Effects of 1,25-Dihydroxy vitamin D3 on TNF-α induced inflammation in human chondrocytes and SW1353 cells: a possible role for toll-like receptors.

The purpose of this study is to evaluate anti-inflammatory and chondro-protective effects of 1,25(OH)2D3 in human chondrocytes and SW1353 cells via investigating expressions of MMPs, TIMPs, VDR, and intracellular signalling pathway mediators such as TLR-2 and -4. The HC and SW1353 cells were treated with 1,25(OH)2D3 at 10, 100, and 1000 nM concentrations in the absence/presence of TNF-α (20 ng/mL) for 48 h. The mRNA expressions of MMP-1, -2, -3, -9, and -13, TIMP-1 and -2, VDR, TLR-2 and -4 in HC and SW1353 cells were detected by qPCR after treatments. The cytotoxicity and cell proliferation analyses were assessed by LDH and WST-1 assay, respectively. Protein levels of MMPs, TIMPs, and VDR were analysed by immunocytochemistry and ELISA methods. TNF-α markedly increased cytotoxicity for 24, 48, 72 h (p < 0.05) and vitamin D treatment was shown to diminish the cytotoxic effect of TNF-α. Cell proliferations increased by Vitamin D in a dose-dependent manner. mRNA expressions of MMP-1, -2, -3, -9, and -13, TLR-2 and -4 genes decreased with 1,25(OH)2D3 treatment (p < 0.05). VDR, TIMP-1 and -2 levels elevated after TNF-α exposure compared with the control group in HC cells (p < 0.05). Protein expression levels were determined using Western blotting, ELISA and immunocytochemistry. 1,25(OH)2D3 via binding to VDR, reversed the effects of TNF-α by inhibiting TLR-2 and 4. Decreased levels of VDR, TIMP-1 and -2 after TNF-α treatment were elevated by 1,25(OH)2D3 proportional with increasing 1,25(OH)2D3 doses. 1,25(OH)2D3 and TNF-α co-treatment decreased MMP-1, -2, -3, -9, and -13 levels were after TNF-α exposure.

Dynamics of viral load in Crimean Congo hemorrhagic fever.

Crimean Congo hemorrhagic fever (CCHF) is a viral zoonotic disease with high mortality rate. There are only a few studies on viral load in CCHF. In our study, we revealed the dynamics of viral load and its relationship with mortality in early phase of the disease. A total of 138 serum samples were collected from 23 patients. All patients had positive PCR for CCHF on admission. Serum samples were obtained daily from all patients for the first 6 days of hospitalization and stored at -80°C for viral load measurement. We found statistically significant difference between mean number of viremic serum samples of fatal and non-fatal patients. Furthermore, non-fatal cases' viral loads demonstrated statistically significant decreases over time; however, we could not observe a similar trend in viral loads of fatal cases. Limited number of studies on CCHF indicate that score of the contest between CCHF virus and immune system determines the survival in CCHF and viral load is found to be the most prognostic factor. In our study, we found that there is a notable decrease trend in viral loads of non-fatal patients over time and this clearance of CCHF virus is significantly related with survival.

Loss of Angiotensin-converting enzyme-related (ACER) peptidase disrupts night-time sleep in adult Drosophila melanogaster.

Drosophila Acer (Angiotensin-converting enzyme-related) encodes a member of the angiotensin-converting enzyme family of metallopeptidases that have important roles in the endocrine regulation of blood homeostasis in mammals. Acer is expressed in the embryonic heart of Drosophila and expression in the adult head appears to be regulated by two clock genes. To study the role of Acer in development and in circadian activity, we have generated Acer null mutants by imprecise excision of a P-element and have compared their development and circadian behaviour with that of wild-type flies with the same genetic background. We show that Acer is not required for normal development, but that night sleep, which is clock regulated, is disrupted in adult flies lacking ACER. Acer null adults have reduced night-time sleep and greater sleep fragmentation, but normal levels of daytime sleep. The quality of night sleep in flies fed inhibitors of ACER is affected in a very similar manner. We have shown, using specific antibodies, that ACER is present in the adult fat body of the head and abdomen, and is secreted into the haemolymph. ACER might therefore have a role in cleaving regulatory peptides involved in metabolism and activity behaviour. There are similarities with mammals, where ACE peptidases are also expressed in adipose tissue and are thought to be part of a signalling system linking metabolism with sleep.

