Monitoring online biomass with a capacitance sensor during scale-up of industrially relevant CHO cell culture fed-batch processes in single-use bioreactors.

In 2004, the FDA published a guideline to implement process analytical technologies (PAT) in biopharmaceutical processes for process monitoring to gain process understanding and for the control of important process parameters. Viable cell concentration (VCC) is one of the most important key performance indicator (KPI) during mammalian cell cultivation processes. Commonly, this is measured offline. In this work, we demonstrated the comparability and scalability of linear regression models derived from online capacitance measurements. The linear regressions were used to predict the VCC and other familiar offline biomass indicators, like the viable cell volume (VCV) and the wet cell weight (WCW), in two different industrially relevant CHO cell culture processes (Process A and Process B). Therefore, different single-use bioreactor scales (50-2000 L) were used to prove feasibility and scalability of the in-line sensor integration. Coefficient of determinations of 0.79 for Process A and 0.99 for Process B for the WCW were achieved. The VCV was described with high coefficients of determination of 0.96 (Process A) and 0.98 (Process B), respectively. In agreement with other work from the literature, the VCC was only described within the exponential growth phase, but resulting in excellent coefficients of determination of 0.99 (Process A) and 0.96 (Process B), respectively. Monitoring these KPIs online using linear regression models appeared to be scale-independent, enabled deeper process understanding (e.g. here demonstrated in monitoring, the feeding profile) and showed the potential of this method for process control.

The role of B lymphocyte stimulator in B cell biology: implications for the treatment of lupus.

B lymphocyte stimulator (BLyS; also known as B cell activating factor (BAFF)) plays a key role in peripheral B cell tolerance. Mounting evidence indicates that B cell tolerance can be either broken or modulated by deliberately manipulating BLyS levels, and belimumab, a BLyS-neutralizing antibody, was recently approved for the treatment of systemic lupus erythematosus (SLE). Thus, intense investigation has focused on understanding how therapeutics targeting BLyS may work, and accumulating evidence suggests multiple points of action. BLyS signaling, in conjunction with B cell receptor (BCR) signaling, determines the size and quality of the mature primary B cell compartment. Moreover, BLyS family members play roles in antigen-experienced B cell selection and differentiation. Together, these findings have implications for the continued development of novel therapeutics that target BLyS.

Effect of propofol on hypoxia re-oxygenation induced neuronal cell damage in vitro*.

Propofol may protect neuronal cells from hypoxia re-oxygenation injury, possibly via an antioxidant actions under hypoxic conditions. This study investigated the molecular effects of propofol on hypoxia-induced cell damage using a neuronal cell line. Cultured human IMR-32 cells were exposed to propofol (30 µm) and biochemical and molecular approaches were used to assess cellular effects. Propofol significantly reduced hypoxia-mediated increases in lactate dehydrogenase, a marker of cell damage (mean (SD) for normoxia: 0.39 (0.07) a.u.; hypoxia: 0.78 (0.21) a.u.; hypoxia+propofol: 0.44 (0.17) a.u.; normoxia vs hypoxia, p<0.05; hypoxia vs hypoxia+propofol, p<0.05), reactive oxygen species and hydrogen peroxide. Propofol also diminished the morphological signs of cell damage. Increased amounts of catalase, which degrades hydrogen peroxide, were detected under hypoxic conditions. Propofol decreased the amount of catalase produced, but increased its enzymatic activity. Propofol protects neuronal cells from hypoxia re-oxygenation injury, possibly via a combined direct antioxidant effect along with induced cellular antioxidant mechanisms.

Ventilatory strategies in septic patients. Results from a nationwide observational trial.

