Clinical diversity and treatment approaches to blastic plasmacytoid dendritic cell neoplasm: a retrospective multicentre study.

BACKGROUND: Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare, aggressive type of haematologic precursor malignancy primarily often manifesting in the skin. We sought to provide a thorough clinical characterization and report our experience on therapeutic approaches to BPDCN. METHODS: In the present multicentric retrospective study, we collected all BPDCN cases occurring between 05/1999 and 03/2018 in 10 secondary care centres of the German-Swiss-Austrian cutaneous lymphoma working group. RESULTS: A total of 37 BPDCN cases were identified and included. Almost 90% of the patients had systemic manifestations (bone marrow, lymph nodes, peripheral blood) in addition to skin involvement. The latter presented with various types of cutaneous lesions: nodular (in more than 2/3) and bruise-like (in 1/3) skin lesions, but also maculopapular exanthema (in circa 1/6). Therapeutically, 22 patients received diverse combinations of chemotherapeutic regimens and/or radiotherapy. Despite initial responses, all of them ultimately relapsed and died from progressive disease. Eleven patients underwent haematopoietic stem cell transplantation (HSCT; autologous HSCT n = 3, allo-HSCT n = 8). The mortality rate among HSCT patients was only 33.33% with a median survival time of 60.5 months. CONCLUSION: Our study demonstrates the clinical diversity of cutaneous BPDCN manifestations and the positive development observed after the introduction of HSCT.

Preclinical Assessment of a Fully Automated Multiplex PCR Panel for Detection of Central Nervous System Pathogens.

We evaluated a multiplexed PCR panel for the detection of 16 bacterial, viral, and fungal pathogens in cerebrospinal fluid. Panel results were compared to routine testing, and discrepancies were resolved by additional nucleic acid amplification tests or sequencing. Overall, the positive and negative agreements across methods were 92.9% and 91.9%, respectively.

Biotin- and Glycoprotein-Coated Microspheres as Surrogates for Studying Filtration Removal of Cryptosporidium parvum in a Granular Limestone Aquifer Medium.

Members of the genus Cryptosporidium are waterborne protozoa of great health concern. Many studies have attempted to find appropriate surrogates for assessing Cryptosporidium filtration removal in porous media. In this study, we evaluated the filtration of Cryptosporidium parvum in granular limestone medium by the use of biotin- and glycoprotein-coated carboxylated polystyrene microspheres (CPMs) as surrogates. Column experiments were carried out with core material taken from a managed aquifer recharge site in Adelaide, Australia. For the experiments with injection of a single type of particle, we observed the total removal of the oocysts and glycoprotein-coated CPMs, a 4.6- to 6.3-log10 reduction of biotin-coated CPMs, and a 2.6-log10 reduction of unmodified CPMs. When two different types of particles were simultaneously injected, glycoprotein-coated CPMs showed a 5.3-log10 reduction, while the uncoated CPMs displayed a 3.7-log10 reduction, probably due to particle-particle interactions. Our results confirm that glycoprotein-coated CPMs are the most accurate surrogates for C. parvum; biotin-coated CPMs are slightly more conservative, while unmodified CPMs are markedly overly conservative for predicting C. parvum removal in granular limestone medium. The total removal of C. parvum observed in our study suggests that granular limestone medium is very effective for the filtration removal of C. parvum and could potentially be used for the pretreatment of drinking water and aquifer storage recovery of recycled water.

A sensitive and robust method for automated on-line monitoring of enzymatic activities in water and water resources.

