Adenovirus infection among pediatric patients with cancer and in pediatric recipients of hematopoietic stem cell: A multicenter nationwide study.

The aim was to evaluate the incidence, clinical course, and outcome of adenoviral infection (AdVI) in pediatric patients diagnosed and treated due to cancer and in pediatric recipients of hematopoietic stem cell. Over a 72-month period, all-in 5599 children with cancer: 2441 patients with hematological malignancy (HM) and 3158 with solid tumors (ST), and 971 patients after transplantation: 741 after allogeneic (allo-HSCT) and 230 after autologous (auto-HSCT) were enrolled into the study. Among cancer patients, 67 episodes of AdVI appeared in 63 (1.1%) children, including 45 (1.8%) with HM and 18 (0.6%; P < .001) with ST. Within transplanted patients, AdVIs were responsible for 88 episodes in 81 (8.3%) children (P < .001), including 78 (10.5%) patients after allo-HSCT and 3 (1.3%) after auto-HSCT. Time to develop AdVI was short, especially after allo-HSCT. The most common clinical manifestation in cancer patients was enteritis diagnosed in 63 (94.0%) cases, while among HSCT recipient asymptomatic adenoviremia was found in 36 (40.9%) cases and the most common clinical manifestation was urinary tract infection. Cancer patients with disseminated disease, as well as HSCT recipients with either asymptomatic viremia or disseminated disease, received antiviral treatment. The most commonly used first-line therapy was cidofovir. None of the cancer patients died due to AdVI, while within HSCT recipients three patients developed disseminated adenoviral disease and died despite antiviral treatment. In cancer patients, AdVIs are rare and associated with very good prognosis even without specific treatment. However, in allo-HSCT recipients, disseminated disease with fatal outcome is more likely to occur.

Dual left anterior descending coronary artery (type III) and the presence of myocardial bridges: a post-mortem examination.

An observational examination of the heart was performed in the Department of Anatomy, during the routine autopsy of an 89-year-old man. The heart was fixed in 10% formalin and an analysis of arterial vasculature was performed (used morphometric abbreviations below [mm]: L - length; D - diameter of origin). Trifurcation of the left main coronary artery (L = 17.4; D = 8.1) was observed during the study, which originated in the left aortic sinus and was followed by three branches: proper left anterior descending artery (pLAD; L = 11.2; D = 7.4), intermediate branch (L = 98.6; D = 3.5) and left circumflex artery (L = 104.2; D = 4.9), respectively. In the pLAD division, there was noted LAD1 (long) which was running in the interventricular septum (L = 32.2) and further in the subepicardial segment (L = 109.3) in the anterior interventricular groove towards the apex (AC) (LAD1; L = 141.4; D = 6.3) and LAD2 (short) running subepicardial in the anterior interventricular groove in the AC direction (LAD2; L = 68.4; D = 3.2). Four diagonal branches (DB) and 9 septal perforators (SP) were observed in the course of LAD1; regarding the LAD2 there were 6 SP only. It is worth noting that the first SP supplying the interventricular septum came from LAD2. Another interesting aspect of the observation was the occurrence of 4 myocardial bridges on the LAD1, LAD2, DB1 arteries and on the second obtuse marginal branch (OM2), respectively. This case describes a rare anatomical anomaly of the LAD course and reminds clinicians of the need for careful planning of cardiac surgeries and percutaneous interventions on the coronary arteries.

Four left renal arteries - a rare variant of kidney arterial supply.

We describe the arterial supply of a human kidney harvested post-mortem from a 75-year-old female volunteer body donor. The kidney was analysed with con- trast-enhanced computed tomography (CT), and corrosion casting was used to reveal the kidney's angio-architecture. In the left kidney, we observed four renal arteries, each originating directly from the abdominal aorta. Three renal arteries, including the main renal artery, coursed through the renal hilum, and the fourth renal artery reached the lower kidney pole. The supply areas of each of the four renal arteries were analysed with a three-dimensional reconstruction of CT images and with corrosion casting. There were no clear boundaries between the areas supplied by the four renal arteries because their branches overlapped in most kidney segments.

