<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT03787160</url>
  </required_header>
  <id_info>
    <org_study_id>28-528 ex 15/16</org_study_id>
    <nct_id>NCT03787160</nct_id>
  </id_info>
  <brief_title>Microbiome and Volatile Organic Compounds in Patients With CDH</brief_title>
  <acronym>CDHVOCS</acronym>
  <official_title>Determining the Effect of Probiotics on Microbiome and Volatile Organic Compounds in Patients After Surgical Repair of Congenital Diaphragmatic Hernia</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Medical University of Graz</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
    <collaborator>
      <agency>University of Rostock</agency>
      <agency_class>Other</agency_class>
    </collaborator>
  </sponsors>
  <source>Medical University of Graz</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
    <is_fda_regulated_drug>No</is_fda_regulated_drug>
    <is_fda_regulated_device>No</is_fda_regulated_device>
  </oversight_info>
  <brief_summary>
    <textblock>
      Despite improved prenatal diagnostics and therapeutic possibilities, congenital diaphragmatic&#xD;
      hernia (CDH) represents a cross-disciplinary challenge. With an incidence of 1:2000-1:5000,&#xD;
      it is a common disease that effects centres of paediatrics and juvenile medicine. The&#xD;
      etiology is still unclear. Patients with this diagnosis are usually affected by other&#xD;
      comorbities such as failure to thrive, gastroesophageal reflux, funnel chest, etc. Depending&#xD;
      on the extent of CDH, a more or less pronounced lung hypoplasia with functional impairment&#xD;
      occurs. The health-relevant importance of the human microbiome is increasingly evident. While&#xD;
      it was previously particularly associated with the gastrointestinal tract, other systems such&#xD;
      as the pulmonary microbiome have become the focus of scientific interest.&#xD;
&#xD;
      Research into changes in the microbiome and volatile organic compounds (VOCs) could provide&#xD;
      new insights into the underlying mechanisms and therapeutic measures of this disease.&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Aim: The aim of this study is to evaluate children who have been enrolled on the basis of a&#xD;
      hernia, the pulmonary microbiome and the volatile organic compounds (VOCs) in the exhaled air&#xD;
      and to compare them with a control group which does not have a chronic or acute lung disease.&#xD;
      Furthermore, the lung function by spirometry, whole body plethysmography and &quot;multiple breath&#xD;
      washout&quot; procedures be precisely recorded and a sports medical examination, including&#xD;
      ergospirometry. In addition, any influence of probiotics on the pulmonary microbiome in&#xD;
      children with diaphragmatic hernia will be investigated.&#xD;
&#xD;
      Study design: This is a prospective study in children who were treated surgically in early&#xD;
      childhood for congenital diaphragmatic hernia. After an initial determination of the&#xD;
      microbiome and the composition of volatile organic substances in breathing air and feces and&#xD;
      a lung function measurement as well as sports medical examination, the study group receive a&#xD;
      probiotic (Omnibiotics 6, obtained from the Allergosan Institute, food supplements) for 3&#xD;
      months. The microbiome and VOCs are observed afterwards. The subjects are divided according&#xD;
      to age, gender, care with or without patch. In addition, a comparison with a control group&#xD;
      that does not show any chronic or acute lung disease is made according to age and gender of&#xD;
      the test group. In addition, in cooperation with the Division of Pediatric Pulmonology and&#xD;
      Allergology of the Department of Pediatrics and Adolescent Medicine at the Medical University&#xD;
      of Graz, the lung function of former diaphragmatic hernias patients (before administration of&#xD;
      probiotics) and a control group is to be measured. In cooperation with the Medical Center of&#xD;
      Sports Medicine the investigator also carry out a sports medical examination including&#xD;
      ergospirometry. The duration of the study is set at 12 months.&#xD;
&#xD;
      Study participants: Study group: Children between 6-16 years of age, who are enrolled at the&#xD;
      Department of Pediatric and Adolescent Surgery at the Medical University of Graz between 2000&#xD;
      and 2010 due to a congenital diaphragmatic hernia have received an operative closure with or&#xD;
      without patch. Control group: Children between 6-16 years who do not have pulmonary disease.&#xD;
      Recruited from the outpatient area at the Department of Pediatric and Adolescent Surgery at&#xD;
      the Medical University of Graz. The aim is to establish contact with the above-mentioned&#xD;
      patients and their parents and to achieve willingness to participate in this study by means&#xD;
      of an information letter. The control group should come from the area of outpatient area in&#xD;
      the Department of Pediatric and Adolescent Surgery, Medical University of Graz, after&#xD;
      appropriate information and possible consent.&#xD;
&#xD;
      Microbiome analysis before treatment with a probiotic: Collection of sample material - in&#xD;
      this case sputum - from the deep respiratory tract by induced sputum after inhalation of&#xD;
      hypersaline saline solution with resulting provocation of cough. The sample is then&#xD;
      deep-frozen. The Microbiome measurement is performed as a comparative 16S rDNA-based profile&#xD;
      via chip-based next-generation sequencing as already published, analyzed using SnowMAn, Qiime&#xD;
      and MOTHUR as well as the own &quot;R&quot;-based software. A sequencing depth of 5,000-10,000 reads&#xD;
      per sample.&#xD;
&#xD;
      VOCs analysis before treatment with a probiotic: I) Taking of the exhaled gas samples: One&#xD;
      sample from inspiration and two from expiration is taken from each subject (n = 3). For&#xD;
      sampling investigator use an automatic sampling system that is directly connected to a&#xD;
      capnometer. This system contains a so-called needle-trap microextraction (NTME) as a&#xD;
      microextraction technique and meets the requirements of an optimal sampling on currently&#xD;
      technically highest level. The exhaled gas samples obtained in this way are then sent to our&#xD;
      cooperation partner, to the Institute for Breathing Gas Analysis at the University of Rostock&#xD;
      for analysis. II) Analysis of exhaled gas samples: There, the exhaled gas samples are&#xD;
      thermally transferred into the inert carrier gas stream (He) in an injector of a&#xD;
      chromatograph. The substances are assigned according to their retention time in the&#xD;
      chromatogram and their mass spectrum. Unknown Compounds in the exhaled gas are identified by&#xD;
      comparison with a reference database based on the mass spectrum. Vital and laboratory data,&#xD;
      as well as microbiological information, are taken from the patients' findings. III)&#xD;
      Identification of biomarkers of exhaled gas samples: From the results of the patient&#xD;
      measurements, those substances and substance concentrations are determined which are specific&#xD;
      for study group 1 and group 2, i.e. compounds which are not present in the comparison group&#xD;
      or only in significantly lower or higher concentrations. The selected volatile markers, as&#xD;
      well as any volatile contaminants that may have been detected in the environment, are stored&#xD;
      in an analytical reference database and, after elimination of the contamination, bundled into&#xD;
