Personalized neoantigen cancer vaccines: An analysis of the clinical and commercial potential of ongoing development programs.

Neoantigen cancer vaccines harbor promise as next-generation immuno-oncology therapies, whereby cancer vaccines are tailored to the patient's tumor antigen and represent the future of personalized cancer therapy. While several biotech companies have ongoing development programs, little has been published about the true commercial potential of these innovative therapies and the challenges these products will face upon regulatory approval. In this paper, we provide an overview of neoantigen cancer vaccine development programs and discuss the commercial environment these therapies will face upon launch.

Values and value in simulated participant methodology: A global perspective on contemporary practices.

This article has been written for the 40th year of the publication of Medical Teacher. While we celebrate the contribution of simulated participants (SPs) to health professions education through values and value-based learning, we also offer critical reflection on elements of our practice, commencing with language. We argue for the use of the term simulated rather than standardized and acknowledge the dominant role of the SP as patient and the origins of the methodology. These shifts in terms and their implications in practice reflect changes in the conceptualization of SP-based methodology. Recently published standards for those who work with SPs (SP practitioners) are noted as an important milestone in our community's development. We consider contemporary practices addressing the complex notions of values and value in SP-based learning. We simultaneously refer to the work of SPs and SP practitioners. Phases of educational design including identifying learning objectives, scenario design, implementation, feedback and debriefing are used to illustrate methodological shifts. Within each of these phases, there are relational issues that have to date often gone unchecked and are under reported in literature. Finally, using the metaphor of a murmuration, we celebrate contemporary practices of the global SP practitioner community.

A cluster of two human cases of tick-borne encephalitis (TBE) transmitted by unpasteurised goat milk and cheese in Germany, May 2016.

In May 2016, two cases of tick-borne encephalitis (TBE) were confirmed by serology (positive IgM and IgG antibodies against TBE virus (TBEV) in serum), with a possible link to raw milk and cheese from a goat farm in a region in Baden-Württemberg, Germany not previously known as TBE-endemic. The outbreak investigation identified 32 consumers of goat dairy products (29 consumers, one farm employee, two owners) of whom none had IgM antibodies against TBEV 3-8 weeks after consumption. Of the 27 notified TBE cases in the State, none reported consumption of raw goat milk or cheese from the suspected farm. Five of 22 cheese samples from 18 different batches were RT-qPCR-positive for TBEV -genome, and two of the five samples were confirmed by virus isolation, indicating viability of TBEV in the cheese. Nine of the 45 goats had neutralising TBEV antibodies, two of them with a high titre indicating recent infection. One of 412 Ixodes ricinus was RT-qPCR-positive, and sequencing of the E gene from nucleic acid extracted from the tick confirmed TBEV. Phylogenetic analyses of tick and cheese isolates showed 100% amino acid homology in the E gene and a close relation to TBEV strains from Switzerland and Austria.

Long-term results after restoring nasal tip support using auricular cartilage as an intercrural columellar strut graft: the "I-Beam" technique.

BACKGROUND: The lack of nasal tip support resulting in poor protection and projection of the nose is a common cause of nasal obstruction and cosmetic impairment. Although septal cartilage is preferred for restoration of nasal tip support, there is often not enough septal cartilage available. We describe an intercrural columellar strut graft in a circumferential soft tissue pocket: the I-Beam technique. METHODOLOGY: In this cohort study over an 8-year period the clinical and biometrical long-term results after restoration of nasal tip support with an autologous auricular cartilage graft, the I-Beam technique, is evaluated. RESULTS: Out of the 72 consecutive septorhinoplasties with the I-Beam technique, we studied 66 patients in the median long-term follow-up of 89 months. In all patients a significant improvement in nasal tip projection was found. A functional improvement in nasal breathing was achieved in 86%. The overall satisfaction with the I-Beam graft was of 83%. No deterioration in smell perception was found after surgery. CONCLUSIONS: The I-Beam technique using back-to-back sutured autologous auricular cartilage is a safe and effective technique and gives long-term structural support and enhances tip projection.

