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Original article
First evidence for the existence of Simkania negevensis in the genitalia of human females
Primera evidencia de la existencia de Simkania negevensis en los genitales femeninos humanos
Raneem A. Abu-Saifa, Asma S. Al-Bashab, Hesham M. Al-Younesa,
Corresponding author
alyounes@ju.edu.jo

Corresponding author.
a Department of Biological Sciences, School of Science, the University of Jordan, Amman, Jordan
b Department of Obstetrics and Gynecology, School of Medicine, the University of Jordan, Amman, Jordan
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    "textoCompleto" => "<span class="elsevierStyleSections"><span id="sec0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Introduction</span><p id="par0005" class="elsevierStylePara elsevierViewall"><span class="elsevierStyleItalic">Simkania negevensis</span>&#44; originally characterized as a <span class="elsevierStyleItalic">Chlamydia</span>-like microorganism&#44; belongs now to family <span class="elsevierStyleItalic">Simkaniaceae</span> within the <span class="elsevierStyleItalic">Chlamydiales</span> order&#46;<a class="elsevierStyleCrossRef" href="#bib0155"><span class="elsevierStyleSup">1</span></a> Similar to other <span class="elsevierStyleItalic">Chlamydiales</span> members&#44; <span class="elsevierStyleItalic">S&#46; negevensis</span> is an obligate intracellular Gram-negative bacterium and possesses a developmental cycle with two distinct phases&#58; the large&#44; non-infectious&#44; intracellular&#44; replicative reticulate body and the small intracellular&#44; infectious elementary body&#46;<a class="elsevierStyleCrossRef" href="#bib0155"><span class="elsevierStyleSup">1</span></a></p><p id="par0010" class="elsevierStylePara elsevierViewall">Seroepidemiological studies from different countries on apparently healthy individuals detected seropositivity rates ranging from 4&#46;3&#37; to 80&#37;&#46;<a class="elsevierStyleCrossRefs" href="#bib0160"><span class="elsevierStyleSup">2&#44;3</span></a> Besides&#44; this agent is considered as a potential emerging pathogen in the respiratory system&#44; causing community-acquired pneumonia &#40;CAP&#41; and bronchiolitis&#44; as confirmed by different techniques&#46;<a class="elsevierStyleCrossRefs" href="#bib0155"><span class="elsevierStyleSup">1&#44;3&#44;4</span></a> However&#44; other reports revealed no link between <span class="elsevierStyleItalic">Simkania</span> and chronic obstructive pulmonary disease&#44; chronic cough&#44; asthma or even CAP and bronchiolitis&#46;<a class="elsevierStyleCrossRefs" href="#bib0175"><span class="elsevierStyleSup">5&#8211;9</span></a> Interestingly&#44; because of the remarkable high and comparable prevalences of <span class="elsevierStyleItalic">Simkania</span> in cases suffering from CAP or bronchitis and in healthy controls &#40;57&#46;1&#37; versus 62&#46;5&#37;&#41;&#44; it was tempting to hypothesize that <span class="elsevierStyleItalic">S&#46; negevensis</span> is not a causative agent of respiratory diseases and may exist as a commensal agent in the respiratory system&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">5</span></a> On the other hand&#44; sporadic studies suggested a role of <span class="elsevierStyleItalic">Simkania</span> in nonrespiratory system conditions&#46; For instance&#44; DNA of <span class="elsevierStyleItalic">Simkania</span> has been amplified from an aortic aneurysm and intestinal biopsies from patients of Crohn&#39;s disease&#46;<a class="elsevierStyleCrossRefs" href="#bib0200"><span class="elsevierStyleSup">10&#44;11</span></a></p><p id="par0015" class="elsevierStylePara elsevierViewall">In general&#44; microorganisms belonging to the order <span class="elsevierStyleItalic">Chlamydiales</span> are medically important&#46; For instance&#44; the very well-known <span class="elsevierStyleItalic">Chlamydia trachomatis</span> and <span class="elsevierStyleItalic">Chlamydia pneumoniae</span> are mainly implicated in human genital and respiratory diseases&#44; respectively&#46;<a class="elsevierStyleCrossRefs" href="#bib0210"><span class="elsevierStyleSup">12&#44;13</span></a> Moreover&#44; recently discovered <span class="elsevierStyleItalic">Chlamydia</span>-like bacteria&#44; including <span class="elsevierStyleItalic">S&#46; negevensis&#44; Waddlia chondrophila</span> and <span class="elsevierStyleItalic">Parachlamydia acanthamoebae</span> are associated with clinical conditions&#46; <span class="elsevierStyleItalic">W&#46; chondrophila</span> has been documented as an agent of adverse pregnancy outcomes&#44; whereas&#44; <span class="elsevierStyleItalic">P&#46; acanthamoebae</span> is an emerging respiratory pathogen and may also be linked to adverse pregnancy outcomes in ruminants and humans&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">4&#44;12&#44;13</span></a></p><p id="par0020" class="elsevierStylePara elsevierViewall">In addition to the biological characteristics of <span class="elsevierStyleItalic">Simkania</span> shared with other <span class="elsevierStyleItalic">Chlamydiales</span> members&#44; tropism to inhabit certain body organs and disease potential of <span class="elsevierStyleItalic">Simkania</span> could be also similar to these related microbes&#46; Thus&#44; it is attractive to hypothesize that <span class="elsevierStyleItalic">Simkania</span> could be implicated in human urogenital infections&#44; in addition to its being a respiratory microbe&#46; Thus&#44; this study aims at investigating the existence of <span class="elsevierStyleItalic">Simkania</span> in genital system of human adult females&#46;</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Study subjects and study design</span><p id="par0025" class="elsevierStylePara elsevierViewall">This study was carried out during the period from July 2018 to August 2019&#46; Here&#44; 169 Jordanian women&#44; who attended for different reasons the Obstetrics and Gynecology clinic of the University of Jordan Hospital&#44; Amman&#44; Jordan&#44; participated in this study and provided genital samples&#46; Females aged between 20 and 91 years with a mean age of 39&#46;5<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>11&#46;5 years&#46; Subjects who had received antibiotics within the last 3 months prior to enrollment or were on menstrual cycle were excluded from the study&#46; Demographic features&#44; available clinical data and laboratory and diagnostic information were recorded for participating females&#46; Moreover&#44; an informed signed consent was obtained from each of them&#46;</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Sample collection and transport</span><p id="par0030" class="elsevierStylePara elsevierViewall">Lower vaginal specimens were collected using sterile cotton-tipped swabs&#44; immediately placed into a sterile tube containing 1<span class="elsevierStyleHsp" style=""></span>ml sterile transport &#40;2-sucrose-phosphate&#41; medium&#44; which consisted of 17&#46;1<span class="elsevierStyleHsp" style=""></span>g sucrose&#44; 0&#46;5<span class="elsevierStyleHsp" style=""></span>g dibasic potassium phosphate&#44; 0&#46;25<span class="elsevierStyleHsp" style=""></span>g