Genomic Characterization of the Novel Bartonella refiksaydamii sp. Isolated from the Blood of a Crocidura suaveolens (Pallas, 1811).

Bartonella species are reemerging infectious agents that are transmitted by arthropod vectors among animals and/or humans. At least 13 of the 35 currently recognized Bartonella species are pathogenic for humans. Most of the pathogenic species, except Bartonella quintana and Bartonella bacilliformis, are zoonotic agents with animal reservoirs, including cats, dogs, coyotes, foxes, cattle, and rodents. In this study, a novel Bartonella species was isolated from the blood of a Crocidura suaveolens (Pallas, 1811) Lesser shrew that was captured in the Bartin region of Northwestern Turkey. The strain, RSKK 19006, was characterized using whole-genome sequencing and comparison, multilocus sequence typing (gltA, rpoB, ssrA, nuoG, and 16S rRNA) and internal transcribed spacer sequencing, electron microscopy scanning, biochemical tests, and MALDI-TOF MS (matrix assisted laser desorption ionization-time of flight mass spectrometry). This novel Bartonella is a Gram-negative, rod-shaped, microaerophilic bacterium and has neither flagella nor pilus. As a consequence, we propose to name this new species Bartonella refiksaydamii sp. nov. in Bartonella genus. The zoonotic potential of this novel Bartonella species is as yet unknown.

Detection and genetic diversity of Bartonella vinsonii subsp. berkhoffii strains isolated from dogs in Ankara, Turkey.

Bartonella vinsonii subsp. berkhoffii has been identified as an important pathogen in dogs and is an emerging pathogen in people with zoonotic potential. This study aimed to isolate Bartonella spp. in 250 blood samples collected from dogs in the province of Ankara, Turkey, between October 2006 and March 2007. The typing of the 23 isolates was carried out by PCR for citrate synthase (gltA) and the 16S-23S intergenic transcribed spacer (ITS). The prevalence of bacteremia was 9.2% in 250 samples. Among 170 shelter dogs, 21 (12.4%) were bacteremic for B. vinsonii subsp. berkhoffii, while the B. vinsonii subsp. berkhoffii bacteremia rate was 5% (2/40) for the stray dogs. B. vinsonii subsp. berkhoffii was not isolated from the pet dogs. The prevalence of bacteremia was found to be 25.5% (13/51) in shelter dogs aged less than one year old. All of the isolates were identified as B.vinsonii subsp. berkhoffii genotype III. In some isolates, MseI digest for gltA was found to be different from the American and European strains due to a single nucleotide change.

[Evaluation of PCR and ELISA-IgM results in the laboratory diagnosis of Crimean-Congo haemorrhagic fever cases in 2008 in Turkey]. / 2008 yili Kirim-Kongo kanamali atesi olgularinin laboratuvar tanisinda PCR ve ELISA-IgM sonuçlarinin irdelenmesi.

Crimean-Congo hemorrhagic fever (CCHF) is a fatal zoonotic viral haemorrhagic infection described in Africa, Asia, Eastern Europe, and the Middle East. CCHF virus (CCHFV) classified in Bunyaviridae family and Nairovirus genus, is transmitted to humans by tick (Hyalomma and Ixodid) bites and human to human transmission may occur by direct contact with blood or other infected tissues. The disease became endemic and a public health problem since 2002 outbreak in Turkey. The specific laboratory diagnosis and confirmation of the disease is performed in Refik Saydam National Public Health Agency, by using molecular and serological methods. For this purpose serum and/or plasma samples from suspected CCHF patients are submitted to the reference laboratory with an official "possible case report form". According to the algorithm in our laboratory, the first samples which were sent from possible acute cases were searched initially by an in-house real time-polymerase chain reaction (PCR) method and those which were found negative with PCR, were then studied by in-house ELISA method in terms of CCHF-IgM antibodies. In 2008, a total of 4634 samples obtained from 2855 CCHF suspected patients have been examined for the positivity of CCHFV, and 1315 (46%) cases were found to be positive by molecular and/or serologic methods. The aim of this study was to evaluate the results of 726 cases whose at least 2 samples were sent to laboratory, with at least 1 positivity in at least 1 clinical sample with either PCR or IgM ELISA, or both, and with complete informations in possible case report form, during 2008 in Turkey. The positive results were also analyzed according to the starting date of the complaints and the date samples received in order to evaluate the positivity rates of molecular and serological methods with regard to the time. The first serum samples in 94.1% (683/726) of cases were found to be positive with PCR and/or ELISA-IgM methods. PCR positivity was found as 78.1% (567/726), while CCHFV-IgM positivity was detected in 116 (72.9%) in the remaining 159 PCR negative samples. In the first sera, PCR and ELISA results were evaluated in relation to the start of complaints and the date samples received. After the onset of symptoms, PCR positivity was determined as 83.4% in the samples taken in the first 5 days, and reduces to 67.5% in the samples between 6-10 days. The detection rate of CCHFV-IgM increases up to 95% when PCR positivity rate decreases after the 5th day. As expected, positivity is determined to be high by PCR in the first days, and ELISA-IgM after the 5th day. In conclusion, recording clinical data such as the onset of disease and the date of sample received ensure the accurate evaluation of the disease and the laboratory results are reliably accomplished in a short time.

