may not reflect the overall results&#46; The strength of this study was that it evaluated the association of <span class="elsevierStyleItalic">MBL2</span> genotype with MBL level and morbidities in preterm infants and simultaneously examined the MBL level in the first 24<span class="elsevierStyleHsp" style=""></span>h of life&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">In conclusion&#44; the presence of <span class="elsevierStyleItalic">MBL2</span> variant-type and low MBL levels are important risk factors for RDS development in preterm infants&#46; Additionally&#44; there is an association between <span class="elsevierStyleItalic">MBL2</span> wild-type and NEC&#46; Considering the importance of showing that <span class="elsevierStyleItalic">MBL2</span> variant-type is an independent predictor of RDS&#44; further prospective randomized studies on this topic are clearly required&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Conflicts of interest</span><p id="par0100" class="elsevierStylePara elsevierViewall">The authors declare no conflicts of interest&#46;</p></span></span>"
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    "fechaRecibido" => "2019-01-03"
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            0 => "Mannose-binding lectin"
            1 => "Preterm"
            2 => "Respiratory distress syndrome"
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          "titulo" => "Palavras-chave"
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          "palabras" => array:3 [
            0 => "Lectina ligante de manose"
            1 => "Prematuro"
            2 => "S&#237;ndrome do desconforto respirat&#243;rio"
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        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Objective</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Mannose-binding lectin&#44; which belongs to the collectin family&#44; is an acute-phase reactant that activates the complement system&#46; This study aimed to investigate the effect of <span class="elsevierStyleItalic">MBL2</span> gene polymorphism on short-term outcomes in preterm infants&#46;</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Method</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Infants of &#60;37 gestational weeks who were admitted to the neonatal intensive care unit during a two-year period were enrolled in this prospective study&#46; The neonates were categorized into two groups according to their <span class="elsevierStyleItalic">MBL2</span> genotypes&#46; Normal <span class="elsevierStyleItalic">MBL2</span> genotype was defined as <span class="elsevierStyleItalic">MBL2</span> wild-type &#40;AA genotype&#41;&#44; whereas mutant <span class="elsevierStyleItalic">MBL2</span> genotype was defined as <span class="elsevierStyleItalic">MBL2</span> variant-type &#40;AO&#47;OO genotype&#41;&#46; The relationship between <span class="elsevierStyleItalic">MBL2</span> genotype and short-term morbidity and mortality was evaluated&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Results</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">During the two-year study period&#44; 116 preterm infants were enrolled in this study&#46; In <span class="elsevierStyleItalic">MBL2</span> variant-type&#44; mannose-binding lectin levels were significantly lower and incidences of mannose-binding lectin deficiency &#40;MBL level<span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>700<span class="elsevierStyleHsp" style=""></span>ng&#47;mL&#41; were higher &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#41;&#46; In this group&#44; the prevalence of respiratory distress syndrome and mortality was significantly higher &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#44; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;03 respectively&#41;&#46; In the <span class="elsevierStyleItalic">MBL2</span> wild-type group&#44; the prevalence of necrotizing enterocolitis &#40;NEC&#41; was higher &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>0&#46;01&#41;&#46; Logistic regression analyses revealed that <span class="elsevierStyleItalic">MBL2</span> variant-type had a significant effect on respiratory distress syndrome development &#40;odds ratio&#44; 5&#46;1&#59; 95&#37; confidence interval&#44; 2&#46;2&#8211;11&#46;9&#59; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;001&#41;&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Conclusions</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">MBL2</span> variant-type and mannose-binding lectin deficiency are important risk factors for respiratory distress syndrome development in preterm infants&#46; Additionally&#44; there is an association between <span class="elsevierStyleItalic">MBL2</span> wild-type and NEC&#46; Further studies on this subject are needed&#46;</p></span>"
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      "pt" => array:3 [
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        "resumen" => "<span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Objetivo</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">A lectina ligante de manose &#40;MBL&#44; do ingl&#234;s <span class="elsevierStyleItalic">mannose-binding lectin</span>&#41;&#44; que pertence &#224; fam&#237;lia das colectinas&#44; &#233; um reagente de fase aguda que ativa o sistema complemento&#46; Este estudo teve como objetivo investigar o efeito do polimorfismo do gene <span class="elsevierStyleItalic">MBL2</span> em desfechos de curto prazo em prematuros&#46;</p></span> <span id="abst0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">M&#233;todo</span><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Este estudo prospectivo incluiu crian&#231;as com menos de 37 semanas de gesta&#231;&#227;o admitidas na unidade de terapia intensiva neonatal durante dois anos&#46; Os neonatos foram categorizados em dois grupos de acordo com os gen&#243;tipos do <span class="elsevierStyleItalic">MBL2</span>&#46; O gen&#243;tipo normal do gene <span class="elsevierStyleItalic">MBL2</span> foi definido como <span class="elsevierStyleItalic">MBL2</span> do tipo selvagem &#40;gen&#243;tipo AA&#41;&#44; enquanto o gen&#243;tipo mutante do gene <span class="elsevierStyleItalic">MBL2</span> foi definido como o gene variante &#40;gen&#243;tipo AO&#47;OO&#41;&#46; Foi avaliada a rela&#231;&#227;o entre o gen&#243;tipo <span class="elsevierStyleItalic">MBL2</span> e a morbidade e mortalidade em curto prazo&#46;</p></span> <span id="abst0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Resultados</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Durante