monobasic potassium phosphate dissolved in 200<span class="elsevierStyleHsp" style=""></span>ml distilled water&#44; followed by the addition of 20&#37; fetal bovine serum and adjustment of pH to 7&#46;2&#46; Tubes containing the swabs were vortexed for 30<span class="elsevierStyleHsp" style=""></span>sec to dislodge any material into the liquid medium and the swabs were then removed and discarded&#46; Next&#44; the medium was transferred into a sterile 1&#46;8-ml cryovial and stored at &#8722;72<span class="elsevierStyleHsp" style=""></span>&#176;C until use&#46;</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">DNA isolation</span><p id="par0035" class="elsevierStylePara elsevierViewall">DNA from 200<span class="elsevierStyleHsp" style=""></span>&#956;l of each specimen was harvested according to a protocol specific for isolating nucleic acids from body fluids&#44; provided with the G-spin&#8482; Total DNA Extraction kit &#40;iNtRON Biotechnology&#44; South Korea&#41;&#46; Extracted DNA was eluted in a final volume of 50<span class="elsevierStyleHsp" style=""></span>&#956;l of a buffer provided with the kit&#44; aliquoted and stored at &#8722;20<span class="elsevierStyleHsp" style=""></span>&#176;C until tested&#46; DNA amounts were estimated spectrophotometrically&#46;</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">PCR assay</span><p id="par0040" class="elsevierStylePara elsevierViewall">To detect the bacterial DNA&#44; a polymerase chain reaction &#40;PCR&#41; assay&#44; amplifying a 311<span class="elsevierStyleHsp" style=""></span>bp fragment of the <span class="elsevierStyleItalic">S&#46;</span><span class="elsevierStyleItalic">negevensis</span><span class="elsevierStyleItalic">chaperonin</span>-60 &#40;<span class="elsevierStyleItalic">cpn</span> 60&#41; gene&#44; was performed according to a protocol of Donati et al&#46;<a class="elsevierStyleCrossRef" href="#bib0220"><span class="elsevierStyleSup">14</span></a> with modification&#46; Forward and reverse primers were 5&#8242;-GATGGAACGATGACTGTTGAA-3&#8242; and 5&#8242;-GCACAAACTTTTAGTCCTGC-3&#8242;&#44; respectively&#46; Conditions for the PCR amplification were&#58; denaturation for 4<span class="elsevierStyleHsp" style=""></span>min at 94<span class="elsevierStyleHsp" style=""></span>&#176;C and 35 cycles&#44; each of which consisted of denaturation step for 1<span class="elsevierStyleHsp" style=""></span>min at 94<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; annealing for 1<span class="elsevierStyleHsp" style=""></span>min at 57<span class="elsevierStyleHsp" style=""></span>&#176;C&#44; extension for 1<span class="elsevierStyleHsp" style=""></span>min at 72<span class="elsevierStyleHsp" style=""></span>&#176;C and a final elongation step for 7<span class="elsevierStyleHsp" style=""></span>min at 72<span class="elsevierStyleHsp" style=""></span>&#176;C&#46; In PCR runs&#44; negative controls were included and contained all components of the amplification mixture and nuclease-free water instead of the purified DNA&#46; Purified <span class="elsevierStyleItalic">Simkania</span> nucleic acids&#44; generously provided by Thomas Rudel and Jo-Ana Herweg&#44; Department of Microbiology&#44; University of W&#252;rzburg&#44; W&#252;rzburg&#44; Germany&#44; and the bacterial DNA isolated previously from <span class="elsevierStyleItalic">Simkania</span>-positive nasopharyngeal samples&#44; served as positive controls&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">5</span></a> For sequencing purposes&#44; some PCR amplicons were shipped with the required primers to MACROGEN &#40;South Korea&#41;&#46;</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Statistical analysis</span><p id="par0045" class="elsevierStylePara elsevierViewall">Statistical analysis of the results was calculated using chi-square test&#46; A probability value &#40;<span class="elsevierStyleItalic">p</span>&#41; &#8804;0&#46;05 was considered statistically significant&#46;</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Ethical statement</span><p id="par0050" class="elsevierStylePara elsevierViewall">This work has been approved by the Institutional Review Board &#40;IRB&#41; of the University of Jordan Hospital&#46; The individuals who participated in this work gave signed and informed consent&#46; Anonymity and confidentiality were guaranteed at all times&#44; as well as data protection&#46;</p></span></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Results</span><p id="par0055" class="elsevierStylePara elsevierViewall">The majority of participating women were married at the time of enrollment &#40;165&#47;169&#59; 97&#46;63&#37;&#41;&#44; whereas 4 women were widowed or divorced &#40;2&#46;37&#37;&#41;&#46; Ninety seven females &#40;57&#46;4&#37;&#41; were middle-aged women &#40;30-49 years&#41;&#46; Sixty one females &#40;36&#46;09&#37;&#41; were apparently healthy subjects with no symptoms&#44; among which 42 women visited the clinic for routine checkup and 19 women were seeking fertility&#44; while the remainder &#40;63&#46;91&#37;&#41; suffered from various symptoms at attendance&#46; About 84&#37; of participants were pregnant for at least one time&#46; In addition&#44; more than half of the enrolled females &#40;58&#37;&#41; did not utilize any birth control method &#40;intrauterine device &#40;IUD&#41;&#44; contraceptive pills or condoms by their husbands&#41;&#46; More sociodemographic and clinical profiles of the entire female population involved in the study are shown in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46; Here&#44; PCR detected <span class="elsevierStyleItalic">Simkania</span> DNA in the genitalia of 41 out of 169 females&#44; suggesting an overall prevalence of 24&#46;26&#37; among individuals examined &#40;<a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41;&#46; Sequencing of some PCR amplicons confirmed the amplification of the desired domain within the bacterial gene <span class="elsevierStyleItalic">cpn</span> 60 &#40;data not shown&#41;&#46; To analyze the age-specific detection rates&#44; subjects were divided into five age groups&#58; 20&#8211;29 years&#44; 30&#8211;39 years&#44; 40&#8211;49 years&#44; 50&#8211;59 years and &#8805;60 years&#46; Only 7 cases were DNA positive out of 35 females in the youngest group aged 20&#8211;29 years&#44; resulting in a prevalence of 20&#46;00&#37; &#40;95&#37; CI&#58; 25&#46;8<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;7&#41;&#46; Clearly&#44; the DNA positivity reached a peak &#40;33&#46;96&#37;&#41; in the age group of 30&#8211;39 years &#40;95&#37; CI&#58; 34&#46;6<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;896&#59; 18&#47;53&#41; and gradually decreased in the subsequent groups&#44; reaching 27&#46;27&#37; in the cohort of 40&#8211;49 years &#40;95&#37; CI&#58; 43&#46;62<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>0&#46;912&#59; 12&#47;44&#41; and 13&#46;33&#37; in the group of 50&#8211;59 years &#40;95&#37; CI&#58; 53&#46;5<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>1&#46;103&#59; 4&#47;30&#41;&#46; Remarkably&#44; none of the participants older than 59 years was DNA positive &#40;95&#37; CI&#58; 