Publications by authors named "Zuzana Cermakova"

24 Publications

  • Page 1 of 1

Annual Incidence and Severity of Acute Episodes in Hereditary Thrombotic Thrombocytopenic Purpura.

Blood 2021 Mar 1. Epub 2021 Mar 1.

Department for BioMedical Research, University of Bern, Bern, Switzerland.

Hereditary thrombotic thrombocytopenic purpura (hTTP) is a rare thrombotic microangiopathy characterized by severe congenital ADAMTS13 deficiency and recurring acute episodes causing morbidity and premature death. Information on the annual incidence and severity of acute episodes in hTTP patients is largely lacking. This study reports prospective data of 87 patients from the Hereditary TTP Registry (ClinicalTrials.gov NCT01257269) for survival, frequency and severity of acute episodes from enrollment until December 2019. The 87 patients, followed for median 4.2 years (range 0.01-15), had a median age at overt disease onset and at clinical diagnosis of 4.6 years and of 18 years (range 0.0-70 for both), respectively. Forty-three patients received regular plasma prophylaxis, while 22 did not, and treatment changed over time or was unknown in the remaining 22. Forty-three patients experienced 131 acute episodes of which 91 (69%) occurred in patients on regular prophylaxis. This resulted in an annual incidence of acute episodes of 0.36 (95%CI 0.29-0.44) with and of 0.41 (95%CI 0.30-0.56) without regular plasma treatment. More than one third of acute episodes (n=51) were documented in children <10 years of age at enrollment and were often triggered by infections. Their annual incidence of acute episodes was significantly higher than in patients >40 years of age (1.18 [95% CI 0.88-1.55] vs. 0.14 [95% CI 0.08-0.23]). Prophylactic plasma infusion regimens used were insufficient to prevent acute episodes in many patients. Such regimens are burdensome, caregivers, patients and their guardians are reluctant to start regular plasma infusions, from which particularly children would benefit.
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http://dx.doi.org/10.1182/blood.2020009801DOI Listing
March 2021

The use of a battery of examination methods for detection of cervical metastases in squamous cell carcinoma of the oral cavity.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2020 Jun 26. Epub 2020 Jun 26.

Department of Oral and Maxillofacial Surgery, University Hospital Ostrava, Czech Republic.

Introduction: In patients with squamous cell carcinoma of the orofacial area, the presence of cervical metastases represents a single most significant prognostic factor. This fact underlines the importance of thorough examination of the cervical lymph nodes for potential tumor involvement. To verify this, the most common investigative methods are physical examination (PE), sonography (US) and computed tomography (CT), which have also been used to assess the stage of the disease in the patients in our research.

Objective: To evaluate the performance of individual methods (physical examination, sonography, computed tomography) and combinations.

Method: Patients with squamous cell carcinoma of the oral cavity, who had undergone physical, US and CT examinations at our department followed by radical neck dissection were included in this retrospective study. A total of 57 patients were included.

Results: The sensitivity of PE, US and CT were 38%, 69% and 61%, respectively, however CT+US combination yielded 83% sensitivity and combination of all these methods 86% sensitivity. The number of false positives was however relatively high with specificity of the 3-way combination at 65%.

Conclusion: A combination of our three widely available inexpensive methods detected 86% of metastases in cervical nodes. The large number of false positives however indicates that the method should rather be used for screening in selecting patients who need additional and more expensive imaging than for diagnosing cervical metastases. Also, as 14% of cervical metastases pass undetected using our method, we would recommend an additional examination at least by US+PE several weeks to a few months after the initial examination.
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http://dx.doi.org/10.5507/bp.2020.026DOI Listing
June 2020

Evaluation of spine structure stability at different locations during SBRT.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2020 Jun 17;164(2):177-182. Epub 2019 Jun 17.

Department of Oncology, University Hospital Ostrava, Ostrava, Czech Republic.

Background And Aims: Modern stereotactic body radiotherapy (SBRT) techniques and systems that use online image guidance offer frameless radiotherapy of spinal tumors and the ability to control intrafraction motion during treatment. These systems allow precise alignment of the patient during the entire treatment session and react immediately to random changes in this alignment. Online tracking data provide information about intrafractional changes, and this information can be useful for designing treatment strategies even if online tracking is not being used. The present study evaluated spine motion during SBRT treatment to assess the risk of verifying patient alignment only prior to starting treatment.

Methods: This study included 123 patients treated with spine SBRT. We analyzed different locations within the spine using system log files generated during treatment, which contain information about differences in the pretreatment reference spine positions by CT versus positions during SBRT treatment. The mean spine motion and intra/interfraction motion was evaluated. We defined and assessed the spine stability and spine significant shifts (SSHs) during treatment.