o per&#237;odo de dois anos&#44; 116 beb&#234;s prematuros foram inclu&#237;dos neste estudo&#46; Os n&#237;veis de lectina ligante de manose foram significativamente menores nos variantes do <span class="elsevierStyleItalic">MBL2</span> e as incid&#234;ncias de defici&#234;ncia de lectina ligante de manose &#40;n&#237;vel de MBL &#60; 700 ng&#47;mL&#41; foram maiores &#40;p &#60; 0&#44;001&#41;&#46; Nesse grupo&#44; a preval&#234;ncia de s&#237;ndrome do desconforto respirat&#243;rio &#40;SDR&#41; e a mortalidade foram significativamente maiores &#40;p &#60; 0&#44;001&#44; p &#61; 0&#44;03&#44; respectivamente&#41;&#46; No grupo <span class="elsevierStyleItalic">MBL2</span> do tipo selvagem&#44; a preval&#234;ncia de enterocolite necrosante foi maior &#40;p &#61; 0&#44;01&#41;&#46; An&#225;lises de regress&#227;o log&#237;stica revelaram que os genes variantes do <span class="elsevierStyleItalic">MBL2</span> apresentaram um efeito significativo no desenvolvimento da s&#237;ndrome do desconforto respirat&#243;rio &#40;<span class="elsevierStyleItalic">odds ratio</span>&#44; 5&#44;1&#59; intervalo de confian&#231;a de 95&#37;&#44; 2&#44;2&#8211;11&#44;9&#59; p &#60; 0&#44;001&#41;&#46;</p></span> <span id="abst0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Conclus&#245;es</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">As variantes do <span class="elsevierStyleItalic">MBL2</span> e a defici&#234;ncia de lectina ligante de manose s&#227;o importantes fatores de risco para o desenvolvimento da s&#237;ndrome do desconforto respirat&#243;rio em neonatos prematuros&#46; Al&#233;m disso&#44; existe uma associa&#231;&#227;o entre <span class="elsevierStyleItalic">MBL2</span> do tipo selvagem e a enterocolite necrosante&#46; Mais estudos s&#227;o necess&#225;rios sobre esse assunto&#46;</p></span>"
        "secciones" => array:4 [
          0 => array:2 [
            "identificador" => "abst0025"
            "titulo" => "Objetivo"
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          1 => array:2 [
            "identificador" => "abst0030"
            "titulo" => "M&#233;todo"
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          2 => array:2 [
            "identificador" => "abst0035"
            "titulo" => "Resultados"
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          3 => array:2 [
            "identificador" => "abst0040"
            "titulo" => "Conclus&#245;es"
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    "NotaPie" => array:1 [
      0 => array:2 [
        "etiqueta" => "&#9734;"
        "nota" => "<p class="elsevierStyleNotepara" id="npar0040">Please cite this article as&#58; Dogan P&#44; Ozkan H&#44; Koksal N&#44; Oral HB&#44; Bagci O&#44; Guney Varal I&#46; Mannose-binding lectin gene polymorphism and its effect on short term outcomes in preterm infants&#46; J Pediatr &#40;Rio J&#41;&#46; 2020&#59;96&#58;520&#8211;6&#46;</p>"
      ]
    ]
    "multimedia" => array:4 [
      0 => array:7 [
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        "etiqueta" => "Figure 1"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
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        "descripcion" => array:1 [
          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">DNA fragments on agarose gel electrophoresis after restriction enzyme digestion of exon 1 of the mannose-binding lectin &#40;<span class="elsevierStyleItalic">MBL2</span>&#41; gene codon 54&#46; In all&#44; 349<span class="elsevierStyleHsp" style=""></span>bp PCR product was digested with <span class="elsevierStyleItalic">BanI</span> for codon 54 polymorphism&#46; The normal allele &#40;allele A&#41; is cut into two fragments with <span class="elsevierStyleItalic">BanI</span> &#40;lanes 2 and 5&#41;&#44; 89 and 260<span class="elsevierStyleHsp" style=""></span>bp&#46; The variant allele &#40;allele O&#41; remains uncut &#40;lanes 1 and 6&#41;&#46; Both uncut and digested fragments are seen in AO heterozygote &#40;lanes 3 and 4&#41;&#46; L&#58; 100<span class="elsevierStyleHsp" style=""></span>bp DNA ladder&#46;</p>"
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      1 => array:8 [
        "identificador" => "tbl0005"
        "etiqueta" => "Table 1"
        "tipo" => "MULTIMEDIATABLA"
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        "detalles" => array:1 [
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        "tabla" => array:3 [
          "leyenda" => "<p id="spar0055" class="elsevierStyleSimplePara elsevierViewall">Values with significance are presented in bold&#46;</p><p id="spar0060" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">MBL2</span>&#44; mannose-binding lectin&#59; GA&#44; gestational age&#59; SGA&#44; small for gestational age&#59; PPROM&#44; preterm premature rupture of membranes&#46;</p>"
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Male&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>Minute 5&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">8 &#40;7&#8211;9&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">0&#46;4<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">c</span></a>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;4<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">c</span></a>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t">0&#46;4<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">c</span></a>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;7<a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">c</span></a>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleItalic">MBL levels&#44; &#40;ng&#47;mL&#41; median &#40;range&#41;</span>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t">993 &#40;257&#8211;1812&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t"><span class="elsevierStyleBold">&#60;0&#46;001</span><a class="elsevierStyleCrossRef" href="#tblfn0005"><span class="elsevierStyleSup">a</span></a>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleBold">&#60;0&#46;001</span><a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">c</span></a>&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">3 &#40;6&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;2&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t">LOS&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">18 &#40;26&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t">Mortality&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">6 &#40;9&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleBold">0&#46;03</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
              ]
