Effect of Texture and Surface Chemistry on Deagglomeration and Powder Retention in Capsule-Based Dry Powder Inhaler.

Pulmonary delivery systems should administer a high dose of the required formulation with the designated dry powder inhaler (DPI) to achieve therapeutic success. While the effects of device geometry and individual components used on powder dispersion are described in literature, potential effects of DPI surface properties on powder retention within the device and deagglomeration have not been adequately studied, but could impact inhalation therapy by modifying the available dose. For this, inner parts of a model DPI were modified by plasma treatment using various processes. Since both the hydrophilic-hydrophobic and structural properties of the surface were altered, conclusions can be drawn for future optimization of devices. The results show that surface topography has a greater influence on powder deposition and deagglomeration than hydrophilic or hydrophobic surface modification. The most important modification was observed with an increased rough surface texture in the mouth piece, resulting in lower powder deposition in this part (from 5 to 1% quantified amount of powder), without any change in powder deagglomeration compared to an untreated device. In summary, increasing the surface roughness of DPI components in the size range of a few nanometers could be an approach for future optimization of DPIs to increase the delivered dose.

State of the Art in Capsule-Based Dry Powder Inhalers: Deagglomeration Techniques and the Consequences for Formulation Aerosolization.

Commercially available dry powder inhalers (DPIs) are usually devices in a fixed combination with the intended formulation, and a change in medication by the physician often forces the patient to use a different device, requiring the patient to relearn how to use it, resulting in lower adherence and inadequate therapy. To investigate whether DPIs can achieve successful outcomes regardless of the formulation and flow rate used, a novel DPI and two commercially available devices were compared in vitro for their deagglomeration behavior for different binary blends and a spray-dried particle formulation. The results demonstrate that the novel device achieved the highest fine particle fraction (FPF) regardless of the formulations tested. In the binary mixtures tested, the highest emitted fraction was obtained by shaking out the powder due to the oscillating motion of the capsule in the novel device during actuation. For DPIs with high intrinsic resistance to airflow, similar FPFs were obtained with the respective DPI and formulation, regardless of the applied flow rate. Additionally, the development and use of binary blends of spray-dried APIs and carrier particles may result in high FPF and overcome disadvantages of spray-dried particles, such as high powder retention in the capsule.

Algorithm-Based Hearing and Speech Therapy Rehabilitation after Cochlear Implantation.

INTRODUCTION: Due to the changes in the indication range for cochlear implants and the demographic development towards an aging society, more and more people are in receipt of cochlear implants. An implantation requires a close-meshed audiological and logopedic aftercare. Hearing therapy rehabilitation currently requires great personnel effort and is time consuming. Hearing and speech therapy rehabilitation can be supported by digital hearing training programs. However, the apps currently on the market are to a limited degree personalized and structured. Increasing digitalization makes it possible, especially in times of pandemics, to decouple hearing therapy treatment from everyday clinical practice. MATERIAL AND METHODS: For this purpose, an app is in development that provides hearing therapy tailored to the patient. The individual factors that influence hearing outcome are considered. Using intelligent algorithms, the app determines the selection of exercises, the level of difficulty and the speed at which the difficulty is increased. RESULTS: The app works autonomously without being connected to local speech therapists. In addition, the app is able to analyze patient difficulties within the exercises and provides conclusions about the need for technical adjustments. CONCLUSIONS: The presented newly developed app represents a possibility to support, replace, expand and improve the classic outpatient hearing and speech therapy after CI implantation. The way the application works allows it to reach more people and provide a time- and cost-saving alternative to traditional therapy.

Per- and polyfluoroalkyl substances in blood plasma - Results of the German Environmental Survey for children and adolescents 2014-2017 (GerES V).