Results: We analyzed 462 fractions. For the cervical (C) spine, the greatest shifts were in the anterior-posterior (AP) direction (2.48 mm) and in pitch rotation (1.75 deg). The thoracic (Th) spine showed the biggest shift in the AP direction (3.68 mm) and in roll rotation (1.66 deg). For the lumbar-sacral (LS) spine, the biggest shift was found for left-right (LR) translation (3.81 mm) and roll rotation (3.67 deg). No C spine case exceeded 1 mm/1 deg for interfraction variability, but 7 of 54 Th spine cases exceeded 1 mm interfraction variability for translations (maximum value, 2.5 mm in the AP direction). The interfraction variability for translations exceeded 1 mm in 2 of 24 LS spine cases (maximum value, 1.7 mm in the LR direction). Only 13% of cases had no SSHs. The mean times to SSH were 6.5±3.9 min, 8.1±5.9 min, and 8.8±7.1 min for the C, Th, and LS spine, respectively, and the mean recorded SSH values were 1.6±0.66, 1.43±0.33, and 1.46±0.47 mm/deg, respectively.

Conclusion: Positional tracking during spine SBRT treatments revealed low mean translational and rotational shifts. Patient immobilization did not improve spine shifts compared with our results for the Th and LS spine without immobilization. For the most precise spine SBRT, we recommend checking the patient's position during treatment.
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http://dx.doi.org/10.5507/bp.2019.027DOI Listing
June 2020

The International Hereditary Thrombotic Thrombocytopenic Purpura Registry: key findings at enrollment until 2017.

Haematologica 2019 10 21;104(10):2107-2115. Epub 2019 Feb 21.

Department of Hematology and Central Hematology Laboratory, Inselspital, Bern University Hospital, Bern, Switzerland

Congenital thrombotic thrombocytopenic purpura is an autosomal recessive inherited disease with a clinically heterogeneous course and an incompletely understood genotype-phenotype correlation. In 2006, the Hereditary TTP Registry started recruitment for a study which aimed to improve the understanding of this ultra-rare disease. The objective of this study is to present characteristics of the cohort until the end of 2017 and to explore the relationship between overt disease onset and ADAMTS13 activity with emphasis on the recurring c.4143_4144dupA mutation. Diagnosis of congenital thrombotic thrombocytopenic purpura was confirmed by severely deficient ADAMTS13 activity (≤10% of normal) in the absence of a functional inhibitor and the presence of mutations on both alleles. By the end of 2017, 123 confirmed patients had been enrolled from Europe (n=55), Asia (n=52, 90% from Japan), the Americas (n=14), and Africa (n=2). First recognized disease manifestation occurred from around birth up to the age of 70 years. Of the 98 different mutations detected, c.4143_4144dupA (exon 29; p.Glu1382Argfs*6) was the most frequent mutation, present on 60 of 246 alleles. We found a larger proportion of compound heterozygous than homozygous carriers of c.4143_4144dupA with overt disease onset at < 3 months of age (50% 37%), despite the fact that ADAMTS13 activity was <1% in 18 of 20 homozygous, but in only 8 of 14 compound heterozygous carriers. An evaluation of overt disease onset in all patients with an available sensitive ADAMTS13 activity assay (n=97) shows that residual ADAMTS13 activity is not the only determinant of age at first disease manifestation. Registered at .
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http://dx.doi.org/10.3324/haematol.2019.216796DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6886414PMC
October 2019

[Pets (dogs/cats) as a possible source of opportunistic pathogenic fungi in humans].

Klin Mikrobiol Infekc Lek 2018 Jun;24(2):41-49

Department of Clinical Microbiology, Faculty of Medicine Charles University and University Hospital Hradec Králové, Czech Repulic, e-mail:

Introduction: The microbiological aspect of a relationship between pets (dogs/cats) and their owners is mainly concerned with the incidence of the shared fungal species that can be potential pathogens. Since sharing homes with pets is very popular in the Czech Republic, there is an increased possibility of communication between microbiota of the two macroorganisms (the pet and the owner). The aim of the study was to determine, based on the close relationship between pets and humans, the biodiversity of shared fungi, also with respect to previous antimicrobial therapy.

Methods: A total of 103 samples were collected from 20 pairs (20 owners, 16 dogs and 4 cats). All owners completed a questionnaire with their pets' veterinarians. In owners, swabs were collected from the nasal mucosa, armpit and interdigital spaces of the foot. In pets, swabs were obtained from the external auditory meatus and nasal mucosa. In individuals with skin lesions, samples were also collected from the affected areas. Fungal species were identified by culture and microscopy methods and confirmed by matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF) mass spectrometry. Statistical methods were used to correlate the closeness of relationship with the number of shared fungal species and to correlate previous antimicrobial therapy with the number of shared species of microscopic fungi.