DAPK2 regulates oxidative stress in cancer cells by preserving mitochondrial function.

Death-associated protein kinase (DAPK) 2 is a serine/threonine kinase that belongs to the DAPK family. Although it shows significant structural differences from DAPK1, the founding member of this protein family, DAPK2 is also thought to be a putative tumour suppressor. Like DAPK1, it has been implicated in programmed cell death, the regulation of autophagy and diverse developmental processes. In contrast to DAPK1, however, few mechanistic studies have been carried out on DAPK2 and the majority of these have made use of tagged DAPK2, which almost invariably leads to overexpression of the protein. As a consequence, physiological roles of this kinase are still poorly understood. Using two genetically distinct cancer cell lines as models, we have identified a new role for DAPK2 in the regulation of mitochondrial integrity. RNA interference-mediated depletion of DAPK2 leads to fundamental metabolic changes, including significantly decreased rate of oxidative phosphorylation in combination with overall destabilised mitochondrial membrane potential. This phenotype is further corroborated by an increase in the production of mitochondrial superoxide anions and increased oxidative stress. This then leads to the activation of classical stress-activated kinases such as ERK, JNK and p38, which is observed on DAPK2 genetic ablation. Interestingly, the generation of oxidative stress is further enhanced on overexpression of a kinase-dead DAPK2 mutant indicating that it is the kinase domain of DAPK2 that is important to maintain mitochondrial integrity and, by inference, for cellular metabolism.

DAPK2 is a novel modulator of TRAIL-induced apoptosis.

Targeting molecules involved in TRAIL-mediated signalling has been hailed by many as a potential magic bullet to kill cancer cells efficiently, with little side effects on normal cells. Indeed, initial clinical trials showed that antibodies against TRAIL receptors, death receptor (DR)4 and DR5, are well tolerated by cancer patients. Despite efficacy issues in the clinical setting, novel approaches to trigger TRAIL-mediated apoptosis are being developed and its clinical potential is being reappraised. Unfortunately, as observed with other cancer therapies, many patients develop resistance to TRAIL-induced apoptosis and there is thus impetuous for identifying additional resistance mechanisms that may be targetable and usable in combination therapies. Here, we show that the death-associated protein kinase 2 (DAPK2) is a modulator of TRAIL signalling. Genetic ablation of DAPK2 using RNA interference causes phosphorylation of NF-κB and its transcriptional activity in several cancer cell lines. This then leads to the induction of a variety of NF-κB target genes, which include proapoptotic DR4 and DR5. DR4 and DR5 protein expression is correspondingly increased on the cell surface and this leads to the sensitisation of resistant cells to TRAIL-induced killing, in a p53-independent manner. As DAPK2 is a kinase, it is imminently druggable, and our data thus offer a novel avenue to overcome TRAIL resistance in the clinic.

Use of artemisinin and its derivatives to treat HPV-infected/transformed cells and cervical cancer: a review.

Cervical cancer and HPV-related diseases remain a burden in the developing world. While much progress has been gained in the detection of HPV and preneoplastic cervical lesions, the rate-limiting step in the prevention of cervical cancer is management of these women. A natural compound, artemisinin, and its derivatives appear to hold promise as a simple means of treatment. Laboratory studies have shown that this compound, and its derivatives, have activity against HPV-infected and -transformed cells and cervical cancer cells. In situations of compassionate use, studies have also demonstrated efficacy in clinical situations. Well-designed clinical trials relating to its use should be undertaken.

Application of biofilm bioreactors in white biotechnology.

