Mirrian Hilbink

Biomarkers and Molecular Analysis to Improve Bloodstream Infection Diagnostics in an Emergency Care Unit

Molecular pathogen detection from blood is still expensive and the exact clinical value remains to be determined. The use of biomarkers may assist in preselecting patients for immediate molecular testing besides blood culture. In this study, 140 patients with ≥ 2 SIRS criteria and clinical signs of infection presenting at the emergency department of our hospital were included. C-reactive protein (CRP), neutrophil-lymphocyte count ratio (NLCR), procalcitonin (PCT) and soluble urokinase plasminogen activator receptor (suPAR) levels were determined. One ml EDTA blood was obtained and selective pathogen DNA isolation was performed with MolYsis (Molzym). DNA samples were analysed for the presence of pathogens, using both the MagicPlex Sepsis Test (Seegene) and SepsiTest (Molzym), and results were compared to blood cultures. Fifteen patients had to be excluded from the study, leaving 125 patients for further analysis. Of the 125 patient samples analysed, 27 presented with positive blood cultures of which 7 were considered to be contaminants. suPAR, PCT, and NLCR values were significantly higher in patients with positive blood cultures compared to patients without (p < 0.001). Receiver operating characteristic curves of the 4 biomarkers for differentiating bacteremia from non-bacteremia showed the highest area under the curve (AUC) for PCT (0.806 (95% confidence interval 0.699–0.913)). NLCR, suPAR and CRP resulted in an AUC of 0.770, 0.793, and 0.485, respectively. When compared to blood cultures, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for SepsiTest and MagicPlex Sepsis Test were 11%, 96%, 43%, 80%, and 37%, 77%, 30%, 82%, respectively. In conclusion, both molecular assays perform poorly when one ml whole blood is used from emergency care unit patients. NLCR is a cheap, fast, easy to determine, and rapidly available biomarker, and therefore seems most promising in differentiating BSI from non-BSI patients for subsequent pathogen identification using molecular diagnostics.

Probiotics for childhood functional gastrointestinal disorders: a systematic review and meta-analysis.

A systematic review and meta‐analysis were performed to investigate the quantity and quality of the current evidence regarding the effect of different probiotic strains in the treatment of functional gastrointestinal disorders (FGID) in children and adolescents.

Dientamoeba fragilis and chronic abdominal pain in children: a case–control study

Background The association between Dientamoeba (D.) fragilis and the aetiology of functional gastrointestinal disorders (FGID) in children is unclear. Aim The aim of this retrospective case–control study is to clarify the clinical relevance of D. fragilis in children with chronic abdominal pain. Methods From April 2011 until April 2013, a total of 132 patients with chronic abdominal pain (AP), aged 8–18 years, referred to a non-academic hospital, and 77 control patients, aged 8–18 years without gastrointestinal symptoms referred to a psychiatric hospital, were included in the study. D. fragilis was diagnosed by real-time PCR in faecal samples. Symptomatic children without a D. fragilis infection fulfilled the ROME III criteria for AP-related FGID (AP-FGID). Clinical data were retrospectively analysed by examining patients’ hospital records from the Jeroen Bosch Hospital and Herlaarhof in The Netherlands. Results D. fragilis was detected in 57 patients with chronic AP (43.2%) and in 39 controls (50.6%) (p=0.255). No significant differences in symptomatology were found between D. fragilis-infected children and children fulfilling the criteria for AP-FGID. Parasitological eradication was achieved in 61.7% of patients after treatment with metronidazole or clioquinol, while clinical improvement occurred in only 40.4% of patients (p=0.435). Conclusions There were no differences in symptoms comparing children with and without D fragilis infection. Furthermore, no relation was found between clinical and microbiological response after treatment for D. fragilis. This retrospective study suggests that there is no association between chronic AP and D. fragilis infection.

Yoga Therapy for Abdominal Pain-Related Functional Gastrointestinal Disorders in Children: A Randomized Controlled Trial.

