Production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans

Rabies kills many people throughout the developing world every year. The murine monoclonal antibody (mAb) 62-71-3 was recently identified for its potential application in rabies postexposure prophylaxis (PEP). The purpose here was to establish a plant-based production system for a chimeric mouse-human version of mAb 62-71-3, to characterize the recombinant antibody and investigate at a molecular level its interaction with rabies virus glycoprotein. Chimeric 62-71-3 was successfully expressed in Nicotiana benthamiana. Glycosylation was analyzed by mass spectroscopy; functionality was confirmed by antigen ELISA, as well as rabies and pseudotype virus neutralization. Epitope characterization was performed using pseudotype virus expressing mutagenized rabies glycoproteins. Purified mAb demonstrated potent viral neutralization at 500 IU/mg. A critical role for antigenic site I of the glycoprotein, as well as for two specific amino acid residues (K226 and G229) within site I, was identified with regard to mAb 62-71-3 neutralization. Pseudotype viruses expressing glycoprotein from lyssaviruses known not to be neutralized by this antibody were the controls. The results provide the molecular rationale for developing 62-71-3 mAb for rabies PEP; they also establish the basis for developing an inexpensive plant-based antibody product to benefit low-income families in developing countries.—Both, L., van Dolleweerd, C., Wright, E., Banyard, A. C., Bulmer-Thomas, B., Selden, D., Altmann, F., Fooks, A. R., Ma, J. K.-C. Production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans

Not Lost in Translation: Multi-Perspective Collaborative Analysis With the Listening Guide, Illustrated by a Case of a Woman in Combat

Der Listening Guide (LG) ist eine qualitative Methodologie, die ursprünglich von GILLIGAN (1982) und Kolleg*innen (BROWN et al. 1988) entwickelt wurde und ihre Wurzeln in der relationalen feministischen Psychologie hat. In diesem Beitrag stellen wir das Potenzial der daraus entwickelten Methoden für eine differenzierte Analyse von Interviewdaten in einer multidisziplinären und multinationalen Zusammenarbeit vor. Dafür beschreiben wir unseren gemeinsamen Prozess als israelische und deutsche Wissenschaftlerinnen, die ein exemplarisches Datenstück aus einem Interview mit einer Soldatin analysierten. Wir zeigen, wie die Verwendung von LG-Methoden mit einem expliziten Fokus auf dem sozialen Kontext des Interviews sowie der Datenanalyse unser Verständnis der Daten verbessert hat. Darüber hinaus wird dargelegt, inwiefern die Anwendung dieser Methoden hilfreich war, um die Aufmerksamkeit auf den kollaborativen Prozess zu richten und eine reflexive Haltung sowie Transparenz bezüglich des Analyseprozesses zu fördern. Da die Arbeit mit übersetzten Daten für ein mehrsprachiges Team eine Notwendigkeit darstellt, diskutieren wir auch, wie die Anwendung der LG-Methoden den Blick auf Sprache, Grammatik und darin eingebettete Bedeutungen lenkt. Wir kommen zu dem Schluss, dass die LG-Methoden für kollaborative Forschungsprozesse von Vorteil sind, insbesondere um die verschiedenen Ebenen der sozialen Einbettung des Erzählten in den Daten, der Datenerhebung und der Datenanalyse zu berücksichtigen.The listening guide (LG) is a qualitative methodology developed by GILLIGAN (1982) and colleagues (BROWN et al., 1988), rooted in relational feminist psychology. In this paper, we present the potential of its methods for nuanced analysis of interview data when working together as a multidisciplinary and multinational team. To this end, we describe our collaborative process as Israeli and German scholars analyzing a sample piece of data from an interview with a female combat soldier. We highlight how using LG methods with an explicit focus on the social context of the interview and the data analysis enhanced our understanding of the data. In addition, we outline the ways in which using these methods was helpful in focusing on the collaborative process and promoting a reflexive stance as well as transparency about the analysis process. Since working with translated data is a necessity for a multilingual collaboration, we also discuss how using LG methods draws attention to language, grammar, and embedded meanings. We conclude that the LG methods we used are beneficial to collaborative research processes, especially when researchers use them to attend to different levels of social embeddedness of the narratives in data, data collection, and data analysis

Pharmacokinetics, pharmacodynamics, and safety of moss-aGalactosidase A in patients with Fabry disease

Moss-aGalactosidase A (moss-aGal) is a moss-derived version of human α-galactosidase developed for enzyme replacement therapy in patients with Fabry disease. It exhibits a homogenous N-glycosylation profile with >90% mannose-terminated glycans. In contrast to mammalian cell produced α-galactosidase, moss-aGal does not rely on mannose-6-phosphate receptor mediated endocytosis but targets the mannose receptor for tissue uptake. We conducted a phase 1 clinical trial with moss-aGal in six patients with confirmed diagnosis of Fabry disease during a 28-day schedule. All patients received a single dose of 0.2 mg/kg moss-aGal by i.v.-infusion. Primary endpoints of the trial were safety and pharmacokinetics; secondary endpoints were pharmacodynamics by analyzing urine and plasma Gb3 and lyso-Gb3 concentrations. In all patients, the administered single dose was well tolerated. No safety issues were observed. Pharmacokinetic data revealed a stable nonlinear profile with a short plasma half-life of moss-aGal of 14 minutes. After one single dose of moss-aGal, urinary Gb3 concentrations decreased up to 23% 7 days and up to 60% 28 days post-dose. Plasma concentrations of lyso-Gb3 decreased by 3.8% and of Gb3 by 11% 28 days post-dose. These data reveal that a single dose of moss-aGal was safe, well tolerated, and led to a prolonged reduction of Gb3 excretion. As previously shown, moss-aGal is taken up via the mannose receptor, which is expressed on macrophages but also on endothelial and kidney cells. Thus, these data indicate that moss-aGal may target kidney cells. After these promising results, phase 2/3 clinical trials are in preparation