68<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>10&#46;311&#59; 0&#47;7&#41;&#46; Further analysis of the results showed the absence of an inverse or a proportional correlation between bacterial nucleic acid positivity and specific ages of females &#40;data not shown&#41;&#46; Interestingly&#44; DNA of <span class="elsevierStyleItalic">Simkania</span> was evident in females with ages &#8804;55 years only&#44; while all females older than 55 years &#40;12 individuals&#41; were found negative&#46; Apparently&#44; more than 50&#37; of positive cases are clustered within the age brackets 29&#8211;35 years and 46&#8211;50 years&#46; These relatively narrow age intervals account for approximately 29&#46;27&#37; &#40;12&#47;41&#41; and 21&#46;95&#37; &#40;9&#47;41&#41; of all PCR-positive cases&#44; respectively&#46; Sociodemographic and clinical information of the 41 PCR-positive women are summarized in <a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>&#46;</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><elsevierMultimedia ident="tbl0010"></elsevierMultimedia><p id="par0060" class="elsevierStylePara elsevierViewall">To demonstrate the presence of any relation between the female reproductive age and some of its correlates with the positivity of <span class="elsevierStyleItalic">S&#46; negevensis</span> in genital system&#44; women were divided into two groups based on criteria set by WHO&#44;<a class="elsevierStyleCrossRef" href="#bib0225"><span class="elsevierStyleSup">15</span></a> which defined the reproductive age of females to be from 15 to 49 years&#46; Accordingly&#44; one group consisted of 132 reproductive age females &#40;20&#8211;49 years old&#41;&#44; while the second group comprised only 37 non-reproductive age females &#40;with ages 50 years or older&#41;&#46; Remarkably&#44; about 2&#46;5-fold significant increase in the prevalence of <span class="elsevierStyleItalic">Simkania</span> was obtained among women of the reproductive age compared to non-reproductive age individuals &#40;28&#46;03&#37; versus 10&#46;81&#37;&#59; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46; Interestingly&#44; <span class="elsevierStyleItalic">Simkania</span> DNA was detected in 30&#46;61&#37; &#40;15&#47;49&#41; of females who experienced spontaneous abortion &#40;miscarriage&#41;&#44; compared to about 22&#46;5&#37; &#40;21&#47;93&#41; of females with no history of miscarriage&#44; suggesting possibly a correlation between the bacterium and this adverse pregnancy outcome &#40;<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0&#46;29&#41;&#46;</p><p id="par0065" class="elsevierStylePara elsevierViewall">In an attempt to correlate <span class="elsevierStyleItalic">S&#46; negevensis</span> to clinical manifestations&#44; detection rates of the bacterium in the females were analyzed based on the reasons for attending the Obstetrics and Gynecology clinic&#46; Reasons were categorized into &#40;1&#41; vaginal inflammation &#40;itching and&#47;or discharge&#41;&#44; &#40;2&#41; menstrual and bleeding complaints&#44; &#40;3&#41; routine checkup&#44; &#40;4&#41; multiple reasons&#44; &#40;5&#41; seeking pregnancy and &#40;6&#41; other reasons&#46; As illustrated in <a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#44; 23&#46;81&#37; &#40;10&#47;42&#41; of symptomless women&#44; visiting the clinic for a regular checkup &#40;pap smear screening&#44; IUD checking&#44; antenatal visit and postpartum follow-up&#41;&#44; were positive for <span class="elsevierStyleItalic">S&#46; negevensis</span>&#46; A slightly less DNA positivity rate&#44; which remained above 20&#37; &#40;4&#47;19&#59; 21&#46;05&#37;&#41;&#44; was reported for women who were planning to conceive and with no detectable symptoms&#46; However&#44; among other pregnancy-seeking females with coincidental complaints &#40;18 cases&#41;&#44; only one case was DNA positive &#40;<a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41;&#46; In females who attended for reasons related to menstrual cycle abnormalities&#44; the prevalence rate was almost the same and did not exceed 25&#37; &#40;13&#47;57&#41;&#46; Surprisingly&#44; the bacterial DNA was evident in 13 out of 30 females with symptoms of vaginal inflammation &#40;itching and&#47;or abnormal discharge&#41;&#44; yielding about 2-fold increase in the nucleic acid positivity &#40;43&#46;33&#37;&#41;&#44; compared to rates reported for females who attended for checkup or menstrual cycle complaints &#40;<a class="elsevierStyleCrossRef" href="#tbl0010">Table 2</a>&#41;&#46; Collectively&#44; these data clearly demonstrate that <span class="elsevierStyleItalic">S&#46; negevensis</span> may be linked to vaginal inflammation and its related signs&#44; such as itching and abnormal secretions&#46;</p></span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Discussion</span><p id="par0070" class="elsevierStylePara elsevierViewall">To the best of our knowledge&#44; this is the first investigation that confirmed the presence of <span class="elsevierStyleItalic">S&#46; negevensis</span> in the female genitalia&#46; During conducting the present research&#44; Greub and his colleagues could not detect this microorganism in 22 urethral samples from both men and women and in 135 cervicovaginal swabs using their newly developed quantitative real-time PCR&#46;<a class="elsevierStyleCrossRef" href="#bib0230"><span class="elsevierStyleSup">16</span></a> Contradictory results using cell culture&#44; PCR and serological methods were also noticed in studies that investigated the prevalence and the association of <span class="elsevierStyleItalic">Simkania</span> with pulmonary conditions&#46;<a class="elsevierStyleCrossRefs" href="#bib0155"><span class="elsevierStyleSup">1&#44;3&#44;5</span></a> The obvious discrepancy between our findings and those of Vouga et al&#46;<a class="elsevierStyleCrossRef" href="#bib0230"><span class="elsevierStyleSup">16</span></a> may reflect variations in the DNA amplification techniques&#44; types of clinical samples and their processing in addition to differences in the populations examined&#46; Many in-house PCR methods are available in the literature for the detection of <span class="elsevierStyleItalic">Simkania</span> and other <span class="elsevierStyleItalic">Chlamydiales</span> members in clinical and environmental samples&#46; In spite of that&#44; the sensitivity and specificity of the majority of these tests remain unknown&#46; Among these protocols are real-time PCR&#44; nested-PCR&#44; multiplex PCR and standard PCR&#46;<a class="elsevierStyleCrossRefs" href="#bib0175"><span class="elsevierStyleSup">5&#44;9&#44;14&#44;17</span></a> Here&#44; we used a standard PCR method to target <span class="elsevierStyleItalic">S&#46; negevensis cpn</span> 60 gene&#44; using outer primers of a previously described nested PCR&#46;<a class="elsevierStyleCrossRef" href="#bib0220"><span class="elsevierStyleSup">14</span></a> This modified test was successfully used to detect <span class="elsevierStyleItalic">Simkania</span> nucleic acids in nasopharyngeal samples&#46;<a class="elsevierStyleCrossRef" href="#bib0175"><span class="elsevierStyleSup">5</span></a> The specificity of this assay was confirmed by sequence analyses of some