      possibly disease-specific marker profiles. IV) Analysis of fecal samples: The fecal samples&#xD;
      are also sent to the Institute for Breathing Gas Analysis of the University of Rostock for&#xD;
      analysis and analyzed there after appropriate preconcentration by solid phase microextraction&#xD;
      (SPME). V) Identification of biomarkers of fecal samples: This is done in analogy to the&#xD;
      exhaled gas samples.&#xD;
&#xD;
      Lung function measurement before treatment with a probiotic: Measurement of lung function&#xD;
      using spirometry and body plethysmography (Fa Jäger spirometer and body plethysmograph) and&#xD;
      nitrogen washout process (N2-multiple breath washout, System Exhalyzer D and Spiroware 3.1,&#xD;
      Eco Medics AG, Duernten, Switzerland). Spirometry and body plethysmography are performed&#xD;
      according to published ERS/ATS Standards. The &quot;multiple breath washout&quot; method is performed&#xD;
      under resting breathing and detects the Ventilation (in)homogeneity at the level of the&#xD;
      functional residual capacity (= FRC = that lung volume that is still in the lungs after a&#xD;
      calm spontaneous exhalation is left behind). The system consists of a flow meter, a fast&#xD;
      analyzing gas measuring system, a gas administration system and the corresponding Computer&#xD;
      analysis software. As a &quot;foreign gas&quot; it will be use 78% of the gas in air occurring nitrogen&#xD;
      (N2). A flow-volume measurement is performed via an Ultrasonic flowmeter performed directly&#xD;
      in the inhalation and exhalation flow of the test person/patient and via a laser O2 sensor&#xD;
      using the side current measurement method and an infrared CO2 sensor in the main current&#xD;
      measuring procedure (= directly in the patient's respiratory flow) the respective gas&#xD;
      concentration. The N2 component is then indirectly measured via the O2 and CO2 concentration&#xD;
      (N2 = 1 - O2 - CO2). During calm spontaneous breathing, the Patient on a snorkel mouthpiece&#xD;
      via a bacterial filter through the flowmeter 100% oxygen and thus &quot;washes&quot; N2 out&#xD;
      (&quot;N2-multiple breath washout&quot;). In doing so, the flow-volume curve of spontaneous breathing&#xD;
      &quot;online&quot; is displayed on the screen and the measurement at Reaching a 1/40 (= 2.5%) of the&#xD;
      initial nitrogen concentration is completed. After that, wait the subject is safe in the&#xD;
      length of twice the duration of the measurement around the oxygen of exhaling. This is&#xD;
      followed by the next measurement. A total of 3 measurements whose mean value serves as a&#xD;
      result. The so-called &quot;lung clearance index&quot;, which indicates the number of functional&#xD;
      residual capacity lung volumes, which can be used to reduce the initial nitrogen&#xD;
      concentration to a 1/40 after oxygenation was required. (LCI = quotient between exhaled&#xD;
      volume and FRC). It expresses how long it takes for the inhaled gas (in our case the&#xD;
      physiologically occurring nitrogen in the air) through inhalation of 100% oxygen. For healthy&#xD;
      persons, this value is on average 7 and is significantly higher in lung patients. Further&#xD;
      measuring parameters, which makes a statement about the peripheral airways proximal to the&#xD;
      terminal bronchioles on the one hand and via the more distal azine airways on the other hand&#xD;
      are calculated.&#xD;
&#xD;
      Sports medical examination: To exclude contraindications for ergometry, a 12-channel resting&#xD;
      ECG and a resting blood pressure measurement (CombynTM Function &amp; Spaces ECG, Academic&#xD;
      Technologies) are performed at the beginning of the examination. After the anthropometric&#xD;
      data (height, weight, BMI) have been collected, the muscle mass and fat mass are determined&#xD;
      by multi-frequency impedance measurement in six body segments (CombynTM Function &amp; Spaces&#xD;
      ECG, Academic Technologies). The lung function is measured by means of small spirometry at&#xD;
      rest and after exercise (Spirometer Oxycon Pro, Reiner). In order to determine&#xD;
      cardiopulmonary performance, ergospirometry is performed on a bicycle (Excalibur Sport&#xD;
      ergometer, Lode company; Oxycon Pro spiroergometry unit, Reiner company) with a gradual&#xD;
      increase in stress up to subjective exhaustion. The evaluation of these data allows&#xD;
      conclusions to be drawn about the performance-limiting system (cardiovascular system, lungs,&#xD;
      musculature) in addition to the determination of aerobic performance.&#xD;
&#xD;
      Microbiome/VOCs analysis after treatment with a probiotic: After sampling for microbiome and&#xD;
      VOC analysis and carrying out lung function measurements and sports medical examination, the&#xD;
      participants in the study group will take a probiotic (Omnibiotic 6, purchased from the&#xD;
      Allergosan Institute, dietary supplement) for a period of 3 months. Immediately afterwards&#xD;
      and another month later, measurements of the pulmonary microbiome and the VOCs in the breath&#xD;
      are taken.&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date type="Actual">March 22, 2018</start_date>
  <completion_date type="Actual">October 1, 2019</completion_date>
  <primary_completion_date type="Actual">October 1, 2019</primary_completion_date>
  <phase>N/A</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>Randomized</allocation>
    <intervention_model>Parallel Assignment</intervention_model>
    <primary_purpose>Basic Science</primary_purpose>
    <masking>Single (Investigator)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Analysis of the pulmonary microbiome in the sputum of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>OTUs (Operational Taxonomic Units) will be visualized as OTU tables, bar charts and PCOA (Principal Coordinates Analysis) plots using the Qiime core microbiome script. For the different groups alpha-diversity (Chao 1 index, Shannon Index etc.) will be compared. Additionally, we will compare beta-diversity by Adonis test. Relative abundances of the bacteria at the different levels (phylum to genus) will be compared between the groups by using Kruskal Wallis Test.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of VOCs in the respiratory air by needle-trap microextraction (NTME) and stool by solid phase microextraction (SPME) of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>From the results of the patient measurements, those substances and substance concentrations are determined which are specific for study group 1 and group 2, i.e. compounds which are not present in the comparison group or only in significantly lower or higher concentrations. The selected volatile markers, as well as any volatile contaminants that may have been detected in the environment, are stored in an analytical reference database and, after elimination of the contamination, bundled into possibly disease-specific marker profiles. The VOCs are recorded and displayed in the following order. The unit in which the VOCs are measured is pars per billion (ppb).&#xD;