The 5th cycle of the German Environmental Survey (GerES V) investigated the internal human exposure of children and adolescents aged 3-17 years in Germany to per- and polyfluoroalkyl substances (PFAS). The fieldwork of the population-representative GerES V was performed from 2014 to 2017. In total, 1109 blood plasma samples were analysed for 12 PFAS including perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), and perfluorohexane sulfonic acid (PFHxS). PFOS was quantified in all and PFOA in almost all samples, demonstrating ubiquitous exposure. The highest geometric mean concentrations measured were 2.49 ng/mL for PFOS, followed by PFOA (1.12 ng/mL) and PFHxS (0.36 ng/mL), while concentrations of other PFAS were found in much lower concentrations. The 95th percentile levels of PFOS and PFOA were 6.00 and 3.24 ng/mL, respectively. The results document a still considerable exposure of the young generation to the phased out chemicals PFOS and PFOA. The observed exposure levels vary substantially between individuals and might be due to different multiple sources. The relative contribution of various exposure parameters such as diet or the specific use of consumer products need to be further explored. Although additional investigations on the time trend of human exposure are warranted, GerES V underlines the need for an effective and sustainable regulation of PFAS as a whole.

EGFR immunohistochemistry as biomarker for antibody-based therapy of squamous NSCLC - Experience from the first ring trial of the German Quality Assurance Initiative for Pathology (QuIP®).

BACKGROUND: EGFR and its downstream signaling pathway are important targets for cancer therapy. Recently, the monoclonal anti-EGFR antibody Necitumumab in combination with gemcitabine and cisplatin was approved (EMA/14106/2016) for first-line treatment of squamous non-small cell carcinoma (SqNSCLC). Eligibility was restricted to cases with positive EGFR expression. In this context, a ring trial of the Quality Assurance Initiative for Pathology (QuIP®) was launched to prepare the German pathology community for a reliable and reproducible, immunohistochemically based biomarker test. MATERIALS AND METHODS: The trial was set up by a three-step approach. Two lead institutes were nominated to organize the trial process and to select appropriate cancer samples. These were first tested by the H-score (range 0-300) to identify positive and negative cases. Seven additional pathology institutes with experience in EGFR immunohistochemistry each tested the selected panel of identical cases (internal ring trial) to confirm the suitability of samples and scoring criteria. Then the open ring trial for all institutes of pathology in German speaking countries was announced. RESULTS: For the internal trial 8 EGFR-positive and 2 negative lung sqNSCLC samples were selected. A cut-off value of cell membranous staining in≥1% of tumor cells was introduced to define a case as EGFR negative or positive. Two points were attainable per correctly assessed sample leading to a maximum of 20 points,≥18 points were required for a successful participation. All 7 panel institute passed this barrier, 5 with the maximum of 20 points and two with one error (18 points) being related to one case with incorrect interpretation of cytoplasmic versus membranous staining and one case with an H-score of 2 as being considered EGFR positive. A second cut-off value (H-score≥3) was therefore introduced. In the open ring trial, 34 institutions participated of which 28 were successful according to the above criteria. The trial revealed a high reproducibility despite the use of different EGFR antibodies and detection systems. There was no association between technical parameters and trial failure. Again, one participant misinterpreted the subcellular EGFR localization. CONCLUSIONS: The first nationwide ring test for determination of EGFR IHC expression in sqNSCLC could be successfully performed in a very tight time frame. By this, the national pathology community was prepared to incorporate this marker in the panel of predictive cancer tests in a quality assessed manner and to initiate and accompany future studies on EGFR pathway pathology.

A weight-of-evidence approach to assess chemicals: case study on the assessment of persistence of 4,6-substituted phenolic benzotriazoles in the environment.

BACKGROUND: One important purpose of the European REACH Regulation (EC No. 1907/2006) is to promote the use of alternative methods for assessment of hazards of substances in order to avoid animal testing. Experience with environmental hazard assessment under REACH shows that efficient alternative methods are needed in order to assess chemicals when standard test data are missing. One such assessment method is the weight-of-evidence (WoE) approach. In this study, the WoE approach was used to assess the persistence of certain phenolic benzotriazoles, a group of substances including also such of very high concern (SVHC). RESULTS: For phenolic benzotriazoles, assessment of the environmental persistence is challenging as standard information, i.e. simulation tests on biodegradation are not available. Thus, the WoE approach was used: overall information resulting from many sources was considered, and individual uncertainties of each source analysed separately. In a second step, all information was aggregated giving an overall picture of persistence to assess the degradability of the phenolic benzotriazoles under consideration although the reliability of individual sources was incomplete. CONCLUSIONS: Overall, the evidence suggesting that phenolic benzotriazoles are very persistent in the environment is unambiguous. This was demonstrated by a WoE approach considering the prerequisites of REACH by combining several limited information sources. The combination enabled a clear overall assessment which can be reliably used for SVHC identification. Finally, it is recommended to include WoE approaches as an important tool in future environmental risk assessments.