The realisation of a novel concept for automated on-line monitoring of enzymatic activities in water was successfully demonstrated by long-term field testing at two remote Austrian ground water resources. The ß-D-glucuronidase (GLUC) activity was selected as a representative enzymatic model parameter for the on-line determination. But the device can be adapted for any enzymatic reaction with diagnostic relevance for microbial water quality monitoring, as demonstrated for the ß-D-galactosidase activity. Automated filtration of volumes up to 5 litres supports sensitive quantification of enzymatic activities. Internet-based data transfer, using internal control parameters for verification and a dynamic determination of the limit of quantification, enabled robust enzymatic on-line monitoring during a 2-year period. A proportion of 5,313 out of 5,506 GLUC activity measurements (96.5%) could be positively verified. Hydrological (discharge, gauge, turbidity, temperature, pH, electric conductivity, spectral absorbance coefficient at 254 nm) as well as microbiological parameters (Escherichia coli, coliforms) were concurrently determined to characterise the investigated ground water resources. The enzymatic on-line measurements closely reflected the different hydrological conditions and contamination patterns of the test sites. Contrary to expectations, GLUC did not qualify as a proxy-parameter for the occurrence of cultivation-based E. coli contamination and warrants further detailed investigations on its indication capacity as a rapid means for microbial faecal pollution detection in such aquatic habitats. Microbial on-line monitoring is likely to become more important in the future, complementing existing surveillance strategies for water safety management. Further perspectives on the application of such analytical on-line technologies, such as their connection with event-triggered sampling and standardised diagnostics, are discussed.

A probabilistic-deterministic approach for assessing climate change effects on infection risks downstream of sewage emissions from CSOs.

The discharge of pathogens into urban recreational water bodies during combined sewer overflows (CSOs) pose a potential threat for public health which may increase in the future due to climate change. Improved methods are needed for predicting the impact of these effects on the microbiological urban river water quality and infection risks during recreational use. The aim of this study was to develop a novel probabilistic-deterministic modelling approach for this purpose building on physically plausible generated future rainfall time series. The approach consists of disaggregation and validation of daily precipitation time series from 21 regional climate models for a reference period (1971-2000, C20), a near-term future period (2021-2050, NTF) and a long-term future period (2071-2100, LTF) into sub-daily scale, and predicting the concentrations of enterococci and Giardia and Cryptosporidium, and infection risks during recreational use in the river downstream of the sewage emissions from CSOs. The approach was tested for an urban river catchment in Austria which is used for recreational activities (i.e. swimming, playing, wading, hand-to-mouth contact). According to a worst-case scenario (i.e. children bathing in the river), the 95th percentile infection risks for Giardia and Cryptosporidium range from 0.08 % in winter to 8 % per person and exposure event in summer for C20. The infection risk increase in the future is up to 0.8 log10 for individual scenarios. The results imply that measures to prevent CSOs may be needed to ensure sustainable water safety. The approach is promising for predicting the effect of climate change on urban water safety requirements and for supporting the selection of sustainable mitigation measures. Future studies should focus on reducing the uncertainty of the predictions at local scale.

Setback distances between small biological wastewater treatment systems and drinking water wells against virus contamination in alluvial aquifers.

Contamination of groundwater by pathogenic viruses from small biological wastewater treatment system discharges in remote areas is a major concern. To protect drinking water wells against virus contamination, safe setback distances are required between wastewater disposal fields and water supply wells. In this study, setback distances are calculated for alluvial sand and gravel aquifers for different vadose zone and aquifer thicknesses and horizontal groundwater gradients. This study applies to individual households and small settlements (1-20 persons) in decentralized locations without access to receiving surface waters but with the legal obligation of biological wastewater treatment. The calculations are based on Monte Carlo simulations using an analytical model that couples vertical unsaturated and horizontal saturated flow with virus transport. Hydraulic conductivities and water retention curves were selected from reported distribution functions depending on the type of subsurface media. The enteric virus concentration in effluent discharge was calculated based on reported ranges of enteric virus concentration in faeces, virus infectivity, suspension factor, and virus reduction by mechanical-biological wastewater treatment. To meet the risk target of <10-4infections/person/year, a 12 log10 reduction was required, using a linear dose-response relationship for the total amount of enteric viruses, at very low exposure concentrations. The results of this study suggest that the horizontal setback distances vary widely ranging 39 to 144m in sand aquifers, 66-289m in gravel aquifers and 1-2.5km in coarse gravel aquifers. It also varies for the same aquifers, depending on the thickness of the vadose zones and the groundwater gradient. For vulnerable fast-flow alluvial aquifers like coarse gravels, the calculated setback distances were too large to achieve practically. Therefore, for this category of aquifer, a high level of treatment is recommended before the effluent is discharged to the ground surface.