Class (for example carbons) VOCs (ppb) CDHV1 (congenital diaphragmatic hernia group visit 1) CDHK (congenital diaphragmatic hernia - control group visit 1) CDHV2 (congenital diaphragmatic hernia group visit 2) CDHV3 (congenital diaphragmatic hernia group visit 3) p-value</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the lung function: Lung clearance index (LCI) is derived from multiple breath washout tests of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>The LCI is about 7 (range from 6.45-7.78) for healthy individuals and is a number without a unit.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the lung function: Forced expiratory volume in one second (FEV1) measured with spirometry of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>The FEV1 is the forced expiratory volume within the first second (liter/second), generated by a maximal voluntary exhalation after maximum inspiration before, usually described as the Tiffeneau-Index in % of FVC (FEV1/FVC).</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Resting ECG of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>A Resting ECG recording the resting heart rate, the rhythm, the PQ duration, the width and height of the QRS complex, the QT duration, and the ST segment is recorded.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Systolic and diastolic blood pressures of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Noninvasiv systolic and diastolic blood pressures are assessed (Unit: mmHg).</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Body height of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Body height is measured in cm.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Body weight of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Body weight is measured in kg.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Body mass index (BMI) of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Body mass index is calculated in kg body weight/body height².</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Muscle mass of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Muscle mass is specified in kg/body height².</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Body fat of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Body fat is specified in percent of body weight.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Aerobic performance of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Aerobic performance is specified in percent of normal values of the Austrian cardiological society.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Maximal oxygen uptake of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Measurements by spiroergometry: Maximal oxygen uptake in ml/kg/min.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Ventilation of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Measurements by spiroergometry: Ventilation in liter/min.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Oxygen pulse of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Measurements by spiroergometry: Oxygen pulse in ml/beats per minute.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Oxygen uptake of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Respiratory exchange ratio = oxygen uptake in ml/carbon dioxide release in ml.</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Analysis of the cardiopulmonary capacity: Breathing reserve of CDH group versus control group.</measure>
    <time_frame>12 months</time_frame>
    <description>Unit: Percent of FEV1 x 35.</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>Alterations of pulmonary microbiome after probiotic treatment for a period of 3 months in patients with CDH.</measure>
    <time_frame>12 months</time_frame>
    <description>After sampling for microbiome and VOC analysis and carrying out lung function measurements and sports medical examination, the participants in the study group will take a probiotic (Omnibiotic 6, purchased from the Allergosan Institute, dietary supplement) for a period of 3 months. Immediately afterwards and another month later, measurements of the pulmonary microbiome are taken.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Alterations of VOCs in the respiratory air after probiotic treatment for a period of 3 months in patients with CDH.</measure>
    <time_frame>12 months</time_frame>
    <description>After sampling for microbiome and VOC analysis and carrying out lung function measurements and sports medical examination, the participants in the study group will take a probiotic (Omnibiotic 6, purchased from the Allergosan Institute, dietary supplement) for a period of 3 months. Immediately afterwards and another month later, measurements of the VOCs in the breath are taken.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Forced expiratory flow (FEF25-75).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography and spirometry: FEF25-75 = Forced expiratory flow 25-75% vital capacity (= MMEF), Unit: l/s.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Forced expiratory flow (FEF25).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography and spirometry: FEF25 = Forced expiratory flow at the time, when 75% of the vital capacity is exhaled (MEF25), Unit: l/s.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Forced expiratory flow (FEF50).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography and spirometry: FEF50 = Forced expiratory flow at 50% of the exhaled vital capacity (= MEF50), Unit: l/s.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Forced expiratory volume (FEV1).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography and spirometry: FEV1 = Forced expiratory volume in 1 s, Unit: l.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Tiffeneau-Index (FEV1%FVC).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography and spirometry: FEV1%FVC = Tiffeneau-Index, described in % of the forced vital capacity, Unit: %.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Functional residual capacity (FRC).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography and spirometry: FRC = Functional residual capacity, Unit: l.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Through &quot;multiple breath washout&quot; acquired FRC (RC(MBW)).</measure>
    <time_frame>12 months</time_frame>
    <description>RC(MBW): Through &quot;multiple breath washout&quot; acquired FRC, Unit: l.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Through body plethysmography acquired FRC (RC(pleth)).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography: RC(pleth): Through body plethysmography acquired FRC (= ITGV, intra thoracic gas volume), Unit: l.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Forced vital capacity (FVC).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by spirometry: FVC = Forced vital capacity, Vital capacity acquired through a forced exhaled manoeuvre, Unit: l.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Intrathoracic gasvolume (ITGV).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by body plethysmography: ITGV = Intrathoracic gasvolume (= FRC(pleth)), Unit: l.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Max. expiratory flow (MEF25).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by spirometry: MEF25 = Max. expiratory flow when 75% of the vital capacity is exhaled (= FEF25), Unit: l/s.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Max. expiratory flow (MEF50).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by spirometry: MEF50 = Max. expiratory flow when 50% of the vital capacity is exhaled (= FEF50), Unit: l/s.</description>
  </secondary_outcome>
  <secondary_outcome>
    <measure>Analysis of the lung function: Max. expiratory flow (MMEF).</measure>
    <time_frame>12 months</time_frame>
    <description>Measured by spirometry: MMEF = Max. expiratory flow (= FEF25-75), Unit: l/s.</description>
  </secondary_outcome>
  <number_of_arms>2</number_of_arms>
  <enrollment type="Actual">18</enrollment>
  <condition>Congenital Diaphragmatic Hernia</condition>
  <arm_group>
    <arm_group_label>CDH Group</arm_group_label>
    <arm_group_type>Active Comparator</arm_group_type>