randomly chosen amplicons produced in the present study&#44; which showed more than 97&#37; homology with the desired target in <span class="elsevierStyleItalic">Simkania</span> genome &#40;data not shown&#41;&#46; Generally&#44; standard PCR test has most likely the advantage of a decreased risk of contamination&#44; compared to nested PCR that is highly sensitive to this drawback by PCR product carryover&#46; To corroborate our novel PCR-based finding that <span class="elsevierStyleItalic">Simkania</span> inhabits the female genitalia&#44; the use of the cell culture approach for isolation and subsequent characterization of the microbe in addition to one of the modern culture-independent molecular techniques&#44; such as next generation sequencing &#40;NGS&#41;&#44; is recommended&#46;</p><p id="par0075" class="elsevierStylePara elsevierViewall">Evaluation of age-related positivity rates clearly showed that about three quarters &#40;73&#46;17&#37;&#41; of <span class="elsevierStyleItalic">Simkania</span>-positive females aged between 30 and 49 years&#46; Importantly&#44; about half of the <span class="elsevierStyleItalic">Simkania</span> DNA-positive females aged between 29&#8211;35 years &#40;12&#47;41&#59; 29&#46;27&#37;&#41; and 46&#8211;50 years &#40;9&#47;41&#59; 21&#46;95&#37;&#41;&#46; Deeper analysis of age-specific prevalences demonstrated that DNA positivity reached the peak in females within the age range 30&#8211;39 years&#44; slightly decreased in the subsequent age group of 40&#8211;49 years and then sharply decreased in females aged 50&#8211;59 years&#46; In contrast&#44; none of the females older than 60 years was positive for <span class="elsevierStyleItalic">Simkania</span> and this may be due to the low number of women investigated in this elderly cohort &#40;only 7 subjects&#41;&#46; Overall&#44; based on these interesting novel findings&#44; further studies with larger female populations are needed to gain a better understanding of the relatedness between the age and the incidence of <span class="elsevierStyleItalic">S&#46; negevensis</span> in genitalia&#46;</p><p id="par0080" class="elsevierStylePara elsevierViewall">Many <span class="elsevierStyleItalic">Simkania</span>-like bacteria have been associated with diseases relevant to the urogenital system and&#47;or respiratory system&#46; Globally&#44; <span class="elsevierStyleItalic">C&#46; trachomatis</span> is considered the most common sexually transmitted pathogen&#44; leading to cervicitis&#44; urethritis&#44; ectopic pregnancy&#44; miscarriage&#44; epididymitis&#44; prostatitis and male infertility&#46;<a class="elsevierStyleCrossRefs" href="#bib0210"><span class="elsevierStyleSup">12&#44;13&#44;18</span></a> The prevalence of this pathogen was estimated at about 100 million new cases in 2005&#46;<a class="elsevierStyleCrossRef" href="#bib0210"><span class="elsevierStyleSup">12</span></a><span class="elsevierStyleItalic">Chlamydia abortus</span> is abortigenic in mammals and is involved in miscarriage and stillbirths&#46;<a class="elsevierStyleCrossRefs" href="#bib0210"><span class="elsevierStyleSup">12&#44;13</span></a> Additionally&#44; <span class="elsevierStyleItalic">W&#46; chondrophila</span> has been associated with miscarriage and ectopic pregnancy in humans and animals&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">4&#44;12&#44;13</span></a><span class="elsevierStyleItalic">P&#46;</span><span class="elsevierStyleItalic">acanthamoebae</span> and <span class="elsevierStyleItalic">C&#46; pneumoniae</span> were initially identified as an emerging respiratory pathogens and they are now linked to adverse pregnancy aspects&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">4&#44;12&#44;13</span></a> Based on the literature and the novel results of this work&#44; <span class="elsevierStyleItalic">Simkania</span> appears to exist in other body parts in addition to the respiratory system&#46; Very recently&#44; <span class="elsevierStyleItalic">S&#46; negevensis</span> DNA was significantly detected in intestinal biopsies taken from patients diagnosed with Crohn&#39;s disease&#46;<a class="elsevierStyleCrossRef" href="#bib0205"><span class="elsevierStyleSup">11</span></a> Consistently&#44; the present study revealed the bacterial nucleic acids in another nonrespiratory site&#44; namely&#44; the female genitalia&#46; Intriguingly&#44; a significant link between the presence of <span class="elsevierStyleItalic">S</span>&#46; <span class="elsevierStyleItalic">negevensis</span> and the reproductive age of women was found&#46; The current work also provides evidence for a possible association of <span class="elsevierStyleItalic">S&#46; negevensis</span> with the vaginal inflammation&#46; However&#44; these results are still immature and need to be backed up by further investigations to verify the ability of this organism to exhibit tropism to the female genital tract and its pathogenic potential in the genitalia&#46;</p><p id="par0085" class="elsevierStylePara elsevierViewall">As mentioned above&#44; this study clearly demonstrated significant higher prevalence of <span class="elsevierStyleItalic">Simkania</span> DNA in vaginal swabs from women of the reproductive age&#46; One of the important correlates of the reproductive age is miscarriage&#46; Among risk factors known to associate with this spontaneous abortion are advancing maternal and paternal age&#44; genetic abnormalities&#44; immunological and implantation disorders&#44; uterine and endocrine abnormalities and lifestyle factors&#46;<a class="elsevierStyleCrossRef" href="#bib0245"><span class="elsevierStyleSup">19</span></a> Another risk factor linked to increasing chance of miscarriage is the infection with certain microbes&#46;<a class="elsevierStyleCrossRef" href="#bib0240"><span class="elsevierStyleSup">18</span></a> For instance&#44; a significant correlation between <span class="elsevierStyleItalic">Waddlia</span> seropositivity in women who experienced miscarriage was demonstrated&#44; compared to those who had no history of miscarriage&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">4&#44;12&#44;13</span></a> Comparably&#44; a correlation between <span class="elsevierStyleItalic">C&#46; trachomatis</span> infection and miscarriage was also documented&#46;<a class="elsevierStyleCrossRefs" href="#bib0210"><span class="elsevierStyleSup">12&#44;13&#44;18&#44;20</span></a> In addition&#44; many studies have linked <span class="elsevierStyleItalic">P&#46; acanthamoebae</span> with miscarriage in ruminants and humans&#46;<a class="elsevierStyleCrossRefs" href="#bib0210"><span class="elsevierStyleSup">12&#44;13&#44;21</span></a> Consistent with these studies&#44; the present research shows a possible involvement of <span class="elsevierStyleItalic">S&#46; negevensis</span> in miscarriage&#46; This result might be inconclusive due to the relatively small size of population examined&#46; Therefore&#44; future research is required to explore if <span class="elsevierStyleItalic">Simkania</span> is able to adversely affect the success of pregnancy in humans&#46;</p><p id="par0090" class="elsevierStylePara elsevierViewall">Possible sources of infection and modes of transmission for <span