Genetic polymorphisms in innate immunity genes influence predisposition to tick-borne encephalitis.

Tick-borne encephalitis (TBE) is a neuroviral disease that ranges in severity from a mild febrile illness to a severe and life-threatening meningoencephalitis or encephalomyelitis. There is increasing evidence that susceptibility to tick-borne encephalitis virus (TBEV)-induced disease and its severity are largely influenced by host genetic factors, in addition to other virus- and host-related factors. In this study, we investigated the contribution of selected single nucleotide polymorphisms (SNPs) in innate immunity genes to predisposition to TBE in humans. More specifically, we investigated a possible association between SNPs rs304478 and rs303212 in the gene Interferon Induced Protein With Tetratricopeptide Repeats 1 (IFIT1), rs7070001 and rs4934470 in the gene Interferon Induced Protein With Tetratricopeptide Repeats 2 (IFIT2), and RIG-I (Retinoic acid-inducible gene I) encoding gene DDX58 rs311795343, rs10813831, rs17217280 and rs3739674 SNPs with predisposition to TBE in population of the Czech Republic, where TBEV is highly endemic. Genotypic and allelic frequencies for these SNPs were analyzed in 247 nonimmunized TBE patients and compared with 204 control subjects. The analysis showed an association of IFIT1 rs304478 SNP and DDX58 rs3739674 and rs17217280 SNPs with predisposition to TBE in the Czech population indicating novel risk factors for clinical TBE but not for disease severity. These results also highlight the role of innate immunity genes in TBE pathogenesis.

Serum matrix metalloproteinase-9 (MMP-9) as a biomarker in paediatric and adult tick-borne encephalitis patients.

Matrix metalloproteinases (MMPs) play an important role in central nervous system infections. We analysed the levels of 8 different MMPs in the cerebrospinal fluid (CSF) of 89 adult patients infected with tick-borne encephalitis (TBE) virus and compared them with the levels in a control group. MMP-9 was the only MMP that showed significantly increased CSF levels in TBE patients. Serum MMP-9 levels were subsequently measured in 101 adult TBE patients at various time points during the neurological phase of TBE and at follow-up. In addition, serum MMP-9 was analysed in 37 paediatric TBE patients. Compared with control levels, both paediatric and adult TBE patients had significantly elevated serum MMP-9 levels. In most adult patients, serum MMP-9 levels peaked at hospital admission, with higher serum MMP-9 levels observed in patients with encephalitis than in patients with meningitis. Elevated serum MMP-9 levels were observed throughout hospitalisation but decreased to normal levels at follow-up. Serum MMP-9 levels correlated with clinical course, especially in patients heterozygous for the single-nucleotide polymorphism rs17576 (A/G; Gln279Arg) in the MMP9 gene. The results highlight the importance of MMP-9 in the pathogenesis of TBE and suggest that serum MMP-9 may serve as a promising bioindicator of TBE in both paediatric and adult TBE patients.

Serum and cerebrospinal fluid phosphorylated neurofilament heavy subunit as a marker of neuroaxonal damage in tick-borne encephalitis.

Extensive axonal and neuronal loss is the main cause of severe manifestations and poor outcomes in tick-borne encephalitis (TBE). Phosphorylated neurofilament heavy subunit (pNF-H) is an essential component of axons, and its detection in cerebrospinal fluid (CSF) or serum can indicate the degree of neuroaxonal damage. We examined the use of pNF-H as a biomarker of neuroaxonal injury in TBE. In 89 patients with acute TBE, we measured CSF levels of pNF-H and 3 other markers of brain injury (glial fibrillary acidic protein, S100B and ubiquitin C-terminal hydrolase L1) and compared the results to those for patients with meningitis of other aetiology and controls. Serum pNF-H levels were measured in 80 patients and compared with findings for 90 healthy blood donors. TBE patients had significantly (P<0.001) higher CSF pNF-H levels than controls as early as hospital admission. Serum pNF-H concentrations were significantly higher in samples from TBE patients collected at hospital discharge (P<0.0001) than in controls. TBE patients with the highest peak values of serum pNF-H, exceeding 10 000 pg ml-1, had a very severe disease course, with coma or tetraplegia. Patients requiring intensive care had significantly higher serum pNF-H levels than other TBE patients (P<0.01). Elevated serum pNF-H values were also observed in patients with incomplete recovery (P<0.05). Peak serum pNF-H levels correlated positively with the duration of hospitalization (P=0.005). Measurement of pNF-H levels in TBE patients might be useful for assessing disease severity and determining prognosis.