[Influenza surveillance results in 2007-2008 winter season in nine provinces of Turkey]. / 2007-2008 kis sezonunda türkiye'nin dokuz ilinde influenza sürveyans sonuçlari.

Influenza virus infections constitute a serious public health problem owing to their epidemic and pandemic potential. Turkish Ministry of Health established the national influenza surveillance programme in two institutes to detect the virus types leading to the illness and the efficiency of the seasonal vaccine. Influenza surveillance is performed by Refik Saydam Hygiene Center, National Influenza Laboratory in nine provinces (which are located at central, northeast, south and east parts of Turkey) and by Istanbul University, Medical Faculty, Virology Laboratory in five provinces (which are located at west and northwest parts of Turkey). These two centers are the members of international information networks. The surveillance was aimed to contribute to the detection of influenza viruses with pandemic potential and also to determine the predominant strain circulating in Turkey. During November 2007-May 2008 period a total of 1157 clinical specimens collected from 90 health centers which were the representatives of nine provinces (Ankara, Samsun, Trabzon, Erzurum, Adana, Konya, Diyarbakir, Malatya and Van) were investigated for the presence of influenza virus and other respiratory viruses (Parainfluenza virus types 1-3, Respiratory Synctial Virus and Adenovirus). Samples were identified and subtyped by both molecular (real-time PCR) and cell culture techniques (MDCK and Hep-2). Influenza virus and at least one of the other respiratory viruses were detected in 321 (27.7%) and two different viruses in 16 of the specimens (total= 337). When all the specimens were considered, the most frequently identified virus was influenza A (n=188, 16.2%), H1N1 being 6.3% and H3N2 9.9%.The rate of identification for influenza B was 7.6% (n=88), for parainfluenza was 2.3% (n=27), for adenovirus was 2% (n=24) and for RSV was 0.9% (n=10). When only the positive specimens (n=337) were evaluated, influenza A was again the most frequently (55.7%) encountered virus, H1N1 being 38.8% and H3N2 61.2% of all. Influenza B was in the second rank with 26.1% frequency among the positive specimens. The results showed that influenza activity started around November and ended around May. When the distribution of influenza viruses were analysed according to months, Influenza A H1N1 predominated in January, influenza A H3N2 in December and February. influenza B viruses started to increase in February, and were also detected in May. The 2007-2008 influenza season in Turkey was characterized by moderate clinical activity, and a predominance of influenza A H3N2. These results indicate good match between the vaccine virus strains and the reported virus strains.

Evaluation estrogen, progesteron and androgen receptor expressions in corneal epithelium in keratoconus.

PURPOSE: To evaluate the role of sex steroid hormone receptors in corneal epithelium in etiopathogenesis of keratoconus (KC). METHODS: Thirty patients with KC who were planned for corneal collagen-crosslinking and 20 patients who were planned for excimer laser for refractive errors included in this study. Corneal epitheliums were curated mechanically during surgeries. Right eyes were evaluated immunohistochemically and left eyes were evaluated by quantitative polymerase chain reaction (qPCR) to investigate estrogenα, estrogenß, progesterone and androgen receptors. RESULTS: Immunohistochemically, staining for progesterone and androgen receptors did not significantly differ between KC and control groups (p > 0.05). None of the cases had staining for estrogenα and estrogenß receptors. qPCR showed that mRNA expressions of estrogenα and androgen receptors were significantly higher in the KC group (p < 0.001). CONCLUSION: A significantly higher rate of estrogenα and androgen receptor expressions in corneal epithelium from patients with KC through qPCR supports a possible relation between KC and sex steroid hormones.

