The importance of serological and molecular analyses for the diagnosis of measles cases and for meeting elimination targets in Turkey from 2007 to 2015.

Measles is an important childhood infection targeted to be eliminated by the World Health Organization (WHO). Virus circulation has not been interrupted in the European Region because high vaccination rates could not be achieved among some countries of the WHO European Region including Turkey. The purpose of this study was to evaluate the laboratory findings of measles cases confirmed in the last nine years, to assess the epidemiological data of the cases, to determine the molecular genotyping studies and to emphasise the importance of laboratory-based surveillance in measles. From 2007 to 2010, only 18 imported cases were detected in Turkey. However, this number increased with a local outbreak of 111 cases in 2011, followed by another outbreak in 2012 in Istanbul that spread countrywide in the following two years; a total of 8661 laboratory-confirmed measles cases were reported from 2012 to 2015. After ELISA detection of a measles IgM-positive result in serum samples of potential measles cases, RT-PCR was performed with urine or nasopharyngeal swab samples of patients, and amplicons were subjected to sequencing. In the samples of 2010 and 2011, D4 and D9 genotypes were mainly detected; as of 2012, the D8 genotype has gained importance. Although D8 was also identified in 2014, in the same year genotype H1 viruses were detected in Turkey for the first time. Therefore, it is important to perform a genotypic analysis of the virus causing the outbreak, analyse epidemiological connections of the contact, determine the source of the outbreak and plan measures based on this information.

Towards measles elimination: Phylogenetic analysis of measles viruses in Turkey (2012-2013) and identification of genotype D8.

Molecular characterization of different measles virus (MV) strains is essential to combat the disease. Sixty measles MV strains were obtained from throat swabs or urine of patients in Turkey between 2012 and 2013 and characterized. MV RNA sequences (n = 60) were analysed for 456 nucleotides representing hypervariable domain of the nucleoprotein (N) gene. Of the 60 strains analysed 53 were the D8 genotype, 6 were B3, 1 was D4, and 1 was A. This report describes MV genotype D8 that was involved in a measles outbreak in Turkey. Sequences of most genotype D8 strains (n = 51) were identical to the sequence of variant D8-Frankfurt-Main, which has been associated with outbreaks throughout Europe. Despite the lack of epidemiologic information, a phylogenetic analysis suggested that the genotype D8 MV may have been brought to Turkey from elsewhere. Phylogenetic and epidemiological findings suggested that strains identified in tourists and associated with importation included one strain of genotype D8, one strain of genotype B3, and one strain of genotype D4. These findings from the 2012 to 2013 outbreak in Turkey confirm that pockets of unimmunised individuals are making the country susceptible to measles outbreaks. To prevent further outbreaks, deliberate and sustained effort must be made to reach, and immunise susceptible age groups. Towards measles elimination process, continued molecular surveillance of measles strains in Turkey will help identify transmission patterns of virus and evaluate vaccination efforts. J. Med. Virol. 88:1867-1873, 2016. © 2016 Wiley Periodicals, Inc.

Cotreatment of congenital measles with vitamin a and intravenous immunoglobulin.

Although the measles vaccine has been part of routine national childhood vaccination programs throughout Europe, measles remains a public health concern. High numbers of cases and outbreaks have occurred throughout the European continent since 2011, and an increasing number of cases have been reported in Turkey since 2012. During a recent measles outbreak in Turkey, 2 pregnant women contracted measles prior to delivering preterm infants at Hacettepe University Hospital. Measles virus genomic RNA and IgM antibodies against measles were detected in the cord blood of infants and mothers in both cases. The infants were treated with intravenous immunoglobulin (IVIG) and vitamin A. Transient thrombocytopenia was present in 1 infant and treated with an additional dose of IVIG and vitamin A. The infants were discharged, without complications, within 10 days of birth. The successful treatment of these cases suggests that infants who have been exposed to, or infected with, measles may benefit from cotreatment of vitamin A and IVIG.

[Evaluation of 44 pediatric measles cases detected in Ankara, Turkey during 2012-2013 epidemic and molecular characterization of the viruses obtained from two cases]. / 2012-2013 kizamik salgini sürecinde Ankara'da saptanan 44 çocuk olgunun degerlendirilmesi ve iki olgudan elde edilen viruslarin moleküler analizi.