Tick-Borne Encephalitis in an 8.5-Month-Old Boy Suspected of Febrile Seizures.

Tick-borne encephalitis (TBE) is a serious viral neuroinfection affecting humans in large areas of Europe and Asia. TBE can occur at any age, but only a few reports of TBE in infants younger than 1 year have been published. Here, we report a case of severe TBE in an 8.5-month-old boy presenting with seizures at the beginning of the neurological phase.

Clinical Characteristics of Patients with Tick-Borne Encephalitis (TBE): A European Multicentre Study from 2010 to 2017.

Tick-borne encephalitis (TBE) virus is a major cause of central nervous system infections in endemic countries. Here, we present clinical and laboratory characteristics of a large international cohort of patients with confirmed TBE using a uniform clinical protocol. Patients were recruited in eight centers from six European countries between 2010 and 2017. A detailed description of clinical signs and symptoms was recorded. The obtained information enabled a reliable classification in 553 of 555 patients: 207 (37.3%) had meningitis, 273 (49.2%) meningoencephalitis, 15 (2.7%) meningomyelitis, and 58 (10.5%) meningoencephalomyelitis; 41 (7.4%) patients had a peripheral paresis of extremities, 13 (2.3%) a central paresis of extremities, and 25 (4.5%) had single or multiple cranial nerve palsies. Five (0.9%) patients died during acute illness. Outcome at discharge was recorded in 298 patients. Of 176 (59.1%) patients with incomplete recovery, 80 (27%) displayed persisting symptoms or signs without recovery expectation. This study provides further evidence that TBE is a severe disease with a large proportion of patients with incomplete recovery. We suggest monitoring TBE in endemic European countries using a uniform protocol to record the full clinical spectrum of the disease.

Unvaccinated child tetanus from nasal toy battery.

We present the case report of an unvaccinated Czech child with tetanus. The child had not received any vaccines due to its parent's refusal. The disease originated from the wound in the nose caused by a small flat battery. The typical onset of tetanus followed after two weeks, rapidly progressing to respiratory failure with the need for mechanic ventilation despite intensive treatment. The child spent five weeks in the hospital. Mild long-term sequelae persisted 5 months.

Identification of positively selected genes in human pathogenic treponemes: Syphilis-, yaws-, and bejel-causing strains differ in sets of genes showing adaptive evolution.

BACKGROUND: Pathogenic treponemes related to Treponema pallidum are both human (causing syphilis, yaws, bejel) and animal pathogens (infections of primates, venereal spirochetosis in rabbits). A set of 11 treponemal genome sequences including those of five Treponema pallidum ssp. pallidum (TPA) strains (Nichols, DAL-1, Mexico A, SS14, Chicago), four T. p. ssp. pertenue (TPE) strains (CDC-2, Gauthier, Samoa D, Fribourg-Blanc), one T. p. ssp. endemicum (TEN) strain (Bosnia A) and one strain (Cuniculi A) of Treponema paraluisleporidarum ecovar Cuniculus (TPeC) were tested for the presence of positively selected genes. METHODOLOGY/PRINCIPAL FINDINGS: A total of 1068 orthologous genes annotated in all 11 genomes were tested for the presence of positively selected genes using both site and branch-site models with CODEML (PAML package). Subsequent analyses with sequences obtained from 62 treponemal draft genomes were used for the identification of positively selected amino acid positions. Synthetic biotinylated peptides were designed to cover positively selected protein regions and these peptides were tested for reactivity with the patient's syphilis sera. Altogether, 22 positively selected genes were identified in the TP genomes and TPA sets of positively selected genes differed from TPE genes. While genetic variability among TPA strains was predominantly present in a number of genetic loci, genetic variability within TPE and TEN strains was distributed more equally along the chromosome. Several syphilitic sera were shown to react with some peptides derived from the protein sequences evolving under positive selection. CONCLUSIONS/SIGNIFICANCE: The syphilis-, yaws-, and bejel-causing strains differed relative to sets of positively selected genes. Most of the positively selected chromosomal loci were identified among the TPA treponemes. The local accumulation of genetic variability suggests that the diversification of TPA strains took place predominantly in a limited number of genomic regions compared to the more dispersed genetic diversity differentiating TPE and TEN strains. The identification of positively selected sites in tpr genes and genes encoding outer membrane proteins suggests their role during infection of human and animal hosts. The driving force for adaptive evolution at these loci thus appears to be the host immune response as supported by observed reactivity of syphilitic sera with some peptides derived from protein sequences showing adaptive evolution.