Hyperbaric Oxygen Therapy Can Induce Neuroplasticity and Significant Clinical Improvement in Patients Suffering From Fibromyalgia With a History of Childhood Sexual Abuse—Randomized Controlled Trial

Background: Fibromyalgia syndrome (FMS), a condition considered to represent a prototype of central sensitization syndrome, can be induced by different triggers including childhood sexual abuse (CSA). Recent studies have demonstrated hyperbaric oxygen therapy (HBOT) can induce neuroplasticity and improve clinical outcome of FMS. The aim of the current study was to evaluate the effect of HBOT on patients suffering from FMS with a history of CSA.Materials and methods: A prospective randomized clinical trial conducted between July 2015 and November 2017 included women with a history of CSA who fulfilled fibromyalgia diagnosis criteria for at least 5 years prior to inclusion. Included participants (N = 30) were randomly assigned to treatment group, treated with 60 HBOT sessions and a control/crossover group received psychotherapy. After the control period, the control/crossover group was crossed to HBOT. Clinical outcomes were assessed using FMS questioners, post-traumatic stress disorder (PTSD) questioners and quality of life questioners. Objective outcome were assessed using brain function and structure imaging.Findings: Following HBOT, there was a significant improvement in all FMS questionnaires (widespread pain index, Fibromyalgia symptoms severity scale, Fibromyalgia functional impairment), most domains of quality of life, PTSD symptoms and psychological distress. The same significant improvements were demonstrated in the control following crossover to HBOT. Following HBOT, brain SPECT imaging demonstrated significant increase in brain activity in the prefrontal cortex, orbital frontal cortex, and subgenual area (p < 0.05). Brain microstructure improvement was seen by MRI-DTI in the anterior thalamic radiation (p = 0.0001), left Insula (p = 0.001), and the right Thalamus (p = 0.001).Conclusion: HBOT induced significant clinical improvement that correlates with improved brain functionality and brain microstructure in CSA related FMS patients.Trial Registration:www.Clinicaltrials.gov, identifier: NCT03376269. url: https://clinicaltrials.gov/show/NCT0337626

Brain and Mind Integration: Childhood Sexual Abuse Survivors Experiencing Hyperbaric Oxygen Treatment and Psychotherapy Concurrently

Due to evidence that traumatic experience impacts the brain, the body (concerning sensory sensitivity), and the mind, a recent study that attempted to answer the question of whether the effects of CSA can be reversed by using a multidisciplinary approach consisting of dual treatments: hyperbaric & psychotherapy, was conducted. Its results showed that in addition to improvement of brain functionality, symptoms of distress were significantly reduced. The current paper aims to present the process as experienced by the 40 female childhood sexual abuse survivor participants. Data included participants' daily journals and drawings, and participants' summaries presented verbally and written, 6-months after the study ended. A phenomenological analysis was used. Results showed three phases, the initial phase—remembering the trauma from both physical and cognitive aspects, the second phase—physiological relaxation as well as positive memories emerge; and, the third phase—bouncing back to life. The results are discussed in light of the study theoretical model and Lev-Wiesel (2015) childhood sexual abuse conceptualization

Serum Apolipoproteins C-I and C-III Are Reduced in Stomach Cancer Patients: Results from MALDI-Based Peptidome and Immuno-Based Clinical Assays

Finding new peptide biomarkers for stomach cancer in human sera that can be implemented into a clinically practicable prediction method for monitoring of stomach cancer. We studied the serum peptidome from two different biorepositories. We first employed a C8-reverse phase liquid chromatography approach for sample purification, followed by mass-spectrometry analysis. These were applied onto serum samples from cancer-free controls and stomach cancer patients at various clinical stages. We then created a bioinformatics analysis pipeline and identified peptide signature discriminating stomach adenocarcinoma patients from cancer-free controls. Matrix Assisted Laser Desorption/Ionization–Time of Flight (MALDI-TOF) results from 103 samples revealed 9 signature peptides; with prediction accuracy of 89% in the training set and 88% in the validation set. Three of the discriminating peptides discovered were fragments of Apolipoproteins C-I and C-III (apoC-I and C-III); we further quantified their serum levels, as well as CA19-9 and CRP, employing quantitative commercial-clinical assays in 142 samples. ApoC-I and apoC-III quantitative results correlated with the MS results. We then employed apoB-100-normalized apoC-I and apoC-III, CA19-9 and CRP levels to generate rules set for stomach cancer prediction. For training, we used sera from one repository, and for validation, we used sera from the second repository. Prediction accuracies of 88.4% and 74.4% were obtained in the training and validation sets, respectively. Serum levels of apoC-I and apoC-III combined with other clinical parameters can serve as a basis for the formulation of a diagnostic score for stomach cancer patients