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                0 => "xTab2367358.png"
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            0 => array:3 [
              "identificador" => "tblfn0020"
              "etiqueta" => "a"
              "nota" => "<p class="elsevierStyleNotepara" id="npar0020">Chi-squared test&#46;</p> <p class="elsevierStyleNotepara" id="npar0025">Values with significance are presented in bold&#46;</p> <p class="elsevierStyleNotepara" id="npar0030"><span class="elsevierStyleItalic">MBL2</span>&#44; mannose-binding lectin&#59; RDS&#44; respiratory distress syndrome&#59; LOS&#44; late onset sepsis&#59; IVH&#44; intraventricular hemorrhage&#59; NEC&#44; necrotizing enterocolitis&#59; BPD&#44; bronchopulmonary dysplasia&#59; ROP&#44; retinopathy of prematurity&#46;</p>"
            ]
          ]
        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0065" class="elsevierStyleSimplePara elsevierViewall">Frequency of early neonatal outcomes according to mannose binding lectin genotypes&#46;</p>"
        ]
      ]
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        "identificador" => "tbl0015"
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        "tipo" => "MULTIMEDIATABLA"
        "mostrarFloat" => true
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          "leyenda" => "<p id="spar0085" class="elsevierStyleSimplePara elsevierViewall">Values with significance are presented in bold&#46;</p><p id="spar0090" class="elsevierStyleSimplePara elsevierViewall">MBL&#44; mannose-binding lectin&#59; RDS&#44; respiratory distress syndrome&#59; LOS&#44; late onset sepsis&#59; IVH&#44; intraventricular hemorrhage&#59; NEC&#44; necrotizing enterocolitis&#59; BPD&#44; bronchopulmonary dysplasia&#59; ROP&#44; retinopathy of prematurity&#46;</p>"
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t" scope="col">&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&#60;150<span class="elsevierStyleHsp" style=""></span>ng&#47;mL&#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>36&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="left" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">150&#8211;700<span class="elsevierStyleHsp" style=""></span>ng&#47;mL&#40;<span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>&#61;<span class="elsevierStyleHsp" style=""></span>36&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="" valign="\n
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t">RDS&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">23 &#40;64&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">1 &#40;3&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
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                  \t\t\t\t">IVH&#44; &#40;Papile grade 3&#8211;4&#41;&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">1 &#40;3&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">2 &#40;5&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;7&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">NEC&#44; &#40;&#62;grade 1&#41;&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">0 &#40;0&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">1 &#40;3&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">8 &#40;20&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t"><span class="elsevierStyleBold">0&#46;002</span>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">BPD&#44; &#40;grade 2&#8211;3&#41;&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">5 &#40;13&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">5 &#40;14&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">12 &#40;29&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">ROP&#44; &#40;&#62;stage 2&#41;&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t">3 &#40;8&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">6 &#40;15&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;06&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">LOS&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">14 &#40;36&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">10 &#40;28&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">10 &#40;24&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">0&#46;5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td-with-role" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t ; entry_with_role_rowhead " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">Mortality&#44; <span class="elsevierStyleItalic">n</span> &#40;&#37;&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">7 &#40;18&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">7 &#40;19&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">3 &#40;7&#41;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">0&#46;3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
              ]
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                0 => "xTab2367360.png"
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0080" class="elsevierStyleSimplePara elsevierViewall">Mannose binding lectin levels in relation to early neonatal outcomes&#46;</p>"
        ]
      ]
    ]
    "bibliografia" => array:2 [
      "titulo" => "References"
      "seccion" => array:1 [
        0 => array:2 [
          "identificador" => "bibs0015"
          "bibliografiaReferencia" => array:25 [
            0 => array:3 [
              "identificador" => "bib0130"
              "etiqueta" => "1"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "MBL2 genotypes and their associations with MBL levels and NICU morbidity in a cohort of Greek neonates"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => true
                          "autores" => array:6 [
                            0 => "M&#46; Speletas"
                            1 => "A&#46; Gounaris"
                            2 => "E&#46; Sevdali"
                            3 => "M&#46; Kompoti"
                            4 => "K&#46; Konstantinidi"
                            5 => "R&#46; Sokou"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1155/2015/478412"
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Journal Information
Vol. 96. Issue 4.
Pages 520-526 (July - August 2020)
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2914
Vol. 96. Issue 4.
Pages 520-526 (July - August 2020)
Original article
Open Access
Mannose-binding lectin gene polymorphism and its effect on short term outcomes in preterm infants
Polimorfismo do gene da lectina ligante de manose e seu efeito em desfechos de curto prazo em bebês prematuros
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2914
Pelin Dogana,
Corresponding author
pelin_akbas@yahoo.com