The neutralizing capacity of Androctonus crassicauda antivenom against Mesobuthus eupeus scorpion venom.

Scorpion envenomations are considerable health problem in tropical and subtropical regions. There are approximately 1,500 species of scorpions worldwide. The number of dangerous species in the Buthidae family is significantly higher than in other families of scorpions. Mesobuthus eupeus is a member of Mesobuthus genus, Buthidae family. In this study, the potency and para-specific activities of Androctonus crassicauda antivenom were investigated against M. eupeus scorpion venom. The median lethal dose of M. eupeus and A. crassicauda scorpion venoms were found to be 0.18 mg/kg, 15.45 microg/kg by i.c.v injection route. The antivenom showed neutralization effect against the venoms of M. eupeus. One milliliter of A. crassicauda antivenom neutralizes 464LD(50) of M. eupeus and 940LD(50)A. crassicauda venom on mice. Western blotting demonstrated immunologic reaction with the venoms. The monovalent antivenom has immunoactivity and neutralizing capacity to the scorpion venoms. This study indicates that the antivenom produced by Refik Saydam Hygiene Center could be used for the treatment of M. eupeus stings in Turkey.

[Evaluation of laboratory diagnosis of the first norovirus outbreak in Turkey in 2008]. / Türkiye'de 2008 yilinda ortaya çikan ilk norovirus salgininin laboratuvar sonuçlarinin degerlendirilmesi.

Norovirus (NoV) is one of the most prominent agents of gastroenteritis and water/food-borne outbreaks affecting all of the age groups in the world. As the identification of the etiologic agent is important during gastroenteritis outbreaks, it is recommended to combine two different methods for rapid and reliable laboratory diagnosis of NoV. Although NoV outbreaks have been observed in many different countries of the world, there was no report on "NoV outbreak" in Turkey till 2008 due to the absence of a regular surveillance system for non-bacterial gastroenteritis. This study aimed to present the laboratory results for "the first NoV outbreak" in Turkey in 2008. A number of cases with diarrhea and nausea/vomiting initially emerged in Aksaray (located at the southern part of central Anatolia) in May 2008, followed by cases from Sereflikochisar, Kirsehir, and Adana provinces (located at central and southern Anatolia; geographically closer regions). However, regional laboratories declared that no known bacterial (Salmonella spp., Shigella spp., enterotoxigenic Escherichia coli), viral (Rotavirus, Adenovirus) and parasitic agents were detected. A total of 50 stool samples were sent to the Virology Reference Laboratory (Refik Saydam Hygiene Center, Ankara) for further investigations including NoV. For the investigation of NoV, the samples were analysed by using antigen-ELISA (Ridascreen, R-Biopharm, Germany) and real-time polymerase chain reaction(PCR) (Roche Diagnostics GmbH, Germany) methods. Of the samples, 26% (13/50) were found antigen positive, whereas 33% (13/40) were positive for viral nucleic acids. The positivity rates determined by ELISA and PCR were as follows, respectively; 57% (4/7) and 71% (5/7) in Aksaray, 25% (1/4) and 25% (1/4) in Sereflikochisar, 28% (7/25) and 40% (6/15) in Kirsehir, 7% (1/14) and 7% (1/14) in Adana. Nine (69.2%), and 4 (30.8%) out of 13 positive samples were genotyped as NoV GI and GII, respectively. The sensitivity and specificity of antigen-ELISA method were found as 61.5% and 100%, respectively, when compared with real-time PCR. In conclusion, further epidemiological studies and genomic analysis are needed for the detection and control of circulating strains in Turkey, since NoV outbreaks spread rapidly and cause serious economical and workforce loss.

Angiotensin-converting enzyme as a target for the development of novel insect growth regulators.

Insect angiotensin converting enzyme (ACE) is a zinc metallopeptidase capable of inactivating a variety of small to medium size peptide hormones by cleavage of C-terminal dipeptides and dipeptideamides. High levels of ACE activity are found in the hemolymph and in reproductive tissues of insects, where the enzyme is considered to have an important role in the metabolism of bioactive peptides. Therefore, inhibiting ACE activity is expected to interfere with the peptidergic endocrine system and to have detrimental effects on growth, development and reproduction. We will review the studies showing that ACE inhibitors do indeed disrupt growth and reproduction in various insect species. We will also present some new genetic and pharmacological data that strengthens our conclusion that ACE should be considered as a potential target for the development of new insect growth regulators.