Clinical and laboratory characteristics of enteroviral meningitis in children, including qRT-PCR and sequencing analysis.

AIMS: Enteroviruses (EVs) are the most common agents of aseptic meningitis. Some serotypes can cause serious neuroinfection leading to death. The aim of this study was to determine the representation of EVs in the etiology of aseptic meningitis in children and to analyze the demographic, clinical, laboratory, and epidemiological characteristics of patients with EV meningitis. PATIENTS AND METHODS: This was a prospective study including 147 patients in three groups: EV meningitis, tick-borne encephalitis, and aseptic meningitis with unidentified agent. RESULTS: Boys with EV meningitis predominated over girls. The average patient age was 11 years. Compared to the control group, these patients suffered more from stiff back (P=0.010), vomiting and nausea (P=0.009). They had shorter symptom duration (P<0.001), higher C-reactive protein in blood (P<0.001), higher predominance of polynuclears (P=0.026), and greater lactate (P=0.003) in cerebrospinal fluid (CSF). The serotype seen most frequently (68%) was ECHO virus (ECV) 30. CONCLUSIONS: Enteroviruses play the most important role in the differential diagnosis of aseptic meningitis. Short symptom duration, slightly higher inflammatory parameters in blood, predominance of polynuclears, and elevated CSF lactate have predictive value in diagnosing this disease. ECV 30 (frequently the agent of epidemics in the Czech Republic) was the aseptic meningitis agent most often seen.

[Guideline for the diagnosis and treatment of Lyme borreliosis].

The national guideline aims to highlight the latest knowledge about clinical manifestations of the infection, to summarize the diagnostic algorithm and to recommend the appropriate antibiotic therapy with respect to evidence-based medicine. The recommendations are consistent with most European guidelines as well as those published by the IDSA. The guideline provides the most recent information on the epidemiology, etiology and pathogenesis of Lyme borreliosis, dermatological, neurological and musculoskeletal involvement, the appropriate diagnostic procedure and prevention. Some information is also provided about post-treatment Lyme disease syndrome. Recommended oral and intravenous antimicrobials are listed in a table showing the doses and duration of therapy. The guideline also mentions diagnostic methods to be avoided or whose results should be interpreted with caution. Although the guideline cannot account for all individual variations among patients, it may provide instructions to physicians in typical and frequent clinical situations.

[Guideline for the diagnosis and treatment of acute bacterial meningitis].

Acute bacterial meningitis is a severe infectious disease of the central nervous system. Its incidence decreases but lethality and sequelae remain high. The early initiation of appropriate treatment is a factor strongly determining the patient´s prognosis. The authors submit the Czech national guideline for diagnosis and treatment of community-acquired acute bacterial meningitis which has to provide clear and simple recommendations for clinicans involved in the care of meningitis in adults and children. The national guideline was based on the European guideline published in 2016 and adapted for the situation in the Czech Republic. It was acknowledged (approved? ratified?) by the Society for Epidemiology and Microbiology and the Society for Medical Microbiology of the Czech Medical Association.

