Assesment of attainment of recommended TSH levels and levothyroxine compliance in differentiated thyroid cancer patients.

OBJECTIVE: Thyroid-stimulating hormone (TSH) suppression treatment can induce signs and symptoms of hyperthyroidism and hypothyroidism due to inappropriate treatment or poor compliance to the treatment. The current study aimed to investigate TSH levels, frequency of being on target TSH, adherence to levothyroxine (LT4) suppression treatment in differentiated thyroid cancer (DTC) patients after surgery in a multicentric setting. DESIGN AND PATIENTS: This multicentric cross-sectional study was conducted at 21 medical centres from 12 cities in Turkey. DTC patients followed at least one year in the same center included in the study. Clinical data, serum TSH, free thyroxine (FT4), thyroglobulin (Tg) and anti-Tg levels were recorded during the most recent visit. Body mass index, systolic and diastolic blood pressures, pulse rate were measured. LT4 doses were recorded and doses per kilogram of bodyweight were calculated. Pill ingestion habits recorded and adherence to the therapy were evaluated using the Morisky Medication Adherence Scale and categorized as good, moderate or poor compliant based on their scores. Risk stratification forpredicting the disease persistance and/or reccurence was assessed using the American Joint Committee on Cancer-7th edition thyroid cancer staging calculator. TSH serum concentrations were classified as severe suppression (TSH < 0.01 mU/L), moderate suppression (TSH: 0.01-0.1 mU/L), mild suppression (TSHL 0.1-0.5 mU/L), euthyroid (TSH: 0.5-4 mU/L) and hypothyroid (TSH > 4 mU/L). TSH levels can also be classified as on being on target, under the target, or beyond over the target, according to the American Thyroid Association recommendations. RESULTS: A group of 1125 patients (F/M: 941/184, 50.7 ± 11.7 years) were included in the study. The mean LT4 daily dosage was 132.4 ± 39.6 mcg/day. TSH levels showed severe suppression in 99 (%8.8) patients, moderate suppression in 277 (%24.6) patients and mild suppression in 315 (%28) patients and euthyroid range in 332 (%29.5) patients and hypothyroid range in 97 (8.6%). TSH levels were in target in 29.2% of the patients 20.4% of the patients were undertreated, 50.4% overtreated. The daily LT4 dose and LT4 dose/kg were significantly higher in the severe suppression group (p < .001, p < .001). According to the Morisky scale, 564 patients (50.1%) were good compliant, 368 patients (32.7%) were moderate compliant, and 193 patients (17.1%) were noncompliant. Patients with poor compliance need a higher dose of LT4 compared to the good compliance group (p < .001). TSH levels of patients with good compliance were 0.67 ± 1.96 mU/L and TSH with poor compliance was 2.74 ± 7.47 mU/L (p < .001). TSH levels were similar in patients on fixed and alternating dosages. CONCLUSION: In 29.2% of the DTC patients, serum TSH levels were at target levels. Remaining of the study group have TSH levels under or over treatment range, exposing the patient to medication side effects. Majorty of the study group 82.8% have good or moderate adherence to LT4 therapy. Reaching TSH targets requires simplified and applicable guidelines and following the guideline recommendations.

Immune responses in multiple hosts to Nucleocapsid Protein (NP) of Crimean-Congo Hemorrhagic Fever Virus (CCHFV).

In 2019, the World Health Organization declared 3 billion to be at risk of developing Crimean Congo Hemorrhagic Fever (CCHF). The causative agent of this deadly infection is CCHFV. The data related to the biology and immunology of CCHFV are rather scarce. Due to its indispensable roles in the viral life cycle, NP becomes a logical target for detailed viral immunology studies. In this study, humoral immunity to NP was investigated in CCHF survivors, as well as in immunized mice and rabbits. Abundant antibody response against NP was demonstrated both during natural infection in humans and following experimental immunizations in mice and rabbits. Also, cellular immune responses to recombinant NP (rNP) was detected in multispecies. This study represents the most comprehensive investigation on NP as an inducer of both humoral and cellular immunity in multiple hosts and proves that rNP is an excellent candidate warranting further immunological studies specifically on vaccine investigations.