Molecular identification of viral agents associated with acute conjunctivitis: a prospective controlled study.

BACKGROUND: Viral conjunctivitis are the most frequent infections in ophthalmology clinics. The diagnosis is usually relying on clinical findings and medical history. However, topical antibiotics are often used unnecessarily addition to symptomatic treatment because of unsure agents. We aimed to detect the Adenovirus, Coxsackievirus and Enterovirus from conjunctiva and pharyngeal samples of patients. METHODS: The conjunctiva and pharyngeal samples of the patients with conjunctivitis were taken by Virocult transport media and kept at -80̊C up to study day. Adenovirus spp, Enterovirus 70 and Enterovirus 71, Coxsackie A24 and Coxsackie A16 were detected by real-time PCR. Samples from healthy health care workers of ophthalmology clinic were used for control group. RESULTS: A total of 176 samples (conjunctival and pharyngeal samples of 62 patient and 26 healthy subjects) were included. The mean age of 34 (55.7%) male and 27 (44.3%) female patients was 34±17. Twenty five (40.3%) of the patients were receiving antibiotic drops at first visit. The main etiologic agent in conjunctival samples was found to be Adenovirus (46/62, 74.2%) followed by Enterovirus 70 (4/62, 6.4%) and Enterovirus 71 (4/62, 6.4%). Coxsackievirus 16 and 24 were also found in 2 patients (1/62 each, 1.6%). Pharyngeal samples were also positive for Adenovirus (20/62, 32.3%), Enterovirus 70 and 71 (7/62, 11.3% and 5/62, 8.1% respectively), Coxsackievirus 16 and 24 (2/62, 3.2% and 1/61, 1.6%). CONCLUSIONS: It is very difficult in viral conjunctivitis to make clinical differentiation caused by different agents because of common clinical signs and symptoms. In routine clinical work, the viral conjunctivitis usually related with Adenovirus. But almost one fourth of the patients' conjunctivitis were not related to Adenovirus, which shows the importance of the laboratory diagnostics. True diagnosis plays an important role on prevention of contamination and unnecessary use of antibiotics in viral conjunctivitis.

Drosophila melanogaster NEP2 is a new soluble member of the neprilysin family of endopeptidases with implications for reproduction and renal function.

The mammalian neprilysin (NEP) family members are typically type II membrane endopeptidases responsible for the activation/inactivation of neuropeptides and peptide hormones. Differences in substrate specificity and subcellular localization of the seven mammalian NEPs contribute to their functional diversity. The sequencing of the Drosophila melanogaster genome has revealed a large expansion of this gene family, resulting in over 20 fly NEP-like genes, suggesting even greater diversity in structure and function than seen in mammals. We now report that one of these genes (Nep2) codes for a secreted endopeptidase with a highly restricted pattern of expression. D. melanogaster NEP2 is expressed in the specialized stellate cells of the renal tubules and in the cyst cells that surround the elongating spermatid bundles in adult testis, suggesting roles for the peptidase in renal function and in spermatogenesis. D. melanogaster NEP2 was found in vesicle-like structures in the syncytial cytoplasm of the spermatid bundles, suggesting that the protein was acquired by endocytosis of protein secreted from the cyst cells. Expression of NEP2 cDNA in D. melanogaster S2 cells confirmed that the peptidase is secreted and is only weakly inhibited by thiorphan, a potent inhibitor of human NEP. D. melanogaster NEP2 also differs from human NEP in the manner in which the peptidase cleaves the tachykinin, GPSGFYGVR-amide. Molecular modelling suggests that there are important structural differences between D. melanogaster NEP2 and human NEP in the S1' and S2' ligand-binding subsites, which might explain the observed differences in inhibitor and substrate specificities. A soluble isoform of a mouse NEP-like peptidase is strongly expressed in spermatids, suggesting an evolutionarily conserved role for a soluble endopeptidase in spermatogenesis.

Crucial parameter of the outcome in Crimean Congo hemorrhagic fever: Viral load.

