Anna Moniuszko-Malinowska

Evaluation of CXCL10, CXCL11, CXCL12 and CXCL13 chemokines in serum and cerebrospinal fluid in patients with tick borne encephalitis (TBE)

PURPOSE The aim of the study was to assess the concentration of chemokines: CXCL10, XCL11, CXCL12, CXCL13 in serum and cerebrospinal fluid (CSF) in patients with tick-borne encephalitis (TBE) before and after treatment. We evaluated also the usefulness of these molecules in diagnosis and monitoring of inflammation in TBE. METHODS Twenty three patients hospitalized in The Department of Infectious Diseases and Neuroinfections of Medical University in Białystok, Poland were included in the study. Patients were divided into 2 groups: TBE group-patients with confirmed TBE and control group (CG): patients with excluded TBE and other inflammatory diseases of CNS. Concentration of CXCL10/IP-10, CXCL11/I-TAC, CXCL12/SDF-1α, CXCL13/BLC/BCA-1 in serum and CSF were measured with ELISA kits (R&D Systems, USA) according to the protocols. RESULTS The analysis of chemokines concentration in TBE patients before treatment and control group using ROC showed that serum CXCL10 and CXCL13 and CSF CXCL10, CXCL11, CXCL12 and CXCL13 differentiate both groups (p<0.05). The analysis of CXCL10, CXCL11, CXCL12 and CXCL13 before and after treatment showed that CXCL10 and CXCL11 in CSF and CXCL13 in serum differentiates both groups with p<0.05. CONCLUSIONS Concentration of CSF CXCL10, CXCL11, CXCL12, CXCL13 and serum CXCL10, CXCL13 may be good biomarkers of CNS inflammation caused by TBEV. Moreover concentration of CXCL10 in CSF and CXCL13 in serum may be used as indicators of patients recovery.

Infection with Babesia microti in humans with non-specific symptoms in North East Poland

Abstract Aim: The aim of the study was to evaluate the clinical course and effectiveness of diagnostics tools for Babesia spp. infection in patients bitten by ticks. Materials and methods: Five hundred and forty-eight patients hospitalised or seen in outpatients department because of various symptoms after a tick bite were included in the study. PCR, nucleotide sequencing of Babesia 18S rRNA gene fragment, blood smears and serological tests for Babesia spp., TBEV, A. phagocytophilum and B. burgdorferi were performed in all patients. Six patients infected with Babesia were included in the final analysis. They had PCR, Babesia 18S rRNA gene fragment nucleotide sequencing, blood smears and serological tests for Babesia spp., TBEV, A. phagocytophilum and B. burgdorferi performed twice. Results: Tick-borne infection with Babesia microti in six immunocompetent patients with non-specific symptoms was confirmed for the first time in Poland. No severe course of the disease was seen. No piroplasm forms were noticed within erythrocytes on blood smear. Three patients developed a serological response. Conclusions: Immunocompetent patients may be unaware of infection with Babesia microti after a tick bite. It must be included in the differential diagnosis after the tick bite. In patients with low parasitaemia PCR and serology seem useful when blood smear is negative. Self-elimination of Babesia spp. is possible, especially in cases with low parasitaemia.

Lipid peroxidation in the pathogenesis of neuroborreliosis.

This study analyzed the onset of lipid peroxidation (LPO) in neuroborreliosis and the effects of ceftriaxone therapy on LPO. Twenty-two patients with early neuroborreliosis and 22 healthy subjects were studied. LPO in the cerebrospinal fluid (CSF), as well as the plasma and urine was estimated by the levels of reactive aldehydes: 4-hydroxynonenal (4-HNE), 4-hydroxyhexenal, malondialdehyde, and 4-oxononenal, F2-isoprostanes and A4/J4-neuroprostanes (NPs). The plasma level of 4-HNE-protein adducts arachidonic acid (AA), docosahexaenoic acid (DHA) and vitamin E was determined. Additionally, enzymatic activities of phospholipase A2 (PLA2), platelet-activating factor acetylhydrolase (PAF-AH) and glutathione peroxidase (GSH-Px) were determined. A decrease of AA, DHA levels and GSH-Px activity in plasma was associated with a significant increase of aldehydes in the CSF, plasma and urine. Similarly, the increase of F2-isoprostanes and NPs in the CSF and plasma was associated with the decreased activity of PLA2 and PAF-AH. Ceftriaxone therapy cured patients and reduced the levels of F2-isoprostanes, NPs and reactive aldehydes. However, the activities of PLA2 and PAF-AH increased. Pathophysiological association of neuroborreliosis with systemic LPO was revealed. Effective antibiotic therapy attenuated LPO. Biomarkers of LPO could be useful to monitor the onset of neuroborreliosis and show the effectiveness of pharmacotherapy.