[Epidemiology, etiology and clinical picture of enteroviral meningitis in children in South Moravia].

OBJECTIVES: Enteroviruses (EVs) are the most common cause of aseptic viral meningitis. In some cases, they can cause severe meningoencephalitis and acute flaccid paralysis - an association with some virulent serotypes. The objectives were to describe the epidemiological situation of EV meningitis in children in South Moravia, to elucidate the etiology including the incidence of virulent serotypes and to evaluate the clinical presentation. MATERIAL AND METHODS: A total of 88 children with EV meningitis were prospectively evaluated. In case of aseptic inflammation in the cerebrospinal fluid, EV was detected using real-time PCR. Genotyping was performed in 56 samples using repeated one-step PCR and partial sequencing on a genetic analyzer in the National Reference Laboratory for Enteroviruses in Prague. RESULTS: The patients' age range was 3-17 years; there were more boys than girls. Two epidemics occurred, one involving 17 Roma children and the other involving 8 swimming pool visitors. The most common symptoms were headache, fever and stiff neck. The most frequently (59%) detected agent was Echovirus 30 identified as the cause of the epidemics. In one boy, EV 71 (virulent serotype) was found. The clinical course did not vary from that in other serotypes. All 88 children recovered without complications. CONCLUSIONS: EVs are an important part of the differential diagnosis of neuroinfections, although most infections are benign aseptic meningitis. The clinical presentation did not vary between infections with various serotypes. Higher incidence rates of virulent serotypes were not reported. Echovirus 30 was detected most frequently and was repeatedly identified as the cause of epidemics throughout the Czech Republic.

[Salmonellosis - a real threat for infants].

Salmonella spp. are a common cause of bacterial gastroenteritis. In infants, Salmonella infection is uncommon, posing a higher risk for a severe course. The authors present a case of a 3-month-old infant with suspected Salmonella sepsis. Also discussed are the correct approach to therapy and potential errors in the treatment of the youngest age groups.

Clinical course and sequelae for tick-borne encephalitis among children in South Moravia (Czech Republic).

UNLABELLED: This study of 170 children in the Czech Republic examines the clinical course and sequelae for tick-borne encephalitis. Evaluated were demographic and epidemiological data, signs and symptoms at admission, clinical course during hospital stay and laboratory findings. Cerebrospinal fluid was analysed for white blood cells, protein, impairment of blood-cerebrospinal fluid (CSF) barrier and tick-borne encephalitis virus (TBEV)-specific antibodies. Subjective complaints and objective neurological deficits were investigated. Tick bites were reported in 74 % of the children. The illness had a biphasic clinical course in 58 % of cases. The second phase was characterized by headache in 98 %, high fever in 86 % (more than 38.5 °C), vomiting in 64 % and meningeal signs in 92 % of children. Meningitis (77 %) dominated over meningoencephalitis (13 %). Inflammatory changes in CSF were found in 90 % of children. Immunoglobulin M (IgM) antibodies against TBEV in serum were found early in the infection in 99 %. IgM positivity lasted up to 1,126 days. Neurocognitive abnormalities were found in 19 (11 %) of children. Acquired aphasia, lasting tremor of the upper extremities, speech impairment, inversion of sleep and wakefulness, abnormal hyperkinetic movements and vertigo were found to be permanent but not progressing. Severe sequelae persisted in two children (1 %) while in three (2 %) were classified as mild or moderate. CONCLUSION: Tick-borne encephalitis in children has a benign course with minimal sequelae. Meningitis with biphasic course is the prevalent involvement and the duration of IgM antibodies in serum and index of positivity are not decisive for postencephalitic disorders.

Detection of Borrelia burgdorferi sensu stricto ospC alleles associated with human lyme borreliosis worldwide in non-human-biting tick Ixodes affinis and rodent hosts in Southeastern United States.