Non-anaplastic peripheral T cell lymphoma in children and adolescents-an international review of 143 cases.

Peripheral T cell lymphomas (PTCL) are rare in children and adolescents, and data about outcome and treatment results are scarce. The present study is a joint, international, retrospective analysis of 143 reported cases of non-anaplastic PTCL in patients <19 years of age, with a focus on treatment and outcome features. One hundred forty-three patients, between 0.3 and 18.7 years old, diagnosed between 2000 and 2015 were included in the study. PTCL not otherwise specified was the largest subgroup, followed by extranodal NK/T cell lymphoma, hepatosplenic T cell lymphoma (HS TCL), and subcutaneous panniculitis-like T cell lymphoma (SP TCL). Probability of overall survival (pOS) at 5 years for the whole group was 0.56 ± 0.05, and probability of event-free survival was (pEFS) 0.45 ± 0.05. Patients with SP TCL had a good outcome with 5-year pOS of 0.78 ± 0.1 while patients with HS TCL were reported with 5-year pOS of only 0.13 ± 0.12. Twenty-five percent of the patients were reported to have a pre-existing condition, and this group had a dismal outcome with 5-year pOS of 0.29 ± 0.09. The distribution of non-anaplastic PTCL subtypes in pediatric and adolescent patients differs from what is reported in adult patients. Overall outcome depends on the subtype with some doing better than others. Pre-existing conditions are frequent and associated with poor outcomes. There is a clear need for subtype-based treatment recommendations for children and adolescents with PTCL.

Extracellular recording of glycine receptor chloride channel activity as a prototype for biohybrid sensors.

There is a continuously growing scientific and technological interest to develop and improve the application of artificial sensors. Biological components which are capable to transduce neutral signals into specific, robust and reproducible indicators frame an attractive alternative to construct biohybrid sensors. Since naturally "occurring" biosensors are only sparsely compatible with artificial devices, genetic engineering of eukaryotic cells provides an attractive approach, where cells can be tailored such to detect target compounds with exquisite specificity and sensitivity. We have developed the prototype for a single-cell-based anion-selective biohybrid sensor. HEK293 cells were stably transfected with a gene encoding glycine receptor alpha(1) subunits. These cells were employed as transducers for glycine-evoked chloride currents in a concentration-dependent way. Cultured on substrate-integrated micro-devices, anionic membrane currents of cells were monitored extracellularly with field-effect transistors (FETs) and gold microelectrode arrays (MEAs). The results supported predictions of state-of-the-art models for cell-sensor coupling mechanisms and confirmed that extracellularly recorded anion currents cause similar signals, regardless whether obtained with field-effect transistors or microelectrodes. The whole-cell sensor successfully tracked glycine concentrations differing by three orders of magnitude. To our knowledge this contribution for the first time marks the functional characterization of an anion-selective biohybrid sensor.

Time-dependent observation of individual cellular binding events to field-effect transistors.

Electrolyte-gate field-effect transistors (EG-FETs) gained continuously more importance in the field of bioelectronics. The reasons for this are the intrinsic properties of these FETs. Binding of analysts or changes in the electrolyte composition are leading to variations of the drain-source current. Furthermore, due to the signal amplification upon voltage-to-current conversion even small extracellular signals can be detected. Here we report about impedance spectroscopy with an FET array to characterize passive components of a cell attached to the transistor gate. We developed a 16-channel readout system, which provides a simultaneous, lock-in based readout. A test signal of known amplitude and phase was applied via the reference electrode. We monitored the electronic transfer function of the FETs with the attached cell. The resulting frequency spectrum was used to investigate the surface adhesion of individual HEK293 cells. We applied different chemical treatments with either the serinpeptidase trypsin or the ionophor amphotericin B (AmpB). Binding studies can be realized by a time-dependent readout of the lock-in amplifier at a constant frequency. We observed cell detachment upon trypsin activity as well as membrane decomposition induced by AmpB. The results were interpreted in terms of an equivalent electrical circuit model of the complete system. The presented method could in future be applied to monitor more relevant biomedical manipulations of individual cells. Due to the utilization of the silicon technology, our method could be easily up-scaled to many output channels for high throughput pharmacological screening.