Corresponding author.
, Hilal Ozkana, Nilgun Koksala, Haluk Barbaros Oralb, Onur Bagcia, Ipek Guney Varala
a Uludag University, Faculty of Medicine, Department of Pediatrics, Bursa, Turkey
b Uludag University, Faculty of Medicine, Department of Immunology, Bursa, Turkey
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Figures (1)
Tables (3)
Table 1. Neonatal and maternal characteristics of the study population.
Table 2. Frequency of early neonatal outcomes according to mannose binding lectin genotypes.
Table 3. Mannose binding lectin levels in relation to early neonatal outcomes.
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Abstract
Objective

Mannose-binding lectin, which belongs to the collectin family, is an acute-phase reactant that activates the complement system. This study aimed to investigate the effect of MBL2 gene polymorphism on short-term outcomes in preterm infants.

Method

Infants of <37 gestational weeks who were admitted to the neonatal intensive care unit during a two-year period were enrolled in this prospective study. The neonates were categorized into two groups according to their MBL2 genotypes. Normal MBL2 genotype was defined as MBL2 wild-type (AA genotype), whereas mutant MBL2 genotype was defined as MBL2 variant-type (AO/OO genotype). The relationship between MBL2 genotype and short-term morbidity and mortality was evaluated.

Results

During the two-year study period, 116 preterm infants were enrolled in this study. In MBL2 variant-type, mannose-binding lectin levels were significantly lower and incidences of mannose-binding lectin deficiency (MBL level<700ng/mL) were higher (p<0.001). In this group, the prevalence of respiratory distress syndrome and mortality was significantly higher (p<0.001, p=0.03 respectively). In the MBL2 wild-type group, the prevalence of necrotizing enterocolitis (NEC) was higher (p=0.01). Logistic regression analyses revealed that MBL2 variant-type had a significant effect on respiratory distress syndrome development (odds ratio, 5.1; 95% confidence interval, 2.2–11.9; p<0.001).

Conclusions

MBL2 variant-type and mannose-binding lectin deficiency are important risk factors for respiratory distress syndrome development in preterm infants. Additionally, there is an association between MBL2 wild-type and NEC. Further studies on this subject are needed.

Keywords:
Mannose-binding lectin
Preterm
Respiratory distress syndrome
Resumo
Objetivo

A lectina ligante de manose (MBL, do inglês mannose-binding lectin), que pertence à família das colectinas, é um reagente de fase aguda que ativa o sistema complemento. Este estudo teve como objetivo investigar o efeito do polimorfismo do gene MBL2 em desfechos de curto prazo em prematuros.

Método

Este estudo prospectivo incluiu crianças com menos de 37 semanas de gestação admitidas na unidade de terapia intensiva neonatal durante dois anos. Os neonatos foram categorizados em dois grupos de acordo com os genótipos do MBL2. O genótipo normal do gene MBL2 foi definido como MBL2 do tipo selvagem (genótipo AA), enquanto o genótipo mutante do gene MBL2 foi definido como o gene variante (genótipo AO/OO). Foi avaliada a relação entre o genótipo MBL2 e a morbidade e mortalidade em curto prazo.

Resultados

Durante o período de dois anos, 116 bebês prematuros foram incluídos neste estudo. Os níveis de lectina ligante de manose foram significativamente menores nos variantes do MBL2 e as incidências de deficiência de lectina ligante de manose (nível de MBL < 700 ng/mL) foram maiores (p < 0,001). Nesse grupo, a prevalência de síndrome do desconforto respiratório (SDR) e a mortalidade foram significativamente maiores (p < 0,001, p = 0,03, respectivamente). No grupo MBL2 do tipo selvagem, a prevalência de enterocolite necrosante foi maior (p = 0,01). Análises de regressão logística revelaram que os genes variantes do MBL2 apresentaram um efeito significativo no desenvolvimento da síndrome do desconforto respiratório (odds ratio, 5,1; intervalo de confiança de 95%, 2,2–11,9; p < 0,001).