    <description>10 patients after surgical closure of CDH will undergo VOC profile analysis (2 breath samples) (initial VOC), fecal sampling for 16S rDNA based pyrosequencing (initial fecal microbiome) and deep induced sputum sampling for 16S rDNA pyrosequencing (initial pulmonary microbiome), bicycle spiroergometry to determine the maximum oxygen uptake (maximum oxygen uptake), body plethysmography, spirometry and N2-multiple breath washout testing to determine the functional residual capacity (functional residual capacity). Thereafter patients will receive probiotic treatment with OmniBiotic6 (R) (Allergosan, Graz, Austria) 1 sachet daily for 3 months (probiotic treatment). Three months after discontinuing probiotic treatment VOC testing (VOC probiotics), fecal microbiome sampling (fecal microbiome probiotics) and deep induced sputum testing (pulmonary microbiome probiotics) will be repeated and compared to the results of the initial tests.</description>
  </arm_group>
  <arm_group>
    <arm_group_label>Control Group</arm_group_label>
    <arm_group_type>Other</arm_group_type>
    <description>10 healthy controls (age and sex matched) will undergo VOC profile analysis (2 breath samples) (initial VOC), fecal sampling for 16S rDNA based pyrosequencing (initial fecal microbiome) and deep induced sputum sampling for 16S rDNA pyrosequencing (initial pulmonary microbiome), bicycle spiroergometry to determine the maximum oxygen uptake (maximum oxygen uptake), body plethysmography, spirometry and N2-multiple breath washout testing to determine the functional residual capacity (functional residual capacity).</description>
  </arm_group>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>initial VOC</intervention_name>
    <description>Difference in VOC profile between patients with CDH and healthy controls (2 samples per patient will be obtained after obtaining informed consent).</description>
    <arm_group_label>CDH Group</arm_group_label>
    <arm_group_label>Control Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>initial fecal microbiome</intervention_name>
    <description>Difference of alpha and beta diversity and relative fecal bacterial abundance between patients with CDH and healthy controls (1 stool sample will be taken per patient after obtaining informed consent)</description>
    <arm_group_label>CDH Group</arm_group_label>
    <arm_group_label>Control Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>initial pulmonary microbiome</intervention_name>
    <description>Difference of alpha and beta diversity and relative pulmonary bacterial abundance between patients with CDH and healthy controls (1 deep induced sputum sample will be taken per patient after obtaining informed consent)</description>
    <arm_group_label>CDH Group</arm_group_label>
    <arm_group_label>Control Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>Maximum oxygen uptake</intervention_name>
    <description>Comparison of the maximum oxygen uptake (corrected for body weight and gender) as determined by bicycle spiroergometry between patients with CDH and healthy controls</description>
    <arm_group_label>CDH Group</arm_group_label>
    <arm_group_label>Control Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>Functional residual capacity</intervention_name>
    <description>FRC will be determined by spirometry, bodyplethysmography and N2-breath wash out method. FRC will be compared between patients after CDH and healthy controls.</description>
    <arm_group_label>CDH Group</arm_group_label>
    <arm_group_label>Control Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Dietary Supplement</intervention_type>
    <intervention_name>Probiotic treatment</intervention_name>
    <description>CDH patients will receive OmniBiotic 6(R) (Allergosan, Graz, Austria) probiotic supplementation 1 sachet daily for 3 months.</description>
    <arm_group_label>CDH Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>VOC probiotic</intervention_name>
    <description>Determination of the VOC profile 3 months after discontinuing probiotic treatment. Comparison to the profiles before the treatment.</description>
    <arm_group_label>CDH Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>Fecal microbiome probiotic</intervention_name>
    <description>Determination of the fecal microbiome from 1 sample per patient (alpha and beta diversity, relative bacterial abundance at the genus level) 3 months after discontinuing probiotic treatment. Comparison to the profiles before the treatment.</description>
    <arm_group_label>CDH Group</arm_group_label>
  </intervention>
  <intervention>
    <intervention_type>Diagnostic Test</intervention_type>
    <intervention_name>Pulmonary microbiome probiotic</intervention_name>
    <description>Determination of the fecal microbiome from 1 deep induced sputum sample per patient (alpha and beta diversity, relative bacterial abundance at the genus level) 3 months after discontinuing probiotic treatment. Comparison to the profiles before the treatment.</description>
    <arm_group_label>CDH Group</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Age from 6-16 years&#xD;
&#xD;
          -  Age 0-6 months at time of CDH-OP (except control group)&#xD;
&#xD;
          -  reliable diagnosis of congenital diaphragmatic hernia (except control group)&#xD;
&#xD;
          -  surgical occlusion with patch (except control group)&#xD;
&#xD;
          -  surgical occlusion without patch (except control group)&#xD;
&#xD;
          -  given approval&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  chronic pulmonary diseases&#xD;
&#xD;
          -  Infection within 4 weeks before the test date&#xD;
&#xD;
          -  unaccepted consent&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>6 Years</minimum_age>
    <maximum_age>16 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Till Holger, MD</last_name>
    <role>Study Director</role>
    <affiliation>Department of Pediatric and Adolescent Surgery, Medical University of Graz</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Ernst Eber, MD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Department of Pediatric and Adolescent Medicine, Medical University of Graz</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Gert Warncke, MD</last_name>
    <role>Principal Investigator</role>
    <affiliation>Department of Pediatric and Adolescent Surgery, Medical University of Graz</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Department of Department of Pediatric and Adolescent Surgery, Medical University of Graz</name>
      <address>
        <city>Graz</city>
        <state>Steiermark</state>
        <zip>8036</zip>
        <country>Austria</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>Austria</country>
  </location_countries>
  <reference>
    <citation>Kotecha S, Barbato A, Bush A, Claus F, Davenport M, Delacourt C, Deprest J, Eber E, Frenckner B, Greenough A, Nicholson AG, Anton-Pacheco JL, Midulla F. Congenital diaphragmatic hernia. Eur Respir J. 2012 Apr;39(4):820-9. doi: 10.1183/09031936.00066511. Epub 2011 Oct 27.</citation>
    <PMID>22034651</PMID>
  </reference>
  <reference>
    <citation>Tracy M, Cogen J, Hoffman LR. The pediatric microbiome and the lung. Curr Opin Pediatr. 2015 Jun;27(3):348-55. doi: 10.1097/MOP.0000000000000212.</citation>
    <PMID>25888147</PMID>
  </reference>
  <reference>
    <citation>Caverly LJ, Zhao J, LiPuma JJ. Cystic fibrosis lung microbiome: opportunities to reconsider management of airway infection. Pediatr Pulmonol. 2015 Oct;50 Suppl 40:S31-8. doi: 10.1002/ppul.23243.</citation>
    <PMID>26335953</PMID>
  </reference>
  <reference>
