Noémi Mihalik

Microarray profiling reveals that placental transcriptomes of early-onset HELLP syndrome and preeclampsia are similar

BACKGROUND The involvement of the placenta in the pathogenesis of preeclampsia and HELLP syndrome is well established, and placental lesions are also similar in these two syndromes. Here we aimed to examine the placental transcriptome and to identify candidate biomarkers in early-onset preeclampsia and HELLP syndrome. METHODS Placental specimens were obtained at C-sections from women with early-onset preeclampsia and HELLP syndrome, and from controls who delivered preterm or at term. After histopathological examination, fresh-frozen placental specimens were used for microarray profiling and validation by qRT-PCR. Differential expression was analysed using log-linear models while adjusting for gestational age. Gene ontology and pathway analyses were used to interpret gene expression changes. Tissue microarrays were constructed from paraffin-embedded placental specimens and immunostained. RESULTS Placental gene expression was gestational age-dependent among preterm and term controls. Out of the 350 differentially expressed genes in preeclampsia and 554 genes in HELLP syndrome, 224 genes (including LEP, CGB, LHB, INHA, SIGLEC6, PAPPA2, TREM1, and FLT1) changed in the same direction (elevated or reduced) in both syndromes. Many of these encode proteins that have been implicated as biomarkers for preeclampsia. Enrichment analyses revealed similar biological processes, cellular compartments and biological pathways enriched in early-onset preeclampsia and HELLP syndrome; however, some processes and pathways (e.g., cytokine-cytokine receptor interaction) were over-represented only in HELLP syndrome. CONCLUSION High-throughput transcriptional and tissue microarray expression profiling revealed that placental transcriptomes of early-onset preeclampsia and HELLP syndrome largely overlap, underlying a potential common cause and pathophysiologic processes in these syndromes. However, gene expression changes may also suggest a more severe placental pathology and pronounced inflammatory response in HELLP syndrome than in preeclampsia.

Activation of villous trophoblastic p38 and ERK1/2 signaling pathways in preterm preeclampsia and HELLP syndrome

Preterm preeclampsia is associated with the failure of trophoblast invasion, placental hypoxic/ischemic injury and the release of toxic substances, which promote the terminal pathway of preeclampsia. In term preeclampsia, factors yet unknown trigger the placenta to induce the terminal pathway. The contribution of the villous trophoblast to these pathologic events has not been fully elucidated. Here we aimed to study how stress and signaling pathways influence trophoblastic functions in various subforms of preeclampsia. Tissue microarrays (TMAs) were constructed from placentas obtained from pregnant women in the following groups: 1–2) preterm preeclampsia with (n = 8) or without (n = 7) HELLP syndrome; 3) late-onset preeclampsia (n = 8); 4–5) preterm (n = 5) and term (n = 9) controls. TMA slides were stained for phosphorylated Akt-1, ERK1/2, JNK, and p38 kinases, and trophoblastic immunostainings were semi-quantitatively evaluated. BeWo cells were kept in various stress conditions, and the expression of FLT1, GCM1, LEP, and PGF was profiled by qRT-PCR, while Akt-1, ERK1/2, JNK, and p38 kinase activities were measured with phospho-kinase immunoassays. We found that: 1) Placental LEP and FLT1 expression was up-regulated in preterm preeclampsia with or without HELLP syndrome compared to controls; 2) Mean pp38 immunoscore was higher in preterm preeclampsia, especially in cases with HELLP syndrome, than in controls. 3) Mean pERK1/2 immunoscore was higher in preterm preeclampsia with HELLP syndrome than in controls. 4) In BeWo cells, ischemia up-regulated LEP expression, and it increased JNK and decreased ERK1/2 activity. 5) Hypoxia up-regulated FLT1 and down-regulated PGF expression, and it increased ERK1/2, JNK and p38 activity. 6) IL-1β treatment down-regulated PGF expression, and it increased JNK and p38 activity. 7) The p38 signaling pathway had the most impact on LEP, FLT1 and PGF expression. In conclusion, hypoxic and ischemic stress, along with unknown factors, activates trophoblastic p38 signaling, which has a key role in villous trophoblastic functional changes in preterm preeclampsia. The activation of ERK1/2 signaling may induce additional trophoblastic functional changes in HELLP syndrome, while distinct mechanisms may promote late-onset preeclampsia.

