Invasive pneumococcal diseases in adults admitted to the Na Bulovce Hospital: Serotype replacement after the implementation of general childhood pneumococcal vaccination.

OBJECTIVE: The aim of this study was to analyse epidemiological and clinical characteristics of invasive pneumococcal disease (IPD) in adults before and after the introduction of the general childhood conjugate pneumococcal vaccination programme in the Czech Republic. MATERIAL AND METHODS: The retrospective observational sentinel study included adults with IPD admitted to the Na Bulovce Hospital in Prague from 1/2000 through 12/2019. A case of IPD was defined as isolation of Streptococcus pneumoniae from a primarily sterile site. RESULTS: A total of 304 IPD cases were diagnosed during the study period, with a male to female ratio of 1.49:1 and age median of 58 years (IQR 43-73). The most prevalent clinical forms were bacteraemic pneumonia (185 cases; 60.9%) and purulent meningitis (90; 29.6%). A total of 157/293 patients (53.6%) required intensive care, and the case fatality rate was 25.3% (n = 77). The serotype was determined in 292 (96.0%) isolates, the most prevalent being serotypes 3 (38; 12.5%), 4 (28; 9.2%), 7F (24; 7.9%), 8 (21; 6.9%), and 1 (18; 5.9%). Both clinical and epidemiological characteristics of IPD caused by the most prevalent serotypes differed considerably. Patients diagnosed with serotype 3 were older, more frequently required intensive care, and showed higher mortality. The proportion of IPD caused by non-PCV13 serotypes increased from 28.8% (19/66) in 2000-2005 to 54.8% (40/70) in 2015-2019 (p = 0.001). CONCLUSION: The study demonstrated that invasive diseases caused by the most prevalent pneumococcal serotypes differ in their epidemiological and clinical characteristics and case fatality rate. During the study period, there was a significant increase in IPD caused by non-PCV 13 serotypes, limiting the effect of vaccination in adults.

Detection of antigen-specific T cells in patients with neuroborreliosis.

There is a lack of laboratory tests in clinical practice that can detect the activity of borrelial infection. This was the reason for testing an antigen-specific T-cell detection method in patients with neuroborreliosis: the ELISPOT method, which is capable of detecting antigen-specific T lymphocytes in clinical conditions. A group of 32 patients (20 diagnosed with neuroborreliosis; NB) was examined using this commercial method (LymeSpot intrerferon-γ Assay Kit®). Of these 20 NB patients, 10 were found to be positive and 10 negative; four of the five persons tested prior to the antibiotic treatment were positive. Eight patients served as the control group, giving four positive and four negative results. The results achieved so far appear to be unequivocal; still, the test could be expected: a) in non-specific clinical symptoms with borderline or negative proof of antibodies; b) in eraly stage of the disease; c) in the case of seropositivity and unequivocal clinical picture. The basic prerequisite for the clinical utilization of the method, however, is that it be thoroughly tested.

[Diagnostic relevance of the chemokine CXCL13 and anti-C6 peptide antibodies in patients with neuroborreliosis]. / Diagnostický význam chemokinu CXCL13 a protilátek proti C6 peptidu u pacientu s neuroborreliózou.

AIM OF THE STUDY: The study was focused on testing the diagnostic value of detection of the chemokine CXCL13 (B lymphocyte chemoattractant) and anti-C6 peptide (synthetic peptide derived from B. burdorferi VlsE protein) antibodies in patients with neuroborreliosis (NB). MATERIAL AND METHODS: One hundred and twenty-nine patients with clinical suspicion of neuroinfection were included in the study. Eighty patients with NB (positive for antibodies in serum and CSF) were subdivided into four groups (A1-A4) based on positivity/negativity of the antibody index (AI) and pleocytosis. The control group was composed of 49 patients with a negative AI and absence of CSF pleocytosis. Chemokine CXCL13 and anti-C6 antibodies were examined by commercial kits (Human CXCL13/BLC/BCA-1 Immunoassay, R&D Systems, INC, USA and C6 B. burgdorferi (Lyme) ELISA, Immunetics Inc. USA). The CXCL13 cut-off values were set to 130 pg/ml for the CSF and 62 pg/ml for the serum. RESULTS: The highest CSF levels of CXCL13 chemokine were found in group A1 (pleocytosis, AI positive), and they were significantly higher (p < 0.001) comparing with other groups except A3 (pleocytosis, AI negative; p = 0.04). Group A3 also showed significantly higher levels of CXCL13 than groups A2 (without pleocytosis, AI positive; p = 0.005), A4 (without pleocytosis, AI negative), and B (p < 0.001). The differences in the serum CXCL13 levels between groups were non-significant. The serum anti-C6 antibodies were detected in all NB groups and the positivity rates did not differ between groups (92%) except for A3 where 55% of the patients were positive. In the CSF, the highest anti-C6 sensitivity was found in the patients with a positive AI (A1 88.6%; A2 76.9%) while in the groups with a negative AI, it was low (A3 25%; A4 0%). In group B, anti-C6 antibodies were not detected. CONCLUSION: The highest CSF CXCL13 levels were found in early stage NB. Elevated CXCL13 concentrations correlate better with pleocytosis than with AI positivity; however, there exist some patients with a positive AI who have low CXCL13 levels. These patients are most probably those in the late - subacute stage of neuroinfection. The CXCL13 testing seems to be the most diagnostically helpful in the acute stage of NB where AI is still negative. The clinical sensitivity of the C6 ELISA test appears to be insufficient for CSF examination under our conditions. On the contrary, the specificity of this test was proven high, because none of the controls tested positive.