Conclusões

As variantes do MBL2 e a deficiência de lectina ligante de manose são importantes fatores de risco para o desenvolvimento da síndrome do desconforto respiratório em neonatos prematuros. Além disso, existe uma associação entre MBL2 do tipo selvagem e a enterocolite necrosante. Mais estudos são necessários sobre esse assunto.

Palavras-chave:
Lectina ligante de manose
Prematuro
Síndrome do desconforto respiratório
Full Text
Introduction

Mannose-binding lectin (MBL) is an acute-phase reactant that activates the complement system. It belongs to the collectin family of proteins, which includes lung surfactant protein A (SP-A) and SP-D.1 MBL plays a key role in first-line immune responses as a component of innate immunity.2 Because adaptive immunity is underdeveloped in preterm infants, innate immunity gains higher importance.2,3 The MBL2 gene is located on the long arm of chromosome 10, and mutant MBL2 alleles occur as a result of three single-point mutations in this gene (B, C, and D). Although functional MBL levels are low in heterozygous polymorphisms, MBL levels in homozygous polymorphisms are so low that they may not even be determinable.4,5 MBL activates the complement system by binding to mannose or sugar motifs, which are found in many microorganisms, and plays an important role in innate immunity and inflammation.2,6 In newborns, an increase in sepsis frequency is observed when MBL levels are low.2,3,7

The mortality and morbidity rates in preterm infants are higher than those in term infants. As gestational week and birth weight decrease, the risk of complications increases. In preterm infants, complications are observed in the early (neonatal period) and late periods (after discharge). Although the survival rate of most preterm infants has improved because of advances in medical care, the incidence of short-term complications remains relatively stable. Short-term complications increase the risk of long-term sequelae.8,9

In recent years, many studies have been conducted on the importance of MBL during the neonatal period, and most of these are associated with sepsis. In sepsis, proinflammatory and anti-inflammatory cytokine ratio is vital in terms of defense against infectious agents. The imbalance in this ratio is manifested by increased morbidity and mortality during the neonatal period.5,6 Increased cytokine levels have a significant role in the pathophysiology of morbidities such as respiratory distress syndrome (RDS), intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), bronchopulmonary dysplasia (BPD), and retinopathy of prematurity (ROP).10 This prospective study aimed to investigate the association of MBL2 polymorphism with short-term outcomes in preterm infants.

Materials and methods

All preterm infants of <37 gestational weeks who were admitted to the neonatal intensive care unit (NICU) of the Uludag University Medical School during a two-year period were enrolled in this prospective study. The neonates were categorized into two groups according to their MBL2 genotypes. Normal MBL2 genotype was defined as MBL2 wild-type (AA genotype), whereas mutant MBL2 genotype was defined as MBL2 variant-type (AO or OO genotype). The exclusion criteria included refusal of parental consent, infants with major congenital abnormalities, and those undergoing a major surgical procedure.

Gestational age, birth weight, gender, mode of delivery, Apgar score at 1 and 5min, prenatal demographics, antenatal steroid administration, premature rupture of membranes, history of chorioamnionitis and durations of invasive mechanical ventilation, total supplemental oxygen, central catheterization, and total parenteral nutrition were recorded. The presence of neonatal morbidities such as RDS, late-onset sepsis (LOS), IVH, NEC, BPD, ROP, and the mortality data of the preterm infants were recorded.

RDS was diagnosed based on clinical findings (tachypnea, retractions, nasal flaring, and cyanosis) or radiological findings (reticular granular pattern or air bronchograms). All neonates underwent the same management according to the NICU protocols and as recommended by European consensus guidelines on the management of neonatal respiratory distress syndrome in preterm infants.11,12 Neonatal sepsis was defined as the presence of clinical signs of sepsis with a positive blood culture. Blood cultures were analyzed using the fully automated BACTEC method in a BACTEC 9240 device (Becton Dickinson, Heidelberg, Germany). LOS was determined by the time at which sepsis occurred between 4 and 30 days after birth.13 IVH was evaluated by cranial ultrasound examinations, which were performed by the same pediatric radiologist and diagnosed using the Papile classification system.14 NEC was diagnosed according to clinical and radiographic findings and classified according to modified Bell's criteria.15 BPD was classified into three groups in terms of BPD severity depending on the duration and level of supplemental oxygen and mechanical ventilatory support at 36 weeks postmenstrual age.16 ROP was classified according to the International Classification of Retinopathy of Prematurity.17

The MBL levels and gene polymorphisms were assessed within 3h in most infants and within 24h after birth in all infants. The blood samples for the measurement of MBL levels were collected in a test tube and these blood samples were centrifuged within thirty minutes after obtaining. After the centrifugation process, serum of the samples were immediately stored at −80°C until analysis.