class="elsevierStyleItalic">Simkania</span>-like agents have been proposed&#46; <span class="elsevierStyleItalic">P&#46; acanthamoebae</span> and <span class="elsevierStyleItalic">C&#46; abortus</span> may have zoonotic nature&#44; leading to disorders in humans after contact with infected cattle&#46;<a class="elsevierStyleCrossRefs" href="#bib0210"><span class="elsevierStyleSup">12&#44;13&#44;22&#44;23</span></a> Cattle may contract <span class="elsevierStyleItalic">P&#46; acanthamoebae</span> from the animal drinking water&#46;<a class="elsevierStyleCrossRefs" href="#bib0170"><span class="elsevierStyleSup">4&#44;21</span></a> Moreover&#44; maternal-fetal transmission of this microbe has also been demonstrated and was postulated to be the result of zoonotic transmission&#46;<a class="elsevierStyleCrossRef" href="#bib0260"><span class="elsevierStyleSup">22</span></a><span class="elsevierStyleItalic">W&#46; chondrophila</span> could be acquired from well water or following contact with infected animals and may be transmitted by sexual intercourse&#46;<a class="elsevierStyleCrossRefs" href="#bib0215"><span class="elsevierStyleSup">13&#44;24</span></a> Similar to <span class="elsevierStyleItalic">Chlamydiales</span> members&#44; postulations on the source of infection&#47;reservoir of <span class="elsevierStyleItalic">Simkania</span> and its mode of transmission have been reported&#46; Different types of water&#44; such as drinking and environmental water and reclaimed wastewater&#44; were suggested to be a major source of the bacterium&#46;<a class="elsevierStyleCrossRefs" href="#bib0165"><span class="elsevierStyleSup">3&#44;14&#44;25&#8211;29</span></a> Ticks were also proposed as a reservoir and a source of infection&#46;<a class="elsevierStyleCrossRef" href="#bib0300"><span class="elsevierStyleSup">30</span></a> Additional modes of transmission could be human-to-human transmission through aerosols or droplets&#44; because of <span class="elsevierStyleItalic">Simkania</span> association with the respiratory system&#46;<a class="elsevierStyleCrossRef" href="#bib0165"><span class="elsevierStyleSup">3</span></a> With respect to the source of infection with <span class="elsevierStyleItalic">Simkania</span> in females tested positive in the current study&#44; zoonotic transmission is very unlikely&#44; as participants inhabit urban areas with no farms and no domestic animals are kept in or between houses&#46; The use of bloodstream for <span class="elsevierStyleItalic">Simkania</span> dissemination from potentially infected or colonized respiratory system or other body organs to the genitalia of females could also be excluded at this point&#44; as blood samples collected from 100 adult males and females were all negative for the bacterium &#40;data not shown&#41;&#46; It might be argued that the tested blood samples did not belong to the cohort of women tested here&#46; Nevertheless&#44; to resolve any dispute on this issue&#44; further work is required to conclusively clarify the exact role of blood and its cells in the spread of <span class="elsevierStyleItalic">Simkania</span> in the human body&#46; The possibility that the household water used for body washing was the source of infection is also ruled out&#46; This seems reasonable because none of the females older than 55 years was found positive&#46; It is&#44; therefore&#44; tempting to hypothesize that sexual transmission might be the source of microbial infection&#46; This could be supported by the fact that all females with confirmed <span class="elsevierStyleItalic">Simkania</span> DNA positivity were married at the time of the study and the incidence of <span class="elsevierStyleItalic">S</span>&#46; <span class="elsevierStyleItalic">negevensis</span> was found significantly associated with reproductive age&#46; This attractive speculation can be explored in future work by detecting and characterizing the bacterium in females along with their sexual partners&#46;</p><p id="par0095" class="elsevierStylePara elsevierViewall">In summary&#44; the present work provides the first evidence for the presence of <span class="elsevierStyleItalic">S&#46; negevensis</span> in the human female genitalia&#44; as confirmed by PCR technique&#46; This study serves as a foundation to verify the causal role of <span class="elsevierStyleItalic">Simkania</span>&#44; if present&#44; in genital system disorders and its association with adverse pregnancy outcomes in addition to its potential being a sexually transmitted agent&#46;</p></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Funding information</span><p id="par0100" class="elsevierStylePara elsevierViewall">This work was funded through a grant from the Deanship of Scientific Research&#44; the University of Jordan&#44; Amman&#44; Jordan&#46;</p></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0125">Conflicts of interest</span><p id="par0105" class="elsevierStylePara elsevierViewall">The authors have no conflicts of interest in relation to this work&#46;</p></span></span>"
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          "titulo" => "Materials and methods"
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          "identificador" => "xack651068"
          "titulo" => "Acknowledgements"
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    "fechaRecibido" => "2021-02-03"
    "fechaAceptado" => "2021-06-02"
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          "clase" => "keyword"
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            0 => "Females"
            1 => "Genitalia"
            2 => "Jordan"
            3 => "PCR"
            4 => "Prevalence"
            5 => "<span class="elsevierStyleItalic">Simkania</span>"
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          "titulo" => "Abbreviations"
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          "palabras" => array:9 [
            0 => "CAP"
            1 => "PCR"
            2 => "Cpn 60"
            3 => "IUD"
            4 => "CI"
            5 => "WHO"
            6 => "NGS"
            7 => "IVF"
            8 => "IUI"
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          "palabras" => array:6 [
            0 => "Mujeres"
            1 => "Genitales"
            2 => "Jordania"
            3 => "PCR"
            4 => "Prevalencia"
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    "resumen" => array:2 [
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        "titulo" => "Abstract"