Effect of hyperparathyroidism on coagulation: a global assessment by modified rotation thromboelastogram (ROTEM)

Background/aim: Hyperparathyroidism is an endocrine disorder characterized by hypercalcemia. Because of calcium's effects on parathyroid glands, bone, intestines, and kidneys, it has an important place in homeostasis. The results of studies regarding hyperparathyroidism hemostasis are conflicting. Thromboelastography helps to evaluate all steps of hemostatic system. Our aim in this study was to investigate the possible role of hemostatic mechanisms in the development of thrombosis in hyperparathyroid patients with the modified rotation thromboelastogram (ROTEM). Materials and methods: Twenty-two patients with primary hyperparathyroidism (PHPT) and 20 healthy controls were involved. This study was conducted in Eskisehir Osmangazi University Faculty of Medicine, Endocrinology and Hematology clinics for 2 years. The complete blood count, fibrinogen, D-dimer levels, prothrombin time, activated prothrombin time, and ROTEM parameters [clot formation time (CFT), clotting time (CT), and maximum clot formation (MCF)] were determined by two activated tests, INTEM and EXTEM analyses. A thromboelastographic evaluation was performed in the preoperative and postoperative (3 months after surgery) periods. Results: In INTEM assay, the CT (p = 0.012) and CFT (p = 0.07) values were increased in preoperative PHPT patients compared with the control group. Although there was a decrease in the postoperative CT and CFT values, no statistical difference was found. Conclusion: The prolongation of the CT and CFT values were consistent with a hypocoagulable state in patients with PHPT. Hyperparathyroidism causes a hypocoagulable state that can be successfully assessed by ROTEM. Hemostatic changes, do not seem to have an effect on increased cardiovascular mortality.

Assessment of fragmented QRS formation and its relationship with left ventricular hypertrophy in nonhypertensive acromegaly patients

Background/aim: It is known that the presence of fragmented QRS (fQRS) on electrocardiography (ECG) is associated with cardiovascular events. The aim of this study was the evaluation of fQRS formation and its relationship with the left ventricular hypertrophy (LVH) parameters in acromegaly patients. Materials and methods: In total, 47 previously diagnosed with non-hypertensive acromegaly patients and 48 control subjects were included in the study. ECG and transthoracic echocardiography (TTE) were performed for each participant. Acromegaly patients were divided into two groups according to the fQRS formation on the ECG. Left ventricular wall thicknesses, and left atrial diameter (LAD), left ventricular mass (LVM), left ventricular mass index (LVMi), and relative wall thickness (RWT) were obtained. Results: In control group 5 (10.4%) and in acromegaly group 17 (36.2%) patients had fQRS on ECG (p = 0.003). LAD [36.0 (34.0­38.0) vs. 38.0 (35.0­41.0) mm, p < 0.001], LVM [155.27 ± 27.00 vs. 173.0 (153.0­235.0) g, p < 0.001], LVMi [83.12 ± 13.19 vs. 92.0 (83.0­118.0) g/m², p < 0.001] and RWT [0.39 ± 0.03 vs. 0.43 (0.41­0.45), p = 0.001] were significantly higher in patients with acromegaly. Disease duration was significantly higher (11.59 ± 1.3 vs. 8.2 ± 1.8 years, p < 0.001) in the fQRS (+) group. LAD [41.0 (39.0­42.5) vs. 37.0 (34.7­38.0) mm, p < 0.001], LVM [219.0 (160.5­254.5) vs. 164.0 (153.0­188.0) g, p = 0.017], LVMi [117.0 (92.5­128.5) vs. 86.0 (82.0­100.2) g/m², p = 0.013] and RWT [0.44 (0.42­0.49) vs. 0.43 (0.40­0.44), p = 0.037] were significantly higher in fQSR (+) acromegaly patients. In multivariate logistic regression analysis, disease duration (odds ratio: 10.05, 95% CI: 1.099­92.012, p = 0.041) and LAD (odds ratio: 2.19, 95% CI: 1.030­4.660, p = 0.042) were found to be the independent predictors of fQRS formation. Conclusion: The results of our study revealed that fQRS (+) acromegaly patients had increased LVH parameters compared to fQRS (-) patients.