Results: Analysis of the questionnaire found that 65 % of owners who participated in the study kept more pets at home than only the tested one. In the previous year, 5 % of pets and 5 % of owners received antimicrobial therapy. As many as 45 % of dogs or cats slept in their owners' beds and 80 % rested on a sofa together with their owners. Also, 45 % of owners had their faces licked by pets. Eighty percent of pets were fed with several types of food (dry food and cooked food). Further, 70 % of pets lived permanently with their owners in the same household. A total of 45 microscopic fungi species were isolated, of which 15 species occurred in both macroorganisms (pets and humans). Thirty-two species were identified from human and 28 species from animal samples. The most frequent species was the yeast Candida albicans, isolated from 30 samples. From the human nasal mucosa, only four species were isolated. The richest biodiversity was observed in interdigital space samples (26 fungal species). Once again, the most frequent fungal species was C. albicans (8 cases). The most numerous animal samples were obtained from the external auditory meatus. There, the most frequent species was Malassezia pachydermatis (17 cases). In seven pairs, microscopic fungi were shared. Of those, two pairs shared two spe-cies and five pairs shared one species. A total of five fungal species were shared, most often the yeasts C. albicans and Geotrichum candidum.

Conclusion: The closeness of the human-pet relationship apparently does not influence the number of shared fungal species. The yeast Candida albicans was most frequently isolated from owners as well as from the nasal mucosa in pets. The lipophilic yeast M. pachydermatis most commonly occurred in the material from the external auditory meatus and skin scales from dogs and cats.
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June 2018

ADAMTS13 kinetics after therapeutic plasma exchange and plasma infusion in patients with Upshaw-Schulman syndrome.

J Clin Apher 2019 Feb 5;34(1):13-20. Epub 2018 Nov 5.

Blood Centre, University Hospital Ostrava, Ostrava, Czech Republic.

Background: Hereditary thrombotic thrombocytopenic purpura, also called Upshaw-Schulman syndrome (USS), is a rare disease caused by genetic mutations in the ADAMTS13 gene, which severely decrease the activity of ADAMTS13, a metalloprotease that cleaves von Willebrand factor multimers (VWF). Genotypically identical patients can show great phenotypic diversity.

Objectives: Comparison of selected laboratory parameters and ADAMTS13 pharmacokinetics among patients with USS was performed.

Patients/methods: Six patients with USS on prophylactic plasma therapy have been reviewed, retrospectively. Blood counts, lactate dehydrogenase (LDH), and ADAMTS13 activity at various time-points before and after different treatment cycles were evaluated.

Results: ADAMTS13 recovery and pharmacokinetics were affected by treatment modality, and also reflected the patients' comorbidities and their current physiological and clinical condition.

Conclusions: Our present findings support a multifactorial contribution to treatment efficacy, and confirm the importance of adaptability and individualization of USS therapy. Therapeutic plasma exchange even in hereditary TTP is an option that can in some patients prolong intervals between plasma administration.
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http://dx.doi.org/10.1002/jca.21664DOI Listing
February 2019

Granulocyte transfusions collected after steroid priming for severe infections during neutropenia: A single center experience.

Transfus Clin Biol 2019 Nov 6;26(4):299-303. Epub 2018 Oct 6.

Department of Hemato-oncology, Faculty Hospital Olomouc and Faculty of Medicine and Dentistry, Palacky University Olomouc, I.P. Pavlova 6, 77520 Olomouc, Czech Republic. Electronic address:

Background: There is no universally accepted opinion on the use of granulocyte transfusions collected using apheresis (GTA) in neutropenic patients and severe infection.

Patients And Methods: The efficacy and safety of GTAs transfused at a single center over 10 years were analyzed retrospectively. GTAs were harvested from voluntary unrelated donors after priming with methylprednisolone using continuous apheresis and hydroxyethylstarch as sedimentation agent.

Results: 41 patients with neutropenia and hematologic malignancy (15 females and 26 males aged 22-69 (median 45.5)) were given a median 3.5 GTAs per patient (range: 1-17) containing a median 1.39×10 granulocyte/GTA (range: 0.65-2.81). The indications for GTA use were soft tissue inflammation, sepsis, and pneumonia in 30, 22, and 14 cases, respectively. After GTA complete (30 patients: 73.2%) or partial (6 patients: 14.6%) healing of the infection was achieved. The success rate was 91.7% in soft tissue infections, 66.7% in invasive fungal infections, and 68% in sepsis. Septic shock (documented in 12 cases) was associated with a poor response (P<0.03; Chi-square test). Clinical worsening was observed in six cases (14.6%); four patients died. No significant short-term side effects of GTA treatment were recorded.

Conclusions: In our study GTAs collected after steroid priming and used for the treatment of infection during severe neutropenia have shown comparable efficacy with several previously reported trials. However retrospective fashion of our study and inhomogeneous group of patients do not allow any firm conclusions. Prospective studies (including patients' registries) are needed for the better clarification of the role and the dose of GTAs necessary for the successful infection management during neutropenia.
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http://dx.doi.org/10.1016/j.tracli.2018.09.001DOI Listing
November 2019

[Concurrent malaria and rickettsiosis in a patient returning from the Republic of South Africa].

Klin Mikrobiol Infekc Lek 2017 12;23(4):142-147

Department of Infectious Diseases, Faculty of Medicine in Hradec Kralove, Charles University, Czech Republic, e-mail:

Definitive diagnosis and therapy proved challenging in the case of a 60-year-old male with malaria and rickettsiosis. Returning travellers who are unwell can present practical difficulties in diagnosis and treatment and the focus here is on conditions relevant to the Republic of South Africa. Malaria, rickettsiosis and Q fever are discussed.
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December 2017

[Sharing bacterial microbiota between owners and their pets (dogs, cats)].