Occurrence of human-associated Bacteroidetes genetic source tracking markers in raw and treated wastewater of municipal and domestic origin and comparison to standard and alternative indicators of faecal pollution.

This was a detailed investigation of the seasonal occurrence, dynamics, removal and resistance of human-associated genetic Bacteroidetes faecal markers (GeBaM) compared with ISO-based standard faecal indicator bacteria (SFIB), human-specific viral faecal markers and one human-associated Bacteroidetes phage in raw and treated wastewater of municipal and domestic origin. Characteristics of the selected activated sludge wastewater treatment plants (WWTPs) from Austria and Germany were studied in detail (WWTPs, n = 13, connected populations from 3 to 49000 individuals), supported by volume-proportional automated 24-h sampling and chemical water quality analysis. GeBaM were consistently detected in high concentrations in raw (median log10 8.6 marker equivalents (ME) 100 ml(-1)) and biologically treated wastewater samples (median log10 6.2-6.5 ME 100 ml(-1)), irrespective of plant size, type and time of the season (n = 53-65). GeBaM, Escherichia coli, and enterococci concentrations revealed the same range of statistical variability for raw (multiplicative standard deviations s* = 2.3-3.0) and treated wastewater (s* = 3.7-4.5), with increased variability after treatment. Clostridium perfringens spores revealed the lowest variability for raw wastewater (s* = 1.5). In raw wastewater correlations among microbiological parameters were only detectable between GeBaM, C. perfringens and JC polyomaviruses. Statistical associations amongst microbial parameters increased during wastewater treatment. Two plants with advanced treatment were also investigated, revealing a minimum log10 5.0 (10th percentile) reduction of GeBaM in the activated sludge membrane bioreactor, but no reduction of the genetic markers during UV irradiation (254 nm). This study highlights the potential of human-associated GeBaM to complement wastewater impact monitoring based on the determination of SFIB. In addition, human-specific JC polyomaviruses and adenoviruses seem to be a valuable support if highly specific markers are needed.

Apoptotic DNA fragmentation is detected by a semi-quantitative ligation-mediated PCR of blunt DNA ends.

Apoptosis is a form of programmed cell death (PCD) characterized by morphological changes and stereotypical DNA degradation described as a nucleosomal ;ladder'. However, nucleosomal ladders have only been clearly demonstrated in vertebrate tissues when large numbers of cells die in synchrony. Their absence may be explained by asynchronous death under physiological conditions, or by distinct molecular mechanisms. In this study, nucleosomal ladders were revealed by a ligation-mediated polymerase chain reaction (LMPCR), that amplifies DNA fragments with blunt, 5' phosphorylated ends. Numerous tissues from different organisms were examined which demonstrated that nucleosomal ladders (a) accompany physiological cell death in mammalian tissues where previously DNA fragmentation has not been detected; (b) are produced during invertebrate cell death; (c) are invariably generated via the production of blunt, 5' phosphorylated double strand breaks. These results suggest that PCD in multicellular organisms consistently involves apoptotic mechanisms and that the endonuclease activity is evolutionarily conserved.

Quantification of sewer exfiltration using the anti-epileptic drug carbamazepine as marker species for wastewater.

The anti-epileptic drug carbamazepine was used as marker species in wastewater to identify and quantify sewer exfiltration. In several studies carbamazepine turned out to be hardly removed in wastewater treatment and not or just slightly attenuated during bank infiltration. Concentrations in wastewater are generally 1000 times higher than the limit of quantification. In contrast to many other marker species a "young" drug as carbamazepine is discharged to the environment only by wastewater. The results from this study carried out in Linz, Austria indicate an average exfiltration rate, expressed as percentage of the dry weather flow that is lost on the city-wide scale, of 1%. This rate is lower than sewage losses reported in most other studies which attempted to quantify exfiltration on the basis of groundwater pollution. However, it was also possible to identify one area with significant higher sewage losses.

Monitoring of carbamazepine concentrations in wastewater and groundwater to quantify sewer leakage.