Comparative analysis of ospC genes from 127 Borrelia burgdorferi sensu stricto strains collected in European and North American regions where Lyme disease is endemic and where it is not endemic revealed a close relatedness of geographically distinct populations. ospC alleles A, B, and L were detected on both continents in vectors and hosts, including humans. Six ospC alleles, A, B, L, Q, R, and V, were prevalent in Europe; 4 of them were detected in samples of human origin. Ten ospC alleles, A, B, D, E3, F, G, H, H3, I3, and M, were identified in the far-western United States. Four ospC alleles, B, G, H, and L, were abundant in the southeastern United States. Here we present the first expanded analysis of ospC alleles of B. burgdorferi strains from the southeastern United States with respect to their relatedness to strains from other North American and European localities. We demonstrate that ospC genotypes commonly associated with human Lyme disease in European and North American regions where the disease is endemic were detected in B. burgdorferi strains isolated from the non-human-biting tick Ixodes affinis and rodent hosts in the southeastern United States. We discovered that some ospC alleles previously known only from Europe are widely distributed in the southeastern United States, a finding that confirms the hypothesis of transoceanic migration of Borrelia species.

[Erythema migrans]. / Erythema migrans.

The article summarizes the latest knowledge about the etiology, epidemiology, pathogenesis, clinical signs, diagnostic procedures and therapy of erythema migrans (EM), a cutaneous form of Borrelia infection. The sole focus on this clinical entity is intentional since EM is the most common clinical involvement in Lyme borreliosis (LB) in the Czech Republic (CR). New genospecies from the Borrelia burgdorferi sensu lato complex, B. lusitaniae and B. spielmanii, as etiologic agents are reported. Although there are no commercial tests available yet for their detection, ticks are demonstrably infected with them in Europe and probably even in the CR. Six pathogenic genospecies circulate among vertebrates and Ixodes ricinus ticks in Europe. The transmission cycle in nature, hosts identified as reservoirs and distribution of borreliae in Europe are presented. Clinical forms of EM are described in detail including accompanying features. Types of erythema in the USA and Europe are compared. In the part on differential diagnosis, allergies, skin and infectious diseases with similar appearance on the skin as EM are mentioned. Although the diagnosis of EM is exclusively clinical and serology is not indicated, methods of direct detection and two-tier testing of antibodies are discussed. There are therapeutic recommendations concerning the choice of antibiotics available in the CR.

[Pertussis in a 4-month-old infant with unrecognized cystic fibrosis]. / Pertusse u kojence s nepoznanou cystickou fibrozou.

We present a case report of a small boy with cystic fibrosis complicated by pertussis and pneumothorax. The child had not been vaccinated against Bordetella pertussis due to the failure to thrive and was infected with the bacterium at the age of 4 months. The course of the disease was severe, with respiratory distress and spontaneous pneumothorax. The diagnosis of pertussis was serologically confirmed. A suspicion for cystic fibrosis increased after an unusually prolonged course of the disease and a history of the failure to thrive. A comprehensive diagnostic procedure revealed Pseudomonas aeruginosa respiratory infection, anemia, pancreatic insufficiency, a positive sweat test and the presence of two CFTR gene mutations. The patient, treated with comprehensive cystic fibrosis therapy, recovered from acute respiratory illness and started to thrive soon.

[Case report - pediatric visceral leishmaniasis]. / Kazuistika -visceralni leishmanioza ditete.

The diagnosis of visceral leishmaniasis (VL) is not an easy task. The clinical presentation of VL is similar to lymphomas which have significantly higher incidence in our country. This fact is one of the complications in the diagnosis of VL. Moreover, diagnostic methods have low sensitivity and bone marrow aspiration is necessary. We present a case report of a patient who fell ill in the first year of life and due to atypical lifestyle of his parents was seriously examined after 11 months from the first symptoms of the disease. Many problems with the diagnostic algorithm leading to the definitive diagnosis of VL were encountered. Thereafter, the patient underwent treatment with amphotericin B lipid complex. The course was complicated by febrile neutropenia and anemia, with the necessity for blood transfusion.

[Bat rabies in Europe and the Czech Republic]. / Vzteklina netopýru v Evrope a v Ceské Republice.