        "resumen" => "<span id="abst0005" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0010">Introduction</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">Simkania negevensis</span> has been linked to some respiratory and non-respiratory diseases&#46; However&#44; there is still a serious lack of clinical investigations that attempt to determine possible body sites that could be inhabited by this microorganism and evaluate its true pathogenic capacity&#46; The goal of this study was to examine the potential presence of <span class="elsevierStyleItalic">Simkania</span> and its prevalence in the genital tract of human adult females&#46;</p></span> <span id="abst0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0015">Methods</span><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">Lower vaginal swabs from 169 Jordanian adult females who attended Obstetrics and Gynecology clinic were collected and tested for <span class="elsevierStyleItalic">Simkania</span> DNA by PCR method&#46;</p></span> <span id="abst0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0020">Results</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">The presence of bacterial nucleic acids was confirmed in the genital system of adult females with an overall prevalence of 24&#46;26&#37; &#40;41&#47;169&#41;&#46; Interestingly&#44; the positivity of <span class="elsevierStyleItalic">Simkania</span> DNA was significantly higher in women of reproductive age than females of non-reproductive age &#40;28&#46;03&#37; versus 10&#46;81&#37;&#59; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#8804;<span class="elsevierStyleHsp" style=""></span>0&#46;05&#41;&#46; Moreover&#44; the presence of <span class="elsevierStyleItalic">S&#46; negevensis</span> was evident in approximately 43&#37; of females suffering from vaginal itching and&#47;or abnormal discharge&#44; exhibiting about two-fold increase in the positivity rate compared to detection rates assessed for women who attended the clinic for routine checkup or menstruation problems&#46; However&#44; the current work failed to find any link between the bacterial agent and spontaneous abortion &#40;miscarriage&#41;&#46;</p></span> <span id="abst0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0025">Conclusions</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">This study showed for the first time the presence of <span class="elsevierStyleItalic">S&#46; negevensis</span> in the genitalia of human females&#46; These novel data could provide a basis to clarify the exact role of <span class="elsevierStyleItalic">S&#46; negevensis</span> in the female genitalia and its potential involvement in genital system disorders&#46;</p></span>"
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            "titulo" => "Introduction"
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            "titulo" => "Methods"
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            "identificador" => "abst0015"
            "titulo" => "Results"
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            "titulo" => "Conclusions"
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        "resumen" => "<span id="abst0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0035">Introducci&#243;n</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall"><span class="elsevierStyleItalic">Simkania negevensis</span> se ha asociado a algunas enfermedades respiratorias y no respiratorias&#46; Sin embargo&#44; todav&#237;a existe una importante carencia de investigaciones cl&#237;nicas para intentar determinar posibles lugares del cuerpo que puedan ser colonizados por este microorganismo&#44; y evaluar su aut&#233;ntica capacidad patog&#233;nica&#46; El objetivo de este estudio era examinar la potencial presencia de <span class="elsevierStyleItalic">Simkania</span> y su prevalencia en el tracto genital de mujeres adultas&#46;</p></span> <span id="abst0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">M&#233;todos</span><p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Se recogieron hisopos vaginales de 169 mujeres adultas de Jordania que acudieron a una cl&#237;nica de obstetricia y ginecolog&#237;a y se analizaron para detectar ADN de <span class="elsevierStyleItalic">Simkania</span> mediante PCR&#46;</p></span> <span id="abst0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Resultados</span><p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Se confirm&#243; la presencia de &#225;cidos nucleicos bacterianos en el aparato genital femenino adulto&#44; con una prevalencia global del 24&#44;26&#37; &#40;41&#47;169&#41;&#46; Una observaci&#243;n interesante fue que la positividad a ADN de <span class="elsevierStyleItalic">Simkania</span> fue significativamente superior en mujeres en edad reproductiva en comparaci&#243;n con las mujeres en edad no reproductiva &#40;28&#44;03 frente al 10&#44;81&#37;&#59; p<span class="elsevierStyleHsp" style=""></span>&#8804;<span class="elsevierStyleHsp" style=""></span>0&#44;05&#41;&#46; Adem&#225;s&#44; la presencia de <span class="elsevierStyleItalic">S&#46; negevensis</span> era evidente en aproximadamente el 43&#37; de las mujeres que sufr&#237;an picor vaginal y&#47;o flujo vaginal anormal&#44; mostrando un incremento cercano al doble en la tasa de positividad en comparaci&#243;n con las tasas de detecci&#243;n evaluadas para mujeres que acud&#237;an a la cl&#237;nica para revisiones de rutina o por problemas menstruales&#46; Sin embargo&#44; la investigaci&#243;n no logr&#243; encontrar una asociaci&#243;n entre el agente bacteriano y el aborto espont&#225;neo &#40;aborto natural&#41;&#46;</p></span> <span id="abst0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Conclusiones</span><p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Este estudio mostr&#243; por primera vez la presencia de <span class="elsevierStyleItalic">S&#46; negevensis</span> en los genitales femeninos humanos&#46; Estos nuevos datos pueden sentar una base para clarificar con precisi&#243;n el papel de <span class="elsevierStyleItalic">S&#46; negevensis</span> en los genitales femeninos humanos y su potencial implicaci&#243;n en trastornos del aparato genital&#46;</p></span>"
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                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black"><span class="elsevierStyleItalic">Simkania</span> DNA-positive females &#40;total number of females in the category&#41;&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">20&#8211;55&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">37&#46;78<span class="elsevierStyleHsp" style=""></span>&#177;<span class="elsevierStyleHsp" style=""></span>9&#46;79&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>20&#8211;29&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">20&#46;00&nbsp;\t\t\t\t\t\t\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>30&#8211;39&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t">13&#46;33&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span>&#8805;60&nbsp;\t\t\t\t\t\t\n
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                  \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">24&#46;85&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"><span class="elsevierStyleHsp" style=""></span>Widowed or divorced&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 &#40;4&#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;0&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">Birth control methods</span></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"><span class="elsevierStyleHsp" style=""></span>IUD<a class="elsevierStyleCrossRef" href="#tblfn0005"><span class="elsevierStyleSup">a</span></a>&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">11 &#40;44&#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">25&#46;00&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"><span class="elsevierStyleHsp" style=""></span>Oral contraception&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">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