    <citation>Bergmann A, Trefz P, Fischer S, Klepik K, Walter G, Steffens M, Ziller M, Schubert JK, Reinhold P, Kohler H, Miekisch W. In Vivo Volatile Organic Compound Signatures of Mycobacterium avium subsp. paratuberculosis. PLoS One. 2015 Apr 27;10(4):e0123980. doi: 10.1371/journal.pone.0123980. eCollection 2015.</citation>
    <PMID>25915653</PMID>
  </reference>
  <reference>
    <citation>Fischer S, Trefz P, Bergmann A, Steffens M, Ziller M, Miekisch W, Schubert JS, Kohler H, Reinhold P. Physiological variability in volatile organic compounds (VOCs) in exhaled breath and released from faeces due to nutrition and somatic growth in a standardized caprine animal model. J Breath Res. 2015 May 14;9(2):027108. doi: 10.1088/1752-7155/9/2/027108.</citation>
    <PMID>25971714</PMID>
  </reference>
  <reference>
    <citation>Pereira J, Porto-Figueira P, Cavaco C, Taunk K, Rapole S, Dhakne R, Nagarajaram H, Camara JS. Breath analysis as a potential and non-invasive frontier in disease diagnosis: an overview. Metabolites. 2015 Jan 9;5(1):3-55. doi: 10.3390/metabo5010003.</citation>
    <PMID>25584743</PMID>
  </reference>
  <reference>
    <citation>Amann A, Costello Bde L, Miekisch W, Schubert J, Buszewski B, Pleil J, Ratcliffe N, Risby T. The human volatilome: volatile organic compounds (VOCs) in exhaled breath, skin emanations, urine, feces and saliva. J Breath Res. 2014 Sep;8(3):034001. doi: 10.1088/1752-7155/8/3/034001. Epub 2014 Jun 19.</citation>
    <PMID>24946087</PMID>
  </reference>
  <reference>
    <citation>Barker M, Hengst M, Schmid J, Buers HJ, Mittermaier B, Klemp D, Koppmann R. Volatile organic compounds in the exhaled breath of young patients with cystic fibrosis. Eur Respir J. 2006 May;27(5):929-36. doi: 10.1183/09031936.06.00085105. Epub 2006 Feb 2.</citation>
    <PMID>16455833</PMID>
  </reference>
  <reference>
    <citation>Miekisch W, Schubert JK, Noeldge-Schomburg GF. Diagnostic potential of breath analysis--focus on volatile organic compounds. Clin Chim Acta. 2004 Sep;347(1-2):25-39. doi: 10.1016/j.cccn.2004.04.023.</citation>
    <PMID>15313139</PMID>
  </reference>
  <reference>
    <citation>Forton J. Induced sputum in young healthy children with cystic fibrosis. Paediatr Respir Rev. 2015 Oct;16 Suppl 1:6-8. doi: 10.1016/j.prrv.2015.07.007. Epub 2015 Sep 26.</citation>
    <PMID>26410282</PMID>
  </reference>
  <reference>
    <citation>Planting NS, Visser GL, Nicol MP, Workman L, Isaacs W, Zar HJ. Safety and efficacy of induced sputum in young children hospitalised with suspected pulmonary tuberculosis. Int J Tuberc Lung Dis. 2014 Jan;18(1):8-12. doi: 10.5588/ijtld.13.0132.</citation>
    <PMID>24365546</PMID>
  </reference>
  <reference>
    <citation>Marri PR, Stern DA, Wright AL, Billheimer D, Martinez FD. Asthma-associated differences in microbial composition of induced sputum. J Allergy Clin Immunol. 2013 Feb;131(2):346-52.e1-3. doi: 10.1016/j.jaci.2012.11.013. Epub 2012 Dec 23.</citation>
    <PMID>23265859</PMID>
  </reference>
  <reference>
    <citation>Gorkiewicz G, Thallinger GG, Trajanoski S, Lackner S, Stocker G, Hinterleitner T, Gully C, Hogenauer C. Alterations in the colonic microbiota in response to osmotic diarrhea. PLoS One. 2013;8(2):e55817. doi: 10.1371/journal.pone.0055817. Epub 2013 Feb 8.</citation>
    <PMID>23409050</PMID>
  </reference>
  <reference>
    <citation>Trefz P, Rosner L, Hein D, Schubert JK, Miekisch W. Evaluation of needle trap micro-extraction and automatic alveolar sampling for point-of-care breath analysis. Anal Bioanal Chem. 2013 Apr;405(10):3105-15. doi: 10.1007/s00216-013-6781-9. Epub 2013 Feb 7. Erratum In: Anal Bioanal Chem. 2013 Jun;405(16):5617.</citation>
    <PMID>23388692</PMID>
  </reference>
  <reference>
    <citation>Miller MR, Hankinson J, Brusasco V, Burgos F, Casaburi R, Coates A, Crapo R, Enright P, van der Grinten CP, Gustafsson P, Jensen R, Johnson DC, MacIntyre N, McKay R, Navajas D, Pedersen OF, Pellegrino R, Viegi G, Wanger J; ATS/ERS Task Force. Standardisation of spirometry. Eur Respir J. 2005 Aug;26(2):319-38. doi: 10.1183/09031936.05.00034805. No abstract available.</citation>
    <PMID>16055882</PMID>
  </reference>
  <reference>
    <citation>Wanger J, Clausen JL, Coates A, Pedersen OF, Brusasco V, Burgos F, Casaburi R, Crapo R, Enright P, van der Grinten CP, Gustafsson P, Hankinson J, Jensen R, Johnson D, Macintyre N, McKay R, Miller MR, Navajas D, Pellegrino R, Viegi G. Standardisation of the measurement of lung volumes. Eur Respir J. 2005 Sep;26(3):511-22. doi: 10.1183/09031936.05.00035005. No abstract available.</citation>
    <PMID>16135736</PMID>
  </reference>
  <verification_date>April 2020</verification_date>
  <study_first_submitted>July 10, 2018</study_first_submitted>
  <study_first_submitted_qc>December 21, 2018</study_first_submitted_qc>
  <study_first_posted type="Actual">December 26, 2018</study_first_posted>
  <last_update_submitted>April 26, 2020</last_update_submitted>
  <last_update_submitted_qc>April 26, 2020</last_update_submitted_qc>
  <last_update_posted type="Actual">April 28, 2020</last_update_posted>
  <responsible_party>
    <responsible_party_type>Principal Investigator</responsible_party_type>
    <investigator_affiliation>Medical University of Graz</investigator_affiliation>
    <investigator_full_name>Warncke Gert, MD</investigator_full_name>
    <investigator_title>Principal Investigator</investigator_title>
  </responsible_party>
  <keyword>congenital diaphragmatic hernia</keyword>
  <keyword>volatile organic compounds</keyword>
  <keyword>microbiome</keyword>
  <keyword>pulmonary function</keyword>
  <keyword>multiple breath washout</keyword>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Hernias, Diaphragmatic, Congenital</mesh_term>
    <mesh_term>Hernia</mesh_term>
    <mesh_term>Hernia, Diaphragmatic</mesh_term>
  </condition_browse>
  <patient_data>
    <sharing_ipd>No</sharing_ipd>
  </patient_data>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT03787160
org study id: 28-528 ex 15/16
nct id: NCT03787160
lead sponsor: 
collaborator: 
has dmc: No
is fda regulated drug: No
is fda regulated device: No
Despite improved prenatal diagnostics and therapeutic possibilities, congenital diaphragmatic
      hernia (CDH) represents a cross-disciplinary challenge. With an incidence of 1:2000-1:5000,
      it is a common disease that effects centres of paediatrics and juvenile medicine. The
      etiology is still unclear. Patients with this diagnosis are usually affected by other
      comorbities such as failure to thrive, gastroesophageal reflux, funnel chest, etc. Depending
      on the extent of CDH, a more or less pronounced lung hypoplasia with functional impairment
      occurs. The health-relevant importance of the human microbiome is increasingly evident. While
      it was previously particularly associated with the gastrointestinal tract, other systems such
      as the pulmonary microbiome have become the focus of scientific interest.

      Research into changes in the microbiome and volatile organic compounds (VOCs) could provide
      new insights into the underlying mechanisms and therapeutic measures of this disease.
Aim: The aim of this study is to evaluate children who have been enrolled on the basis of a
      hernia, the pulmonary microbiome and the volatile organic compounds (VOCs) in the exhaled air
      and to compare them with a control group which does not have a chronic or acute lung disease.
      Furthermore, the lung function by spirometry, whole body plethysmography and "multiple breath
      washout" procedures be precisely recorded and a sports medical examination, including
      ergospirometry. In addition, any influence of probiotics on the pulmonary microbiome in
      children with diaphragmatic hernia will be investigated.