MRI and planimetric CT follow-up study of patients with severe tick-borne encephalitis.

Background: The aim of the study was to evaluate the magnetic resonance imaging (MRI) and planimetric computed tomography (CT) of brain lesions in patients with a history of tick-borne encephalitis (TBE); to assess the influence of steroid treatment on the brain and whether lesions were age-dependent. Methods: A total of 19 patients with abnormal initial imaging in the acute stage of the disease had a follow-up MRI after 1 year; 34 patients hospitalized for TBE encephalitis/encephalomyelitis had planimetric CT after 10 years. Results: On MRI cortico-subcortical atrophy with widening of anterior horns of the lateral ventricles and vascular changes was more marked on follow-up examination. Virchow–Robin spaces dilatation, widening of the lateral ventricles, periventricular lesions, and cortico-subcortical atrophy correlated with age. Results of planimetric CT study showed increased percentage of tracings, widened anterior horns, lateral ventricles, and III ventricle, which suggest new non-age-related atrophic lesions. Conclusions: Radiological lesions in the acute phase of TBE and after recovery are non-specific. Cortico-subcortical atrophy with widening of the anterior horns of the lateral ventricles and vascular changes are most common. Long-term follow-up confirms the formation of new non-age-related cerebral atrophic lesions due to TBE. The logit model may serve as a background for the hypothesis concerning an accelerated local atrophy of the brain tissues in patients with a history of severe TBE.

Tick-borne infections and co-infections in patients with non-specific symptoms in Poland

AIM The aim of the study was the evaluation of the frequency of infections and co-infections among patients hospitalized because of non-specific symptoms after a tick bite. MATERIALS AND METHODS Whole blood, serum and cerebrospinal fluid samples from 118 patients hospitalised for non-specific symptoms up to 8 weeks after tick bite from 2010 to 2013 were examined for tick-borne infections. ELISA, Western blot and/or molecular biology (PCR; fla gene; 16S rRNA; sequencing) and thin blood smears (MDD) were used. Control group included 50 healthy blood donors. All controls were tested with PCR and serology according to the same procedure as in patients. RESULTS Out of 118 patients 85 (72%) experienced headaches, 15 (13%) vertigo, 32 (27%) nausea, 17 (14%) vomiting, 37 (31%) muscle pain, 73 (62%) fever and 26 (22%) meningeal signs. 47.5% were infected with at least one tick-borne pathogen. Borrelia burgdorferi sensu lato infection was confirmed with ELISA, Western blot in serum and/or (PCR (fla gene) in whole blood in 29.7% cases. In blood of 11.9% patients Anaplasma phagocytophilum DNA (16S rRNA gene) was detected; in 0.9% patients 1/118 Babesia spp. DNA (18S rRNA gene) was also detected. Co-infections were observed in 5.1% of patients with non-specific symptoms. B. burgdorferi s.l. - A. phagocytophilum co-infection (5/118; 4.2%) was most common. In 1/118 (0.8%) A. phagocytophilum - Babesia spp. co-infection was detected. All controls were negative for examined pathogens. CONCLUSIONS Non-specific symptoms after tick bite may be caused by uncommon pathogens or co-infection, therefore it should be considered in differential diagnosis after tick bite.