BACKGROUND: Mortality in intensive care unit (ICU) patients is affected by multiple variables. The possible impact of the mode of ventilation has not yet been clarified; therefore, a secondary analysis of the "epidemiology of sepsis in Germany" study was performed. The aims were (1) to describe the ventilation strategies currently applied in clinical practice, (2) to analyze the association of the different modes of ventilation with mortality and (3) to investigate whether the ratio between arterial partial pressure of oxygen and inspired fraction of oxygen (PF ratio) and/or other respiratory variables are associated with mortality in septic patients needing ventilatory support. METHODS: A total of 454 ICUs in 310 randomly selected hospitals participated in this national prospective observational 1-day point prevalence of sepsis study including 415 patients with severe sepsis or septic shock according to the American College of Chest Physicians/Society of Critical Care Medicine criteria. RESULTS: Of the 415 patients, 331 required ventilatory support. Pressure controlled ventilation (PCV) was the most frequently used ventilatory mode (70.6 %) followed by assisted ventilation (AV 21.7 %) and volume controlled ventilation (VCV 7.7 %). Hospital mortality did not differ significantly among patients ventilated with PCV (57 %), VCV (71 %) or AV (51 %, p=0.23). A PF ratio equal or less than 300 mmHg was found in 83.2 % of invasively ventilated patients (n=316). In AV patients there was a clear trend to a higher PF ratio (204±70 mmHg) than in controlled ventilated patients (PCV 179±74 mmHg, VCV 175±75 mmHg, p=0.0551). Multiple regression analysis identified the tidal volume to pressure ratio (tidal volume divided by peak inspiratory airway pressure, odds ratio OR=0.94, 95 % confidence interval 95% CI=0.89-0.99), acute renal failure (OR=2.15, 95% CI=1.01-4.55) and acute physiology and chronic health evaluation (APACHE) II score (OR=1.09, 95% CI=1.03-1.15) but not the PF ratio (univariate analysis OR=0.998, 95 % CI=0.995-1.001) as independent risk factors for in-hospital mortality. CONCLUSIONS: This representative survey revealed that severe sepsis or septic shock was frequently associated with acute lung injury. Different ventilatory modes did not affect mortality. The tidal volume to inspiratory pressure ratio but not the PF ratio was independently associated with mortality.

How visual information links to multijoint coordination during quiet standing.

The link between visual information and postural control was investigated based on a multi-degree-of-freedom model using the framework of the uncontrolled manifold (UCM) hypothesis. The hypothesis was that because visual information specifies the position of the body in space, it would couple preferentially into those combinations of degrees of freedom (DOFs) that move the body in space and not into combinations of DOFs that do not move the body in space. Subjects stood quietly in a virtual reality cave for 4-min trials with or without a 0.2, 2.0 Hz, or combined 0.2 and 2.0 Hz visual field perturbation that was below perceptual threshold. Motion analysis was used to compute six sagittal plane joint angles. Variance across time of the angular motion was partitioned into (1) variance associated with motion of the body and (2) variance reflecting the use of flexible joint combinations that keep the anterior-posterior positions of the head (HD(POS)) and center of mass (CM(POS)) invariant. UCM analysis was performed in the frequency domain in order to link the sensory perturbation to each variance component at different frequencies. As predicted, variance related to motion of the body was selectively increased at the 0.2-Hz drive frequency but not at other frequencies of sway for both CM(POS) and HD(POS). The dominant effect with the 2.0-Hz visual drive also was limited largely to variance related to motion of the body.

Evidence for multiple potential drivers of increased phosphorus in high-elevation lakes.

Total phosphorus (TP) concentrations have increased in many remote mountain waterbodies across the western United States, and reports of algal blooms in these systems have increased in frequency. Explanations for observed TP increases are uncertain, and typical landscape drivers, such as agricultural/urban runoff, are implausible. We investigated multiple atmospheric and terrestrial-P loading mechanisms to explain the observed decadal increase in TP, including a novel hypothesis that warming soils may lead to elevated P fluxes to receiving water bodies. Using northern Utah mountains ranges as a case study, we measured prospective inputs of total and bioavailable P via dust deposition. Terrestrial loading was evaluated through soil leaching experiments designed to simulate soil acidification and recovery, as well as observed decadal increases in soil temperatures and extended growing season. In the Uinta Mountains, dust-P flux appears to be one of the most plausible mechanisms for P increases where we estimated bioavailable dust-P loading ranged from 1.6 to 23.1 mg P m-2 yr-1. However, our results revealed that an increase of soil pH by 0.5 units could lead to a rise in leached P, ranging from 4.7 to 65 mg P m-2. Rising temperatures also showed the potential to increase soil P leaching; Observed average historical (~ +3 °C) and future (+2 °C) increases in temperature led to a prospective increase in leached P from 2 to 264 mg SRP m-2. While we found that pH shifts can mobilize significant amounts of P in some locations, we found no evidence of pH changes through time in the Uinta Mountains. However, summer soil temperatures increased at most locations. The mechanisms evaluated in this study can help explain the widespread observed increases in P across Western US lakes, but the mechanisms that dominate in any given region are likely to vary based on local to regional factors.