In 2005, six children were treated in our department who had been in contact with a bat infected with European bat lyssavirus 1 (EBL1). In the Czech Republic, this was the first confirmed case of rabies since 2002, but the fourth case of bat rabies since 1994. All the cases were related to Southern Moravia and bat species which almost do not migrate. This suggests only endemic prevalence of EBL1 in this country. Bat rabies is different from rabies in terrestic mammals. Based on genetic analyses, the lyssavirus genus may be divided into 6 genotypes of which genotypes 1 (rabies virus), 5 (EBL1) and 6 (EBL2) are found in Europe. The infectious cycles of bat lyssaviruses are limited solely to bat populations, in particular the Eptesicus serotinus species and the Myotis genus. Transmission to other mammals is rare. In Europe, four cases of human infection and death due to rabies caused by any of bat lyssaviruses have been reported. Immunologically, bat lyssaviruses are very similar to the common rabies virus. Therefore, the standard prophylactic methods are sufficient. Because of a different natural reservoir, the Czech Republic may be still considered a rabies-free country. However, contacts with bats always pose a risk of rabies.

[Eye involvement of borrelia aetiology]. / Ocní postizení borreliové etiologie.

We present a case of eye involvement -- intermediate uveitis -- during tick-borne borreliosis in a 10-year-old boy. Ophthalmologic examination revealed impaired vision, apparent thick floating whitish opacity in the vitreous humour of the left eye and fine fibres in the vitreous humour of the right eye. Sonographic examination confirmed hyperechogenic opacity in the vitreous humour. An autoimmune process was suspected but not confirmed. Serological examination showed IgG antibodies against three pathogenic borreliae and borderline values of IgM antibodies against Borrelia garinii were found by immunoblot. The boy was treated with intravenous ceftriaxone for 21 days. The subsequent sonographic examination showed only minute sporadic echogenicity. Biomicroscopically, only residual opacity in the vitreous humour was found. Isolated eye involvement of borrelia aetiology is rare. The discussion provides a review of similar cases of uveitis including diagnosis of the eye form as published in literature.

[Comparison of EIA Borrelia recombinant IgM of the 3rd generation with EIA test based on whole cell antigen in the diagnosis of Lyme borreliosis]. / Srovnání soupravy III. generace EIA Borrelia recombinant IgM s vysetrením EIA testem s celobunecným antigenem v diagnostice Lymeské borreliózy.

OBJECTIVES: Evaluation of a new kit of the 3rd generation, EIA Borrelia recombinant IgM in the serodiagnosis of Lyme borreliosis (LB) and comparison with the EIA test based on the whole cell antigen in the detection of IgM antibodies. MATERIAL AND METHODS: In total 123 children (147 sera) were examined, 71 children (93 sera) of whom showed clinically defined stage of LB. The other 54 sera of 52 children were tested for suspicion for LB which was later excluded, and for cross-reacting antibodies. They represented the control group of children with other infectious or autoimmune diseases. The EIA B. recombinant IgM kit (Test-Line, CR) is based on the selected antigen fragments: outer surface protein C (OspC) and internal flagellin (p41i). The fragments were selected according to the most frequent Borrelia spp. in the Czech Republic. RESULTS: Samples compared 101/147 (69 %) with both tests showed correspondent reactions. 46 samples reacted inconsistently, 26 from children with LB and 20 from the control group. The total specificity and sensitivity of the recombinant kit in IgM antibody class was 88,9 % and 26,9 %, respectively. In the case of early disseminated LB infection the sensitivity was 35,6 % and it is comparable with the sensitivity of immunoblot. Statistically, there is no difference in the sensitivity (p = 0,101) and specificity (p = 0,383) of EIA B. recombinant IgM and immunoblot. CONCLUSIONS: The EIA B. recombinant IgM kit shows comparable diagnostic sensitivity in children with acute LB together with significantly higher specificity in children with non-specific response in kits based on the whole cell antigen and at the same time high conformity with results of IgM class immunoblot. This kit of the 3rd generation is reliable for screening of IgM antibodies.