Klin Mikrobiol Infekc Lek 2017 06;23(2):48-57

Department of Clinical Microbiology, Faculty of Medicine Charles University and University Hospital Hradec Králové, Czech Republic, e-mail:

Introduction: The microbiological aspect of a relationship between pets (dogs/cats) and their owners is mainly concerned with the incidence of shared bacterial species, in particular potential pathogens. Given the great popularity of sharing homes with pets (dogs/cats) in the Czech Republic, there is an increased possibility of communication between microbiota of the two macroorganisms (pet and owner). The aim of the study was to determine the biodiversity of shared bacteria and possibility of exchange of genes of resistance to antimicrobial agents between potential pathogens based on the close relationship between pets and humans.

Methods: A total of 103 samples were collected from 20 pairs (20 owners, 16 dogs and 4 cats). All owners completed a questionnaire with their pets' veterinarians. In owners, swabs were collected from the nasal mucosa, armpit and interdigital spaces of the foot. In pets, swabs were obtained from the external auditory meatus and nasal mucosa. In individuals with skin lesions, samples were also collected from the affected areas. Bacterial species were identified by culture and matrix-assisted laser desorption/ionization - time of flight (MALDI-TOF) mass spectrometry. In shared species, susceptibility to antibiotics was tested by the disk diffusion method. Statistical methods were used to correlate the closeness of relationship with the number of shared bacterial species and to correlate previous antimicrobial therapy with shared resistance of the common bacteria.

Results: Analysis of the questionnaires showed that 65 % of owners who participated in the study kept more pets at home than only the tested one. In the previous year, 5 % of pets and 5 % of owners received antimicrobial therapy. As many as 45 % of dogs or cats slept in their owners' beds and 80 % rested on a sofa together with their owners. Also, 45 % owners had their faces licked by pets. Eighty percent of pets were fed with several types of food (dry food and cooked food). Further, 70 % of pets lived permanently with their owners in the same household. A total of 76 bacterial species of 33 genera were identified. The most frequently isolated species (29 samples) was S. intermedius. Seventeen bacterial species occurring in both humans and animals were found and identified. At least one bacterial species was shared by 11 pairs and two shared species were found in two pairs. The shared species were S. intermedius, E. coli, E. faecalis, A. lwoffii, P. putida and S. aureus. Antimicrobial susceptibility was tested in the shared species. Common antimicrobial resistance was found in four pairs. In one pair, shared E. faecalis showed identical resistance to co-trimoxazole; in another pair, S. intermedius was resistant to gentamycin, erythromycin, clindamycin and co-trimoxazole. The third resistant bacterial species was E. coli; in one pair, it showed borderline resistance to colistin; in the second case, it was fully resistant to this antimicrobial agent. The other pairs with shared bacteria did not show any common resistance.

Conclusion: The study results showed that there was an association between closeness of the human-pet relationship and the prevalence of shared bacterial species. Pairs with a close relationship were 37.5 % more likely to share bacteria than pairs with a less close relationship. The study suggests that antimicrobial therapy in at least one pair member may increase the risk of shared bacterial resistance.
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June 2017

Evaluation of anti-inflammatory activity of prenylated substances isolated from Morus alba and Morus nigra.

J Nat Prod 2014 Jun 5;77(6):1297-303. Epub 2014 Jun 5.

Department of Natural Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences Brno , CZ-612 42 Brno, Czech Republic.

Chromatographic separation of root extracts of Morus alba and M. nigra led to the identification of the 2-arylbenzofurans moracin C (1), mulberrofuran Y (2), and mulberrofuran H (3), and the prenylated flavonoids kuwanon E (4), kuwanon C (5), sanggenon H (6), cudraflavone B (7), and morusinol (8), and the Diels-Alder adducts soroceal (9), and sanggenon E (10). The cytotoxicity and their antiphlogistic activity, determined as the attenuation of the secretion of TNF-α and IL-1β and the inhibition of NF-κB nuclear translocation in LPS-stimulated macrophages, were evaluated for compounds 1-10.
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http://dx.doi.org/10.1021/np401025fDOI Listing
June 2014

Evaluation of biochemical markers and bone mineral density in patients with chronic kidney disease stage 5D at the start of hemodialysis treatment.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2015 Mar 29;159(1):93-9. Epub 2013 Nov 29.

Clinic of Internal Medicine, University Hospital Ostrava, Ostrava, Czech Republic.

Background: Patients with chronic kidney disease (CKD) have significant disorders of bone and mineral metabolism. In addition, they can also develop other bone disorders including osteoporosis. This study evaluated the bone mineral density (BMD) of patients at the start of hemodialysis treatment as well as the relationship between BMD and possible risk factors or biochemical markers.