Unpredictable elbow joint perturbation during reaching results in multijoint motor equivalence.

Motor equivalence expresses the idea that movement components reorganize in the face of perturbations to preserve the value of important performance variables, such as the hand's position in reaching. A formal method is introduced to evaluate this concept quantitatively: changes in joint configuration due to unpredictable elbow perturbation lead to a smaller change in performance variables than expected given the magnitude of joint configuration change. This study investigated whether motor equivalence was present during the entire movement trajectory and how magnitude of motor equivalence was affected by constraints imposed by two different target types. Subjects pointed to spherical and cylindrical targets both with and without an elbow joint perturbation produced by a low- or high-stiffness elastic band. Subjects' view of their arm was blocked in the initial position, and the perturbation condition was randomized to avoid prediction of the perturbation or its magnitude. A modification of the uncontrolled manifold method variance analysis was used to investigate how changes in joint configuration on perturbed vs. nonperturbed trials (joint deviation vector) affected the hand's position or orientation. Evidence for motor equivalence induced by the perturbation was present from the reach onset and increased with the strength of the perturbation after 40% of the reach, becoming more prominent as the reach progressed. Hand orientation was stabilized more strongly by motor equivalent changes in joint configuration than was three-dimensional position regardless of the target condition. Results are consistent with a recent model of neural control that allows for flexible patterns of joint coordination while resisting joint configuration deviations in directions that affect salient performance variables. The observations also fit a general scheme of synergic control with referent configurations defined across different levels of the motor hierarchy.

Motor equivalence and self-motion induced by different movement speeds.

This study investigated pointing movements in 3D asking two questions: (1) Is goal-directed reaching accompanied by self-motion, a component of the joint velocity vector that leaves the hand's movement unaffected? (2) Are differences in the terminal joint configurations among different speeds of reaching motor equivalent (i.e., terminal joint configurations differ more in directions of joint space that do not produce different pointer-tip positions than in directions that do) or non-motor equivalent (i.e., terminal joint configurations differ equally or more in directions of joint space that lead to different pointer-tip positions than in directions that do not affect the pointer-tip position). Subjects reached from an identical starting joint configuration and pointer-tip location to targets at slow, moderate, and fast speeds. Ten degrees of freedom of joint motion of the arm were recorded. The relationship between changes in the joint configuration and the three-dimensional pointer-tip position was expressed by a standard kinematic model, and the range- and null subspaces were computed from the associated Jacobian matrix. (1) The joint velocity vector and (2) the difference vector between terminal joint configurations from pairs of speed conditions were projected into the two subspaces. The relative length of the two components was used to quantify the amount of self-motion and the presence of motor equivalence, respectively. Results revealed that reaches were accompanied by a significant amount of self-motion at all reaching speeds. Self-motion scaled with movement speed. In addition, the difference in the terminal joint configuration between pairs of different reaching speeds revealed motor equivalence. The results are consistent with a control system that takes advantage of motor redundancy, allowing for flexibility in the face of perturbations, here induced by different movement speeds.

The effect of real-time gait retraining on hip kinematics, pain and function in subjects with patellofemoral pain syndrome.