Evaluation of the cell culture based and the mouse brain derived inactivated vaccines against Crimean-Congo hemorrhagic fever virus in transiently immune-suppressed (IS) mouse model.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus in the Nairoviridae family within the Bunyavirales order of viruses. Crimean-Congo hemorrhagic fever (CCHF) is the most widespread among tick-borne human viral diseases. It is endemic in many areas of Africa, Asia, the Middle East, in the Balkans, Russia and countries of the former Soviet Union. The confirmed CCHF cases were seen in Spain in 2016 to signify expansion of the virus into new geographical areas. CCHFV causes a viral human disease characterized by sudden onset of fever, headache, abdominal pain, nausea, hypotension, hemorrhage, and hepatic dysfunction with fatality rates up to 30%. Currently, there are no spesific treatments or licensed vaccines available for CCHFV. The absence of a susceptible animal model for CCHFV infection was severely hindered work on the development of vaccines. However, several animal models of CCHFV infection have been recently developed and used to assess vaccine efficacy. In this study, we have used the transiently immune-suppressed (IS) mouse model that MAb-5A3 was used to block IFN-I signaling in immune intact, wild-type mice at the time of CCHFV infection to evaluate the immune response and efficacy of the cell culture based and the mouse brain derived inactivated vaccines against CCHFV. Both vaccine preparations have provided complete protection but the cell culture based vaccine more effectively induced to CCFHV spesific antibodies and T cell responses. This is the first comparison of the cell culture based and the mouse brain derived vaccines for assessing the protective efficacy and the immunogenicity in the IS mouse CCHFV model.

[Measles outbreak in the adult age group: evaluation of 28 cases]. / Eriskin yas grubunda kizamik salgini: 28 olgunun degerlendirilmesi.

Nowadays, the age group affected from measles has widened and the disease has become more common among adolescents and young adults. The number of measles case reports have increased in our country, particularly from 2010-2011, and measles outbreaks occurred in various regions in 2012 and 2013. The aim of this study was to analyze the demographical and epidemiological characteristics, clinical and laboratory findings, and complications of adult patients with measles who were affected during the outbreak. A total of 28 patients (25 male, 3 female; age range: 19-39 years, median age: 24) who were hospitalized and followed-up in our clinic between January 2013 and June 2013, were evaluated. In the serum sample of the index case, measles-specific IgM antibodies were detected by ELISA, and measles virus RNA by real-time polymerase chain reaction (RT-PCR), then genotyping was performed to detect the epidemiological relationship. In all of the other cases, measles IgM and IgG antibodies were screened by ELISA. The most common symptoms on admission included high fever (n= 28, 100%), malaise (n= 25, 89%), sore throat (n= 25, 89%), headache (n= 20, 71%) and cough (n= 18, 64%). At physical examination, rash (n= 28, 100%), lymphadenopathy (n= 11, 39%) and conjunctivitis (n= 10, 36%) were in the foreground, and Koplik spots were detected in five (18%) cases. The most common laboratory findings were; increased level of C-reactive protein (n= 15, 54%), leukopenia (n= 12, 43%) and increased serum levels of aminotransferases (n= 12, 43%), and thrombocytopenia was detected in five (18%) patients. One or more complications (secondary bacterial pneumonia in 5, diarrhea in 4, hepatitis in 3 and otitis in 2 cases) developed in the eight (29%) patients. Measles RT-PCR and IgM tests yielded positive results for the index case, and the isolate was identified as D8 strain by genotyping. Measles lgM antibodies were also positive in all of the other cases. The hospitalization period was estimated as 3-7 days (median: 5 days), while all the patients were discharged with recovery. It appeared that, our index case had come from a troop in Amasya province three days ago and he had a history of contact with suspected measles patients. In addition, the D8 strain determined in the index case was found to be related with the strain that caused the outbreak in Amasya province. Of the cases, 20 (71.4%) were military personnel, and eight (28.6%) were civilian who had histories of contact with military personnel. Regardless of immunity status in the outbreak period, all of the healthcare staff in our hospital, especially in risky departments, was recommended to be vaccinated. Personnel vaccination was provided at a high rate, however nosocomial transmission occurred in two unvaccinated cases. In conclusion, measles is an important health problem, especially in the adult age group, because of the complications and labour loss. For outbreak management; the awareness of health personnel should be increased following the identification of index case, proper isolation measures should be taken for the hospitalized patients, and routine reporting should be carried out timely and accurately.

