Lenka Zdražilová Dubská

Differential role of passerine birds in distribution of Borrelia spirochetes, based on data from ticks collected from birds during the postbreeding migration period in Central Europe.

ABSTRACT Borrelia spirochetes in bird-feeding ticks were studied in the Czech Republic. During the postbreeding period (July to September 2005), 1,080 passerine birds infested by 2,240 Ixodes ricinus subadult ticks were examined. Borrelia garinii was detected in 22.2% of the ticks, Borrelia valaisiana was detected in 12.8% of the ticks, Borrelia afzelii was detected in 1.6% of the ticks, and Borrelia burgdorferi sensu stricto was detected in 0.3% of the ticks. After analysis of infections in which the blood meal volume and the stage of the ticks were considered, we concluded that Eurasian blackbirds (Turdus merula), song thrushes (Turdus philomelos), and great tits (Parus major) are capable of transmitting B. garinii; that juvenile blackbirds and song thrushes are prominent reservoirs for B. garinii spirochetes; that some other passerine birds investigated play minor roles in transmitting B. garinii; and that the presence B. afzelii in ticks results from infection in a former stage. Thus, while B. garinii transmission is associated with only a few passerine bird species, these birds have the potential to distribute millions of Lyme disease spirochetes between urban areas.

Metronomic Chemotherapy with the COMBAT Regimen in Advanced Pediatric Malignancies: A Multicenter Experience

Background: The outcome of children with refractory/relapsed malignancies remains poor and novel therapies are urgently required. One of the promising approaches is metronomic chemotherapy. We present the clinical results of 74 children with advanced solid tumors treated according to treatment recommendation with data registry in three European pediatric centers. Methods: COMBAT (Combined Oral Metronomic Biodifferentiating Antiangiogenic Treatment) included low-dose daily temozolomide, etoposide, celecoxib, vitamin D, fenofibrate and retinoic acid. From 2004 to 2010, 74 children were enrolled. Results: The 2-year overall survival (OS) was 43.1% (median 15.4, range 1.3–69.9 months). Of the 74 patients, 50 patients (68%) died and 24 are alive: 6 (8%) with progressive disease, 7 (9%) with stable disease/partial response and 11 (15%) in complete response. Median time to response was 6 months. Of 62 patients with initially measurable disease, 25 (40%) had radiological response or stable disease. Fourteen of 25 showing clinical benefit responded within the first 6 months. The treatment was well tolerated on an outpatient basis. Regarding non-hematological toxicity of grade ≥2, hepatotoxicity of grade 3 occurred in 8 children and grade 3 cheilitis in 16 children. Conclusion: COMBAT is a feasible and effective treatment option for patients with relapsing/refractory malignancies. The treatment is well tolerated with a low acute toxicity profile.

HER2 signaling downregulation by trastuzumab and suppression of the PI3K/Akt pathway: an unexpected effect on TRAIL-induced apoptosis.

We investigated whether HER2 downregulation by trastuzumab modulates the responsiveness of breast cancer cells to TNF‐related apoptosis‐inducing ligand (TRAIL). Interestingly, in contrast to increased response to TRAIL in SKBr3 cells, trastuzumab decreased the susceptibility of BT474 cells to TRAIL. This decrease was also observed after exogenous inhibition of PI3‐K/Akt kinase, but not MAPK/ERK kinase (MEK)/mitogen‐activated protein kinase (MAPK). In BT474 cells, but not SKBr3 cells, inhibition of the HER2/phosphatidylinositol 3′ kinase (PI3K)/Akt pathway resulted in downregulation of the pro‐apoptotic receptors TRAIL‐receptor 1 (TRAIL‐R1) and TRAIL‐R2. TRAIL‐induced caspase‐8 activation, Bid processing, drop of ΔΨ m, and poly ADP‐ribose polymerase (PARP) cleavage but not in caspase‐9 activation, and these events were inhibited in HER2/PI3K/Akt‐suppressed BT474 cells, which on the other hand exhibited downregulation of Bcl‐xL and increased response to mitomycin C. We show that HER2/PI3K/Akt pathway may play a specific pro‐apoptotic role in certain cell type by inducing TRAIL‐R1 and ‐R2 expression and thereby enhancing responsiveness to TRAIL.

Synanthropic Birds Influence the Distribution of Borrelia Species: Analysis of Ixodes ricinus Ticks Feeding on Passerine Birds

ABSTRACT Ixodes ricinus ticks collected from 835 birds and from vegetation in the Czech Republic were analyzed. Host-seeking ticks (n = 427) were infected predominantly by Borrelia afzelii (25%). Ticks (n = 1,012) from songbirds (Passeriformes) were infected commonly by Borrelia garinii (12.1%) and Borrelia valaisiana (13.4%). Juveniles of synanthropic birds, Eurasian blackbirds (Turdus merula) and song thrushes (Turdus philomelos), were major reservoir hosts of B. garinii.

The effect of cellular environment and p53 status on the mode of action of the platinum derivative LA-12

SummaryIn this study, we characterized the effects of LA-12 on tumor cell lines possessing wild type p53 and on p53-deficient/mutant cell lines and the results were compared to those obtained using cisplatin. We have determined changes of p53 levels, of its transcriptional activity, of its posttranscriptional modifications and the effect of the treatment on the cell cycle, on the induction of apoptosis and on gene expression. LA-12 induces weak accumulation of both transcriptionally active p53 tumor suppressor and of p21WAF1/CIP1 protein. LA-12 and cisplatin also significantly differ in their effects on apoptosis and cell cycle and on gene expression spectra in studied cell lines. LA-12 induces higher apoptosis levels in comparison with those induced by cisplatin, especially in p53-deficient H1299 cells and in MCF-7DD cells with transcriptionally inactive p53. We suggest that LA-12-mediated apoptosis is not fully dependent on p53. This confirms the therapeutic potential of LA-12 as a more potent cytostatic agent for both tumor cells expressing wild type p53 and for p53-deficient or mutant cells.