BACKGROUND: Patellofemoral pain syndrome (PFPS) is the most common overuse injury in runners. Recent research suggests that hip mechanics play a role in the development of this syndrome. Currently, there are no treatments that directly address the atypical mechanics associated with this injury. OBJECTIVE: The purpose of this study was to determine whether gait retraining using real-time feedback improves hip mechanics and reduces pain in subjects with PFPS. METHODS: Ten runners with PFPS participated in this study. Real-time kinematic feedback of hip adduction (HADD) during stance was provided to the subjects as they ran on a treadmill. Subjects completed a total of eight training sessions. Feedback was gradually removed over the last four sessions. Variables of interest included peak HADD, hip internal rotation (HIR), contralateral pelvic drop, as well as pain on a verbal analogue scale and the lower-extremity function index. We also assessed HADD, HIR and contralateral pelvic drop during a single leg squat. Comparisons of variables of interest were made between the initial, final and 1-month follow-up visit. RESULTS: Following the gait retraining, there was a significant reduction in HADD and contralateral pelvic drop while running. Although not statistically significant, HIR decreased by 23% following gait retraining. The 18% reduction in HADD during a single leg squat was very close to significant. There were also significant improvements in pain and function. Subjects were able to maintain their improvements in running mechanics, pain and function at a 1-month follow-up. An unexpected benefit of the retraining was an 18% and 20% reduction in instantaneous and average vertical load rates, respectively. CONCLUSIONS: Gait retraining in individuals with PFPS resulted in a significant improvement of hip mechanics that was associated with a reduction in pain and improvements in function. These results suggest that interventions for PFPS should focus on addressing the underlying mechanics associated with this injury. The reduction in vertical load rates may be protective for the knee and reduce the risk for other running-related injuries.

Association between DNA ploidy and micronucleus frequency in chronic smokers and impact of smoking cessation.

Tobacco use may initiate the process of oral carcinogenesis with clinically undetectable changes. Smoking cessation may prevent its progression. The objective of this study was to evaluate the association between DNA ploidy and micronucleus (MN) frequency in chronic smokers. Three groups were evaluated: Smoker Group, Former Smoker Group and Control Group. Exfoliative cytology was performed on the lateral border of the tongue and mouth floor. MN and DNA ploidy analyses were performed, as well as the correlation between the variables. The data showed a difference between the groups for the total MN (p = 0.0227), and the Smoker group had the highest mean (4.22 ± 4.12). The three groups did not differ statistically from each other on ploidy evaluation (p-value > 0.05). There was also an association between aneuploidy and increased MN frequency in the Former Smoker group (p = 0.0036). In conclusion, these results point out that there is a relationship between the frequency of MN and aneuploidy in former smokers. Moreover, smoking cessation, even for a short period of time, may promote the decrease of MN frequency caused by tobacco use.

Potential Pitfalls in the Analysis and Structural Interpretation of Seismic Data from the Mars InSight Mission.

The Seismic Experiment for Interior Structure (SEIS) of the InSight mission to Mars, has been providing direct information on Martian interior structure and dynamics of that planet since it landed. Compared to seismic recordings on Earth, ground motion measurements acquired by SEIS on Mars are made under dramatically different ambient noise conditions, but include idiosyncratic signals that arise from coupling between different InSight sensors and spacecraft components. This work is to synthesize what is known about these signal types, illustrate how they can manifest in waveforms and noise correlations, and present pitfalls in structural interpretations based on standard seismic analysis methods. We show that glitches, a type of prominent transient signal, can produce artifacts in ambient noise correlations. Sustained signals that vary in frequency, such as lander modes which are affected by variations in temperature and wind conditions over the course of the Martian Sol, can also contaminate ambient noise results. Therefore, both types of signals have the potential to bias interpretation in terms of subsurface layering. We illustrate that signal processing in the presence of identified nonseismic signals must be informed by an understanding of the underlying physical processes in order for high fidelity waveforms of ground motion to be extracted. While the origins of most idiosyncratic signals are well understood, the 2.4 Hz resonance remains debated and the literature does not contain an explanation of its fine spectral structure. Even though the selection of idiosyncratic signal types discussed in this paper may not be exhaustive, we provide guidance on best practices for enhancing the robustness of structural interpretations.

Posaconazole salvage treatment in paediatric patients: a multicentre survey.