Blood samples were analyzed using enzyme-linked immunosorbent assay. PCR and restriction fragment length polymorphism were used for MBL2 genotyping. Serum MBL levels were measured using an immunoassay kit (Oligomer ELISA kit; Antibody Shop, Copenhagen, Denmark) according to the manufacturer instructions. The lowest detectable MBL concentration was 10ng/mL. For the definition of the functional MBL deficiency, this study used two different cut-off values of MBL concentration. An MBL level<700ng/mL was determined as deficiency and <150ng/mL as severe deficiency.1–3,18 DNA was extracted from the blood samples using a commercially available kit (Puregene, Gentra, MN, United States), and MBL2 genotyping was performed using these samples. DNA samples were maintained at −20°C until use. All genotypes were detected using PCR and restriction enzyme digestion. Exon 1 of MBL2 was amplified by PCR. The primer sequences were 5′-GTA GGA CAG AGG GCA TGC TC-3′ and 5′-CAG GCA GTT TCC TCT GGA AGG-3′. In all, a 349-bp PCR product was digested with BanI and MboII for codon 54 and codon 57, respectively. The normal allele (allele A) was cut into two fragments with BanI, 260 and 89bp. The variant allele B (rs1800450) and allele D (rs5030737) remained uncut. MboII cleaved the variant allele C (rs1800451) into 270 and 79bp fragments. The fragments were visualized using electrophoresis on 2% agarose gel. At electrophoresis, the dual band at the restriction site was defined as a heterozygous mutation, whereas the single band was defined as a homozygous mutation. As stated, the normal structural MBL2 allele was named A, whereas alleles B, C and D (mutation in codons 54, 57 and 52) were named O. A representative gel electrophoresis of the MBL2 exon 1 codon 54 polymorphisms is shown in Fig. 1.

Figure 1.

DNA fragments on agarose gel electrophoresis after restriction enzyme digestion of exon 1 of the mannose-binding lectin (MBL2) gene codon 54. In all, 349bp PCR product was digested with BanI for codon 54 polymorphism. The normal allele (allele A) is cut into two fragments with BanI (lanes 2 and 5), 89 and 260bp. The variant allele (allele O) remains uncut (lanes 1 and 6). Both uncut and digested fragments are seen in AO heterozygote (lanes 3 and 4). L: 100bp DNA ladder.

(0.08MB).

This study was approved by the Ethics Committee of Uludag University Medical School and conformed to the standards set by the Declaration of Helsinki (15.01.2013-1/20). All parents provided informed consents prior to the inclusion of their children in the study.

Statistical analysis

Statistical analysis was performed using SPSS v. 20.0 software (SPSS Inc., Chicago, IL, United States). The results are presented as median (interquartile range) for the variables showing non-Gaussian distribution and mean±standard deviation for data showing normal distribution. Student's t-test was used for group comparisons of normal distributions, and the Mann–Whitney U test was used for group comparisons of non-normal distributions. The chi-squared test and Fisher's exact test were used for the comparison of categorical variables. Logistic regression analysis was performed to investigate the effect of MBL2 genotype on RDS. The analysis included factors that were demonstrated in the literature to have an effect on RDS: gestational age, birth weight, sex, antenatal steroid administration, and MBL2 genotype were included in the analysis. A p-value of <0.05 was considered statistically significant.

Results

Overall, 131 preterm infants were included in this study. Ten were excluded because of blood sample insufficiency, four because of major congenital abnormalities, and one because of major surgery. In the final analysis, a total of 116 preterm infants were included: 69 with MBL2 wild-type (AA genotype) and 47 with MBL2 variant-type (AO/OO genotype). Overall, the rate of MBL2 variant-type in preterm infants was 41%. MBL levels were significantly lower and MBL deficiency and severe deficiency were higher in MBL2 variant-type than in MBL2 wild-type (p<0.001). Table 1 shows the demographic features of the study population. Codon 57 and 52 polymorphisms were not detected in any of the 116 preterm infants during the genetic evaluation. MBL2 codon 54 genotype and allele frequencies were 59% for MBL2 wild-type (AA genotype) and 41% for MBL2 variant-type (AO and OO genotype).

Table 1.

Neonatal and maternal characteristics of the study population.