Second complete remission of relapsed medulloblastoma induced by metronomic chemotherapy.

Prognosis for children with relapsed medulloblastoma remains poor. Metronomic chemotherapy may offer some benefit to patients treated initially with intensive regimens. However, dosing and duration of such palliative treatment have not been systematically studied. Here we describe a child with medulloblastoma relapsing after initial high‐dose chemotherapy and standard radiotherapy. The patient was then treated with metronomic chemotherapy and achieved second complete remission after 21 months of treatment. Three months off therapy he relapsed again and died from progressive disease. This case illustrates the potential benefit of metronomic chemotherapy but also shows the uncertainty of when to stop metronomic chemotherapy while balancing toxicity. Pediatr Blood Cancer 2010;54:616–617.

Avipoxvirus in blackcaps (Sylvia atricapilla)

From July to September 2005, 1075 wild birds of 37 species were mist-netted at a location in the north-eastern part of the Czech Republic. The birds were examined for the presence of avipoxvirus lesions. This was demonstrated by electron microscopy in skin lesions in nine of 244 blackcaps (Sylvia atricapilla) examined (4% prevalence). Blackcaps skin bioptates were processed using the ultrathin section method. In skin bioptates, avipoxviruses were demonstrated in intracytoplasmic inclusions where, in addition to mature viruses, lipids and filamentous structures concentrated into large circular formations were found. The so-called additional inclusions were also found. These did not contain any virus components, and they served as the precursor of A-type intracytoplasmic inclusions. Blackcap avipoxvirus was isolated by passage on the chorioallantoic membrane of 9-day-old chicken embryos. The virus was successfully adapted after 11 passages (each passage lasted 5 to 7 days), at which time a marked changes in the form of tiny nodules 2 to 3 mm in diameter were observed on the chorioallantoic membrane. Further identification of field isolates and of the cultured virus was carried out using polymerase chain reaction and sequencing. Sequences were compared with consensus sequences of both canarypoxviruses and fowlpoxviruses. Our sequence was found to be 98.8% identical to the canarypox consensus sequence, but only 63% identical to the fowlpox consensus sequence. Our avipoxvirus sequence was proven to be significantly more closely related to canarypoxviruses than to fowlpoxviruses also by phylogenetic analysis.

An Outbreak of the Polyomavirus Infection in Budgerigars and Cockatiels in Slovakia, Including a Genome Analysis of an Avian Polyomavirus Isolate

Abstract In winter 2003–04, large numbers of budgerigars (Mellopsitacus undulatus) and cockatiels (Nymphicus hollandicus) fell ill and died in a large parrot-breeding aviary in Slovakia. In budgerigars, the disease outbreak occurred at the age of 2–3 weeks; cockatiels died within their first 7 days of life. In budgerigars, symptoms of the disease included delayed growth, tremor, darkish discoloration of skin, quill bleeding, and feathering defects. cockatiels often died without any symptoms and with a full crop; feathering defects occurred sporadically. Electron microscopy with negative staining of aqueous lysates of the affected skin and of bleeding quills showed isolated or clustered polyomavirus particles 45–50 nm in size. Long filamentous forms of the virus were also found in virion clusters of skin lysates from the budgerigars. In ultrathin sections through the pathologically altered skin tissue of budgerigars, virus particles were present in both nuclei and cytoplasm of epidermal cells, often in crystalline form. In infected cells, enlarged nuclei showed an extensive chromatin margination. On the DNA level, presence of a polyomavirus infection was conclusively proved by the polymerase chain reaction using avian polyomavirus (APV)-specific primers. A sequence analysis of the gene encoding viral protein (VP)1 and of the combined region for VP2 and VP3 proteins revealed a previously undescribed synonymous mutation in this isolate. This report extended the knowledge of the area of APV occurrence and of the spectrum of hosts in the context of genomic and morphologic variability of APV isolates.

Surface-enhanced laser desorption ionization/time-of-flight (SELDI-TOF) mass spectrometry (MS) as a phenotypic method for rapid identification of antibiotic resistance.

Based on experiments with 10 defined strains of Escherichia coli, we present a new method for bacterial phenotyping using SELDI-TOF mass spectrometry. Changes in bacterial protein profiles in the context of the time of cultivation and the antibiotic environment were minimal. Proteom subprofiling may further distinguish between strains with specific susceptibility to antimicrobials. Mass spec-based methods may become common in the future of bacterial pathogen identification in clinical microbiology diagnostics.

The cell type-specific effect of TAp73 isoforms on the cell cycle and apoptosis

Abstractp73, a member of the p53 family, exhibits activities similar to those of p53, including the ability to induce growth arrest and apoptosis. p73 influences chemotherapeutic responses in human cancer patients, in association with p53. Alternative splicing of the TP73 gene produces many p73 C- and N-terminal isoforms, which vary in their transcriptional activity towards p53-responsive promoters. In this paper, we show that the C-terminal spliced isoforms of the p73 protein differ in their DNA-binding capacity, but this is not an accurate predictor of transcriptional activity. In different p53-null cell lines, p73β induces either mitochondrial-associated or death receptor-mediated apoptosis, and these differences are reflected in different gene expression profiles. In addition, p73 induces cell cycle arrest and p21WAF1 expression in H1299 cells, but not in Saos-2. This data shows that TAp73 isoforms act differently depending on the tumour cell background, and have important implications for p73-mediated therapeutic responses in individual human cancer patients.