While a paediatric dosage has not been defined, posaconazole is occasionally being used in children. We conducted a multicentre retrospective survey and identified 15 patients (median age 10 years [range 3.6-17.5]) who received posaconazole salvage therapy for proven (9 patients) or probable (6 patients) invasive fungal infections. Posaconazole was administered for a median of 32 days (range 4-262) at a median dosage of 21 mg/kg (range 4.8-33.3). None of the patients discontinued therapy due to adverse events, which were mostly mild and observed in 11 patients. Complete or partial responses were observed in 4/7 patients with zygomycosis, 3/4 patients with invasive mould infection, 1/2 patients with invasive aspergillosis and 1/2 patients with chronic disseminated candidiasis. We conclude from the data that posaconazole displays favourable safety and tolerance and may be useful for management of individual paediatric patients with invasive infections.

Correlation and agreement between the bispectral index vs. state entropy during hypothermic cardio-pulmonary bypass.

BACKGROUND: The bispectral index (BIS) and spectral entropy enable monitoring the depth of anaesthesia. Mild hypothermia has been shown to affect the ability of electroencephalography monitors to reflect the anaesthetic drug effect. The purpose of this study was to investigate the effect of hypothermia during a cardio-pulmonary bypass on the correlation and agreement between the BIS and entropy variables compared with normothermic conditions. METHODS: This prospective clinical study included coronary artery bypass grafting patients (n=25) evaluating correlation and agreement (Bland-Altman analysis) between the BIS and both spectral and response entropy during a hypothermic cardio-pulmonary bypass (31-34 degrees C) compared with nomothermic conditions (34-37.5 degrees C). Anaesthesia was maintained with propofol and sufentanil and adjusted clinically, while the anaesthetist was blinded to the monitors. RESULTS: The BIS and entropy values decreased during cooling (P<0.05), but the decrease was more pronounced for entropy variables compared with BIS (P<0.05). The correlation coefficients (bias+/-SD; percentage error) between the BIS vs. spectral state entropy and response entropy were r(2)=0.56 (1+/-11; 42%) and r(2)=0.58 (-2+/-11; 43%) under normothermic conditions, and r(2)=0.17 (10+/-12; 77%) and r(2)=0.18 (9+/-11; 68%) under hypothermic conditions, respectively. Bias was significantly increased under hypothermic conditions (P<0.001 vs. normothermia). CONCLUSION: Acceptable agreement was observed between the BIS and entropy variables under normothermic but not under hypothermic conditions. The BIS and entropy variables may therefore not be interchangeable during a hypothermic cardio-pulmonary bypass.

CaRdiac Arrest Survival Score (CRASS) - A tool to predict good neurological outcome after out-of-hospital cardiac arrest.

AIM: The aim of this study was to develop a score to predict the outcome for patients brought to hospital following out-of-hospital cardiac arrest (OHCA). METHODS: All patients recorded in the German Resuscitation Registry (GRR) who suffered OHCA 2010-2017, who had ROSC or ongoing CPR at hospital admission were included. The study population was divided into development (2010-2016: 7985) and validation dataset (2017: 1806). Binary logistic regression analysis was used to derive the score. The probability of hospital discharge with good neurological outcome was defined as 1/(1 + e-X), where X is the weighted sum of independent variables. RESULTS: The following variables were found to have a significant positive (+) or negative (-) impact: age 61-70 years (-0·5), 71-80 (-0·9), 81-90 (-1·3) and > = 91 (-2·3); initial PEA (-0·9) and asystole (-1·4); presumable trauma (-1·1); mechanical CPR (-0·3); application of adrenalin > 0 - < 2 mg (-1·1), 2 - <4 mg (-1·6), 4 - < 6 mg (-2·1), 6 - < 8 mg (-2·5) and > = 8 mg (-2·8); pre emergency status without previous disease (+0·5) or minor disease (+0·2); location at nursing home (-0·6), working place/school (+0·7), doctor's office (+0·7) and public place (+0·3); application of amiodarone (+0·4); hospital admission with ongoing CPR (-1·9) or normotension (+0·4); witnessed arrest (+0·6); time from collapse until start CPR 2 - < 10 min (-0·3) and > = 10 min (-0·5); duration of CPR <5 min (+0·6). The AUC in the development dataset was 0·88 (95% CI 0·87-0·89) and in the validation dataset 0·88 (95% CI 0·86-0·90). CONCLUSION: The CaRdiac Arrest Survival Score (CRASS) represents a tool for calculating the probability of survival with good neurological function for patients brought to hospital following OHCA.