  MBL2 wild-type  MBL2 variant -type  p 
  (n=69)  (n=47)   
GA at birth, median in weeks (range)  30 (29–33)  31 (29–33)  0.6a 
Birth weight, g (mean±SD)  1539±574  1459±556  0.5b 
Sex, n (%)
Male  40 (58)  29 (62)   
Female  29 (42)  18 (38)  0.7c 
SGA, n (%)  13 (19)  14 (30)  0.1c 
Cesarian delivery, n (%)  58 (84)  36(77)  0.3c 
Apgar score, median (range)
Minute 1  7 (5–8)  6 (4–7)  0.1a 
Minute 5  8 (7–9)  8 (7–9)  0.1a 
Antenatal steroid, n (%)
None    33 (48)  30 (64) 
Single course    18 (26)  9 (19) 
Repeat course    18 (26)  8 (17) 
Maternal preeclampsia, n (%)  17 (25)  15 (32)  0.4c 
Maternal infection, n (%)  3 (4)  4 (9)  0.4c 
PPROM, n (%)  13 (19)  6 (13)  0.4c 
Chorioamnionitis, n (%)  4 (6)  2 (4)  0.7c 
MBL levels, (ng/mL) median (range)  993 (257–1812)  10 (10–473)  <0.001a 
MBL deficiency, (MBL level <700ng/mL), n (%)  30 (44)  45 (96)  <0.001c 

Values with significance are presented in bold.

MBL2, mannose-binding lectin; GA, gestational age; SGA, small for gestational age; PPROM, preterm premature rupture of membranes.

a

Mann–Whitney U test.

b

Student's t-test.

c

Chi-squared test.

Evaluation of short-term morbidity based on MBL2 genotype revealed that RDS and mortality rates were significantly higher in the MBL2 variant-type group (p<0.001, p=0.03; respectively). NEC was found to be more prevalent in the MBL2 wild-type group (p=0.01). There was no difference between the MBL2 wild-type and variant-type groups in terms of IVH, BPD, ROP, and LOS (Table 2). Consideration of short-term morbidity based on MBL levels revealed that RDS was significantly higher in both the MBL deficient and severely deficient groups (p<0.001). NEC was found to be more common with normal levels of MBL (p=0.002). There was no significant difference in infants with or without MBL deficiency with respect to IVH, BPD, ROP, LOS, and mortality (Table 3). Further, because univariate analyses revealed that RDS development was more common in the MBL2 variant-type, the effect of gestational age, birth weight, gender, antenatal steroid use, and MBL2 genetics, which are factors that may affect RDS development, was investigated by logistic regression analysis. MBL2 variant-type was found to be an independent factor for the development of RDS (OR: 5.1, 95% CI: 2.2–11.9, p<0.001).

Table 2.

Frequency of early neonatal outcomes according to mannose binding lectin genotypes.

  MBL2 wild-type(n=69)  MBL2 variant-type(n=47)  pa 
RDS, n (%)  21 (30)  31 (66)  <0.001 
IVH, (Papile grade 3–4), n (%)  3 (4)  2 (4)  0.9 
NEC, (>grade 1), n (%)  9 (13)  0 (0)  0.01 
BPD, (grade 2–3), n (%)  16 (23)  6 (13)  0.2 
ROP, (>stage 2), n (%)  10 (15)  3 (6)  0.2 
LOS, n (%)  18 (26)  16 (34)  0.4 
Mortality, n (%)  6 (9)  11 (23)  0.03 
a

Chi-squared test.

Values with significance are presented in bold.

MBL2, mannose-binding lectin; RDS, respiratory distress syndrome; LOS, late onset sepsis; IVH, intraventricular hemorrhage; NEC, necrotizing enterocolitis; BPD, bronchopulmonary dysplasia; ROP, retinopathy of prematurity.

Table 3.

Mannose binding lectin levels in relation to early neonatal outcomes.

  MBL deficiencyNormal MBL>700ng/mL(n=41)  pa 
  <150ng/mL(n=36)  150–700ng/mL(n=36)     
RDS, n (%)  28 (72)  23 (64)  1 (3)  <0.001 
IVH, (Papile grade 3–4), n (%)  2 (5)  1 (3)  2 (5)  0.7 
NEC, (>grade 1), n (%)  0 (0)  1 (3)  8 (20)  0.002 
BPD, (grade 2–3), n (%)  5 (13)  5 (14)  12 (29)  0.1 
ROP, (>stage 2), n (%)  3 (8)  4 (11)  6 (15)  0.06 
LOS, n (%)  14 (36)  10 (28)  10 (24)  0.5 
Mortality, n (%)  7 (18)  7 (19)  3 (7)  0.3 

Values with significance are presented in bold.

MBL, mannose-binding lectin; RDS, respiratory distress syndrome; LOS, late onset sepsis; IVH, intraventricular hemorrhage; NEC, necrotizing enterocolitis; BPD, bronchopulmonary dysplasia; ROP, retinopathy of prematurity.

a

Chi-squared test.