BACKGROUND: Crimean Congo hemorrhagic fever (CCHF) is a fatal disease with a mortality rate of 5-30%. CCHF can be asymptomatic or it may progress with bleeding and cause mortality. OBJECTIVES: To evaluate relation of viral load with mortality, clinical and laboratory findings in CCHF. STUDY DESIGN: A total of 126 CCHF patients were included. Serum samples obtained from all patients on admission for measurement of viral load. RESULTS: In our study, mortality rate was 11.1%. The most important prognostic factor was viral load. Mean viral load was 8.3×10(7)copy/ml and 4.6×10(9)copy/ml in survived and dead patients, respectively (p<0.005). Probability of survival is found to be significantly reduced where AST >1130U/l, ALT >490U/l, CPK >505U/l, LDH >980U/l, platelet count <23×10(3)/l, creatinine >1.4mg/dl, INR >1.3, d-dimer >7100ng/dl, and viral load >1.03×10(8)copy/ml. Patients with 10(8)copy/ml or higher viral load had diarrhea, headache, unconsciousness, bleeding, and seizure significantly more frequently (p<0.05). WBC, hemoglobin, platelet counts were significantly lower whereas AST, ALT, CPK, LDH, creatinine levels, PT and aPTT time, d-dimer levels, and INR were found to be significantly higher in these group. CONCLUSIONS: There are several severity criteria for prognosis of CCHF. In addition to these parameters, we introduce creatinine as a predictive factor for prognosis. Our study, which has the largest number of patients among studies that evaluate viral load on CCHF shows that viral load is the most effective parameter on mortality.

The First Identification of Encephalitozoon cuniculi Infection in an Animal Care Worker in Turkey.

BACKGROUND: As a zoonotic pathogen, Encephalitozoon cuniculi is a cause of serious disease in animals and people. The present study was to evaluate the health status examination of this seropositive animal care worker in our previous study. METHODS: Blood samples were taken from five workers. CIA test was applied to detect antibodies against E. cuniculi in blood serum. The indirect immunofluorescence antibody test was used as confirmation test. Seropositive worker had a complete medical examination. RESULTS: Only one worker was found to be seropositive according to the results of the serological test. Sera positive to E. cuniculi was confirmed with IFAT and spores were detected in the urine sample of the worker. The worker was treated with albendazole. CONCLUSION: Rabbits should be examined routinely for the presence of anti-E. cuniculi antibody. People working with laboratory animal should avoid contact with urine and faeces of infected or pay attention to personal hygiene.

Mutation in slowmo causes defects in Drosophila larval locomotor behaviour.

We have identified a mutant slowmotion phenotype in first instar larval peristaltic behaviour of Drosophila. By the end of embryogenesis and during early first instar phases, slowmo mutant animals show a marked decrease in locomotory behaviour, resulting from both a reduction in number and rate of peristaltic contractions. Inhibition of neurotransmitter release, using targeted expression of tetanus toxin light chain (TeTxLC), in the slowmo neurons marked by an enhancer-trap results in a similar phenotype of largely absent or uncoordinated contractions. Cloning of the slowmo gene identifies a product related to a family of proteins of unknown function. We show that Slowmo is associated with mitochondria, indicative of it being a mitochondrial protein, and that during embryogenesis and early larval development is restricted to the nervous system in a subset of cells. The enhancer-trap marks a cellular component of the CNS that is seemingly required to regulate peristaltic movement.

Molecular identification of viral agents associated with acute conjunctivitis: a prospective controlled study

Abstract Background: Viral conjunctivitis are the most frequent infections in ophthalmology clinics. The diagnosis is usually relying on clinical findings and medical history. However, topical antibiotics are often used unnecessarily addition to symptomatic treatment because of unsure agents. We aimed to detect the Adenovirus, Coxsackievirus and Enterovirus from conjunctiva and pharyngeal samples of patients. Methods: The conjunctiva and pharyngeal samples of the patients with conjunctivitis were taken by Virocult transport media and kept at -80 ºC up to study day. Adenovirus spp, Enterovirus 70 and Enterovirus 71, Coxsackie A24 and Coxsackie A16 were detected by real-time PCR. Samples from healthy health care workers of ophthalmology clinic were used for control group. Results: A total of 176 samples (conjunctival and pharyngeal samples of 62 patient and 26 healthy subjects) were included. The mean age of 34 (55.7%) male and 27 (44.3%) female patients was 34 ± 17. Twenty five (40.3%) of the patients were receiving antibiotic drops at first visit. The main etiologic agent in conjunctival samples was found to be Adenovirus (46/62, 74.2%) followed by Enterovirus 70 (4/62, 6.4%) and Enterovirus 71 (4/62, 6.4%). Coxsackievirus 16 and 24 were also found in 2 patients (1/62 each, 1.6%). Pharyngeal samples were also positive for Adenovirus (20/62, 32.3%), Enterovirus 70 and 71 (7/62, 11.3% and 5/62, 8.1% respectively), Coxsackievirus 16 and 24 (2/62, 3.2% and 1/61, 1.6%). Conclusions: It is very difficult in viral conjunctivitis to make clinical differentiation caused by different agents because of common clinical signs and symptoms. In routine clinical work, the viral conjunctivitis usually related with Adenovirus. But almost one fourth of the patients' conjunctivitis were not related to Adenovirus, which shows the importance of the laboratory diagnostics. True diagnosis plays an important role on prevention of contamination and unnecessary use of antibiotics in viral conjunctivitis.