Methods: The study was performed in 82 patients (28 females, 54 males). BMD was measured by dual-energy X-ray absorptiometry (DXA) at the lumbar spine and the proximal femur.

Results: We found a high prevalence of 25-hydroxyvitamin D deficiency (96%; mean levels 30.0 ± 17.7 nmol/L) and a reduction of BMD in comparison with gender- and age-matched normal population values at the total hip (Z-score = -0.31 ± 1.11) and the femoral neck (Z-score = -0.48 ± 1.16), but not at the lumbar spine (Z-score = 0.68 ± 1.81). The prevalence of T-scores ≤ -2.5 SD in the group of patients over 50 years was 52.0% in females and 33.3% in males. BMD positively correlated: with male gender and calcium levels at all measured sites, with age at the lumbar spine and with weight or BMI at the proximal femur.

Conclusion: CKD patients at the start of hemodialysis treatment had a high prevalence of low T-score values, corresponding to values for osteoporosis in the general population. BMD at the proximal femur was below the expected average for age and gender, but at the lumbar spine, BMD in hemodialysis patients was above average in persons without known CKD.
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http://dx.doi.org/10.5507/bp.2013.087DOI Listing
March 2015

Real-time PCR method for the detection of the gene encoding surface lipoprotein LipL32 of pathogenic Leptospira: use in the laboratory diagnosis of the acute form of leptospirosis.

Scand J Infect Dis 2013 Aug;45(8):593-9

Department of Clinical Microbiology, Charles University in Prague, Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove, Hradec Kralove, Czech Republic.

Background: The aims of this work were to replace the obsolete PCR method for the laboratory diagnosis of the acute form of leptospirosis using the G1, G2 and B64 I, B64 II primers, and to improve the PCR detection time.

Methods: We introduced a real-time PCR method for the detection of the gene encoding the surface lipoprotein LipL32 of pathogenic Leptospira into our laboratory diagnosis of the acute form of leptospirosis. The positive and negative analytical specificities of the real-time PCR method were both equal to 100%; the detection limit was determined to be 1-5 genome copies/1 ml of liquid biological material. The method was further validated on 230 laboratory strains of leptospires.

Results: All laboratory strains of pathogenic Leptospira were evaluated as LipL32-positive and all non-pathogenic strains as LipL32-negative. In addition, 455 biological materials (253 plasma, 121 urine, 72 cerebrospinal fluid (CSF), 7 bronchoalveolar lavage, and 2 sputum) from 295 patients with suspected leptospirosis were examined. From this set of patients, 9 were evaluated to be LipL32-positive, from 15 positive biological materials (10 urine, 4 blood plasma, and 1 CSF).

Conclusions: This real-time PCR method for the detection of the gene encoding the surface lipoprotein LipL32 is a reliable, sensitive, and rapid method for the detection of the acute form of leptospirosis.
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http://dx.doi.org/10.3109/00365548.2013.795656DOI Listing
August 2013

[Question marks concerning serological tests in congenital toxoplasmosis - a case report].

Klin Mikrobiol Infekc Lek 2012 Dec;18(6):192-5

Department of Infectious Diseases, Charles University, Czech Republic.

Two weeks after delivery of a healthy term neonate, the mother developed lymph node syndrome, which corresponded serologically to acute toxoplasmosis. The blood of the newborn showed positive IgM, IgG and IgA antibody titers against Toxoplasma gondii with a low avidity of IgG. The newborn did not show any clinical signs or organ damage connected to toxoplasmosis either at the beginning or during the follow-up. The IgA and IgM titers exhibited a decrease over time, while the KFR, IgG antibody titers and avidity had an increasing trend. A sharp increase of KFR, IgE and IgA antibody titers was detected during the sixth month of life, probably due to maturation of the immune system in the setting of an asymptomatic infection with T. gondii. This short increase was followed by a subsequent decrease in titers of these antibodies until they reached negative levels during the 21st month of life. The evaluation of serological results in newborns infected with T. gondii is always difficult and should be performed by an expert physician. Children at risk should be placed under a long-term follow-up to avoid potential development of toxoplasma chorioretinitis.
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December 2012

Detection of nano- and micro-sized particles in routine biopsy material - pilot study.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2015 Mar 10;159(1):87-92. Epub 2012 Dec 10.

Faculty of Medicine, University of Ostrava, Syllabova 19, 703 00 Ostrava, Czech RepublicbFaculty Hospital Ostrava, 17.listopadu 1790, 708 52 Ostrava cNanotechnology Center, VSB - Technical University Ostrava, 17. listopadu 15, 70833 Ostrava.

Background: Nanotechnology is receiving enormous funding. Very little however is known about the health dangers of this technology so far. Chronic tonsillitis is one of a number of diseases called idiopathic. Among other factors, the tonsils are exposed to suspended particles in inhaled air including nano particles. The objective of this study was to detect and evaluate metallic particles in human tonsil tissue diagnosed with chronic tonsillitis and in amniotic fluid as a comparison.

Methods: . Scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDX) was used for identification of solid particles in a total of 64 samples of routinely analyzed biopsy and cytologic material.