[West Nile virus fever]. / Virus západonilské horecky (west Nile virus).

West Nile virus (WNV) is a member of the Flaviviridae family, genus Flavivirus. Its reservoir hosts are wild birds. Infection is transmitted to humans by infected mosquitoes of the genus Culex. In most cases, it is either asymptomatic or manifests itself as mild fever. Typically, WNV illnesshas a sudden onset with fever above 39 degrees C and accompanying symptoms such as chills, headache, arthralgia, myalgia, back ache, cough and sore throat. Gastrointestinal symptoms are frequently reported. Generalized lymphadenopathy and conjunctivitis may develop. In some patients the infection can progress to meningoencephalitis. Diagnosis is currently based on detection of IgM antibodies in blood and cerebrospinal fluid or direct detection of WNV RNA.

Prospective study on the chemokine CXCL13 in neuroborreliosis and other aseptic neuroinfections.

The study evaluates the clinical significance of CXCL13 (leukocyte chemoattractant synthesized in CSF ) in Lyme neuroborreliosis (LNB) and other aseptic CNS infections. 244 patients with symptoms of neuroinfection and/or LNB were divided into groups: A - patients with LNB-positive antibodies in serum and CSF (96) or CSF only (14); B - patients with aseptic non-borrelial neuroinfections (82); C - negative controls (52). Group A was divided into A1-A4 according to pleocytosis in CSF and AIIgG positivity. The highest CSF CXCL13 concentrations (max. 81,287.60pg/ml; median 1766.90pg/ml) were in A1 (positive AI, pleocytosis) and A3 (negative AIIgG, pleocytosis; max. 7201,60pg/ml, median 56.22pg/ml). A2 (positive AI without pleocytosis) and A4 (negative AI without pleocytosis) had low CXCL13 levels - A2 max. 650.50pg/ml (median<7.80pg/ml); A4 max. 118.56pg/ml (median<7.8pg/ml). In B the median was 28.10pg/ml (max. 595.87pg/ml). In C the CXCL13 concentrations were the lowest (max. 83.83pg/ml; median<7.80pg/ml). The lowest cut-off was 29pg/ml (sensitivity 90.0%, specificity 72.2%), the highest one 400pg/ml (sensitivity 59.6%, specificity 94.0%). The group differences of serum CXCL13 were insignificant. The highest concentrations were at the beginning of the disease. In LNB CXCL13 correlates better with the CSF pleocytosis than AI positivity.

A continuous method for enzymatic assay of sucrose synthase in the synthetic direction.

An appropriate method was developed for the continuous assay of sucrose synthase (SS) (EC 2.4.1.13) by spectrophotometry. The uridine 5'-diphosphate derived from sucrose synthesis was stoichiometrically coupled to oxidation of beta-nicotinamide adenine dinucleotide by the enzymes nucleoside-5'-diphosphate kinase (NDPK), pyruvate kinase, and lactate dehydrogenase. Utilization of crude extracts led to a complete masking of SS assay by adenylate kinase, adenosine 5'-triphosphatase (ATPase), and phosphoenolpyruvate phosphatase found in the crude extracts. These interfering enzymes were mostly removed from the crude extracts by using a combination of gel filtration, centrifugation through a selectively permeable membrane (Biomax-100 Ultrafree centrifugal device), and inhibition by the addition of K(2)HPO(4) to the assay buffer. Sensitivity of the SS assay was significantly increased by the inclusion of NDPK and ATP, which are essential to the reaction in the coupling system.