Elimination of measles and rubella until the end of 2015 in parallel with the "World Health Organization (WHO) Europe Region's Measles Elimination" work-up has been targetted and "Measles Elimination Program'' has been carried out since 2002 in Turkey. Due to the routine vaccination programmes the number of the vaccinated children have increased and epidemic incidences have been falling. However, imported measles cases from Europe and other neighboring countries have been observed in Turkey in the recent two years. Patients who applied to Dr. Sami Ulus Maternity and Children's Training and Research Hospital with a pre-diagnosis of measles between December 2012 and April 2013 were screened in this study. Seventy-eight patients who match the clinical definition of the disease (> 38°C fever and maculopapular rash and cough or nasal discharge or conjunctivitis) were evaluated. Forty-four children (25 male, 19 female; age range: 4-191 months, mean age: 58.6 ± 59.5 months) with a positive measles IgM test result were taken into consideration and the epidemiological and clinical features of these children were evaluated. In addition to fever and rash, cough, nasal discharge and conjunctivitis were seen in 36 (82%), 24 (55%), and 18 (41%) patients, respectively. Thirty five (80%) patients were diagnosed in December, 6 (14%) in January, 2 (4%) in February, and 1 (2%) in March. All patients included in the study were unvaccinated or too young to be vaccinated according to the routine vaccination calendar. The index case was a three-year old unvaccinated girl who had a history of contact with the Syrian neighborhoods. During the study period; following contact with the index case, two doctors (born in 1986 with a history of single dose of vaccination at ninth month) and three children (without vaccination) were also diagnosed as measles. Eight (18%) patients were hospitalized because of complications. Four (50%) of them had pneumonia and the other four (50%) had lack of oral feeding and dehydration. Average duration of hospitalization for patients was 4 ± 1.7 (range: 2-6) days and all patients were discharged with full recovery. For molecular typing, viral RNAs were isolated from urine samples of two of the measles IgM positive patients, subjected to sequence analysis of 450 nucleotides comprising the most variable C-terminal region of the nucleoprotein (N) gene. Phylogenetic analysis revealed that those two strains belonged to genotype D8. This study represented the involvement of measles virus genotype D8 in an outbreak in Turkey for the first time. During a measles epidemic, following the index case; medical personnel should be informed about possible, probable, and definite case definitions and should apply for appropriate triage or fast-track (rapidly examination) if necessary, and routine announcements should be made precisely and accurately at proper times and unvaccinated medical personnel and any people in touch with the patient should be vaccinated. In order to reach the elimination goal declared by European WHO for 2015, susceptible populations should be identified and vaccinated in Turkey to obtain sufficient herd immunity for preventing outbreaks.

Effects of local and sustained release of FGF, IGF, and GH on germ cells in unilateral undescended testis in rats.

OBJECTIVES: To evaluate the effects of the local release of fibroblast growth factor (FGF), insulin-like growth factor (IGF), and growth hormone (GH) on a germ cell population of ipsilateral undescended and contralateral descended testes of rats with a surgically created unilateral abdominal testis for 12 weeks and after the application of growth factors after orchidopexy. METHODS: Forty 4-week old male Wistar albino rats were divided into 5 groups as follows: group 1, sham; group 2, gelatin; group 3, FGF; group 4, IGF; and group 5, GH. In the sham group, the right testis was exposed and sutured with 5-0 silk sutures. In groups 2-5, a right intra-abdominal testis was surgically created. After 12 weeks, orchidopexy was performed, and 1 cm(2) gelatin films were sutured to the right testes, either unloaded (group 2), or containing 2.5 microg FGF (group 3), 5 microg IGF (group 4), or 5 microg GH (group 5). After 30 days, both testes were removed for histopathologic investigation and DNA flow cytometry. The mean seminiferous tubular diameters (MSTDs), mean testicular biopsy scores (MTBSs), and percentages of haploid (1n) cells were calculated. RESULTS: Ipsilateral MSTD and MTBS significantly decreased in the gelatin and FGF groups compared with the sham, IGF, and GH groups. Contralateral MSTDs and MTBSs did not differ among groups. The haploid cell percentage significantly decreased in the ipsilateral and contralateral testes of the gelatin group compared with the sham, FGF, IGF, and GH groups. CONCLUSIONS: Local release of IGF and GH resulted in the preservation of germ cell histology in the ipsilateral testes of rats with a surgically created unilateral undescended testis for 12 weeks and after orchidopexy. The administration of IGF, GH, and FGF increased the haploid germ cell population in both ipsilateral undescended and contralateral descended testes.