Immunization of knock-out α/ß interferon receptor mice against high lethal dose of Crimean-Congo hemorrhagic fever virus with a cell culture based vaccine.

Crimean-Congo hemorrhagic fever (CCHF) is an acute tick-borne zoonotic disease. The disease has been reported in many countries of Africa, Asia, the Middle East, and in Eurasia. During the past decade, new foci of CCHF have emerged in the Balkan Peninsula, southwest Russia, the Middle East, western China, India, Africa, and Turkey. CCHF virus produces severe hemorrhagic manifestations in humans with fatality rates up to 30%. Vaccine development efforts have been significantly hampered by a lack of animal models and therefore, no protective vaccine has been achieved. Lately, IFN α/ß receptor deficient (IFNAR-/-) mice have been established as a novel small animal model of CCHF virus infection. In the present study, we found that IFNAR-/- mice highly susceptible to CCHF virus Turkey-Kelkit06 strain. Immunization with the cell culture based vaccine elicited a significant level of protection against high dose challenge (1,000 PPFU) with a homologous CCHF virus in IFNAR-/- mice.

In vitro activity of Brucella melitensis isolates to various antimicrobials in Turkey.

BACKGROUND: Brucellosis is a systemic infectious disease caused by Brucella bacteria. A successful treatment requires antibiotics that can penetrate into the cell at high concentrations. The aim of this study was to assess the biotype and in vitro activity of 80 Brucella isolates obtained from blood against various antimicrobials for human brucellosis in Turkey. METHODS: Identification of the types of the species designated Brucella species was made using the polymerase chain reaction (PCR), with type-specific primers. Serotyping was performed using mono-specific A and M antisera. The minimum inhibitory concentrations (MICs) of antibiotics known to have good intracellular penetration (doxycycline, rifampicin, ofloxacin, levofloxacin, moxifloxacin, clarithromycin, and azithromycin) were determined by the agar dilution method. RESULTS: All of the 80 Brucella isolates were determined to be Brucella melitensis: 75 B. melitensis biotype 3 (93.7%) and 5 B. melitensis biotype 1 (6.3%). Doxycycline was the most effective among the tested antibiotics against Brucella species (MIC(50)-MIC(90), 0.25-0.5 µg/ml), and it was followed by levofloxacin (MIC(50)-MIC(90), 0.5-1 µg/ml), moxifloxacin (MIC(50)-MIC(90), 1-1 µg/ml), ofloxacin (MIC(50)-MIC(90), 1-1 µg/ml), rifampicin (MIC(50)-MIC(90), 2-4 µg/ml), azithromycin (MIC(50)-MIC(90), 4-8 µg/ml), and clarithromycin (MIC(50)-MIC(90), 8-32 µg/ml), respectively. CONCLUSIONS: The in vitro activity of doxycycline and rifampicin, which are used in the classic treatment of brucellosis, was found to be very good. Quinolones were found to have in vitro activity against Brucella isolates. Among the macrolides, azithromycin had a higher level of activity compared with clarithromycin. A combination of quinolones and azithromycin could be an alternative to doxycycline and rifampicin in the treatment of brucellosis.