Syphilis and HIV coinfection — Hungarian Sexually Transmitted Infection Centre Experience between 2005 and 2013

BACKGROUND STIs like HIV and syphilis are acquired as comorbidities by high risk populations and may influence their original course and prognosis. METHODS Between January of 2005 and 2013 data of syphilis and HIV patients were collected at the Department of Dermatology of Semmelweis University, Budapest. Diagnostic procedures included clinical analysis and screening of serum samples for Treponema pallidum and HIV antibodies. RESULTS A total of 1,401 new syphilitic and 338 new HIV infections were diagnosed. In syphilis patients 86.58% had monoinfection,7.92% already had an HIV infection and 5.5% had acquired syphilis and HIV infection simultaneously, so 22.78% of the new HIV patients acquired the infection with syphilis together. Male gender, MSM (men who had sex with men) orientation and positive past venerological history were dominant in all groups. Most patients were diagnosed in a latent infectious stage based on the result of a serological check-up. Secondary stage and neurosyphilis were more common in coinfections. CONCLUSION (i) male gender, MSM orientation, and positive venerological history are risk factors for acquiring new STIs, (ii) clinical course were different in HIV infected patients, (iii) but their timely and regular check-ups resulted in earlier diagnosis of syphilis, suggesting the necessity for frequent screening.

Clinical observations in cutan mastocytosis

INTRODUCTION Mastocytosis is a clonal mast cell proliferative disease, divided into cutaneous and systemic forms. The characteristic symptoms are caused by neoplastic mast cell infiltrations in different organs and/or the release of mediators. AIM The aim of the authors was to summarize their clinical observations in patients with mastocytosis. METHOD 22 adult patients diagnosed consecutively with mastocytosis were enrolled in the study. Skin and bone marrow biopsies were taken to establish the diagnosis and perform c-KIT mutation (D816V) analysis. RESULTS One of the 22 patients had teleangiectasia macularis eruptiva perstans, while 20/22 patients had urticaria pigmentosa. All patients had cutaneous lesions. In 12 patients iliac crest biopsy was performed and 9 of them had bone marrow involvement, classified as indolent systemic mastocytosis. The c-kit mutation D816V was found in one subject both in skin and bone marrow samples. The patients were treated with antihistamine, PUVA, interferon-α or imatinib. CONCLUSIONS The authors draw attention to this rare disease in order to help recognition of relevant signs and symptoms and establish an early diagnosis.

Purulent keratoconjunctivitis due to Neisseria gonorrhoeae and Chlamydia trachomatis coinfection

Gonococcal conjunctivitis is a rare infection induced by Neisseria gonorrhoeae and it usually manifests as a hyperacute purulent conjunctivitis. Ocular access of the infectious secretion during sexual intercourse is the way of transmission among adults. Inclusion conjunctivitis caused by the serovars D-K of Chlamydia trachomatis also affects the sexually active population. Authors present a case of a 33-year-old homosexual man who was treated for late latent syphilis formerly. Clinical symptoms were yellow purulent discharge for 3 weeks without any urological or upper respiratory tract symptoms. Conjunctival Neisseria gonorrhoeae and Chlamydia trachomatis infection was identified using cultures and polymerase chain reaction; pharyngeal swab culture and polymerase chain reaction showed positive results for both pathogens. The patient was probably under influence of party drugs at the time of sexual abuse when he became infected. After parenteral and oral cephalosporin and azithromycin therapy the patient had complete recovery within three weeks.

Antimicrobial susceptibility and genotyping analysis of Hungarian Neisseria gonorrhoeae strains in 2013

Emergence and spread of antimicrobial resistance in Neisseria gonorrhoeae is a major public health concern worldwide. The current study aims to determine the antimicrobial resistance in N. gonorrhoeae and associated molecular typing to enhance gonococcal antimicrobial surveillance in Hungary. In the National N. gonorrhoeae Reference Laboratory of Hungary 187 N. gonorrhoeae infections were detected in 2013, antibiograms were determined for all the isolated strains, and 52 (one index strain from every sexually contact related group) of them were also analysed by the N. gonorrhoeae multi-antigen sequence typing (NG-MAST) method. Twenty-two different NG-MAST sequence types (STs) were identified, of which 8 STs had not been previously described. In Hungary, the highly diversified gonococcal population displayed high resistance to penicillin, ciprofloxacin and tetracycline (the antimicrobials previously recommended for gonorrhoea treatment). Resistance to the currently recommended extended spectrum cephalosporines were rare: only two of the expected strains, an ST 1407 and an ST 210, had cefixime MIC above the resistance breakpoint. By the revision of our National Treatment Guideline, it must be considered, that the azithromycin resistance is about 60% among the four most frequently isolated STs in Hungary.

Yeast species in vulvovaginitis candidosa

INTRODUCTION Vulvovaginal candidiasis is the most common mycosis, however, the available information about antifungal susceptibilities of these yeasts is limited. AIM To compare the gold standard fungal culture with a new molecular identification method and report the incidence of yeast species in vulvovaginitis candidosa. METHOD The authors studied 370 yeasts isolated from vulvovaginal candidiasis and identified them by phenotypic and molecular methods. RESULTS The most common species was Candida albicans (85%), followed by Candida glabrata, and other Candida species. CONCLUSION At present there are no recommendations for the evaluation of antifungal susceptibility of pathogenic fungal species occurring in vulvovaginal candidiasis and the natural antifungal resistance of the different species is known only. Matrix Assisted Laser Desorption Ionization Time of Flight identification can be used to differentiate the fluconazole resistant Candida dubliniensis and the sensitive Candida albicans strains.