      Study design: This is a prospective study in children who were treated surgically in early
      childhood for congenital diaphragmatic hernia. After an initial determination of the
      microbiome and the composition of volatile organic substances in breathing air and feces and
      a lung function measurement as well as sports medical examination, the study group receive a
      probiotic (Omnibiotics 6, obtained from the Allergosan Institute, food supplements) for 3
      months. The microbiome and VOCs are observed afterwards. The subjects are divided according
      to age, gender, care with or without patch. In addition, a comparison with a control group
      that does not show any chronic or acute lung disease is made according to age and gender of
      the test group. In addition, in cooperation with the Division of Pediatric Pulmonology and
      Allergology of the Department of Pediatrics and Adolescent Medicine at the Medical University
      of Graz, the lung function of former diaphragmatic hernias patients (before administration of
      probiotics) and a control group is to be measured. In cooperation with the Medical Center of
      Sports Medicine the investigator also carry out a sports medical examination including
      ergospirometry. The duration of the study is set at 12 months.

      Study participants: Study group: Children between 6-16 years of age, who are enrolled at the
      Department of Pediatric and Adolescent Surgery at the Medical University of Graz between 2000
      and 2010 due to a congenital diaphragmatic hernia have received an operative closure with or
      without patch. Control group: Children between 6-16 years who do not have pulmonary disease.
      Recruited from the outpatient area at the Department of Pediatric and Adolescent Surgery at
      the Medical University of Graz. The aim is to establish contact with the above-mentioned
      patients and their parents and to achieve willingness to participate in this study by means
      of an information letter. The control group should come from the area of outpatient area in
      the Department of Pediatric and Adolescent Surgery, Medical University of Graz, after
      appropriate information and possible consent.

      Microbiome analysis before treatment with a probiotic: Collection of sample material - in
      this case sputum - from the deep respiratory tract by induced sputum after inhalation of
      hypersaline saline solution with resulting provocation of cough. The sample is then
      deep-frozen. The Microbiome measurement is performed as a comparative 16S rDNA-based profile
      via chip-based next-generation sequencing as already published, analyzed using SnowMAn, Qiime
      and MOTHUR as well as the own "R"-based software. A sequencing depth of 5,000-10,000 reads
      per sample.

      VOCs analysis before treatment with a probiotic: I) Taking of the exhaled gas samples: One
      sample from inspiration and two from expiration is taken from each subject (n = 3). For
      sampling investigator use an automatic sampling system that is directly connected to a
      capnometer. This system contains a so-called needle-trap microextraction (NTME) as a
      microextraction technique and meets the requirements of an optimal sampling on currently
      technically highest level. The exhaled gas samples obtained in this way are then sent to our
      cooperation partner, to the Institute for Breathing Gas Analysis at the University of Rostock
      for analysis. II) Analysis of exhaled gas samples: There, the exhaled gas samples are
      thermally transferred into the inert carrier gas stream (He) in an injector of a
      chromatograph. The substances are assigned according to their retention time in the
      chromatogram and their mass spectrum. Unknown Compounds in the exhaled gas are identified by
      comparison with a reference database based on the mass spectrum. Vital and laboratory data,
      as well as microbiological information, are taken from the patients' findings. III)
      Identification of biomarkers of exhaled gas samples: From the results of the patient
      measurements, those substances and substance concentrations are determined which are specific
      for study group 1 and group 2, i.e. compounds which are not present in the comparison group
      or only in significantly lower or higher concentrations. The selected volatile markers, as
      well as any volatile contaminants that may have been detected in the environment, are stored
      in an analytical reference database and, after elimination of the contamination, bundled into
      possibly disease-specific marker profiles. IV) Analysis of fecal samples: The fecal samples
      are also sent to the Institute for Breathing Gas Analysis of the University of Rostock for
      analysis and analyzed there after appropriate preconcentration by solid phase microextraction
      (SPME). V) Identification of biomarkers of fecal samples: This is done in analogy to the
      exhaled gas samples.

      Lung function measurement before treatment with a probiotic: Measurement of lung function
      using spirometry and body plethysmography (Fa Jäger spirometer and body plethysmograph) and
      nitrogen washout process (N2-multiple breath washout, System Exhalyzer D and Spiroware 3.1,
      Eco Medics AG, Duernten, Switzerland). Spirometry and body plethysmography are performed
      according to published ERS/ATS Standards. The "multiple breath washout" method is performed
      under resting breathing and detects the Ventilation (in)homogeneity at the level of the
      functional residual capacity (= FRC = that lung volume that is still in the lungs after a
      calm spontaneous exhalation is left behind). The system consists of a flow meter, a fast
      analyzing gas measuring system, a gas administration system and the corresponding Computer
      analysis software. As a "foreign gas" it will be use 78% of the gas in air occurring nitrogen
      (N2). A flow-volume measurement is performed via an Ultrasonic flowmeter performed directly
      in the inhalation and exhalation flow of the test person/patient and via a laser O2 sensor
      using the side current measurement method and an infrared CO2 sensor in the main current
      measuring procedure (= directly in the patient's respiratory flow) the respective gas
      concentration. The N2 component is then indirectly measured via the O2 and CO2 concentration
      (N2 = 1 - O2 - CO2). During calm spontaneous breathing, the Patient on a snorkel mouthpiece
      via a bacterial filter through the flowmeter 100% oxygen and thus "washes" N2 out
      ("N2-multiple breath washout"). In doing so, the flow-volume curve of spontaneous breathing
      "online" is displayed on the screen and the measurement at Reaching a 1/40 (= 2.5%) of the
      initial nitrogen concentration is completed. After that, wait the subject is safe in the
      length of twice the duration of the measurement around the oxygen of exhaling. This is
      followed by the next measurement. A total of 3 measurements whose mean value serves as a
      result. The so-called "lung clearance index", which indicates the number of functional
      residual capacity lung volumes, which can be used to reduce the initial nitrogen
      concentration to a 1/40 after oxygenation was required. (LCI = quotient between exhaled
      volume and FRC). It expresses how long it takes for the inhaled gas (in our case the
      physiologically occurring nitrogen in the air) through inhalation of 100% oxygen. For healthy
      persons, this value is on average 7 and is significantly higher in lung patients. Further
      measuring parameters, which makes a statement about the peripheral airways proximal to the
      terminal bronchioles on the one hand and via the more distal azine airways on the other hand
      are calculated.

      Sports medical examination: To exclude contraindications for ergometry, a 12-channel resting
      ECG and a resting blood pressure measurement (CombynTM Function & Spaces ECG, Academic
      Technologies) are performed at the beginning of the examination. After the anthropometric
      data (height, weight, BMI) have been collected, the muscle mass and fat mass are determined
      by multi-frequency impedance measurement in six body segments (CombynTM Function & Spaces
      ECG, Academic Technologies). The lung function is measured by means of small spirometry at
      rest and after exercise (Spirometer Oxycon Pro, Reiner). In order to determine
      cardiopulmonary performance, ergospirometry is performed on a bicycle (Excalibur Sport
      ergometer, Lode company; Oxycon Pro spiroergometry unit, Reiner company) with a gradual
      increase in stress up to subjective exhaustion. The evaluation of these data allows
      conclusions to be drawn about the performance-limiting system (cardiovascular system, lungs,
      musculature) in addition to the determination of aerobic performance.