                  \t\t\t\t\ttop\n
                  \t\t\t\t">40&#46;00&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"><span class="elsevierStyleHsp" style=""></span>Condoms&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">4 &#40;12&#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">33&#46;33&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"><span class="elsevierStyleHsp" style=""></span>None&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">20 &#40;98&#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">20&#46;41&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">Previous pregnancies &#40;births&#41;</span></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"><span class="elsevierStyleHsp" style=""></span>Yes&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">36 &#40;142&#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">25&#46;35&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"><span class="elsevierStyleHsp" style=""></span>No&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;27&#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">18&#46;52&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">Type of previous pregnancies</span></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"><span class="elsevierStyleHsp" style=""></span>No pregnancies at all&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;27&#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">18&#46;52&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"><span class="elsevierStyleHsp" style=""></span>Full-term delivery and no abortion history&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">21 &#40;92&#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">22&#46;83&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"><span class="elsevierStyleHsp" style=""></span>Spontaneous abortion &#40;miscarriage&#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">15 &#40;49&#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">30&#46;61&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"><span class="elsevierStyleHsp" style=""></span>Ectopic pregnancy&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 &#40;1&#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;00&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"><span class="elsevierStyleHsp" style=""></span>Stillbirth&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 &#40;0&#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;00&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">Number of previous pregnancies</span></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"><span class="elsevierStyleHsp" style=""></span>Zero&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;27&#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">18&#46;52&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"><span class="elsevierStyleHsp" style=""></span>One&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">4 &#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
                  \t\t\t\t\ttop\n
                  \t\t\t\t">20&#46;00&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"><span class="elsevierStyleHsp" style=""></span>Two&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">7 &#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
                  \t\t\t\t\ttop\n
                  \t\t\t\t">29&#46;17&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"><span class="elsevierStyleHsp" style=""></span>Three&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">8 &#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">28&#46;57&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"><span class="elsevierStyleHsp" style=""></span>Four&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">6 &#40;19&#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">31&#46;58&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"><span class="elsevierStyleHsp" style=""></span>&#8805;five&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">11 &#40;51&#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">21&#46;57&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleBold">Self-reported symptoms at attendance</span></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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">Vaginal itching and&#47;or discharge</span>&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">13 &#40;44&#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">29&#46;54&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="3" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">Menstrual and bleeding complaints&#58;</span></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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleHsp" style=""></span>Bleeding between periods and after intercourse&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">2 &#40;17&#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">11&#46;76&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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleHsp" style=""></span>Heavy cycles&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">7 &#40;18&#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">38&#46;89&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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleHsp" style=""></span>Irregular cycles&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">3 &#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">23&#46;08&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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleHsp" style=""></span>Missed &#40;infrequent&#41; cycles&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">2 &#40;11&#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">18&#46;18&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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">Mixed symptoms</span><a class="elsevierStyleCrossRef" href="#tblfn0010"><span class="elsevierStyleSup">b</span></a>&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 &#40;2&#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;00&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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">Other symptoms</span><a class="elsevierStyleCrossRef" href="#tblfn0015"><span class="elsevierStyleSup">c</span></a>&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 &#40;4&#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;00&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"><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">None</span>&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">14 &#40;60&#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">23&#46;33&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
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            1 => array:3 [
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                0 => """
                  <table border="0" frame="\n