Evaluation of NF-κB concentration in patients with tick-borne encephalitis, neuroborreliosis, anaplasmosis and Anaplasma phagocythophilum with tick-borne encephalitis virus co-infection

Objectives The aim of the study was the evaluation of NF‐&kgr;B concentration in serum and cerebrospinal fluid (CSF) of patients with diagnosis of tick‐borne diseases: tick‐borne encephalitis (TBE), neuroborreliosis (NB), anaplasmosis (ANA) and patients co‐infected with tick‐borne encephalitis virus and Anaplasma phagocythophilum (TBE + ANA). Additionally NF‐&kgr;B concentration during acute and convalescent period was compared. Methods Sixty‐seven patients with diagnosis of tick‐borne diseases were included in the study. The control group (CG) consisted of 18 patients hospitalized because of headaches and had lumbar puncture performed. The NF‐&kgr;B was measured by human inhibitory subunit of NF‐&kgr;B ELISA Kit during acute and convalescent period. Results In serum the significant differences were observed only in patients with TBE + ANA co‐infection. In CSF the concentration of NF‐&kgr;B was significantly higher in patients with TBE, TBE + ANA co‐infection, and patients with NB than in CG. Receiver operating characteristic (ROC) curves analysis showed that NF‐&kgr;B concentration in CSF differentiated patients with NB with CG; patients co‐infected with TBE and ANA with CG and patients with TBE with CG. NF‐&kgr;B concentration in serum differentiated patients co‐infected with TBE and ANA with NB and with ANA, with TBE and with CG. In TBE group the serum NF‐&kgr;B concentration significantly decreased in convalescent period, while in NB and TBE groups significant CSF decrease of NF‐&kgr;B concentration was observed. ConclusionsInflammatory process in the CNS results in the increase of NF‐&kgr;B concentration in CSF, due to the damage of blood‐brain barrier.Co‐infection of TBE + ANA increases NF‐&kgr;B concentration in serum.NF‐&kgr;B concentration in serum may be useful in the differentiation of TBE + ANA co‐infection with TBE and NB.NF‐&kgr;B concentration may be used to monitor TBE and NB treatment effectiveness. HighlightsNF‐&kgr;B increases in cerebrospinal fluid (CSF) during nervous system inflammation.Tick‐borne encephalitis with anaplasmosis (TBE + ANA) increases NF‐&kgr;B in serum.Serum NF‐&kgr;B concentration differentiates TBE + ANA with TBE and neuroborreliosis.Serum and CSF NF‐&kgr;B concentration may monitor tick‐borne encephalitis treatment.Cerebrospinal fluid NF‐&kgr;B concentration may monitor neuroborreliosis treatment.

Intrathecal expression of IL-5 and humoral response in patients with tick-borne encephalitis

AIM The aim of the study was to assess the role of an early specific humoral response in human infection with a tick-borne encephalitis virus (TBEV) and the role of IL-5 as its potential mediator and marker. MATERIALS AND METHODS The retrospective study involved a cohort of 199 patients diagnosed with TBE, in whom anti-TBEV IgM and IgG antibody titers were analyzed on admission and compared with clinical presentation and basic laboratory parameters. The prospective study included 50 TBE patients in whom IL-5 serum and CSF concentration was measured with ELISA on admission in the TBE neurologic phase and in selected patients before discharge, at follow-up or in samples obtained before the neurologic phase onset. RESULTS The serum anti-TBEV IgM correlated with good clinical outcome and the CSF anti-TBEV IgM with more pronounced CSF inflammation on admission, but also with its more complete resolution on follow-up. The serum anti-TBEV IgG correlated with milder presentation and better outcome. Concentration of IL-5 was increased in CSF but not in the serum of TBE patients. IL-5 concentration index on admission favored its intrathecal synthesis. IL-5 did not correlate significantly with clinical presentation and specific IgM and IgG titers. CONCLUSIONS Specific anti-TBEV IgM systemic and intrathecal response and IgG systemic response are protective, together favoring milder presentation, better outcome and resolution of central nervous system (CNS) inflammation. IL-5 is expressed intrathecally in TBE, but its pathogenetic role remains unclear.