Expression of cell surface markers on methicillin resistant Staphylococcus aureus stimulated lymphocytes.

BACKGROUND & OBJECTIVE: Methicillin-resistant Staphylococcus aureus (MRSA) has gradually been increasing, new strategies in the treatment of MRSA infections are required. This depends on the understanding of the infection pathogenesis and the immune response. This study was therefore designed to determine the immune response which develops during MRSA infection and the role of chemokines in this response, and also to compare the results with the changes occurring after MSSA infection. METHODS: The expression of the surface markers of human lymphocytes stimulated by heat-killed MRSA or MSSA was analysed by flow cytometry. The chemokine levels in the lymphocytes culture supernatants stimulated or not stimulated by microorganisms were determined by ELISA. RESULTS: Human peripheral blood mononuclear cells (PBMCs) stimulated by MRSA the levels of CD4(+)CD25(+) regulatory T cells, CD69 expressions in the activated T lymphocytes, CD3(-)CD16(+)CD56(+) NK cells and the levels of MIP-1alpha, MIP-1beta, MCP-1 chemokines increased as compared to the cells not stimulated by MRSA. Although stimulation by MSSA caused an increase in CD25 expression after 24 h, the increase was found to be lower than the one caused by MRSA stimulation. The increase in CD69 expression was statistically significant compared to the cells stimulated by MRSA. Different from the cells stimulated by MRSA, no change was observed in the expressions of CD54 and CD3(-)CD16(+)CD56(+) NK cells in the cells stimulated by MSSA. INTERPRETATION & CONCLUSION: Our findings showed that cellular as well as humoral immunity are critical in MRSA infection and that T cell activation and the increase in chemokines may play a role in the regulation of immune response.

The efficiency of Viscum album ssp. album and Hypericum perforatum on human immune cells in vitro.

Viscum album L. ssp. album and Hypericum perforatum L. are used for the treatment of different diseases. In this study, the effects of these herbals on immune cells were assessed in vitro. The phagocytosis, candidacidal activity of neutrophils and adhesion function of epithelial cells were investigated. Also, the expression of the surface markers of lymphocytes was analyzed by flow cytometry. It was observed that V. album ssp. album increased phagocytic activity and candidacidal activity of neutrophils and decreased adhesion function of epithelial cells. We also observed that in human peripheral blood mononuclear cells stimulated by Viscum album L. ssp. album the levels of CD4(+)CD25(+) and CD8(+)CD25(+) T cells, CD69 expressions in the activated T lymphocytes and CD3(-)CD16(+)CD56(+) NK cells increased compared to the cells that were not stimulated by this herbal. Whereas CD4(+)CD25(+), CD8(+)CD25(+) T cells, CD 69 expression and CD3(-)CD16(+)CD56(+) Natural killer cells did not show any significant differences with the presence of Hypericum perforatum L. compared to the control group. Hypericum perforatum L. increased candidacidal activity of neutrophils and decreased adhesion function of epithelial cells. In the light of these findings, it is considered that these extracts may be used as an adjuvant treatment option for immune activation in immunosuppressed patients.

Expression of the surface antigens of lymphocytes and the levels of cytokines in mice infected with Aspergillus fumigatus.

BACKGROUND: Aspergillus fumigatus (A. fumigatus) is an opportunistic fungus that causes invasive aspergillosis. Determining the immune changes during A. fumigatus infection and the factors leading to such changes clearly will make it possible to prevent the spread of the infection and to provide new strategies in the treatment of infection. Thus, the present study aims at determining the changes of lymphocyte surface antigens which develop during A. fumigatus infection and the role of cytokines in immune response. METHODOLOGY: The expression of the surface antigens of lymphocytes was analysed by flow cytometry and the cytokine levels were determined by ELISA in a mouse model of aspergillosis. RESULTS: It was observed that in mice infected by A. fumigatus the percentage of CD19+ B cells and the levels of IL-4 and IL-10 increased when compared to those in noninfected mice cells. CONCLUSIONS: These results suggest that Th2 type cytokines are important in the pathogenesis of A. fumigatus infection. Humoral immunity is considered to be effective during A. fumigatus infection because of the increase in Th2 type response.