Effect of preeclampsia on human milk cytokine levels

Introduction: Preeclampsia (PE) is a systemic inflammatory disease, and its effect on human milk immune components is poorly understood.

Objective: To investigate whether PE affects human milk cytokine levels.

Methods: This was a prospective observational study involving mothers diagnosed with PE and with singleton pregnancy with no fetal malformation. The following cases were excluded: diabetes, chorioamnionitis, use of illicit drugs and alcohol, mastitis and congenital infection. In total, 228 mothers were studied and divided into two groups matched by gestational age: PE (n?=?114) and normotensive (control, n?=?114). Colostrum was collected from 24–72?hours postpartum, and mature milk was collected at the end of the first month. Cytokines (IL-1β, IL-6, IL-8, IL-10, IL-12, and TNF-α) were measured using flow cytometry. A generalized linear model with a gamma distribution was used to analyze the differences between groups versus time interaction.

Results: The mean gestational age was 36 weeks. Increased IL-1 and IL-6 levels and reduced IL-12 levels in the colostrum were detected in PE, while in the mature milk, the IL-6 and IL-8 levels were lower than those of the control group.

Conclusions: PE is associated with increased levels of inflammatory cytokines in colostrum and decreased levels in mature milk.     

Abdominal pain of cardiovascular origin

Infective endocarditis is a microbial infection of the endocardium and it is rare in the pediatric population. In children, congenital heart disease is one of the most important risk factors for developing infective endocarditis and can involve other structures in addition to cardiac valves. The prognosis is generally better than in other forms of endocarditis, although the average mortality rate in the pediatric population is 15-25%. Clinical manifestations can mimic other diseases such as meningitis and collagen-vascular disease or vasculitis. Therefore, a high degree of suspicion is required to make an early diagnosis. Gram-positive bacteria, specifically alpha-hemolytic streptococci, Staphylococcus aureus and coagulase-negative staphylococci, are the most commonly involved bacteria. Diagnosis is based on the modified Duke criteria, which rely mostly on clinical assessment, echocardiography and blood cultures. Antibacterial treatment should ideally be targeted. However, if no specific bacteria have been identified, patients should promptly be treated empirically with multiple drug regimens based on local resistance and the most common etiologies.The authors describe a case of a seven-year-old girl with classic clinical signs of endocarditis, with a clinical twist.     

Performance comparison of two whole genome amplification techniques in frame of multifactor preimplantation genetic testing

Purpose

To compare multiple displacement amplification and OmniPlex whole genome amplification technique performance during array comparative genome hybridization (aCGH), Sanger sequencing, SNaPshot and fragment size analysis downstream applications in frame of multifactor embryo preimplantation genetic testing.

Methods

Preclinical workup included linked short tandem repeat (STR) marker selection and primer design for loci of interest. It was followed by a family haplotyping, after which an in vitro fertilization preimplantation genetic testing (IVF-PGT) cycle was carried out. A total of 62 embryos were retrieved from nine couples with a confirmed single gene disorder being transmitted in their family with various inheritance traits—autosomal dominant (genes—ACTA2, HTT, KRT14), autosomal recessive (genes—ALOX12B, TPP1, GLB1) and X-linked (genes—MTM1, DMD). Whole genome amplification (WGA) for the day 5 embryo trophectoderm single biopsies was carried out by multiple displacement amplification (MDA) or polymerase chain reaction (PCR)-based technology OmniPlex and was used for direct (Sanger sequencing, fragment size analysis, SNaPshot) and indirect mutation assessment (STR marker haplotyping), and embryo aneuploidy testing by array comparative genome hybridization (aCGH).

Results

Family haplotyping revealed informative/semi-informative microsatellite markers for all clinical cases for all types of inheritance. Indirect testing gave a persuasive conclusion for all embryos assessed, which was confirmed through direct testing. The overall allele dropout (ADO) rate was higher for PCR-based WGA, and MDA shows a better genomic recovery scale. Five euploid embryos were subjected to elective single embryo transfer (eSET), which resulted in four clinical pregnancies and birth of two healthy children, which proved free of disease causative variants running in the family postnataly.

Conclusions

A developed multifactor PGT protocol can be adapted and applied to virtually any genetic condition and is capable of improving single gene disorder preimplantation genetic testing in a patient-tailored manner thus increasing pregnancy rates, saving costs and increasing patient reliability.

     

Granulomatous mycosis fungoides - A diagnostic challenge

Granulomatous mycosis fungoides is a rare subtype of T-cell cutaneous lymphoma. Due to its clinical heterogenicity the diagnosis is delayed and based on histopathological and immuno-histochemical findings, sometimes requiring gene rearrangement studies for confirmation. We report the case of a patient who was submitted to several biopsies before diagnostic conclusion.     

Hepatitis C virus (HCV)‐specific T‐cell responses among recombinant immunoblot assay‐3–indeterminate blood donors: a confirmatory evidence of HCV exposure

BACKGROUND: Blood donors are routinely screened for hepatitis C virus (HCV) infection. Some show weak anti‐HCV responses, often restricted to a single antigen on confirmatory immunoblot (recombinant immunoblot assay [RIBA]) testing. The aim of this study was to investigate the extent to which such RIBA‐indeterminate donors had previously been exposed to HCV. STUDY DESIGN AND METHODS: T‐cell responses to HCV recombinant proteins (core, NS3, and NS3 helicase) were analyzed using an interferon‐γ (IFN‐γ) enzyme‐linked immunospot (ELISpot) assay and quantification of cytokines in culture supernatants in 27 RIBA‐indeterminate donors, 60 RIBA‐confirmed donors (48 with and 12 without HCV RNA), and 30 RIBA‐negative donors. RESULTS: HCV‐specific T‐cell responses were identified in 13 (48%) RIBA‐indeterminate donors, 33 (55%) RIBA‐confirmed donors, and 4 (13%) RIBA‐negative controls (p = 0.008 and p < 0.001, respectively). The magnitude of the T‐cell response among indeterminate donors was similar to that of RIBA‐confirmed donors for all HCV antigens and the specificity of the ELISpot results was confirmed by antigen‐specific cytokine production (interleukin‐2 and IFN‐γ) in short‐term culture supernatants. CONCLUSIONS: These findings confirm that approximately half of RIBA‐indeterminate donors have resolved a previous HCV infection and suggest that ELISpot might be a useful tool to clarify the status of such donors and help in their counseling and management.