Pseudo-plaque reduction neutralization test (PPRNT) for the measurement of neutralizing antibodies to Crimean-Congo hemorrhagic fever virus.

BACKGROUND: Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus of the genus Nairovirus family Bunyaviridae, which are enveloped viruses containing tripartite, negative polarity, single-stranded RNA. CCHF is characterized by high case mortality, occurring in Asia, Africa, the Middle East and Europe. Currently, there are no specific treatments or licensed vaccines available for CCHFV. Recently, two research groups have found adult mice with defective interferon responses allowed to lethal CCHFV infection. These mouse models could provide invaluable information for further studies. Efforts to develop a vaccine against CCHFV are being made. To determine the efficacy of vaccine candidates it is important to conduct serological studies that can accurately measure levels of protective antibodies. In the present study, a pseudo-plaque reduction neutralization test (PPRNT) based on enzyme-catalyzed color development of infected cells probed with anti-CCHFV antibodies was used to measure neutralization antibody of CCHFV. METHODS: Sixty-nine human serum samples (20 acute and 49 convalescent) were tested. The presence of CCHFV antibodies was determined and confirmed by a commercial ELISA kit. CCHFV RNA was determined by RT-PCR. All the samples were analyzed by PPRNT and fluorescent focus reduction neutralization test (FFRNT) to measure of CCHFV-neutralizing antibodies. RESULTS: Pseudo-plaque reduction neutralization test showed a high sensitivity (98%), specificity (100%) and agreement (96,6%) in qualitative comparison with those of the FFRNT. There was a high correlation between the titers obtained in PPRNT and FFRNT (R2 = 0.92). The inter- and intra-assay variation of PPRNT revealed good reproducibility and positive cut-off of PPRNT was defined as 1:4 by the geometric mean titers for the individual samples distributed. CONCLUSION: The pseudo-plaque reduction neutralization test described in this study is a fast, reproducible and sensitive method for the measurement of CCHF neutralizing antibodies. This novel assay could serve as useful tools for CCHF research in epidemiology, vaccine development and other studies of immunity. It also provides an alternative to PRNT when viruses with no or poor CPE in cell culture.

Treatment of Gram-positive left-sided infective endocarditis with daptomycin.

The aim of this study was to evaluate the effectiveness of daptomycin in left-sided infective endocarditis (IE) patients. Fourteen patients with left heart endocarditis, monitored with a diagnosis of IE based on modified Duke criteria between July 2010 and May 2011, and receiving daptomycin as monotherapy, were enrolled. The success of daptomycin in these patients was revealed with improvements in microbiological, biochemical, and radiologic findings, as well as physical examination findings. Patient average age was 63.5 ± 14.2 years (36-80 years); 8 (57 %) were men and 6 (43 %) women. The pathogens methicillin-resistant Staphylococcus aureus (71.5 %), Streptococcus mutans (21.5 %), and methicillin-sensitive Staphylococcus aureus (7 %) were isolated from our patients. Daptomycin was used in initial treatment in 5 (36 %) patients; treatment was subsequently modified to daptomycin in 9 (64 %) patients as a consequence of drug serum level insufficiency, agent sensitivity to the drug administered, or drug side effects. Thirteen patients were discharged in a healthy condition, with successful surgical treatment in 5 (36 %). Only 1, an 80-year-old IE patient, was lost from advanced cardiac failure. No significant side effects were seen in any patient receiving daptomycin. The most frequent side effects were minimal rises in serum CPK levels during treatment; these values returned to normal after treatment. Daptomycin can be used successfully in left heart endocarditis with no significant side effects. Studies involving a wider patient series are now needed to support the use of daptomycin in left heart endocarditis.

Application of the pseudo-plaque assay for detection and titration of Crimean-Congo hemorrhagic fever virus.