      Microbiome/VOCs analysis after treatment with a probiotic: After sampling for microbiome and
      VOC analysis and carrying out lung function measurements and sports medical examination, the
      participants in the study group will take a probiotic (Omnibiotic 6, purchased from the
      Allergosan Institute, dietary supplement) for a period of 3 months. Immediately afterwards
      and another month later, measurements of the pulmonary microbiome and the VOCs in the breath
      are taken.
allocation: Randomized
intervention model: Parallel Assignment
primary purpose: Basic Science
masking: Single (Investigator)
measure: Analysis of the pulmonary microbiome in the sputum of CDH group versus control group.
time frame: 12 months
description: OTUs (Operational Taxonomic Units) will be visualized as OTU tables, bar charts and PCOA (Principal Coordinates Analysis) plots using the Qiime core microbiome script. For the different groups alpha-diversity (Chao 1 index, Shannon Index etc.) will be compared. Additionally, we will compare beta-diversity by Adonis test. Relative abundances of the bacteria at the different levels (phylum to genus) will be compared between the groups by using Kruskal Wallis Test.
measure: Analysis of VOCs in the respiratory air by needle-trap microextraction (NTME) and stool by solid phase microextraction (SPME) of CDH group versus control group.
time frame: 12 months
description: From the results of the patient measurements, those substances and substance concentrations are determined which are specific for study group 1 and group 2, i.e. compounds which are not present in the comparison group or only in significantly lower or higher concentrations. The selected volatile markers, as well as any volatile contaminants that may have been detected in the environment, are stored in an analytical reference database and, after elimination of the contamination, bundled into possibly disease-specific marker profiles. The VOCs are recorded and displayed in the following order. The unit in which the VOCs are measured is pars per billion (ppb).
Class (for example carbons) VOCs (ppb) CDHV1 (congenital diaphragmatic hernia group visit 1) CDHK (congenital diaphragmatic hernia - control group visit 1) CDHV2 (congenital diaphragmatic hernia group visit 2) CDHV3 (congenital diaphragmatic hernia group visit 3) p-value
measure: Analysis of the lung function: Lung clearance index (LCI) is derived from multiple breath washout tests of CDH group versus control group.
time frame: 12 months
description: The LCI is about 7 (range from 6.45-7.78) for healthy individuals and is a number without a unit.
measure: Analysis of the lung function: Forced expiratory volume in one second (FEV1) measured with spirometry of CDH group versus control group.
time frame: 12 months
description: The FEV1 is the forced expiratory volume within the first second (liter/second), generated by a maximal voluntary exhalation after maximum inspiration before, usually described as the Tiffeneau-Index in % of FVC (FEV1/FVC).
measure: Analysis of the cardiopulmonary capacity: Resting ECG of CDH group versus control group.
time frame: 12 months
description: A Resting ECG recording the resting heart rate, the rhythm, the PQ duration, the width and height of the QRS complex, the QT duration, and the ST segment is recorded.
measure: Analysis of the cardiopulmonary capacity: Systolic and diastolic blood pressures of CDH group versus control group.
time frame: 12 months
description: Noninvasiv systolic and diastolic blood pressures are assessed (Unit: mmHg).
measure: Analysis of the cardiopulmonary capacity: Body height of CDH group versus control group.
time frame: 12 months
description: Body height is measured in cm.
measure: Analysis of the cardiopulmonary capacity: Body weight of CDH group versus control group.
time frame: 12 months
description: Body weight is measured in kg.
measure: Analysis of the cardiopulmonary capacity: Body mass index (BMI) of CDH group versus control group.
time frame: 12 months
description: Body mass index is calculated in kg body weight/body height².
measure: Analysis of the cardiopulmonary capacity: Muscle mass of CDH group versus control group.
time frame: 12 months
description: Muscle mass is specified in kg/body height².
measure: Analysis of the cardiopulmonary capacity: Body fat of CDH group versus control group.
time frame: 12 months
description: Body fat is specified in percent of body weight.
measure: Analysis of the cardiopulmonary capacity: Aerobic performance of CDH group versus control group.
time frame: 12 months
description: Aerobic performance is specified in percent of normal values of the Austrian cardiological society.
measure: Analysis of the cardiopulmonary capacity: Maximal oxygen uptake of CDH group versus control group.
time frame: 12 months
description: Measurements by spiroergometry: Maximal oxygen uptake in ml/kg/min.
measure: Analysis of the cardiopulmonary capacity: Ventilation of CDH group versus control group.
time frame: 12 months
description: Measurements by spiroergometry: Ventilation in liter/min.
measure: Analysis of the cardiopulmonary capacity: Oxygen pulse of CDH group versus control group.
time frame: 12 months
description: Measurements by spiroergometry: Oxygen pulse in ml/beats per minute.
measure: Analysis of the cardiopulmonary capacity: Oxygen uptake of CDH group versus control group.
time frame: 12 months
description: Respiratory exchange ratio = oxygen uptake in ml/carbon dioxide release in ml.
measure: Analysis of the cardiopulmonary capacity: Breathing reserve of CDH group versus control group.
time frame: 12 months
description: Unit: Percent of FEV1 x 35.
measure: Alterations of pulmonary microbiome after probiotic treatment for a period of 3 months in patients with CDH.
time frame: 12 months
description: After sampling for microbiome and VOC analysis and carrying out lung function measurements and sports medical examination, the participants in the study group will take a probiotic (Omnibiotic 6, purchased from the Allergosan Institute, dietary supplement) for a period of 3 months. Immediately afterwards and another month later, measurements of the pulmonary microbiome are taken.
measure: Alterations of VOCs in the respiratory air after probiotic treatment for a period of 3 months in patients with CDH.
time frame: 12 months
description: After sampling for microbiome and VOC analysis and carrying out lung function measurements and sports medical examination, the participants in the study group will take a probiotic (Omnibiotic 6, purchased from the Allergosan Institute, dietary supplement) for a period of 3 months. Immediately afterwards and another month later, measurements of the VOCs in the breath are taken.
measure: Analysis of the lung function: Forced expiratory flow (FEF25-75).
time frame: 12 months
description: Measured by body plethysmography and spirometry: FEF25-75 = Forced expiratory flow 25-75% vital capacity (= MMEF), Unit: l/s.
measure: Analysis of the lung function: Forced expiratory flow (FEF25).
time frame: 12 months
description: Measured by body plethysmography and spirometry: FEF25 = Forced expiratory flow at the time, when 75% of the vital capacity is exhaled (MEF25), Unit: l/s.
measure: Analysis of the lung function: Forced expiratory flow (FEF50).
time frame: 12 months
description: Measured by body plethysmography and spirometry: FEF50 = Forced expiratory flow at 50% of the exhaled vital capacity (= MEF50), Unit: l/s.
measure: Analysis of the lung function: Forced expiratory volume (FEV1).