Keratinocytic epidermal nevus syndrome with Schwann cell proliferation, lipomatous tumour and mosaic KRAS mutation.

BACKGROUND: Keratinocytic epidermal nevus syndrome (KENS) is a complex disorder not only characterized by the presence of epidermal nevi but also by abnormalities in the internal organ systems. A small number of cases with KENS are molecularly characterized and reported in the literature with somatic activating RAS, FGFR3 and PIK3CA mutations. CASE PRESENTATION: In this study we present a patient with hyper- and hypopigmented regions, verrucous pigmented skin lesions and a paravertebral conglomerate tumour at the level of the cervical and thoracic spine. A large lipomatous dumbbell tumour caused atrophy of the spinal cord with progressive paraparesis. We identified a mosaic c.35G > A (p.Gly12Asp) KRAS mutation in the pigmented verrucous epidermal nevus tissue, the intraneural schwann cells and the lipoma. The c.35G > A (p.Gly12Asp) KRAS mutation was absent in the peripheral blood leukocytes. CONCLUSION: We conclude that KENS, the intraneural Schwann cell proliferation and the lipoma in this individual were caused by a postzygotic and mosaic activating c.35G > A (p.Gly12Asp) KRAS mutation.

Generic expansion of the Jastrow correlation factor in polynomials satisfying symmetry and cusp conditions.

Jastrow correlation factors play an important role in quantum Monte Carlo calculations. Together with an orbital based antisymmetric function, they allow the construction of highly accurate correlation wave functions. In this paper, a generic expansion of the Jastrow correlation function in terms of polynomials that satisfy both the electron exchange symmetry constraint and the cusp conditions is presented. In particular, an expansion of the three-body electron-electron-nucleus contribution in terms of cuspless homogeneous symmetric polynomials is proposed. The polynomials can be expressed in fairly arbitrary scaling function allowing a generic implementation of the Jastrow factor. It is demonstrated with a few examples that the new Jastrow factor achieves 85%-90% of the total correlation energy in a variational quantum Monte Carlo calculation and more than 90% of the diffusion Monte Carlo correlation energy.

Retrospective monitoring of perfluorocarboxylates and perfluorosulfonates in human plasma archived by the German Environmental Specimen Bank.

Due to the increased awareness of the ubiquitous contamination of all environmental compartments and of human beings with perfluoroalkyl substances (PFAS), voluntary withdrawals and shifts in products and manufacturing technologies, as well as in regulatory measures, have been made. To investigate whether these activities are reflected in the human exposure to PFASS, we examined human blood archived by the German Environmental Specimen Bank. Plasma samples (n=258, age range 20-29 years) covering the observation period 1982-2010 were analyzed for eleven perfluoroalkylcarboxylates (C4-C14) and five perfluoroalkylsulfonates (C4-C10) by HPLC-MS-MS. We detected perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), and perfluorohexanesulfonate (PFHxS) most often of all PFASs. Following a sharp increase from 1982 to 1986, median PFOS concentrations remained in the range of 20-24ng/mL until the end of the 1990s. Between 2001 and 2010, PFOS concentrations decreased steadily to 4ng/mL in plasma. Except for a similar strong increase from 1982 to 1986, we observed PFOA concentrations fluctuating between 4.8 and 6.3ng/mL in the following years. Since 2008, ESB data suggest a decreasing trend of PFOA. PFHxS concentrations increased continuously between 1982 and 2001 from about 1-2ng/mL. After nearly unchanged concentrations until 2005, a downward trend of PFHxS in plasma became apparent and in 2010 resulted in levels which were about 20% lower than those observed in the early 1980s. In the case of shorter and longer chained PFASs, quantification frequencies were between 0 and 60% and we found no indication of any temporal trends in human plasma concentrations.