Detection of Crimean-Congo hemorrhagic fever virus genome in saliva and urine.

BACKGROUND: The Crimean-Congo hemorrhagic fever (CCHF) virus is transmitted by tick bites and by contact with the blood or tissues of infected patients and livestock. This study was designed to investigate the genome of CCHF virus in saliva and urine samples of patients with CCHF. METHODS: Eight patients with laboratory-confirmed CCHF were included in the study. The diagnosis was made by detection of viral RNA in blood by real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR). Samples of saliva from six patients and samples of urine from three patients were collected at the same time as the blood samples and analyzed for viral RNA. RESULTS: The genome of CCHF virus was detected in the saliva from five of the six patients and in the urine from two of the three patients. The levels of viral load in the saliva and urine samples were similar to those in the blood samples in all but one patient, in whom higher levels were detected in blood compared to saliva or urine. CONCLUSIONS: This study shows that during human infection with CCHF virus, viral genomes are present in the saliva and urine. Further studies to isolate infectious viruses from these fluids and to study whether they represent an infectious risk are underway.

Characterization of a sandfly fever Sicilian virus isolated during a sandfly fever epidemic in Turkey.

BACKGROUND: Phleboviruses cause sandfly fever but isolates are rare. OBJECTIVES: To analyse samples from concurrent outbreaks of suspected sandfly fever in the Mediterranean provinces of Adana, Izmir and the central province of Ankara, Turkey. STUDY DESIGN: Samples from acute cases were analysed by immunofluorescence assay (IFA). Virus isolation was attempted and pyrosequencing performed. RESULTS: In IFA 38% of 106 samples tested scored IgM positive for sandfly fever Sicillian virus (SFSV), 12% for SFSV/sandfly fever Cyprus Virus (SFCV) and only 4% for SFCV. A sandfly fever Sicilian type virus designated sandfly fever Turkey virus (SFTV) was isolated. The S-segment sequence of SFTV had a homology of 98% to that of SFCV. The M-segment sequence showed a 91.1% homology to the only SFSV sequence available. The L-segment sequence showed a homology of 58% and 60.3% to Toscana virus and Rift Valley Fever virus sequences, a partial 201nt sequence showed 95.5% homology to the SFSV Sabin strain. CONCLUSION: A new phlebovirus related to sandfly fever Sicilian virus, SFTV was isolated and characterized from acute patient material. The sandfly fever Sicilian virus activity seems to be changing in Turkey. Entomological studies are needed.

Molecular epidemiology of Crimean-Congo hemorrhagic fever virus in Turkey: occurrence of local topotype.

The goal of this study was to investigate the molecular epidemiology of Crimean-Congo hemorrhagic fever virus (CCHFV) in Turkey. The study was performed on a total of 48 confirmed human CCHF cases from 2006 to 2008. The majority of the CCHF viral strains in Turkey were found to belong to the European lineage. Local CCHF viral strains are grouped into two main clusters, which can be further divided into two sub-groups. We also identified an AP92-like virus causing clinical disease in Corum (a mid-Anatolian province). Phylogenetic analysis revealed that the most recent CCHFV infections were caused by intrinsic (or native) CCHF viral strains, which we identified as the local topotype. Comparison of deduced amino acid sequences of S-segment RNAs indicated that the local topotype was derived from viruses of previous years, most likely by a low rate recombination. No genetic differences, based on S- and M-segment RNA sequences, were found between human and tick viral isolates. This data suggest that replication of CCHFV in the tick vector, whether Rhiphicephalus spp. or Hyalomma spp., has no effect on the viral genomic structure.