Objectives: The aim of the present study was to compare effects of 10 weeks of yoga therapy (YT) and standard medical care (SMC) on abdominal pain and quality of life (QoL) in children with abdominal pain–related functional gastrointestinal disorders (AP-FGIDs). Methods: Sixty-nine patients, ages 8 to 18 years, with AP-FGIDs, were randomized to SMC complemented with YT or SMC alone. YT is a mixture of yoga poses, meditation, and relaxation exercises and was given once a week in group sessions. SMC consisted of education, reassurance, dietary advice, and fibers/mebeverine, if necessary. Pain intensity (pain intensity score [PIS] 0–5) and frequency (pain frequency score [PFS] 0–4) were scored in a pain diary, and QoL was measured with KIDSCREEN-27. Follow-up was 12 months. Treatment response was defined as ≥50% reduction of weekly pain scores. Results: At 1-year follow-up, treatment response was accomplished in 58% of the YT group and in 29% of the control group (P = 0.01); no significant differences for other time points were found. YT, and not SMC, resulted in a significant reduction of PIS (P < 0.01) and PFS (P < 0.01) after 12 months. During the study, however, YT was not significantly superior compared with SMC. Subanalyses for time points demonstrated a significant greater reduction of PIS at 12 months in favor of YT. No differences were found for QoL. YT was more effective in the reduction of reported monthly school absence (P = 0.03). Conclusion: At 1-year follow-up, YT in addition to standard care was superior compared with SMC according to treatment success, PIS, and reduction of school absence. YT, however, was not significantly more effective in improving PFS or QoL, compared with SMC.

Emergence of methicillin resistance and Panton-Valentine leukocidin positivity in hospital- and community-acquired Staphylococcus aureus infections in Beira, Mozambique

The objective of this study was to investigate the antibiotic resistance patterns, including methicillin resistance, inducible macrolide–lincosamide–streptogramin B (MLSB) resistance and Panton‐Valentine leukocidin (PVL) toxin gene carriage among hospital‐acquired Staphylococcus aureus (HA‐SA) and community‐acquired S. aureus (CA‐SA), in Beira, Mozambique.

Single Nucleotide Polymorphism (SNP)-Based Loss of Heterozygosity (LOH) Testing by Real Time PCR in Patients Suspect of Myeloproliferative Disease

During tumor development, loss of heterozygosity (LOH) often occurs. When LOH is preceded by an oncogene activating mutation, the mutant allele may be further potentiated if the wild-type allele is lost or inactivated. In myeloproliferative neoplasms (MPN) somatic acquisition of JAK2V617F may be followed by LOH resulting in loss of the wild type allele. The occurrence of LOH in MPN and other proliferative diseases may lead to a further potentiating the mutant allele and thereby increasing morbidity. A real time PCR based SNP profiling assay was developed and validated for LOH detection of the JAK2 region (JAK2LOH). Blood of a cohort of 12 JAK2V617F-positive patients (n = 6 25–50% and n = 6>50% JAK2V617F) and a cohort of 81 patients suspected of MPN was stored with EDTA and subsequently used for validation. To generate germ-line profiles, non-neoplastic formalin-fixed paraffin-embedded tissue from each patient was analyzed. Results of the SNP assay were compared to those of an established Short Tandem Repeat (STR) assay. Both assays revealed JAK2LOH in 1/6 patients with 25–50% JAK2V617F. In patients with >50% JAK2V617F, JAK2LOH was detected in 6/6 by the SNP assay and 5/6 patients by the STR assay. Of the 81 patients suspected of MPN, 18 patients carried JAK2V617F. Both the SNP and STR assay demonstrated the occurrence of JAK2LOH in 5 of them. In the 63 JAK2V617F-negative patients, no JAK2LOH was observed by SNP and STR analyses. The presented SNP assay reliably detects JAK2LOH and is a fast and easy to perform alternative for STR analyses. We therefore anticipate the SNP approach as a proof of principle for the development of LOH SNP-assays for other clinically relevant LOH loci.