Chronic lymphocytic leukemia-associated chromosomal abnormalities and miRNA deregulation.

Chronic lymphocytic leukemia is the most common leukemia in adults. By cytogenetic investigations major subgroups of the disease can be identified that reflect different routes of tumor development. Of these chromosomal deviations, trisomy 12 and deletions of parts of either the long arm of chromosome 13, the long arm of chromosome 11, or the short arm of chromosome 17 are most commonly detected. In some of these aberrations the molecular target has been identified as eg, ataxia telangiectasia mutated (ATM) in case of deletions of chromosomal region 11q22~23 and the genes encoding microRNAs miR-15a/16-1 as likely targets of deletions of chromosomal band 13q14.3. Of note, these aberrations do not characterize independent subgroups but often coexist within the metaphases of one tumor. Generally, complex aberrations are associated with a worse prognosis than simple karyotypic alterations. Due to smaller sizes of the missing segment the detection of recurrent deletions is not always possible by means of classical cytogenetics but requires more advanced techniques as in particular fluorescence in situ hybridization (FISH). Nevertheless, at this time it is not recommended to replace classical cytogenetics by FISH because this would miss additional information given by complex or secondary karyotypic alterations. However, the results of cytogenetic analyses allow the stratification of prognostic and predictive groups of the disease. Of these, the group characterized by deletions involving TP53 is clinically most relevant. In the future refined methods as eg, array-based comparative genomic hybridization will supplement the existing techniques to characterize CLL.

Problems faced when evaluating the bioaccumulation potential of substances under REACH.

The identification and regulation of substances that combine persistence, bioaccumulation potential, and toxicity ("PBT" substances) is one central aspect of the European chemical legislation REACH (Registration, Evaluation, Authorisation, and Restriction of Chemicals), because these substances may elicit adverse long-term effects after release to the environment. The determination of a substance that has persistence, bioaccumulation potential, and toxicity is based on a set of distinct cutoff criteria identified in Annex XIII of the REACH regulation. Regarding the bioaccumulation potential, the evaluation is focused on the substance's bioconcentration factor as single decisive criterion. In addition, the REACH guidelines provide a selection of standardized test procedures for measuring bioconcentration factor and guidance in appraising test results. However, alternative test results like bioaccumulation factors and biomagnification as well as additional indications for a bioaccumulation potential such as trophic magnification are only allowed for supporting evidence. The currently used test systems with aquatic exposure have been demonstrated to generate reliable results for the majority of neutral, lipophilic organic substances, which facilitate clear decision-making by means of the crucial bioconcentration factor cutoff criteria of Annex XIII. However, certain substance groups such as highly hydrophobic organic substances and amphiphilic and nonlipophilic compounds are difficult to evaluate with common test strategies due to inappropriate test systems or accumulation mechanisms not based on lipophilicity. Recent scientific progress has already been made to establish alternative test systems and to refine the bioaccumulation assessment by consideration of additive accumulation mechanisms and indications. This article gives an overview on actual shortcomings in the current bioaccumulation assessment under REACH and also provides suggestions for a refinement of evaluation.

High expression of the immature laminin receptor protein correlates with mutated IGVH status and predicts a favorable prognosis in chronic lymphocytic leukemia.

The immature laminin receptor (iLR) is a tumor-associated antigen. We analyzed the expression of iLR on malignant B cells of 134 unselected patient samples with CLL and hypothesized that iLR expression would have prognostic significance due to a differential expression pattern. High ILR expression (cut-off value 30%) was correlated with mutated IGVH status (p<0.0001). Patients with high iLR-expression had a significantly longer time to progression (p=0.039). Combination of CD38, ZAP-70, and iLR by flow cytometry can be used to construct a diagnostic score identifying patients with a median progression free survival of 80 months, if no adverse marker is present.