Discussion

It was observed that MBL levels were lower in preterm infants with MBL2 variant-type than in those with MBL2 wild-type. RDS was significantly more common in the MBL2 variant-type group and also in the MBL deficient group. Additionally, the mortality rates were higher in preterm infants with MBL2 variant-type. In the study model, MBL2 variant-type was a significant independent factor for RDS after adjusting for the effects of other factors. Besides, the prevalence of NEC was higher in the MBL2 wild-type group and with normal MBL levels. It is believed that these findings will contribute toward accumulating evidence on the effect of MBL in preterm morbidities.

The collectin family and MBL play an important role in the primary immune elimination of invasive microorganisms in the innate immune response as well as in the regulation of ongoing immune responses against microbial invasion. Studies have reported an association between MBL deficiency or variant genotype as well as infection and pulmonary pathologies.19 Pulmonary function impairment has been reported in patients with MBL deficiency and cystic fibrosis. Patients with bronchiectasis with MBL deficiency or variant-type have a higher rate of chronic microbial colonization and frequent recurrence of pulmonary problems.19,20 In some studies, similar to the results obtained in the present study, it has been shown that MBL deficiency or variant-type cause respiratory morbidity independent of infection.1,21 There is high sequence homology between MBL and SP-A and SP-D. The genes encoding these proteins are located on the long arm of chromosome 10 and belong to a similar lineage.22 SP-A and SP-D are involved in the removal of many pathogens in the lungs, and although SP-A is particularly known for its immune functions, RDS is associated with decreased SP-A levels.23 Mutant MBL2 genetics are associated with insufficient surfactant protein-A production, which may facilitate the development of RDS. Similarly, the present study also found a significant increase in RDS prevalence and mortality in patients with mutant MBL2 genetics. Early selective surfactant therapy in RDS has been reported to reduce pulmonary injury and mortality.24 It is believed that during the evaluation of MBL2 genotype at the time of delivery in premature infants with high RDS risk and ≤32 gestational weeks and in borderline cases with an indication for surfactant, the early administration of surfactants to patients with mutant MBL2 genetics will reduce mortality and pulmonary morbidities.

In recent years, there has been an increased interest in the association between MBL and inflammatory morbidities. It has been reported that MBL activates the lectin pathway of complement, resulting in ischemia-perfusion damage. In patients with MBL2 wild-type, higher MBL levels have been reported and associated with NEC, resulting in reperfusion injury after intestinal ischemia.10 In agreement with these findings, in the present study the prevalence of NEC was higher in preterm infants with MBL2 wild-type and normal MBL levels. However, some studies have reported that there is no association between MBL2 genotype and NEC.5,25 In the present work, the development of NEC may have been relatively more common because of significantly higher mortality rates in patients with MBL2 wild-type. Because there are debatable opinions in the literature on MBL2 genotype and NEC development, additional studies are needed to clarify this issue.

In this study, in agreement with previous data, no correlation was found between MBL2 genotype and MBL levels with inflammation-associated pathologies, BPD, IVH, and ROP.5,25 The evaluation of the association between MBL2 genotype and morbidity was the common aspect of these studies. It would be misleading to evaluate the association of MBL2 genotype and MBL value obtained at the time of delivery with morbidities alone. Because MBL levels increases as the gestational week increases in the MBL2 wild-type group, the evaluation of morbidity development with MBL levels obtained at different postnatal weeks may provide more accurate results to demonstrate the association between lectin pathway and inflammatory morbidities.2 There is a clear need for extensive studies to investigate the association between MBL and inflammatory morbidities in premature infants.

Although the association of MBL2 genotype with culture-proven sepsis has not been reported in the literature, the association between MBL2 genotype and early clinical sepsis has been reported.4,5,25 In contrast, the association between MBL deficiency and sepsis has been reported in many studies.3,18 In the present study, no association was found between MBL2 genotype and MBL levels with LOS. The authors believe that inadequate immune response to infection is observed because of low MBL levels in the early postnatal weeks in preterm infants even if the MBL2 genotype is wild-type. Therefore, future studies should evaluate MBL values at the time of sepsis together with genotype.

This study had some limitations. Morbidity was evaluated based on only MBL2 genotype and with the levels of MBL within 24h after birth because the MBL levels of the infants were not reassessed during the subsequent postnatal days. Additionally, the results obtained with a limited number of cases may not reflect the overall results. The strength of this study was that it evaluated the association of MBL2 genotype with MBL level and morbidities in preterm infants and simultaneously examined the MBL level in the first 24h of life.

In conclusion, the presence of MBL2 variant-type and low MBL levels are important risk factors for RDS development in preterm infants. Additionally, there is an association between MBL2 wild-type and NEC. Considering the importance of showing that MBL2 variant-type is an independent predictor of RDS, further prospective randomized studies on this topic are clearly required.

Conflicts of interest

The authors declare no conflicts of interest.

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Please cite this article as: Dogan P, Ozkan H, Koksal N, Oral HB, Bagci O, Guney Varal I. Mannose-binding lectin gene polymorphism and its effect on short term outcomes in preterm infants. J Pediatr (Rio J). 2020;96:520–6.

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