A pseudo-plaque assay was developed for detection and quantitation of Crimean-Congo hemorrhagic fever virus Turkey-Kelkit06. Enzyme-catalyzed color development of infected cells probed with anti-Crimean-Congo hemorrhagic fever virus antibodies was used for determining the titer of Crimean-Congo hemorrhagic fever Turkey-Kelkit06 and for its detection in samples from persons infected with the Crimean-Congo hemorrhagic fever virus. The pseudo-plaque assay accuracy was confirmed by comparing pseudo-plaque assay titers with fluorescent immunofocus assay and focus formation assay titers using three stocks of virus. No significant difference in virus titers of Crimean-Congo hemorrhagic fever Turkey-Kelkit06 among the three methods was observed. The pseudo-plaque assay is more sensitive than the fluorescent immunofocus assay for detecting the virus in primary isolates of Crimean-Congo hemorrhagic fever virus collected from humans, but no difference in sensitivity between the two methods was observed in the cell-adapted strain of Crimean-Congo hemorrhagic fever Turkey-Kelkit06. The pseudo-plaque assay is suitable for titration of Crimean-Congo hemorrhagic fever Turkey-Kelkit06, which does not develop plaques, suggesting it may also be suitable for the detection of other viruses.

The complete genome analysis of Crimean-Congo hemorrhagic fever virus isolated in Turkey.

Crimean-Congo hemorrhagic fever virus (CCHFV) is the causative agent of a tick-borne disease with high mortality rates in humans. The distribution of CCHFV includes over 30 countries in Asia, the Middle East, southeastern Europe, and Africa. It was first recognized in Turkey in 2002, with an increasing number of cases reported between 2002 and 2009. Recent analysis of complete genome sequences of CCHFV isolates has revealed that the genomic plasticity of the virus is surprisingly high for an arthropod-borne virus. We have determined the complete nucleotide and deduced amino acid sequences of strain CCHFV Turkey-Kelkit06 isolated from the blood of a patient in an endemic region of Turkey in 2006. The complete sequence length of the CCHFV Turkey-Kelkit06 strain is 19,186 nt, consisting of a 1673 nt S segment, a 5364 nt M segment, and a 12,149 nt L segment. Based on the analysis of S, M, and L segments, CCHFV Turkey-Kelkit06 clustered in Group V, which represents the Europe/Turkey geographic lineage. Although glycoproteins encoded by the M gene are the most variable part of the CCHFV Turkey-Kelkit06 strain, some functional domains of the glycoproteins are well conserved. Here, we report the complete sequence and genome organization of the CCHFV Turkey-Kelkit06 strain and its phylogenetic relationship to other strains of CCHFV. Collecting data on viral sequences among isolates from CCHF epidemics may provide valuable information regarding the molecular basis of the epidemic potential of the virus.

Molecular detection and identification of Ehrlichia and Anaplasma species in ixodid ticks.

A polymerase chain reaction (PCR) assay followed by partial sequencing of the 16S ribosomal RNA gene was performed for the presence of Ehrlichia and/or Anaplasma. A total of 242 ixodid ticks were collected from domestic ruminants and their shelters, as well as humans, and their individual salivary glands were dissected out for DNA. From the 242 ticks analyzed, six (2.47%), comprising three Hyalomma anatolicum anatolicum, one Rhipicephalus bursa, and two Rhipicephalus sanguineus, were positive. Of these sequenced samples directly obtained from the PCR products, three sequences from H. a. anatolicum were identical to that of the gene of Ehrlichia spp. strains. One sequence identified in R. bursa was closely related to Anaplasma platys. The remaining two sequences detected in R. sanguineus were similar to that of the gene of Anaplasma ovis. The study presented here provides preliminary data regarding the presence of rickettsial pathogens in ticks in Turkey.

Toxocara seroprevalence in schizophrenic patients in Turkey.