Results: Almost all samples were found to contain solid particles of various metals. The most frequent, regardless of diagnosis, were iron, chromium, nickel and aluminium. The size, determined using SEM, varied from around 500 nm to 25 µm. The majority formed aggregates of several micrometers in size but there were a significant number of smaller (sub-micrometer or nano-sized) particles present. The incidence of metallic particles was similar in child and adult tissues. The difference was in composition: the presence of several metals in adults was due to occupational exposure.

Conclusions: The presence of metallic particles in pathologically altered tissues may signal an alternative causation of some diseases. The ethiopathogenic explanation of these diseases associated with the presence of nano-sized particles in the organism has emerged into a new field of pathology, nanopathology.
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http://dx.doi.org/10.5507/bp.2012.104DOI Listing
March 2015

Our experience using real-time PCR for the detection of the gene that encodes the superficial lipoprotein LipL32 of the pathogenic leptospires to confirm the acute form of human leptospirosis.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2013 Dec 7;157(4):387-91. Epub 2012 Dec 7.

Department of Clinical Microbiology, Faculty of Medicine in Hradec Kralove, Charles University in Prague and University Hospital in Hradec Kralove, Hradec Kralove, Czech Republic.

Aims: To examine biological materials (blood, urine, cerebrospinal fluid) of patients with suspected leptospirosis using real-time PCR for detecting the gene that codes the superficial LipL32 lipoprotein, and to evaluate the contribution of the real-time PCR method for the laboratory diagnosis of the acute form of leptospirosis.

Material And Methods: During the monitored period of April 2010 - December 2011, a total of 340 biological materials samples were examined (177x blood plasma, 88x urine, 68x, cerebrospinal fluid, 6x bronchoalveolar lavage and 1x sputum) from 216 patients with suspected leptospirosis using real-time PCR LipL32 gene detection.

Results: From the mentioned 216 patients suspected of leptospirosis, 8 patients were evaluated as being PCR LipL32 positive, from which 14 positive biological materials originated (9 x urine, 4x blood and 1x liquor).

Conclusion: As demonstrated in the study, the real-time PCR method for detecting the gene for the superficial lipoprotein LipL32 is an appropriate, quick and reliable method for the diagnosis of the acute form of leptospirosis.
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http://dx.doi.org/10.5507/bp.2012.109DOI Listing
December 2013

Procedure for granulocyte collection performed at the Blood Centre of the Faculty Hospital Ostrava.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2014 Jun 1;158(2):309-12. Epub 2012 Nov 1.

Department of Biomedical Sciences, Faculty of Medicine, University Ostrava, Czech Republic.

Background: Granulocyte apheresis is a safe and effective method for granulocyte collection. We present a five year experience (2006-2010) of the Blood Center, Faculty Hospital Ostrava, Czech Republic. Donor granulocyte transfusion is one treatment option for haemato-oncology patients with severe neutropenia complicated by bacterial/fungal infections unresponsive to standard antibiotic/antifungal treatment. In this study, we describe the experiences of the Blood Centre at the Faculty Hospital in Ostrava of granulocyte apheresis.

Methods And Results: A total of 149 granulocyte units were collected for 33 patients from the Department of Haemato-oncology, University Hospital Olomouc, over a 5-year period (2006-2010). Information on donor selection, laboratory screening, mobilization regimen and granulocyte yield was obtained and analyzed. All mandatory screening tests for infection markers, AB0 and Rh and abnormal erythrocyte antibodies were performed. The cytomegalovirus status of the donors was not investigated. Steroids were the only mobilization regimen used, and were generally well accepted. The mean granulocyte yield was 12.6×10(9)/per transfusion unit (range 5.4-30.3). All granulocyte concentrates were irradiated and transfused according to AB0 Rh compatibility within 24 h after collection.

Conclusion: Based on our experience, granulocytapheresis is a safe and effective method for obtaining granulocytes but the yield can be significantly influenced by other variables. From the recipients' perspective, the use of donor granulocytes supports an effective therapeutic modality.
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http://dx.doi.org/10.5507/bp.2012.071DOI Listing
June 2014

[Comparison of results of two serological methods for diagnosing leptospirosis - microagglutination test and ELISA].

Klin Mikrobiol Infekc Lek 2011 Oct;17(5):173-8

Department of Microbiology, Charles University Hradec Králové, Czech Republic.

Background: The aim of this study was to evaluate diagnostic sensitivity and specificity of the SERION ELISA classic IgM and SERION ELISA classic IgG kits and to confirm the results by the microagglutination test (MAT).

Material And Methods: A total of 45 blood serum samples from 45 patients, 30 from males and 15 from females (mean age 44.24 ± 15.56 range 19-82 year), were included in our study. Blood serum samples were examined using the ELISA and MAT methods and diagnostic sensitivity and specificity of both methods were calculated.

Results: The MAT was shown to have 100 % diagnostic sensitivity and specificity. The ELISA kits for detecting IgM and IgG antibodies against pathogenic leptospires had diagnostic sensitivity of 100 % and diagnostic specificity of 88.6 % and 54.3 %, respectively.