Solution of the Poisson-Nernst-Planck equations in the cell-substrate interface.

Electrogenic cells are able to generate electrical signals which can be measured by various invasive electrophysiological methods such as patch-clamp or sharp microelectrode recordings. Growing cells on the surfaces of e.g. metal microelectrodes or field-effect transistors allows the recording of an extracellular component of these signals. For an understanding of such extracellular signals it is mandatory to get detailed topographical as well as electrical information about the cell-sensor interface. In a first approximation, this interface can be described by a flat disk between cell membrane and sensor surface. For a correct description of the signals, the electrodiffusion of ions in this interface is modeled by using the stationary Poisson-Nernst-Planck equations. We solve the equations analytically, and derive expressions for the potential, the ionic charge densities, and the seal resistance. The results provide a method for determining the distance h between sensor surface and cell membrane. For human embryonic kidney cells, we receive h approximately 70 nm. Comparison with literature shows good agreement.

Drug profiling using planar microelectrode arrays.

Microelectrode arrays (MEAs) with evenly distributed multiple sensor spots have been designed for specific applications. Using the MEAs, we determined the relative profiles of potassium channel openers (KCOs) on cultured embryonic Sprague-Dawley rat cardiac myocytes. KCO, pinacidil (PIN), cromakalim (CROM), SDZ PCO400 (SDZ), or its vehicle, was added to the myocytes cumulatively. The action potential signal shapes in the presence of PIN and SDZ show that the changes in voltage over time and the magnitudes of the associated voltage change were reduced concentration-dependently. CROM affected sodium influx more than PIN and SDZ. The comparisons of changes in the rate of beating and propagation speed in the presence of KCOs were made using their corresponding pD(2) values (the negative log of EC(50)). All KCOs caused concentration-dependent reductions in the rate of beating and propagation speed, with SDZ being the most potent. In addition to the signal shapes, rate of beating, and propagation speed, the origin of excitation and the excitation pattern inside the culture can be also extracted. The results show that the present system can differentiate the effects of different KCOs on myocytes. It might be possible to utilise the MEA as a means to classify drug action based upon a combined interpretation of the three different datasets gained from the extracellular recordings. The combination of these observations might be used as 'drug signatures' when profiling drugs in the future.

Influence of the first amplifier stage in MEA systems on extracellular signal shapes.

Recording of extracellular signals with planar metal microelectrodes (ME) has already been presented more than 30 years ago. To date, microelectrode array (MEA) systems are able to measure extracellular signals at about 64 sites, simultaneously. This enables monitoring of electrical activity of many cells in a large area. The extracellular recording technique has become a widely used method for neurological, toxicological or pharmacological studies. It already proved its potential to supplement the classical methods in electrophysiology. The interpretation of the recorded signal shapes in order to extract electrophysiological meaningful data--however--is still under discussion. In this article, we analyse the preamplifier circuit for extracellular recording of cardiac myocyte signals. We use a circuit model for the cell-electrode contact including the first amplification stage. In test experiments, we observe different signal shapes, when different shunt resistors are introduced at the input of the preamplifier. According to the frequency spectra of the recordings, we evaluate the transfer function between the source signal and the readout signal. As a result of our studies, an optimum readout electronics for originally, preserved extracellular signal shapes is proposed. Our amplifier design will be most valuable, if the use of small microelectrodes with high input impedances for in vitro as well as for in vivo experiments is desired.

UV/VIS spectrophotometric methods for determination of caffeine and phenylephrine hydrochloride in complex pharmaceutical preparations. Validation of the methods.