Influence of different remifentanil concentrations on the performance of the surgical stress index to detect a standardized painful stimulus during sevoflurane anaesthesia.

BACKGROUND: Although measurement of cerebral hypnotic drug effect and muscle relaxation is common clinical routine during anaesthesia, a reliable measurement of the neurophysiological effects evoked by a painful stimulus is still missing. Recently, the surgical stress index (SSI) has been introduced as a surrogate measure of 'nociception'. The present study aimed to examine the influence of increasing remifentanil concentrations on the ability of SSI to detect a standardized painful stimulus during sevoflurane anaesthesia. METHODS: Twenty-four patients received incremental or decremental doses of 0, 2, and 4 ng ml(-1) remifentanil effect-site concentration (Ce(remi)) during 0.7 MAC sevoflurane. Painful tetanic stimulation was applied at least 5 min after changing Ce(remi). SSI, heart rate (HR), response entropy (RE), state entropy (SE), RE-SE difference, and bispectral index (BIS) were obtained in each patient before and after stimulation. Further prediction of an author-defined response to painful stimulus was analysed. RESULTS: SSI and BIS, but not HR, SE, RE, or RE-SE difference were significantly altered after stimulation. Change in SSI (Delta SSI) was significantly dependent on Ce(remi), as Delta SSI was [median (inter-quartile range)] 20 (15-31), 10 (1-19), and 3 (1-10) at 0, 2, and 4 ng ml(-1) Ce(remi). In 10 out of 63 cases, SSI detected response to stimulation, not detected by another variable. SSI was unable to predict movement after stimulation as P(K) value is 0.59 (0.09). CONCLUSIONS: The SSI response to tetanic stimulation was dependent on the remifentanil concentration.

A comparison of the i-gel with the LMA-Unique in non-paralysed anaesthetised adult patients.

SUMMARY: This study assessed two disposable devices; the newly developed supraglottic airway device i-gel and the LMA-Unique in routine clinical practice. Eighty patients (ASA 1-3) undergoing minor routine gynaecologic surgery were randomly allocated to have an i-gel (n = 40) or LMA-Unique (n = 40) inserted. Oxygen saturation, end-tidal carbon dioxide, tidal volume and peak airway pressure were recorded, as well as time of insertion, airway leak pressure, postoperative sore-throat, dysphonia and dysphagia for each device. Time of insertion was comparable with the i-gel and LMA-Unique. There was no failure in the i-gel group and one failure in the LMA-Unique group. Ventilation and oxygenation were similar between devices. Mean airway pressure was comparable with both devices, whereas airway leak pressure was significantly higher (p < 0.0001) in the i-gel group (mean 29 cmH(2)O, range 24-40) compared with the LMA-Unique group (mean 18 cmH(2)O, range 6-30). Fibreoptic score of the position of the devices was significantly better in the i-gel group. Post-operative sore-throat and dysphagia were comparable with both devices. Both devices appeared to be simple alternatives to secure the airway. Significantly higher airway leak pressure suggests that the i-gel may be advantageous in this respect.

Influence of anaesthesia resident training on the duration of three common surgical operations.

We investigated the influence of resident training on anaesthesia workflow of three standard procedures--laparoscopic cholecystectomy, diagnostic gynaecological laparoscopy and transurethral prostate gland resection (TURP)--comparing 259 non-emergency resident vs 341 consultant cases from 20 German hospitals. Each hospital provided 10 random cases for each procedure, yielding 600 cases for analysis. Standard time intervals as documented in the hospital information system were: 'Case Time' (the time from the start of anaesthesia induction to discharge of the patient to the recovery area) and 'Anaesthesia Control Time' (which was the Case Time minus the time from the start of surgery to the end of surgical closure). Case Time was significantly shorter for consultants in all three procedures (p < 0.05, analysis of variance) and Anaesthesia Control Time shorter for consultants only in gynaecological laparoscopy and TURP. Patient comorbidity, patient age and geographical location of the hospital were not influential factors in the analysis of variance. We conclude that resident training significantly increases duration of elective operative times.