Conclusion: The above results suggest that the MAT with diagnostic sensitivity and specificity of 100 % remains the gold standard for detection of specific antibodies against pathogenic leptospires. The diagnostic sensitivity of both ELISA kits is high but due to their low diagnostic specificity, especially in the case of IgG antibodies, the kits are inappropriate for use in routine clinical practice.
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October 2011

[Incidence of leptospirosis in Pardubice and Hradec Králové region and a part of Vysočina region in the years 2002-2009].

Klin Mikrobiol Infekc Lek 2011 Oct;17(5):168-72

Department of Microbiology, Medical Faculty of Charles University Hradec Králové, Czech Republic.

Background: The aim of this study was to evaluate the incidence of leptospirosis in Pardubice and Hradec Králové regions and a part of Vysočina region in 2002-2009. A group of patients was statistically analyzed and the incidence rates of leptospirosis in individual months and years were calculated.

Material And Methods: A group of 4,813 patients with suspected leptospirosis from Pardubice and Hradec Králové regions and a part of Vysočina region were examinated in our centre in 2002-2009. A total of 5,366 biological samples were studied. From 4,625 blood samples, 3,958 were examinated with the serological microagglutination-lysis method and the remaining 667 with the PCR method.

Results: From 4,813 patients suspected to have leptospirosis, 89 were found to be leptospirosis-positive. Leptospirosis was diagnosed in 63 males (70.7 %) a 26 females (29.3 %); the mean age were 39.52 ± 17.67 years in males and 46.85 ± 18.34 years in females; the mean age men + women was 41.66 ± 18.13 years (range 3-78 years). Most positive male patients (n = 28) were in 26-45 age group; the findings were similar in females (n = 10). The highest number of leptospirosis cases was noted in 2005. Most frequently, infection caused by pathogenic leptospires was diagnosed in November.

Conclusion: The incidence rates of leptospirosis in Pardubice and Hradec Králové regions and a part of Vysočina region in 2002-2009 (with the exception of 2008) exceeded the overall reported levels 0.4/100,000 population.
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October 2011

[Considering the role of the PCR method in routine diagnosis of toxoplasmosis in peripheral blood tests in immunocompetent patients].

Klin Mikrobiol Infekc Lek 2009 Aug;15(4):138-42

Department of Clinical Microbiology, University Hospital Hradec Králové, Charles University, Czech Republic.

Infections caused by pathogenic protozoan Toxoplasma gondii in our geographic area is the most frequent parasitic infection; Czech Republic declares seroprevalence approx. 30 %. Diagnosis of toxoplasmosis is mostly based on serological methods are used (EIA IgM, IgA, IgE, IgG and avidity in IgG, Western Blot, complement fixation). According to positive results of these tests diagnosis of acute toxoplasmosis is established. In our retrospective study we tried to evaluate results of T. gondii DNA positivity from blood samples by PCR compared with positive markers of acute infection in patients before specific therapy was initiated. In accordance with literature we concluded, that in routine examination of immunocompetent outpatients of Clinic of Infectious Diseases from the moment of 4 weeks after lymphatic nodes swelling protozoan DNA detection in blood sample is not possible.
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August 2009

[Malaria-an unusual cause of fever in the infant.].

Klin Mikrobiol Infekc Lek 2005 Dec;11(6):229-32

Department of Clinical Microbiology, University Hospital Hradec Králové,Czech republic.

Malaria is the fourth most frequent cause of death in African children. Connected with perinatal diseases as well as gastrointestinal and respiratory infections malaria has been still a serious health problem of that region. Occurrence of tropical malaria in infants reported in European countries is relatively rare. Not only from that reason, the assesment of diagnosis in children under one year of age seems to be obviously more difficult. The authors report the malaria in five-month-old infant from Cameroon who became ill during his stay in the Czech Republic. Non-specific symptoms, high level of parasitemia and impairment of blood coagulation were the main features of the emergent infection. On conclusion, the lack of suitable forms of childrens antimalarial drugs both for profylaxis and treatment is mentioned.
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December 2005

[Method of polymerase chain reaction in toxoplasmosis diagnosis].

Acta Medica (Hradec Kralove) Suppl 2004 ;47(2):71-3

Fakultní nemocnice v Hradci Králové, Ustav klinické mikrobiologie.

Since introduction of polymerase chain reaction for the detection of DNA Toxoplasma gondii 398 biological samples from 301 patients were examined from 2000 to 2003. Positive finding of toxoplasmosis DNA we noted in 23 cases. Polymerase chain reaction enables exact and fast diagnosis of the actual parasitemia Toxoplasma gondii, which is important especially in the pregnant patients, new-born with suspicion on congenital toxoplasmosis and in the patients with immunosupression.
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October 2005

Diagnosis of Lyme borreliosis using enzyme immunoanalysis.

Med Sci Monit 2005 Apr 24;11(4):BR121-5. Epub 2005 Mar 24.