The production of valuable compounds in industrial biotechnology is commonly done by cultivation of suspended cells or use of (immobilized) enzymes rather than using microorganisms in an immobilized state. Within the field of wastewater as well as odor treatment the application of immobilized cells is a proven technique. The cells are entrapped in a matrix of extracellular polymeric compounds produced by themselves. The surface-associated agglomerate of encapsulated cells is termed biofilm. In comparison to common immobilization techniques, toxic effects of compounds used for cell entrapment may be neglected. Although the economic impact of biofilm processes used for the production of valuable compounds is negligible, many prospective approaches were examined in the laboratory and on a pilot scale. This review gives an overview of biofilm reactors applied to the production of valuable compounds. Moreover, the characteristics of the utilized materials are discussed with respect to support of surface-attached microbial growth.

Release of ANP and fat oxidation in overweight persons during aerobic exercise in water.

Exercise in water compared to land-based exercise (LE) results in a higher release of natriuretic peptides, which are involved in the regulation of exercise-induced adipose tissue lipolysis. The present study was performed to compare the release of atrial natriuretic peptide (ANP) and free fatty acids (FFA) during prolonged aerobic water-based exercise (WE) with the release after an identical LE. 14 untrained overweight subjects performed 2 steady state workload tests on the same ergometer in water and on land. Before and after exercise, venous blood samples were collected for measuring ANP, FFA, epinephrine, norepinephrine, insulin and glucose. The respiratory exchange ratio (RER) was determined for fat oxidation.The exercises resulted in a significant increase in ANP in LE (61%) and in WE (177%), and FFA increased about 3-fold in LE and WE with no significant difference between the groups. Epinephrine increased, while insulin decreased similarly in both groups. The RER values decreased during the exercises, but there was no significant difference between LE and WE. In conclusion, the higher ANP concentrations in WE had no additional effect on lipid mobilization, FFA release and fat oxidation. Moderate-intensity exercises in water offer no benefit regarding adipose tissue lipolysis in comparison to LE.

Gd-based single-ion magnets with tunable magnetic anisotropy: molecular design of spin qubits.

We report ac susceptibility and continuous wave and pulsed EPR experiments performed on GdW10 and GdW30 polyoxometalate clusters, in which a Gd3+ ion is coordinated to different polyoxometalate moieties. Despite the isotropic character of gadolinium as a free ion, these molecules show slow magnetic relaxation at very low temperatures, characteristic of single molecule magnets. For T≲200 mK, the spin-lattice relaxation becomes dominated by pure quantum tunneling events, with rates that agree quantitatively with those predicted by the Prokof'ev and Stamp model [Phys. Rev. Lett. 80, 5794 (1998)]. The sign of the magnetic anisotropy, the energy level splittings, and the tunneling rates strongly depend on the molecular structure. We argue that GdW30 molecules are also promising spin qubits with a coherence figure of merit Q(M)≳50.

From incus bypass to malleostapedotomy: technical improvements and results.

OBJECTIVE: To assess results of malleostapedotomy using a Fisch Storz titanium piston with at least 10 months' follow up. METHODS: Using a prospective database, the indications, surgical technique, and pre- and post-operative audiometric data for 60 patients undergoing malleostapedotomy between 2002 and 2010 were evaluated. Diagnoses and primary and revision surgeries were compared with reference to the literature. RESULTS: Sixty endaural malleostapedotomies were performed, 28 as a primary intervention and 32 as revision surgery. In 68 per cent, the underlying pathology was otosclerosis. The most common reason for revision surgery (i.e. in 59 per cent) was prosthesis dysfunction. Overall, the mean air-bone gap (0.5-3 kHz) for the primary intervention and revision surgery groups was 9.4 and 11.3 dB, respectively; an air-bone gap of less than 20 dB was obtained in 100 and 81 per cent of patients, respectively. There was no significant audiological difference between the primary and revision surgeries groups, and no deafness. CONCLUSION: Malleostapedotomy shows comparable results to standard incus-stapedotomy and may be preferable in the presented situations.

The Agger Nasi cell and uncinate process, the keys to proper access to the nasolacrimal drainage system.