Changes in invertase activities and reducing sugar content in sweetpotato stored at different temperatures.

Cured sweetpotato roots were stored at different temperatures (4.5, 15.6, and 24 degrees C) for 7 weeks and assayed for invertase activities and reducing sugar levels during two separate years. Invertase activities and reducing sugar concentration significantly increased in the roots kept at low temperature. Of the three types of invertases assayed, acid invertase specific activity was the highest. Acid invertase was the most influential in determining reducing sugar levels in stored sweetpotato. Cultivar differences were found in invertase specific activities and reducing sugar concentration. Reducing sugar content was highly correlated to acid and total invertase activity, regardless of cultivar.

Infective endocarditis caused by unusual gram-positive pathogens.

Of a total of 81 patients hospitalized in the infectious diseases department in 1990-2000 with infectious endocarditis caused by Gram-positive pathogen, unusual etiological agents were found in several cases: Streptococcus pyogenes, Streptococcus pneumoniae, Corynebacterium diphtheriae, and Gemella morbillorum. Cardiac defects were present in the latter two patients: bicuspid aortic valve and tetralogy of Fallot. Two patients were successfully treated with antibiotics only and one patient with antibiotics and surgery. The patient with C. diphtheriae endocarditis died due to progressive sepsis and multiple organ failure.

[Immunologic study of Lyme borreliosis]. / Imunologická studie lymeské borreliózy.

In 84 patients in the second or third stage of serologically proved Lyme borreliosis suffering from different forms of central and/or peripheral nervous system involvement the participation of autoimmunological mechanisms was investigated. The cellular hypersensitivity to encephalitogenic basic protein and to antigens from peripheral myelin was evaluated on the cytopherometer Opton according to the influence of macrophage slowing factor--liberated during the short-term incubation of the sensibilized lymphocytes with specific antigen--on the mobility of tanned sheep red blood cells in the electrical field. The frequency of positive findings in comparison with the results in other types of neuroinfections, examined previously, put the Lyme borreliosis on the first place. The explanation of this fact may be seen in the immunoregulatory disturbances, even though the changes of the total and active T lymphocytes were not significant. The knowledge of immunological changes is very important for the indication of immunomodulating procedures suitably complementing the treatment by antibiotics.

DNA persistence after treatment of Lyme borreliosis.

One hundred twenty-four patients-53 with neuroborreliosis, 48 with erythema migrans, and 23 with Lyme arthritis-were tested in a prospective study for the presence of the DNA of Borrelia burgdorferi sensu lato in plasma, cerebrospinal fluid (CSF), urine, and synovial fluid by nested polymerase chain reaction (PCR). Specific DNA was detected using five amplification systems simultaneously: three targeted chromosomal genes encoding 16S rDNA, flagellin, and p66; and two plasmid sequences of OspA and OspC. Patients were examined clinically and by PCR before and after treatment and again after 3 and 6 months. Before treatment, the specific DNA was detected in 78 patients (62.9 %). Forty-one neuroborreliosis patients were DNA-positive (77.4 %), with CSF positivity in 26 patients, urine in 25, and plasma in 16. Twenty-six erythema migrans patients were DNA-positive (54.2 %), with plasma positivity in 18 cases and urine in 14. Eleven Lyme arthritis cases (47.8 %) were DNA positive (six in urine, five in plasma, and four in synovial fluid). The frequency of PCR positives was comparable in CSF and urine, and it was lower by approximately 50 % in plasma. Specific DNA was also found in a significant number of patients in later testing periods: 48 patients after treatment, 29 patients after 3 months, and 6 patients after 6 months. The prolonged PCR positivity was not explainable by persistent infection according to the clinical manifestations of the disease. Possible explanations of the problem are discussed.

Methodology optimization for quantification of total phenolics and individual phenolic acids in sweetpotato (Ipomoea batatas L.) roots.