Dynamics of peripheral blood lymphocyte subpopulations in the acute and subacute phase of Legionnaires' disease

Study Objective Absolute lymphocytopenia is recognised as an important hallmark of the immune response to severe infection and observed in patients with Legionnaires’ disease. To explore the immune response, we studied the dynamics of peripheral blood lymphocyte subpopulations in the acute and subacute phase of LD. Methods and Results EDTA-anticoagulated blood was obtained from eight patients on the day the diagnosis was made through detection of L. pneumophila serogroup 1 antigen in urine. A second blood sample was obtained in the subacute phase. Multiparametric flow cytometry was used to calculate lymphocyte counts and values for B-cells, T-cells, NK cells, CD4+ and CD8+ T-cells. Expression of activation markers was analysed. The values obtained in the subacute phase were compared with an age and gender matched control group. Absolute lymphocyte count (×109/l, median and range) significantly increased from 0.8 (0.4–1.6) in the acute phase to 1.4 (0.8–3.4) in the subacute phase. B-cell count showed no significant change, while T-cell count (×106/l, median and range) significantly increased in the subacute phase (495 (182–1024) versus 979 (507–2708), p = 0.012) as a result of significant increases in both CD4+ and CD8+ T-cell counts (374 (146–629) versus 763 (400–1507), p = 0.012 and 119 (29–328) versus 224 (107–862), p = 0.012). In the subacute phase of LD, significant increases were observed in absolute counts of activated CD4+ T-cells, naïve CD4+ T-cells and memory CD4+ T-cells. In the CD8+ T-cell compartment, activated CD8+ T-cells, naïve CD8+ T-cell and memory CD8+ T-cells were significantly increased (p<0.05). Conclusion The acute phase of LD is characterized by absolute lymphocytopenia, which recovers in the subacute phase with an increase in absolute T-cells and re-emergence of activated CD4+ and CD8+ T cells. These observations are in line with the suggested role for T-cell activation in the immune response to LD.

Declining antibody levels after hepatitis B vaccination in Down syndrome: A need for booster vaccination?

We determined the anti‐HBs titer in 227 children of all ages with Down syndrome (DS). Only 48.1% (95%CI: 35.1‐61.3) of the DS children aged 7‐10 years and 31.9% (95%CI: 22.1‐43.6) of the DS children aged >10 years had a protective anti‐HBs titer (≥10 IU/L). The geometric mean anti‐HBs titer was significantly lower in the DS children; this suggests booster vaccination for HBV may be needed.

Lower percentage of allergic sensitization in children with Down syndrome

from increased vascular permeability.5,6 The episodes are variable in terms, severity, location, and frequency, as they may occur spontaneously or may be triggered by curtain stimuli, including trauma, stress, or infections.6 The associated clinical symptoms include noninflammatory swellings of the skin and mucous membranes, vomiting, diarrhea, paroxysmal colicky pain, laryngeal edema, and upper airway obstruction, while erythema marginatum may precede the occurrence of edema in 25% of the cases.5 We present a case of acquired ACM type 1 in a 16yearold male patient, with longlasting hereditary angioedema type 1, treated with recombinant C1 esterase inhibitors for 9 years. To the best to our knowledge, this is the first report in the literature of such association. Although it has been postulated that ACM could be acquired at later ages, as a result of trauma, injury, or toxic substance exposure,3 the limited literature data on this issue hinder the hypothesis for eventual pathogenetic relationship between this two rare conditions. In the presented case, the impaired cerebrospinal fluid, resulting in clinical manifestation of ACM at later age, could be provoked from by the chronic increased production of bradykinin, formed as insufficient C1 inhibitor activity in patient with HAE, with further fail of the restriction of the action of factor XII and kallikrein. We could not find any literature data, supporting or denying the idea for triggering the acquired malformation by chronic increased vascular permeability or chronic exposure to curtain drugs. The presented case of association of ACM in patient with hereditary angioedema remains this question open for further investigations.

Sputum Induction in Children Is Feasible and Useful in a Bustling General Hospital Practice

We prospectively studied the feasibility and effectiveness of sputum induction in obtaining good quality sputum and its subsequent bacterial yield in children with clinically suspected acute lower-respiratory-tract infection (aLRTI). Good quality sputum was collected in 89/98 (91%) patients. Sputum cultures revealed ≥1 bacterial pathogens in 22 cases (25%). Adverse events were infrequent and mild (6%). Sputum induction is feasible in young children and leads to an increased number of etiological diagnoses of aLRTI.