BACKGROUND: Profound knowledge of the surgical anatomy is required to perform any dacryo-cysto-rhinostomy (DCR) procedure. In endoscopic endonasal DCR surgery, the uncinate process (UP) and/or agger nasi (AN) cell are frequently found to obstruct access to the lacrimal fossa. The aim of this study was to analyse the radiological anatomy of the UP and the AN cell in relation to the lacrimal sac. METHODS: Preoperative computed tomography (CT) scans of patients undergoing DCR and other endoscopic sinus surgeries were analysed regarding their anatomical variations concerning the UP and the AN cell at the level of the common canaliculus. The covering of the lacrimal fossa by the UP was graded and a surgically relevant AN cell was sought after. RESULTS: CT datasets of 60 sides were included. Analysis showed an AN cell overlying the upper parts of the lacrimal sac in 55% of patients. An anteriorly attaching UP covering at least 50% of the lacrimal fossa was found in 63% of individuals. CONCLUSION: In this study more than 80% of all patients presented with either a very anteriorly attached UP or an AN cell that would have to be removed to perform effective endoscopic DCR. The AN cell and the UP need to be included in the endonasal surgical concept.

Broadband electron spin resonance at 4-40 GHz and magnetic fields up to 10 T.

A broadband electron spin resonance spectrometer is described which operates at frequencies between 4 and 40 GHz and can be used in superconducting magnets. A tunable cylindrical cavity is connected to a vector network analyzer via coaxial cables, and the radiation is fed into the cavity by a coupling loop. No field modulation is employed. Resonance frequencies below 14 GHz are obtained by inserting dielectrics with different permittivities into the cavity. The setup allows for measurements with the microwave magnetic field either parallel or perpendicular to the external field.

Direct observation of quantum coherence in single-molecule magnets.

Direct evidence of quantum coherence in a single-molecule magnet in a frozen solution is reported with coherence times as long as T{2}=630+/-30 ns. We can strongly increase the coherence time by modifying the matrix in which the single-molecule magnets are embedded. The electron spins are coupled to the proton nuclear spins of both the molecule itself and, interestingly, also to those of the solvent. The clear observation of Rabi oscillations indicates that we can manipulate the spin coherently, an essential prerequisite for performing quantum computations.

[The consequences of subcutaneous India ink injection to produce factitial disease. Bilateral mastectomy, lymph node dissection, irradiation, and continued suspicion of neoplasia]. / Folgen von subkutaner Tuscheinjektion bei artifizieller Störung. Mastektomie, Lymphknotenausräumung, Radiatio und fortdauernder Malignomverdacht.

A 68-year-old woman with insulin-dependent diabetes mellitus presented with blue nodules on the ventral aspect of the thorax. According to the past history, these lesions had developed repeatedly. She had already had bilateral mastectomies and lymph node dissection. The histologic diagnosis was always mastitis with plasma cells and no neoplasia. Yet another biopsy was taken; the subcutis was stained blue-black. Histology revealed exogenous black pigment and mastitis. With Raman spectroscopy the pigment was identified as carbon black, which is a component of India ink. These findings together with the unusual course of the disease suggested the diagnosis of an artificial disorder. The likely conclusion is that our patient, over years, used her own (insulin) syringe to inject India ink into her skin and subcutaneous tissue; the damaging effect and tissue reaction was probably caused by preservatives such as phenol.

[Cost-of-illness Study for the Treatment of COPD in Germany]. / Krankheitskosten von COPD in Deutschland.

The present cost-of-illness study is focused on the costs of COPD in Germany. In a pre-study, data on 814 randomly selected patients were collected to achieve reliable figures for the distribution of COPD severity grades and the frequencies of exacerbations. The main study was performed on 321 randomly selected patients from the pre-study. Data on resource use were collected in a face-to-face interview with the respective physicians using the patient records as a basis. Costs associated with resource consumption were weighted with the frequencies of COPD severity grades as assessed in the pre-study to determine the costs of COPD. Annual COPD-related costs per patient were 3,027 from the societal perspective. Main cost components were hospitalisations (26 %), medication (23 %) and early retirement (17 %). Annual COPD-related costs from the perspective of the German health insurance system (GKV) were 1,944 euros per patient.