Effect of a single dose of mannitol on hydration status and electrolyte concentrations in patients with tick-borne encephalitis:

Objective This study was performed to assess the effect of a single dose of 15% mannitol on the hydration status and electrolyte balance in patients with tick-borne encephalitis (TBE). Methods Forty-one patients with TBE were treated with 0.25 g/kg of 15% mannitol. The electrolyte concentrations (Na, K, and Cl), creatinine concentration, and hydration status were measured before and after mannitol infusion. Results After mannitol administration, 7 patients had hyponatremia, 3 had hypokalemia, 1 had hyperkalemia, and 17 had hypochloremia. The total body water volume (TBW) changed by 0.44% ± 0.55%, the external body water volume (EBW) changed by 0.12% ± 0.15%, and the internal body water volume (IBW) changed by 0.19% ± 0.40%. The mean ECW/ICW ratio was 0.7694 ± 0.07 before treatment and 0.7699 ± 0.07 after treatment. Age was correlated with the TBW change in men (R = 0.42, p < 0.05) and with the potassium change in women (R = 0.66, p < 0.05). Conclusions Patients with TBE should receive mannitol two to four times daily depending on the clinical manifestation. Administration of a single dose of mannitol (0.25 g/kg) requires at least 300 mL of fluid supplementation. Bioimpedance might be useful for individual evaluation of dehydration. Additionally, patients require monitoring for potential hyponatremia. Older men may be more prone to dehydration after receiving mannitol.

Assessment of HMGB-1 concentration in tick-borne encephalitis and neuroborreliosis

BACKGROUND The aim of this study was to determine the concentration of HMGB-1 (high mobility group box 1) in the serum and cerebrospinal fluid (CSF) of patients suffering from tick-borne encephalitis (TBE) and neuroborreliosis (NB). Focus was placed on HMGB-1 measurement in the CSF or serum in order to establish whether this could help to differentiate between NB and TBE. METHODS Eighty patients with meningitis and meningoencephalitis were enrolled in the study. The patients were divided into two groups: group I comprised patients with NB (n=40) and group II comprised patients with TBE (n=40). The diagnosis was made based on the clinical picture, CSF examination, and the presence of specific antibodies in the serum and CSF. The control group for the evaluation of the parameters in serum were healthy blood donors (n=25), while the control group for the evaluation of parameters in the CSF were patients in whom a central nervous system (CNS) inflammatory process was excluded. The concentrations of HMGB-1 were measured by ELISA method using a commercial kit (HMGB-1 ELISA Kit; EIAab, China). The results were analyzed using Statistica 10, Gretl, receiver operating characteristics curve (ROC), and the Pearson correlation coefficient. RESULTS AND CONCLUSIONS HMGB-1 is associated with the development of inflammatory processes in the CNS caused by both tick-borne pathogens: viral (TBE) and bacterial (Lyme borreliosis). Measurement of the serum HMGB-1 concentration in the early stages of both diseases of the CNS may contribute to the differentiation between TBE and NB, which may have a clinical impact for patients bitten by ticks.

Brain perfusion alterations in tick-borne encephalitis—preliminary report

BACKGROUND Magnetic resonance imaging (MRI) changes in tick-borne encephalitis (TBE) are non-specific and the pathophysiological mechanisms leading to their formation remain unclear. This study investigated brain perfusion in TBE patients using dynamic susceptibility-weighted contrast-enhanced magnetic resonance perfusion imaging (DSC-MRI perfusion). METHODS MRI scans were performed for 12 patients in the acute phase, 3-5days after the diagnosis of TBE. Conventional MRI and DSC-MRI perfusion studies were performed. Cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT), and time to peak (TTP) parametric maps were created. The bilateral frontal, parietal, and temporal subcortical regions and thalamus were selected as regions of interest. Perfusion parameters of TBE patients were compared to those of a control group. RESULTS There was a slight increase in CBF and CBV, with significant prolongation of TTP in subcortical areas in the study subjects, while MTT values were comparable to those of the control group. A significant increase in thalamic CBF (p<0.001) and increased CBV (p<0.05) were observed. Increased TTP and a slight reduction in MTT were also observed within this area. CONCLUSIONS The DSC-MRI perfusion study showed that TBE patients had brain perfusion disturbances, expressed mainly in the thalami. These results suggest that DSC-MRI perfusion may provide important information regarding the areas affected in TBE patients.