time frame: 12 months
description: Measured by body plethysmography and spirometry: FEV1 = Forced expiratory volume in 1 s, Unit: l.
measure: Analysis of the lung function: Tiffeneau-Index (FEV1%FVC).
time frame: 12 months
description: Measured by body plethysmography and spirometry: FEV1%FVC = Tiffeneau-Index, described in % of the forced vital capacity, Unit: %.
measure: Analysis of the lung function: Functional residual capacity (FRC).
time frame: 12 months
description: Measured by body plethysmography and spirometry: FRC = Functional residual capacity, Unit: l.
measure: Analysis of the lung function: Through "multiple breath washout" acquired FRC (RC(MBW)).
time frame: 12 months
description: RC(MBW): Through "multiple breath washout" acquired FRC, Unit: l.
measure: Analysis of the lung function: Through body plethysmography acquired FRC (RC(pleth)).
time frame: 12 months
description: Measured by body plethysmography: RC(pleth): Through body plethysmography acquired FRC (= ITGV, intra thoracic gas volume), Unit: l.
measure: Analysis of the lung function: Forced vital capacity (FVC).
time frame: 12 months
description: Measured by spirometry: FVC = Forced vital capacity, Vital capacity acquired through a forced exhaled manoeuvre, Unit: l.
measure: Analysis of the lung function: Intrathoracic gasvolume (ITGV).
time frame: 12 months
description: Measured by body plethysmography: ITGV = Intrathoracic gasvolume (= FRC(pleth)), Unit: l.
measure: Analysis of the lung function: Max. expiratory flow (MEF25).
time frame: 12 months
description: Measured by spirometry: MEF25 = Max. expiratory flow when 75% of the vital capacity is exhaled (= FEF25), Unit: l/s.
measure: Analysis of the lung function: Max. expiratory flow (MEF50).
time frame: 12 months
description: Measured by spirometry: MEF50 = Max. expiratory flow when 50% of the vital capacity is exhaled (= FEF50), Unit: l/s.
measure: Analysis of the lung function: Max. expiratory flow (MMEF).
time frame: 12 months
description: Measured by spirometry: MMEF = Max. expiratory flow (= FEF25-75), Unit: l/s.
arm group label: CDH Group
arm group type: Active Comparator
description: 10 patients after surgical closure of CDH will undergo VOC profile analysis (2 breath samples) (initial VOC), fecal sampling for 16S rDNA based pyrosequencing (initial fecal microbiome) and deep induced sputum sampling for 16S rDNA pyrosequencing (initial pulmonary microbiome), bicycle spiroergometry to determine the maximum oxygen uptake (maximum oxygen uptake), body plethysmography, spirometry and N2-multiple breath washout testing to determine the functional residual capacity (functional residual capacity). Thereafter patients will receive probiotic treatment with OmniBiotic6 (R) (Allergosan, Graz, Austria) 1 sachet daily for 3 months (probiotic treatment). Three months after discontinuing probiotic treatment VOC testing (VOC probiotics), fecal microbiome sampling (fecal microbiome probiotics) and deep induced sputum testing (pulmonary microbiome probiotics) will be repeated and compared to the results of the initial tests.
arm group label: Control Group
arm group type: Other
description: 10 healthy controls (age and sex matched) will undergo VOC profile analysis (2 breath samples) (initial VOC), fecal sampling for 16S rDNA based pyrosequencing (initial fecal microbiome) and deep induced sputum sampling for 16S rDNA pyrosequencing (initial pulmonary microbiome), bicycle spiroergometry to determine the maximum oxygen uptake (maximum oxygen uptake), body plethysmography, spirometry and N2-multiple breath washout testing to determine the functional residual capacity (functional residual capacity).
intervention type: Diagnostic Test
intervention name: initial VOC
description: Difference in VOC profile between patients with CDH and healthy controls (2 samples per patient will be obtained after obtaining informed consent).
arm group label: CDH Group
arm group label: Control Group
intervention type: Diagnostic Test
intervention name: initial fecal microbiome
description: Difference of alpha and beta diversity and relative fecal bacterial abundance between patients with CDH and healthy controls (1 stool sample will be taken per patient after obtaining informed consent)
arm group label: CDH Group
arm group label: Control Group
intervention type: Diagnostic Test
intervention name: initial pulmonary microbiome
description: Difference of alpha and beta diversity and relative pulmonary bacterial abundance between patients with CDH and healthy controls (1 deep induced sputum sample will be taken per patient after obtaining informed consent)
arm group label: CDH Group
arm group label: Control Group
intervention type: Diagnostic Test
intervention name: Maximum oxygen uptake
description: Comparison of the maximum oxygen uptake (corrected for body weight and gender) as determined by bicycle spiroergometry between patients with CDH and healthy controls
arm group label: CDH Group
arm group label: Control Group
intervention type: Diagnostic Test
intervention name: Functional residual capacity
description: FRC will be determined by spirometry, bodyplethysmography and N2-breath wash out method. FRC will be compared between patients after CDH and healthy controls.
arm group label: CDH Group
arm group label: Control Group
intervention type: Dietary Supplement
intervention name: Probiotic treatment
description: CDH patients will receive OmniBiotic 6(R) (Allergosan, Graz, Austria) probiotic supplementation 1 sachet daily for 3 months.
arm group label: CDH Group
intervention type: Diagnostic Test
intervention name: VOC probiotic
description: Determination of the VOC profile 3 months after discontinuing probiotic treatment. Comparison to the profiles before the treatment.
arm group label: CDH Group
intervention type: Diagnostic Test
intervention name: Fecal microbiome probiotic
description: Determination of the fecal microbiome from 1 sample per patient (alpha and beta diversity, relative bacterial abundance at the genus level) 3 months after discontinuing probiotic treatment. Comparison to the profiles before the treatment.
arm group label: CDH Group
intervention type: Diagnostic Test
intervention name: Pulmonary microbiome probiotic
description: Determination of the fecal microbiome from 1 deep induced sputum sample per patient (alpha and beta diversity, relative bacterial abundance at the genus level) 3 months after discontinuing probiotic treatment. Comparison to the profiles before the treatment.
arm group label: CDH Group
criteria: 
gender: All
minimum age: 6 Years
maximum age: 16 Years
healthy volunteers: Accepts Healthy Volunteers
last name: Till Holger, MD
role: Study Director
affiliation: Department of Pediatric and Adolescent Surgery, Medical University of Graz
last name: Ernst Eber, MD
role: Principal Investigator
affiliation: Department of Pediatric and Adolescent Medicine, Medical University of Graz
last name: Gert Warncke, MD
role: Principal Investigator
affiliation: Department of Pediatric and Adolescent Surgery, Medical University of Graz
facility: 
country: Austria
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responsible party type: Principal Investigator
investigator affiliation: Medical University of Graz
investigator full name: Warncke Gert, MD
investigator title: Principal Investigator
mesh term: Hernias, Diaphragmatic, Congenital
mesh term: Hernia
mesh term: Hernia, Diaphragmatic
sharing ipd: No