The contents of active substances were determined in a preparation TP-4 (tablets) containing paracetamol, ascorbic acid, caffeine and phenylephrine hydrochloride. For determination of caffeine and phenylephrine hydrochloride, UV/VIS spectrophotometric method was used. The VIS spectrophotometric method based on the reaction of phenylephrine with ninhydrine in sulphuric acid (1.127 kg/l). Validation of methods performed for model mixtures proved those methods were accurate, precise, repeatable and linear in the range from 50% to 150% of the amount declared in the preparation. The content of caffeine and phenylephrine hydrochloride in TP-4, Thompyrin, Panadol Extra, Ring N satisfies the FP V demands.

Redox methods validation of paracetamol and ascorbic acid in pharmaceutical preparations.

The contents of active substances were determined in a preparation TP-4 (tablets) containing paracetamol, ascorbic acid, caffeine and phenylephrine hydrochloride. For the determination of paracetamol and ascorbic acid a non-specific (cerometric and titration of 2,6-dichloroindophenol) method based on a redox reaction was used. Validation of the methods, performed on model mixtures, proved those methods to be accurate, precise, reproducible and linear within the range from 50% to 150% of the amount declared in the preparation. The content of paracetamol and ascorbic acid in TP-4, Thompayrin, Panadol Extra, Ring N, Polopiryna C, Efferalgan Vitamin C and Vitaminum C 0.2 satisfies the FP V demands (+/- 10% of the declared amount).

[Adverse reactions associated with the use of L-asparaginase during therapy of patients with nB non-Hodgkin's lymphoma. Report of the Polish Paediatric LeuKaemia/Lymphoma Study Group]. / Objawy niepozadane leczenia preparatami L-asparaginazy u chorych z nieziarniczym chloniakiem zlosliwym typu NB w oparciu o material Polskiej Pediatrycznej Grupy ds. Leczenia Bialaczek i Chloniaków Zlosliwych u dzieci.

The aim of the study was to determine the side effects of asparaginase administration during treatment protocol for childhood non-Hodgkin's lymphoma (NHL). Drug adverse reactions occurred in 20/66 of patients (30,3%) treated in 9 centres in Poland between 1993 and 1998. The most common side effects were coagulation disturbances in 12/66 of the children (18,2%), which occurred due to reduced production of important coagulation factors. Six patients (9,1%) developed impairment of liver function (9,1%). Drug toxicity caused the modifications of treatment protocol in 12/66 (18,2%) of patients, mainly in the induction phase; 3 children died due to relapse of disease.

[Abdominal presentation of B-cell non-Hodgkin's lymphoma (B-NHL) - surgical treatment and its results. Report of the Polish Paediatric Leukaemia/Lymphoma Study Group]. / Manifestacja brzuszna chloniaka B-komórkowego u dzieci - interwencja chirurgiczna i wyniki leczenia. Raport PPGLBC.

The aim of this study was to analyse the effect of LMB-89 protocol and surgical procedure at initial laparotomy on the outcome in children with abdominal B-cell NHL. The initial surgery intervention was: complete resection (20% pts), subtotal resection (20%), partial resection (4%), biopsy (36%). Postoperative complications occurred in 5 children. Complete recovery (CR) was achieved in 92% pts. There were 4% non responder patients. Two patients died before CR evaluation (tumour lysis syndrome; bleeding and multi organ failure after initial surgery). One patient died in CCR from sepsis probably influenced by the previous local operation. 10.8% patients relapsed. The estimate EFS for all patients with AB-NHL is 81%, 85% for stage III and 73% for stage IV. Major surgery in advanced stages is not recommended since it delays chemotherapy and fails to improve overall survival.

[The analysis of failures in the treatment of children with non-B non-Hodgkin's lymphoma. Polish pediatric leukemia/lymphoma study group report]. / Analiza niepowodzen w leczeniu dzieci z nieziarniczym chloniakiem zlosliwym typu NB. Raport Polskiej Pediatrycznej Grupy ds. Leczenia Bialaczek i Chloniaków zlosliwych u dzieci.