Department of Clinical Microbiology, Faculty of Medicine in Hradec Králové, Charles University of Prague, Czech Republic.

Background: Antiborrelia antibodies in Lyme borreliosis (LB) are mostly detected by enzyme immunoassay (EIA), confirmed by immunoblot (the "two-step system"). In indicated cases, direct evidence of Borrelia burgdorferi is obtained with the PCR method, electron microscopy and cultivation. The "one-step system" of testing for IgM and IgG antibodies in LB is economically preferably, but it requires an EIA kit with more than 90% sensitivity and specificity.

Material/methods: 90 blood samples were collected, 54 from patients with clinically defined LB and 36 samples from individuals free of LB. IgM and IgG antibodies against Borrelia burgdorferi were detected in parallel with five different EIA kits from various producers. The results were verified clinically in all cases, in disputable cases with additional immunoblot (BAG-Med), and analyzed statistically.

Results: Specificity and sensitivity were calculated from the measured values, and diagnostic efficiency was determined for each EIA kit. EIA kits for antiborrelia antibody assay with high specificity have low sensitivity and vice versa. In 9 samples from patients with clinical diagnoses (multiple sclerosis, Parkinson disease, epilepsy, rheumatoid arthritis) we found false positives in EIA and WB tests.

Conclusions: The best results for a "one-step system" of examinations for antiborrelia antibodies were obtained with the Abbot and Euroimmun EIA kits in our set. A "two-step system" of serological examination could be composed from the basic IgM and IgG examination with a high sensitivity EIA kit (Viroimmun, Test-Line) followed with confirmation of positives by specific immunoblot.
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April 2005

[Diagnostics of larval toxocariasis].

Klin Mikrobiol Infekc Lek 2004 Aug;10(4):181-5

Institute of Clinical Microbiology, Faculty of Medicine, Charles University, Hradec Králové, Czech Republic.

Purpose Of The Study: The paper, conceived as a retrospective study, resumes the results of our own investigations of the presence of serum IgG antibodies against Toxocara canis (as a crossover also against T. cati) in the sera of patients suspected of presenting larval toxocariasis in the region of Eastern Bohemia of the Czech Republic in the years 1997-2002. Our aim was to determine the immunity rate of IgG, its relationship to age and sex, further the whole range of diagnoses, the frequency of demands for such an examination by physicians and the evaluation of our experience with the detection of low avidity IgG.

Material And Methods: With a commercial diagnostic kit the ELISA method was used to detect the presence of IgG antibodies against T. canis in 2 994 sera. IgG avidity was determined in 92 positive samples with an avidity solution, which is included in the diagnostic kit and has been used in our laboratory since 2001.

Results: The immunity rate of larval toxocariasis in our set was 20.4 % (612 samples). With age this rate increases (in individual decades of life - 11.9; 8.3; 11.4; 17.9; 26.1; 29.8 and 34.2%). Up to adolescence positive findings predominate among boys (males 0-10 years 84.2 %; 11-20 years 76 %). In the third decade we see a dramatic change in favour of women (women 21-30 years - 74.6 %). In absolute figures the number of positive findings stagnates in men after the age of 30 years, while among women positive findings continue to grow - yet expressed in percentages there is signifinct change in the male-female ratio (beginning with the 4th decade we find among the women 61.2; 69.7; 72.6 and 64.8 %). Physicians' interest in the diagnosis of toxocariasis in the years 1997-2002 is confirmed by the significantly higher number of investigated samples (135; 446; 617; 422; 558; 816). This, however, does not go hand in hand with a higher proportion of IgG positivity. IgG avidity was investigated in 92 positive samples - low avidity was confirmed in six of them (6.5 %); the appropriate case histories are not a part of the study.

Conclusions: Our results tally with the data published by other departments in the Czech Republic. They confirm that the methods used have, as before, their place in the diagnostic range of microbiological laboratories.
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August 2004

CC chemokine receptor (CCR)2 polymorphism in Czech patients with myocardial infarction.

Immunol Lett 2003 Jul;88(1):53-5

Department of Immunology, Palacký University, I.P. Pavlova str. 6, Olomouc, Czech Republic.

Examining an association between myocardial infarction (MI) and the Val/Ile polymorphism in the gene for CC chemokine receptor (CCR)2 at the position 64 (CCR2-V64I), 122 MI Czech patients and 277 unrelated control (C) subjects were genotyped by PCR-SSP. The frequency of the VI genotype of CCR2-V64I was increased in MI patients in comparison with the control population (P=0.03). Further analysis revealed that relationship between the VI genotype and MI is specific only for females and, strikingly, this genotype was associated to an early MI onset (before or at the age of 50 years). Females with the VI genotype were seven times more prone to suffer from MI before 50 years than those with the VV genotype (P<0.01). If the VI genotype of the CCR2-V64I is indeed a risk factor for an earlier MI onset in females must be checked by independent studies in other centres and/or populations.
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http://dx.doi.org/10.1016/s0165-2478(03)00053-1DOI Listing
July 2003