Phenolic acids are one of the several classes of naturally occurring antioxidant compounds found in sweetpotatoes. Simplified, robust, and rapid methodologies were optimized to quantify total and individual phenolic acids in sweetpotato roots. Total phenolic acid content was quantified spectrophotometrically using both Folin-Denis and Folin-Ciocalteu reagents. The Folin-Ciocalteu reagent gave an overestimation of total phenolic acids due to the absorbance of interfering compounds (that is, reducing sugars and ascorbic acid). Individual phenolic acids were quantified by high-performance liquid chromatography (HPLC) using the latest in column technology. Four reversed-phase C18 analytical columns with different properties (dimensions, particle size, particle shape, pore size, and carbon load) were compared. Three different mobile phases using isocratic conditions were also evaluated. A column (4.6 x 150 mm) packed with 5-microm spherical silica particles of pore size 110 A combined with 14% carbon load provided the best and fast separation of individual phenolic acids (that is, chlorogenic acid, caffeic acid, and 3 isomers of dicaffeoylquinic acid) with a total analysis time of less than 7 min. Among the 3 mobile phases tested, a mobile phase consisting of 1% (v/v) formic acid aqueous solution: acetonitrile: 2-propanol, pH 2.5 (70:22:8, v/v/v) gave adequate separation. Among the solvents tested, aqueous mixtures (80:20, solvent:water) of methanol and ethanol provided higher phenolic acid extraction efficiency than the aqueous mixture of acetone.

Neuroborreliosis in an HIV-1 positive patient.

Simultaneous co-infections of Borrelia burgdorferi sensu lato and HIV-1 are rare events, with only six published cases. A case of acute neuroborreliosis with facial palsy, meningoradiculitis (Bannwarth's syndrome) in an HIV-1 positive individual is described. Diagnosis was confirmed by Western immunoblot analysis of serum and CSF and by proof of intrathecal production of antibodies against B. garinii. The patient was successfully treated with cefotaxime. In all published HIV+ cases, the course of borreliosis did not differ from that of the HIV negative population and the prognosis in properly treated patients was good.

PCR in lyme neuroborreliosis: a prospective study.

OBJECTIVES: DNA proof is the only widely available direct diagnostic tool in Lyme borreliosis. Sensitive PCR detecting of spirochetal DNA was prepared and a prospective study in neuroborreliosis was performed. MATERIALS AND METHODS: 57 hospitalised patients with active neuroborreliosis and proved CSF antibodies synthesis were examined. Nested-PCR (utilizing three targets) was used for the detection of specific DNA in plasma, CSF and urine. RESULTS: Before treatment 36 positive patients (63.1%) were found in all tested specimens in parallel, 28 patients (49.1%) were positive in urine, 20 in CSF (35.0%) and 16 in plasma 28.0%). Later only urine was tested and the following results were obtained: 17 positive patients (30.0%) immediately after treatment, 8 (14.0%) after 3 months and one patient persisted positivity after 6 months. CONCLUSIONS: The highest sensitivity of PCR was achieved in the acute period of neuroborreliosis - 63.1% in three body fluids comparing with CSF antibody synthesis.

Disseminated Nocardia asteroides infection in an immunocompetent woman following an arm injury.

A case of disseminated infection with Nocardia asteroides in a 55-year-old immunocompetent woman after mild trauma to the arm is reported. Secondary dissemination was identified in the skin, right kidney, liver, peritoneal cavity, lungs and thigh. The patient was successfully treated with surgical drainage and a 9-week course of antibiotics including cefotaxime, amikacin, chloramphenicol, trimethoprim/sulfamethoxazole (TMP/SMX) and doxycycline. The administration of TMP/SMX in combination with doxycycline was clinically beneficial despite in vitro resistance.

An immunological study of Lyme disease.

84 patients in the second or third stages of serologically diagnosed Lyme disease suffering from different forms of central as well as peripheral nervous system involvement were tested for the participation of autoimmune mechanisms. Cell hypersensitivity to the encephalitogenic basic protein and to antigens from peripheral myelin was evaluated on an Opton cytopherometer according to the effect of the macrophage slowing factor--liberated during short-term incubation of sensibilized lymphocytes by the respective antigen--on the mobility of tannin-treated sheep red blood cells in an electric field. Judged by the presence of autoimmune reactions. Lyme disease has come to occupy a prominent position compared with previously examined other types of neuroinfections. An explanation can be sought in disordered immunoregulating mechanisms, even though the changes in total and active T lymphocytes were not statistically significant. Data on immunological changes are relevant for the indication of immunomodulating therapy as a suitable complement to antibiotic treatment.