Vulvovaginitis candidosában előforduló sarjadzógomba-speciesek

INTRODUCTION Vulvovaginal candidiasis is the most common mycosis, however, the available information about antifungal susceptibilities of these yeasts is limited. AIM To compare the gold standard fungal culture with a new molecular identification method and report the incidence of yeast species in vulvovaginitis candidosa. METHOD The authors studied 370 yeasts isolated from vulvovaginal candidiasis and identified them by phenotypic and molecular methods. RESULTS The most common species was Candida albicans (85%), followed by Candida glabrata, and other Candida species. CONCLUSION At present there are no recommendations for the evaluation of antifungal susceptibility of pathogenic fungal species occurring in vulvovaginal candidiasis and the natural antifungal resistance of the different species is known only. Matrix Assisted Laser Desorption Ionization Time of Flight identification can be used to differentiate the fluconazole resistant Candida dubliniensis and the sensitive Candida albicans strains.

P2.190 Neurosyphilis Cases in the Hungarian National STD Centre in the Last 5 Years

Background The incidence of syphilis has increased in Hungary (6.3/100000 in 2012), with simultaneous increase in neurosyphilis incidece. The aim of this study is to summarise our experience on clinical and serological characteristics, the treatment results, and association with HIV-infection. Methods clinical, serological and cerebrospinal fluid (CSF) analysis: RPR/VDRL, TPPA/TPHA, TP ELISA, TP IgM/IgG Western blot, albumin, mononuclear cell count of 8 patients with neurosyphilis. The diagnosis of neurosyphlis was based on clinical symptoms, syphilis serology, the positive results of VDRL and/or TPHA tests, and the increased number of mononuclear cells in CSF. Results The 7 male and 1 female patients were between 25 and 84 years of age. 4 male patients were HIV-positive, 3 of them were MSM, one was bisexual. 5 patients had neurosyphilis with symptoms such as headache, dizziness, central facial palsy, visual impairment, sensory loss, diminished tendon reflexes. Asyptomatic patients had schizoaffective disorder, visual impairment, syphilitic reinfection respecively, neurological symptoms were obsereved more frequently in patients with HIV-infection. TPPA/TPHA test in 7 patients’, VDRL test in 3 patients’ and increased number of mononuclear cells in 7 patients’ CSF were positive. All patients were treated with high dose intravenous benzyl penicillin (24 million units iv. daily for 14 days), the effectiveness of treatment was documented by the improvement in clinical symptoms and by the decrease of RPR titer. Conclusion The clinical course was similar in patients with HIV and without it. One third of patients with neurosyphilis was symptoms free, the remaining of them presented clinical symptomps of neurosyphilis. The adequate indication of CSF examination is essential for the diagnosis of neurosyphilis. The long-term penicillin therapy was effective in our cases.

P2.114 Coinfection of Treponema Pallidum and Cytomegalovirus (CMV): A Complicated Case of a Newborn in Hungary

Background Although congenital syphilis and congenital CMV infections are preventable they are still the major causes of perinatal mortality and morbidity. Expectant mothers from lower socioeconomical status and intravenous drug users belong to the highest risk groups for vertical transmission of infections. Here we present a coexistence of congenital syphilis and CMV infection complicated with multiplex jejunal atresia. Methods Clinical analysis, laboratory, serological and PCR examinations. Case presentation 990 grams, 38 cm height (pc.75–91) girl was born to an intravenous drug user mother on the 26th gestational week. The expectant mother did not participated in the prenatal care and early latent syphilis (RPR 1:128 positive, TPPA and TpELISA positive), genital Streptococcus agalactiae and fungal infectionwere detected shortly before delivery. The preterm and immature girl had jaundice, oedema, gluteal haematome and petechiae. Extremely enlarged liver and spleen (reaching the hip bone) and increased muscle tone with rigid joints were found. Multiple jejunal atresia was detected by bedside X-ray examination. Anisocytosis, thrombocytopenia, elevated liver enzymes (ASAT: 3850, ALAT: 558, GGT: 292, ALP: 436) and elevated LDH (38180) and CK (7.1) with direct hyperbilirubinaemia were found. During microbiological examinations high copy of CMV virus number was detected by quantitative real-time PCR and syphilis serology was positive (RPR: 1:16 positive, TPPA, Tp ELISA, T. pallidum IgM immunoblot positive). Intravenous penicilline-G (100.000 IU/kg/dose for 10 days) and intravenous ganciclovir was administered. Gancyilovir was stopped after 4 weeks because of progressing thrombocytopenia. The multiplex jejunal atresia was fixed by operation resulting in satisfactory intestinal passage. Conclusions Although syphilis screening test within the prenatal care is mandatory in Hungary, congenital syphilis cases do occur. Immature immune system is predisposing factor for coinfections of a newborn. The symptoms of T. pallidum and CMV infection is very similar, presenting a diagnostic challenge.