Monitoring of carbamazepine concentrations in wastewater and groundwater enables us to identify and quantify sewer exfiltration. The antiepileptic drug carbamazepine is hardly removed in wastewater treatment plants and not or just slightly attenuated during bank infiltration and subsoil flow. Concentrations in wastewater are generally 1000 times higher than the limit of quantification. In contrast to . many other wastewater tracers carbamazepine is discharged to the environment only via domestic wastewater. The results from this study carried out in Linz, Austria indicate an average exfiltration rate of 1%, expressed as percentage of the dry weather flow that is lost to the groundwater on the city-wide scale. This rate is lower than sewage losses reported in most other studies which attempted to quantify exfiltration on the basis of groundwater pollution. However, it was also possible to identify one area with significantly higher sewage losses. This method seems to be very suitable for the verification of leakage models used to assess sewer exfiltration on a regional scale.

Lessons learned from investigations on case study level for modelling of nutrient emissions in the Danube basin.

In the framework of the project daNUbs (Nutrient Management in the Danube Basin and its Impact on the Black Sea) the MONERIS emission model is used for the basin wide calculation of nutrient (nitrogen and phosphorus) emissions in the Danube Basin. The MONERIS model was developed and successfully applied for German river catchments. Based on investigations in selected test regions (case studies) the daNUbs approach is to check the applicability of the MONERIS emission model for the specific conditions of the Danube Basin in more detail than is possible with a basin wide application. Six case studies with areas of 400-3,500 km2 and several subcatchments have been selected in order to represent different conditions along the Danube Basin. In this study region intensive data collection and enhanced monitoring has been performed in order to raise the database significantly above the generally available data. Water balance as well as nutrient balance calculations have been performed with the MONERIS model as well as with other approaches. Results are compared to each other and to data from monitoring. Results up till now showed the applicability and sensitivity of the MONERIS approach in different conditions of the Danube Basin (e.g. emissions via groundwater). They indicated that the nitrogen retention in the catchments is well described with the MONERIS model.

Programmed cell death is a universal feature of embryonic and postnatal neuroproliferative regions throughout the central nervous system.

During central nervous system (CNS) development, programmed cell death (PCD) has been viewed traditionally as a fate reserved for differentiating neurons that are in the process of making synaptic connections. Recent studies in the embryonic cerebral cortex (Blaschke et al. [1996] Development 122:1165-1174), however, have shown that many neuroblasts in the proliferative ventricular zone undergo PCD as well and that this likely represents a novel form distinct from that found in regions of postmitotic neurons. To determine the commonality of this form of PCD throughout the CNS, the prevalence of dying cells identified by in situ end labeling plus (ISEL +; Blaschke et al. [1996]) was determined within populations of proliferating neuroblasts that were identified by rapid bromodeoxyuridine incorporation. Based on this approach, dying cells were observed to be a common feature of all proliferative neuroblast populations examined. In addition, when ISEL+ was combined with in situ hybridization for postmitotic neural gene-1 (png-1; Weiner and Chun [1997] J. Comp. Neurol. 381:130-142), which identifies newly postmitotic neurons, a positive correlation was found between the start of differentiation and the onset of PCD. These data indicate that PCD in neuroblast proliferative zones is a universal feature of nervous system development. Moreover, cell death represents a prominent cell fate that may be linked to mechanisms of differentiation.

Decreased apoptosis in proliferative and postmitotic regions of the Caspase 3-deficient embryonic central nervous system.

Caspase 3 (CPP32/Yama/apopain), a mammalian homolog of the Caenorhabditis elegans pro-cell death gene ced-3, is required for normal programmed cell death (PCD) in the nematode. Its prior deletion by homologous recombination in mice resulted in embryonic/early postnatal lethality associated with dramatic central nervous system (CNS) hyperplasia, yet a reported subtle decrease in cell death (Kuida et al. [1996] Nature 384:368-372). By comparison, the magnitude and distribution of dying cells identified using a DNA end-labeling technique, in situ end-labeling plus (ISEL+) (Blaschke et al. [1996] Development 122:1165-1174; Blaschke et al. [1998] J. Comp. Neurol. 396:39-50), supported an alternative explanation where the loss of caspase 3 function produces a more pervasive block in cell death, particularly among neuroblasts. To determine the relationship between loss of caspase 3 and dying cells identified by ISEL+, we analyzed caspase 3 +/+, +/-, and -/- embryos for normal caspase 3 expression and ISEL+ labeling. Both caspase 3 mRNA and active caspase 3 protein are present throughout the +/+ embryonic CNS, and both are absent from -/- embryonic cortices. Quantitation of dying cells identified by ISEL+ reveals a 30% reduction of labeled cells throughout the caspase 3 -/- embryonic cortices relative to +/+ littermates. Associated with this decrease is marked expansion of the total population of actively proliferating neuroblasts identified by 5-bromo-2;-deoxyuridine incorporation that nevertheless appears to maintain histological features of normal neurogenesis rather than dysregulated, neoplastic growth. These data indicate that caspase 3 deficiency results in a pervasive, albeit partial, decrease in embryonic neuroblast apoptosis that can account for the observed phenotypic hyperplasia in -/- embryos, and support the additional operation of caspase 3-independent PCD mechanisms during embryonic CNS development.