                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Reasons for visit&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">No&#46; examined&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">No&#46; positive&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">Positivity &#37;&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th><th class="td" title="\n
                  \t\t\t\t\ttable-head\n
                  \t\t\t\t  " align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t" scope="col" style="border-bottom: 2px solid black">95&#37; CI<a class="elsevierStyleCrossRef" href="#tblfn0020"><span class="elsevierStyleSup">a</span></a>&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t\t\t</th></tr></thead><tbody title="tbody"><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"><span class="elsevierStyleItalic">Vaginal inflammation&#58; itching and&#47;or discharge</span>&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">30&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">13&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">43&#46;33&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;254&#8211;0&#46;625&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"><span class="elsevierStyleItalic">Menstrual and bleeding complaints</span>&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">57&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">13&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">22&#46;81&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;127&#8211;0&#46;358&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"><span class="elsevierStyleItalic">Routine checkup &#40;without symptoms&#41;</span>&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">42&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&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">23&#46;81&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;120&#8211;0&#46;394&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"><span class="elsevierStyleItalic">Multiple reasons &#40;vaginal itching and&#47;or discharge with menstrual complaint&#41;</span>&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">2&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&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;0&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="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="5" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleVsp" style="height:0.5px"></span></td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\n
                  \t\t\t\t  " colspan="5" align="left" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t"><span class="elsevierStyleItalic">Seeking pregnancy &#40;n</span><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">&#61;</span><span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">37&#41;</span> &#40;<span class="elsevierStyleItalic">IVF</span><a class="elsevierStyleCrossRef" href="#tblfn0025"><span class="elsevierStyleSup">b</span></a>&#44; <span class="elsevierStyleItalic">IUI</span><a class="elsevierStyleCrossRef" href="#tblfn0030"><span class="elsevierStyleSup">c</span></a>&#41;</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"><span class="elsevierStyleHsp" style=""></span>Without symptoms&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">19&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">4&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">21&#46;05&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;060&#8211;0&#46;455&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"><span class="elsevierStyleHsp" style=""></span>With coincidental vaginal discharge&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">11&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&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;0&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="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&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"><span class="elsevierStyleHsp" style=""></span>With coincidental menstrual and bleeding complaints&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&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">1&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">20&#46;00&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;005&#8211;0&#46;716&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"><span class="elsevierStyleHsp" style=""></span>With coincidental painful intercourse&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">1&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
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                  \t\t\t\t">0&nbsp;\t\t\t\t\t\t\n
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                  \t\t\t\t\ttable-entry\n
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                  \t\t\t\t">0&#46;0&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="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&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"><span class="elsevierStyleHsp" style=""></span>With coincidental blood in urine&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">0&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;0&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="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="\n
                  \t\t\t\t\ttable-entry\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">Other reasons</span><a class="elsevierStyleCrossRef" href="#tblfn0035"><span class="elsevierStyleSup">d</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  " align="char" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">1&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&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;0&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="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&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
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                  \t\t\t\t  " align="char" valign="\n
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                  \t\t\t\t">169&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">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">24&#46;26</span>&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="" valign="\n
                  \t\t\t\t\ttop\n
                  \t\t\t\t">&nbsp;\t\t\t\t\t\t\n
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                      "titulo" => "Infections with the <span class="elsevierStyleItalic">Chlamydia</span>-like microorganism <span class="elsevierStyleItalic">Simkania negevensis</span>&#44; a possible emerging pathogen"
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                      "doi" => "10.1016/s1286-4579(03)00188-6"
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                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/12941393"
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                      "titulo" => "High seroprevalence of <span class="elsevierStyleItalic">Simkania negevensis</span> in Jordan"
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                          "etal" => false
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Article information
ISSN: 2529993X
Original language: English
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