Phase II trial of intralesional therapy with interleukin-2 in soft-tissue melanoma metastases.

The objective of the present study was to validate the use of intralesional injection of interleukin-2 (IL-2) in patients with skin and soft-tissue melanoma metastases. A total of 24 patients with AJCC stage III or IV melanoma and single or multiple skin and soft-tissue metastases were included. Interleukin-2 injections were administered intralesionally into the total number of cutaneous and soft-tissue metastases accessible from the skin, 2-3 times weekly, over 1-57 weeks. Single doses varied from 0.6 to 6 x 10(6) IU, depending on lesion size. The clinical response was monitored by sonography and confirmed by histopathology; response evaluation was confined to the intralesionally treated tumours. Complete response (CR) of the treated metastases was achieved in 15 patients (62.5%), the longest remission lasting 38 months to date. In five patients, partial response (PR) was achieved (21%) and in another three patients, progressive disease was observed (one patient not assessable). A total of 245 metastases were treated with CR in 209 (85%), and PR in 21 (6%). The therapy was generally well tolerated; the observed adverse events were mainly of grade 1-2 severity. Immunohistochemical studies showed the tumour cells undergoing apoptosis and revealed a mixed character of the inflammatory infiltrate. The unusual high CR rate in metastatic melanoma of 62.5% and the limited toxicity suggest that treatment of skin and soft-tissue melanoma metastases with intralesional injection of IL-2 may be a safe and effective alternative to conventional therapies. The optimal dosage and duration of this therapy still remain to be defined in larger prospective multicentre trials.

Localization of functional calcitonin gene-related peptide binding sites in a subpopulation of cultured dorsal root ganglion neurons.

In this study we investigated whether cultured dorsal root ganglion (DRG) neurons from the adult rat express binding sites for calcitonin gene-related peptide (CGRP). These were identified on fixed cells by using CGRP labeled at the N-terminal site with 1.4-nm gold particles. After 1 day in culture, about 20% of small to medium-sized DRG neurons showed CGRP-gold binding. Binding of CGRP-gold was dose-dependently reduced by coadministration of CGRP. The calcium imaging technique in living cells revealed that the bath administration of CGRP evoked an increase of the intracellular calcium in up to 30% of the DRG neurons tested. Both depletion of intracellular calcium stores by thapsigargin or using a calcium-free medium blocked the CGRP-mediated increase of cytosolic calcium in most neurons. Thus intracellular and extracellular sources of calcium are relevant for the CGRP response. Using the whole-cell patch-clamp technique, about 30% of the neurons were found to exhibit an inward current and a depolarization upon administration of CGRP close to the neurons. Immunocytochemical double-labeling techniques showed that most of the CGRP-gold binding sites were expressed in unmyelinated (neurofilament 200-negative) DRG neurons. Most of the neurons with CGRP-gold binding sites also expressed the tyrosine kinase A receptor, and all of them showed CGRP-like immunoreactivity. This study shows, therefore, that a subpopulation of unmyelinated, peptidergic primary afferent neurons express CGRP binding sites that can be activated by CGRP in an excitatory direction. The binding sites may serve as autoreceptors because all of these neurons also synthesize CGRP. The activation of CGRP binding sites may sensitize primary afferent neurons and influence the release of mediators.

Photoneutron cross sections for unstable neutron-rich oxygen isotopes.

The dipole response of stable and unstable neutron-rich oxygen nuclei of masses A = 17 to A = 22 has been investigated experimentally utilizing electromagnetic excitation in heavy-ion collisions at beam energies about 600 MeV/nucleon. A kinematically complete measurement of the neutron decay channel in inelastic scattering of the secondary beam projectiles from a Pb target was performed. Differential electromagnetic excitation cross sections d sigma/dE were derived up to 30 MeV excitation energy. In contrast to stable nuclei, the deduced dipole strength distribution appears to be strongly fragmented and systematically exhibits a considerable fraction of low-lying strength.