Enumerating Microorganism Surrogates for Groundwater Transport Studies Using Solid-Phase Cytometry.

Investigations on the pollution of groundwater with pathogenic microorganisms, e.g. tracer studies for groundwater transport, are constrained by their potential health risk. Thus, microspheres are often used in groundwater transport studies as non-hazardous surrogates for pathogenic microorganisms. Even though pathogenic microorganisms occur at low concentrations in groundwater, current detection methods of microspheres (spectrofluorimetry, flow cytometry and epifluorescence microscopy) have rather high detection limits and are unable to detect rare events. Solid-phase cytometry (SPC) offers the unique capability of reliably quantifying extremely low concentrations of fluorescently labelled microorganisms or microspheres in natural waters, including groundwater. Until now, microspheres have been used in combination with SPC only for instrument calibration purposes and not for environmental applications. In this study, we explored the limits of the SPC methodology for its applicability to groundwater transport studies. The SPC approach proved to be a highly sensitive and reliable enumeration system for microorganism surrogates down to a minimum size of 0.5 µm, in up to 500 ml of groundwater, and 0.75 µm, in up to 1 ml of turbid surface water. Hence, SPC is proposed to be a useful method for enumerating microspheres for groundwater transport studies in the laboratory, as well as in the field when non-toxic, natural products are used.

Effects of fluctuations in river water level on virus removal by bank filtration and aquifer passage--a scenario analysis.

Riverbank filtration is an effective process for removing pathogenic viruses from river water. Despite indications that changing hydraulic conditions during floods can affect the efficacy of riverbank filtration to remove viruses, the impact on advection and dispersion of viruses in the riverbank is not well understood. We investigated the effects of fluctuations in river water level on virus transport during riverbank filtration, considering 3-D transient groundwater flow and virus transport. Using constant removal rates from published field experiments with bacteriophages, removal of viruses with distance from the riverbank was simulated for coarse gravel, fine gravel and fine sandy gravel. Our simulations showed that, in comparison with steady flow conditions, fluctuations in river water level cause viruses to be transported further at higher concentrations into the riverbank. A 1-5 m increase in river water levels led to a 2- to 4-log (log10 reduction in concentration relative to the initial concentration in the river) increase in virus concentration and to up to 30% shorter travel times. For particular cases during the receding flood, changing groundwater flow conditions caused that pristine groundwater was carried from further inland and that simulated virus concentrations were more diluted in groundwater. Our study suggests that the adverse effect of water level fluctuations on virus transport should be considered in the simulation of safe setback distances for drinking water supplies.

Identification of neural programmed cell death through the detection of DNA fragmentation in situ and by PCR.

A universal feature in the development of multicellular organisms is a physiological form of cell death called programmed cell death (PCD). A subset of PCD is apoptosis, which is defined by characteristic morphological changes and genomic DNA fragmentation producing what are referred to as nucleosomal ladders. To understand how PCD operates in a developing tissue or in a tissue following an experimental procedure, dying cells must be identified in relation to their surviving neighbors. One way to accomplish this is to visualize fragmented DNA in situ, in conjunction with gel electrophoresis of isolated DNA to visualize the nucleosomal ladders associated with apoptosis. Two approaches are presented in this unit: in situ end-labeling plus (ISEL+), a technique to identify dying cells in tissue sections or cell cultures of central nervous system (CNS) tissue (optimized for embryonic samples); and the use of ligation-mediated polymerase chain reaction (LMPCR) to identify nucleosomal ladders from intact tissues. Also included are procedures for preparing thymocyte cell cultures for use as controls in the ISEL+ procedure and for isolating genomic DNA for LMPCR.