PURPOSE: To investigate the seroprevalence of toxocariasis in patients diagnosed as schizophrenia. PATIENTS AND METHODS: Ninety-eight schizophrenic patients hospitalized at The Elazig Psychiatric Hospital were included in the study. Anti-Toxocara IgG and/or IgM antibodies were determined by using commercial Toxocara canis IgG and/or IgM ELISA kit. RESULTS: Seropositivity for T. canis was detected in 45 (45.9%) of 98 patients and 2 (2.0%) of 100 control subjects the difference was statistically significant (p<0.001). The seroprevalence was 40.4% (19 cases) and 51.0% (26 cases) for female and male subjects, respectively (p=0.3). When the seropositive and seronegative schizophrenic patients were compared with respect to the age group environment they were living in, occupation period of follow up and number of hospitalizations, there were no differences between the two groups (all, p>0.05). CONCLUSION: In conclusion, the schizophrenic state seems to present a high risk for Toxocara infection in Turkey.

The relationship of graft survival and herpes simplex virus latency in recipient corneal buttons.

PURPOSE: To demonstrate relationship between herpes simplex virus (HSV) corneal latency and graft survival. METHODS: Prospective case control study. 28 recipient corneal buttons and donor cornea-scleral remnants were examined for HSV DNA with polymerase chain reaction (PCR). None of the recipient had a history of HSV infection. Serum samples of graft recipients were analyzed for the presence of anti-HSV IgG and IgM with enzyme-linked immunosorbent assay technique. All corneas were free of stromal scarring or epithelial defect before sampling and had an endothelial cell density of >2000 cells/mm(2). RESULTS: In twenty three patients (82%) anti-HSV IgG was detected in serum. In none of the recipients anti-HSV IgM was positive. HSV DNA was positive in six out of twenty eight (21%) of the recipient corneal buttons and none of the donor cornea-scleral remnants. In eighteen-months follow-up period three out of six (50%) HSV DNA positive and one out of twenty-two (4.5%) HSV DNA negative patients experienced late endothelial failure that was statistically significantly different (p = 0.022). CONCLUSION: Even without a history of HSV keratitis, presence of latent HSV virus in recipient cornea is an important risk factor for subsequent graft survival.

[Expression of the cathepsin L1 gene of Fasciola hepatica eucaryotic cells]. / Fasciola hepatica cathepsin L1 geninin ökaryotik hücrelerde belirlenmesi.

The parasitic trematode Fasciola hepatica is the causative agent of fasciolosis that is common in ruminants especially sheep and cattle and is occasionally found in humans. Fasciolosis has a worldwide distribution including Turkey and causes major economic losses in agricultural industry. Cathepsin L1 is one of the major molecules in the excretory-secretory products of F. hepatica and is involved in tissue penetration, immune evasion and feeding and therefore may be used in vaccination and serological diagnosis. The aim of this study was to evaluate cloning and expression of the cathepsin L1 gene of F. hepatica eucaryotic cells. For this purpose, total RNA was extracted from adult F. hepatica. Cathepsin L1 DNA amplicons were obtained with the reverse transcription polymerase chain reaction (RT-PCR). The 981 base-coding gene region of cathepsin L1 was amplified using specific primers to the cathepsin L1 gene. Then, the cathepsin L1 gene was cloned into the pCI-neo mammalian expression vector. The presence of the cathepsin L1 gene was confirmed by PCR screening and enzyme digestion assays. So, the resulting recombinant plasmid was named pFhCL1. Afterwards, the pFhCL1 vector was transiently transfected into Vero cells. The presence of the cathepsin L1 proteins was shown by Western immunoblotting.

Crimean-Congo hemorrhagic fever virus: genetic analysis and tick survey in Turkey.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus in the family Bunyaviridae, genus Nairovirus. The virus is transmitted to humans through infected tick bites or from direct contact with viremic animals or humans. In the present study, a total of 1,015 adult ticks were collected from cattle (603 specimens), sheep (17 specimens), and goats (395 specimens) in the Kelkit Valley in Turkey. Four tick species were recognized on the animals in the surveyed region. The most abundant species were Rhipicephalus bursa and Hyalomma marginatum marginatum, at 47.68% (484/1,015) and 46.40% (471/1,015), respectively. Reverse transcriptase PCR was used to recover partial sequences of the CCHFV small (S) genome segment. The presence of CCHFV was determined in 3 of 33 (9.09%) R. bursa pools and in 1 of 31 (3.22%) H. m. marginatum pools. Virus sequences from R. bursa were extremely different from those of the Greek CCHFV strain (U04958) isolated from an R. bursa tick. Phylogenetic analysis indicated that the CCHFV isolates obtained in this study clustered in group 5, whose range encompasses southwestern Russian and Kosovo. This is the first evidence of CCHFV in ticks from Turkey. Even though Hyalomma is the main vector for CCHFV, R. bursa may play a role in CCHFV transmission.