[The gynecological TURP syndrome. Severe hyponatremia and pulmonary edema during hysteroscopy]. / Das gynäkologische "TUR-Syndrom". Schwere Hyponatriämie und Lungenödem während operativer Hysteroskopie.

Fluid overload and dilutional hyponatremia are rare but typical complications of endoscopic transurethral resection of the prostate gland (TURP syndrome). Less commonly, this complication caused by excessive absorption of hypotonic, electrolyte-free non-conductive distention solution, may also occur during endoscopic surgery of the uterus, e.g. operative hysteroscopy. A case report is presented of a 44-year-old woman scheduled for operative hysteroscopy for intracavital myoma under general anaesthesia, suffering from severe absorption syndrome leading to hyponatremia of 106 mEq/l and pulmonary edema necessitating subsequent admission to ICU. Anatomical and physiological as well as technical aspects of this syndrome are discussed. Recommendations for anesthesiology management are offered and a possible treatment of acute hyponatremia is discussed.

Architecture of Anoteropora latirostris (Bryozoa, Cheilostomata) and implications for their biomineralization.

Cheilostome Bryozoa Anoteropora latirostris, a colonial marine invertebrate, constructs its skeleton from calcite and aragonite. This study presents firstly correlated multi-scale electron microscopy, micro-computed tomography, electron backscatter diffraction and NanoSIMS mapping. We show that all primary, coarse-grained platy calcitic lateral walls are covered by fine-grained fibrous aragonite. Vertical lateral walls separating autozooid chambers have aragonite only on their distal side. This type of asymmetric mineralization of lateral walls results from the vertical arrangement of the zooids at the growth margins of the colony and represents a type of biomineralization previously unknown in cheilostome bryozoans. NanoSIMS mapping across the aragonite-calcite interface indicates an organic layer between both mineral phases, likely representing an organic template for biomineralization of aragonite on the calcite layer. Analysis of crystallographic orientations show a moderately strong crystallographic preferred orientation (CPO) for calcite (7.4 times random orientation) and an overall weaker CPO for aragonite (2.4 times random orientation) with a high degree of twinning (45%) of the aragonite grains. The calculated Young's modulus for the CPO map shows a weak mechanical direction perpendicular to the colony's upper surface facilitating this organism's strategy of clonal reproduction by fragmentation along the vertical zooid walls.

An automatable platform for genotoxicity testing of nanomaterials based on the fluorometric γ-H2AX assay reveals no genotoxicity of properly surface-shielded cadmium-based quantum dots.

The large number of nanomaterial-based applications emerging in the materials and life sciences and the foreseeable increasing use of these materials require methods that evaluate and characterize the toxic potential of these nanomaterials to keep safety risks to people and environment as low as possible. As nanomaterial toxicity is influenced by a variety of parameters like size, shape, chemical composition, and surface chemistry, high throughput screening (HTS) platforms are recommended for assessing cytotoxicity. Such platforms are not yet available for genotoxicity testing. Here, we present first results obtained for application-relevant nanomaterials using an automatable genotoxicity platform that relies on the quantification of the phosphorylated histone H2AX (γ-H2AX) for detecting DNA double strand breaks (DSBs) and the automated microscope system AKLIDES® for measuring integral fluorescence intensities at different excitation wavelengths. This platform is used to test the genotoxic potential of 30 nm-sized citrate-stabilized gold nanoparticles (Au-NPs) as well as micellar encapsulated iron oxide nanoparticles (FeOx-NPs) and different cadmium (Cd)-based semiconductor quantum dots (QDs), thereby also searching for positive and negative controls as reference materials. In addition, the influence of the QD shell composition on the genotoxic potential of these Cd-based QDs was studied, using CdSe cores as well as CdSe/CdS core/shell and CdSe/CdS/ZnS core/shell/shell QDs. Our results clearly revealed the genotoxicity of the Au-NPs and its absence in the FeOx-NPs. The genotoxicity of the Cd-QDs correlates with the shielding of their Cd-containing core, with the core/shell/shell architecture preventing genotoxicity risks. The fact that none of these nanomaterials showed cytotoxicity at the chosen particle concentrations in a conventional cell viability assay underlines the importance of genotoxicity studies to assess the hazardous potential of nanomaterials.