[Granular cell tumor of the breast]. / Granularzelltumor der Mamma.

Report about a 46 year old women with granular cell tumor of the breast. Clinical features, mammography and sonography has appearances highly suspicious for breast cancer. Tumorectomy and axillary lymphonodectomy were performed. Estrogen and progesterone receptor both were histochemically and biochemically negative. Estradiol and prolactin levels in serum were normal. Cathepsin D in cytosol was slightly elevated, DNA-flow-cytometry revealed no signs of malignancy. Evident was the high content of protein S100 in cytosol of tumor cells.

Widespread programmed cell death in proliferative and postmitotic regions of the fetal cerebral cortex.

A key event in the development of the mammalian cerebral cortex is the generation of neuronal populations during embryonic life. Previous studies have revealed many details of cortical neuron development including cell birthdates, migration patterns and lineage relationships. Programmed cell death is a potentially important mechanism that could alter the numbers and types of developing cortical cells during these early embryonic phases. While programmed cell death has been documented in other parts of the embryonic central nervous system, its operation has not been previously reported in the embryonic cortex because of the lack of cell death markers and the difficulty in following the entire population of cortical cells. Here, we have investigated the spatial and temporal distribution of dying cells in the embryonic cortex using an in situ endlabelling technique called 'ISEL+' that identifies fragmented nuclear DNA in dying cells with increased sensitivity. The period encompassing murine cerebral cortical neurogenesis was examined, from embryonic days 10 through 18. Dying cells were rare at embryonic day 10, but by embryonic day 14, 70% of cortical cells were found to be dying. This number declined to 50% by embryonic day 18, and few dying cells were observed in the adult cerebral cortex. Surprisingly, while dying cells were observed throughout the cerebral cortical wall, the majority were found within zones of cell proliferation rather than in regions of postmitotic neurons. These observations suggest that multiple mechanisms may regulate programmed cell death in the developing cortex. Moreover, embryonic cell death could be an important factor enabling the selection of appropriate cortical cells before they complete their differentiation in postnatal life.

Hypoxaemia in acute respiratory and non-respiratory illnesses in neonates and children in a developing country.

AIMS: To determine, in sick neonates and children requiring admission to a hospital in the highlands of Papua New Guinea: (1) the incidence and severity of hypoxaemia; (2) the proportion with hypoxaemia who do not fulfil criteria for acute lower respiratory infection (ALRI); and (3) the power of clinical signs to predict hypoxaemia, according to age and disease category. METHODS: Age dependent normal values for transcutaneous oxygen saturation (SpO(2)) were established in 218 well neonates and children in Goroka. A total of 491 sick neonates and children were then studied on presentation to the paediatric department at Goroka Hospital. RESULTS: A total of 257 sick neonates and children (52%) were hypoxaemic. Hypoxaemia was present in 179/245 (73%) with clinical criteria for ALRI; 79/246 (32%) with non-ALRI illnesses (including meningitis, septicaemia, severe malnutrition, low birth weight, birth asphyxia, and congenital syphilis) were also hypoxaemic. For children aged 1 month to 5 years with ALRI, the clinical signs best predicting hypoxaemia were cyanosis, respiratory rate >60, poor feeding, or reduced spontaneous activity; in those without ALRI the best predictors were cyanosis, respiratory rate >60 per minute, and inability to feed, but the positive predictive value was much lower than for children with ALRI. For neonates cyanosis was predictive of hypoxaemia, but tachypnoea or inability to feed were not. CONCLUSIONS: Hypoxaemia is an under recognised complication of non-ALRI illnesses in children and in sick neonates in developing countries. Use of algorithms with high sensitivity for the recognition of hypoxaemia, and protocols for administration of oxygen to neonates, and to children with non-ALRI illnesses, might substantially reduce case fatality.