Integrated approach to PBT and POP prioritization and risk assessment.

This article summarizes discussions at the SETAC Pellston Workshop on "Science-Based Guidance and Framework for the Evaluation and Identification of PBTs and POPs" and provides an overview of other articles from that workshop that are also published in this issue. Identification of persistent, bioaccumulative, and toxic substances (PBTs) and persistent organic pollutants (POPs) and evaluation of their impact are more complicated than those for other chemicals and remain a challenge. The main reason for this is that PBT substance and POP assessment is associated with higher uncertainty and generally requires more data. However, for some data-rich PBTs and POPs, that identification and assessment of impact are feasible has been clearly demonstrated. New scientific developments and techniques are able to significantly increase the certainty of the various elements of PBT and POP assessment, and the current scientific literature provides many successful and illustrative examples that can be used as methodologies to build on. Applying multiple approaches for assessment is advisable, because it will reduce uncertainty and may increase confidence and improve the quality of decision-making.

Humoral immune responses against the immature laminin receptor protein show prognostic significance in patients with chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) is characterized by a highly variable clinical course. The role of an autologous tumor-specific immune control contributing to the variable length of survival in CLL is poorly understood. We investigated whether humoral immunity specific for the CLL-associated Ag oncofetal Ag/immature laminin receptor (OFA/iLR) has a prognostic value in CLL. Among sera of 67 untreated patients with CLL, 23 (34.3%) had detectable OFA/iLR Abs that were reactive for at least one specific OFA/iLR epitope. Patients with humoral responses compared with patients with nonreactive sera had a longer progression-free survival (p = 0.029). IgG subclass analyses showed a predominant IgG1 and IgG3 response. OFA/iLR Abs were capable of recognizing and selectively killing OFA/iLR-expressing CLL cells in complement-mediated and Ab-dependent cellular cytotoxicity assays. In the analysis of 11 CLL patients after allogeneic hematopoietic stem cell transplantation, 8 showed high values for OFA/iLR Abs that specifically recognized the extracellular domain of the protein, suggesting a potential role of anti-OFA/iLR-directed immune responses to the graft-vs-leukemia effect in CLL. Our data suggest that spontaneous tumor-specific humoral immune responses against OFA/iLR exist in a significant proportion of CLL patients and that superior progression-free survival in those patients could reflect autologous immune control.

Activated coagulation factors in human malignant effusions and their contribution to cancer cell metastasis and therapy.

We have shown that the thrombin G-protein coupled receptors (GPCR) designated as protease-activated receptors (PAR-1) are expressed in primary cancer cells isolated from peritoneal and pleural malignant effusions. Here, our main goal was to evaluate several coagulation and thrombin activation effectors and markers in a series of 136 malignant effusions from cancer patients with gastrointestinal, lung and mammary carcinomas. All these patients present a highly activated coagulation system in blood and their malignant effusions, as indicated by high levels of prothrombin F1.2 fragments and D-dimers. Notably, we detected in the effusions all the coagulation factors of the tissue factor pathway inducing thrombin activation, namely factors VII, V, X and II, as well as high VEGF levels and IGF-II in mature and precursor forms. Fibrin clot formation also correlated with higher levels of free ionized calcium (iCa), suggesting that iCa and its binding protein albumin are regulatory factors for fibrinogenesis in effusions. Consequently, thrombin, VEGF and IGFII appear to converge in the promotion of survival and invasivity of the metastatic cancer cells from blood to the malignant effusions. Thus, we add new insights on the interconnections between blood coagulation disorders in cancer patients and thrombin activation in malignant effusions, including their functional interaction with PAR in metastatic cancer cells. Based on these data we propose to counteract the metastatic cascades by targeted invalidation of key effectors of the coagulation system. Therefore, potential therapeutic approaches include the application of thrombin protease inhibitors, VEGF-blocking antibodies, and drugs targeting the VEGF and thrombin signaling pathways, such as tyrosine kinase or GPCR inhibitors.