The aim of the study was to evaluate the results of treatment of 46 children with non B non-Hodgkin's Lymphoma registered in 7 centers of Polish Paediatric Leukaemia/Lymphoma Study Group from 1993 to 1998. The patients were treated according to BFM-90 protocol based on German regimen. The overall probability of event-free survival for the all children after 4 years of follow-up was 71%, for patients in stage III--65%, stage IV--70%. The achieved results were not as positive as in the BFM Study Group, what was related to: the great number of children with advanced stage of disease (54.3%), the late final diagnosis, the great number of recurrences (22.5%) and deaths caused by the toxicity of medication (6.5%) (infections, drug toxicity).

Openness in adoption and the level of child participation.

There is great controversy regarding the impact of openness in adoption, especially the impact of such an arrangement on adopted children. Three indicators of the level of child participation in the openness arrangement were examined: (a) level of openness reported by adoptive parents, (b) level of information adopted children reported having about their birthparents, and (c) whether adoptive parents have withheld any pertinent information gained through communication with the birthmother from the adopted child. 171 children (90 males, 81 females; mean age = 7.99) wee studied to assess how that participation influenced their conceptual understanding of what adoption means, general self-worth, satisfaction with level of openness, and curiosity about birthparents. Overall it does not appear that providing information about a child's birthparents will confuse the child about the meaning of adoption or lower the child's self-esteem, but neither will it move them to levels of understanding that are beyond their cognitive capabilities to reach.

Opsonic activity of sera in children with hematological malignancies treated with Sandoglobulin.

Twenty seven children with hematological malignancies were treated with Sandoglobulin for life threatening infections due to severe granulocytopenia. We have studied the opsonic activity of sera in patients before and 7,14 and 21 days after the infusion of Sandoglobulin. Before the therapy a decrease of serum opsonic activity at the stage of ingestion and intracellular killing of bacteria has been shown. It was due to a deficiency of opsonizing factors. After treatment with Sandoglobulin the significant improvement of the opsonic activity of tested sera was found, but only at the stage of the ingestion of bacteria. The optimal interrelationship between opsonizing capacity of sera at the ingestion and intracellular killing phase was observed in the group of children treated with the relatively low Sandoglobulin dose (0.3-0.6 g/kg). In patients with the longest infection duration, who received the high Sandoglobulin doses (> 0.6 g/kg), the largest percentage of sera containing immune complexes was detected. These data demonstrate that high doses of globulins should be administered with certain care.

Determining consanguinity by oligonucleotide fingerprinting with (GTG)5/(CAC)5.

Simple tandemly organized (GTG)n/(CAC)n sequences are spread throughout the human chromosomes. The most informative DNA fingerprints for the testing of pedigrees and/or paternity were obtained with the simple triplet repeat probe (GTG)5 or its complement (CAC)5. These hypervariable simple-repeat fragments are stably inherited in a Mendelian fashion. Using these highly discriminating probes, all human individuals could, theoretically, be differentiated, except for genetically identical monozygotic twins. Examples from actual case work are reported and pertinent advantages of this methodology are discussed.

[Oligonucleotide fingerprinting using (GTG)5 and (GACA)4 probes for the differentiation of body fragments]. / Oligonucleotid-Fingerprinting mit (GTG)5 und (GACA)4 für die Zuordnung von Leichenteilen.

For the detection of postmortem stability of DNA and for the identification of parts of dead bodies of unknown origin the oligonucleotide probes (GTG)5 and (GACA)4 can be used. (GTG)5 is a highly discriminating probe which allows to differentiate in the 4 to 25 kilobase range of DNA fragments. DNA fingerprints obtained by (GACA)4 show useful results especially in the short fragment range. The (GACA)4 probe can therefore be used to analyze partially degraded DNA.