Paradoxically developed optochiasmatic tuberculoma and tuberculous lymphadenitis: a case report with 18-month follow up by MRI.

We report the first case ofextracranial tuberculous lymphadenitis which paradoxically developed during treatment of intracranial tuberculoma. Our patient, a 15-year-old girl who initially presented with meningitis and intracranial tuberculomas, developed extracranial tuberculomas during treatment for central nervous system tuberculosis. She was followed clinically with cerebrospinal fluid (CSF) studies and magnetic resonance imaging (MRI) at three monthly intervals. Within 18 months of specific antituberculous treatment, the patient had fully recovered. The course and response to therapy are discussed in light of the current literature.

Detection of Molluscum contagiosum virus (MCV) subtype I as a single dominant virus subtype in Molluscum lesions from a Turkish population.

BACKGROUND: Molluscum contagiosum has a worldwide occurrence and its primary mode of transmission is via direct human contact including sexual means. The aim of the study was to implement a polymerase chain reaction-based assay for detection and subtyping of Molluscum contagiosum virus (MCV) in skin lesions diagnosed with molluscum contagiosum in a large regional teaching hospital in Turkey. METHODS: For this purpose, a total of 61 patients were included in the study. Randomly selected single lesion from each patient was used to extract DNA material and a specific PCR reaction amplifying 393-bp- and 575-bp-long regions from MCV genome was used in the detection. Subtyping was carried out by digestion of the amplified 575-bp product with restriction endonuclease enzyme BamHI. Both amplified and restriction enzyme digested products visualized on agarose gel electrophoresis. RESULTS: All 61 molluscum cases (100%) included in the study contained MCV genetic material as demonstrated by the presence of 393- and 575-bp-long PCR amplified products. Restriction enzyme BamHI digestion of the 575-bp-long amplicon indicated that the infecting subtype in all the cases (100%) was MCV subtype I. CONCLUSIONS: Results of this study demonstrate that subtype I is the only infecting strain dominant in our region. Because the only consecutive molluscum patients admitted to our hospital were included in the study, our data do not rule out the possibility that other genotypes might be present in the Turkish population. However, it is not unreasonable to conclude that similar trends exist in the rest of the country. Results also show that a molecular-based diagnostic assay would be feasible in cases where diagnosis was deemed necessary.

Combined branched-DNA and conventional HBV PCR assays for detection of serum HBV-DNA in hepatitis B e antigen-positive chronic hepatitis B patients.

BACKGROUND/AIMS: Monitoring of HBV replication level is very useful for the management of patients with chronic HBV. However, the use of the correct tools to quantify HBV-DNA levels in serum and monitor the replication of HBV is of paramount importance in terms of diagnosis, and antiviral treatment of patients with chronic HBV infection. The aim of this study was to combine the bDNA assay and HBV PCR to improve detection of viremia the patients with HBeAg-positive chronic hepatitis B infection. METHODOLOGY: In this study, 67 HBeAg-positive chronic hepatitis B patients were analyzed to determine viremia level using bDNA and HBV PCR assays. RESULTS: Sixty-four patients with HBeAg-positive chronic hepatitis B showed positivity by conventional HBV PCR, whereas 56 subjects with HBeAg-positive chronic hepatitis B showed HBV-DNA levels by bDNA. CONCLUSIONS: The results indicated that it is reasonable to use the bDNA assay to determine HBV replicative activity first, and use conventional HBV-PCR for HBeAg-positive chronic hepatitis B patient samples that are negative in bDNA assay.