The MURAL collection of prostate cancer patient-derived xenografts enables discovery through preclinical models of uro-oncology.

Preclinical testing is a crucial step in evaluating cancer therapeutics. We aimed to establish a significant resource of patient-derived xenografts (PDXs) of prostate cancer for rapid and systematic evaluation of candidate therapies. The PDX collection comprises 59 tumors collected from 30 patients between 2012-2020, coinciding with availability of abiraterone and enzalutamide. The PDXs represent the clinico-pathological and genomic spectrum of prostate cancer, from treatment-naïve primary tumors to castration-resistant metastases. Inter- and intra-tumor heterogeneity in adenocarcinoma and neuroendocrine phenotypes is evident from bulk and single-cell RNA sequencing data. Organoids can be cultured from PDXs, providing further capabilities for preclinical studies. Using a 1 x 1 x 1 design, we rapidly identify tumors with exceptional responses to combination treatments. To govern the distribution of PDXs, we formed the Melbourne Urological Research Alliance (MURAL). This PDX collection is a substantial resource, expanding the capacity to test and prioritize effective treatments for prospective clinical trials in prostate cancer.

Lipid Droplets Accumulate in the Hypothalamus of Mice and Humans with and without Metabolic Diseases.

BACKGROUND: In peripheral tissues, the lipid droplet (LD) organelle links lipid metabolism, inflammation, and insulin resistance. Little is known about the brain LDs. OBJECTIVES: We hypothesized that hypothalamic LDs would be altered in metabolic diseases. METHODS: We used immunofluorescence labeling of the specific LD protein, PLIN2, as the approach to visualize and quantify LDs. RESULTS: LDs were abundant in the hypothalamic third ventricle wall layer with similar heterogeneous distributions between control mice and humans. The LD content was enhanced by high-fat diet (HFD) in both wild-type and in low-density lipoprotein receptor deficient (Ldlr -/- HFD) mice. Strikingly, we observed a lower LD amount in type 2 diabetes mellitus (T2DM) patients when compared with non-T2DM patients. CONCLUSIONS: LDs accumulate in the normal hypothalamus, with similar distributions in human and mouse. Moreover, metabolic diseases differently modify LD content in mouse and human. Our results suggest that hypothalamic LD accumulation is an important target to the study of metabolism.

HSP90 and Its Novel Co-Chaperones, SGT1 and CHP-1, in Brain of Patients with Parkinson's Disease and Dementia with Lewy Bodies.

BACKGROUND: Parkinson's disease (PD) is a neurodegenerative disorder characterized by the presence of inclusions known as Lewy bodies in some brain regions. Lewy bodies consist of α-synuclein and many other proteins including chaperones. OBJECTIVE: To learn more about the role of chaperone complexes in PD and a related disorder, i.e., dementia with Lewy bodies (DLB), in this work we analyzed the expression of HSP90 and its two quite recently identified co-chaperones, SGT1 and CHP-1, in selected brain regions from patients suffering from these diseases. METHODS: To fulfill the aim of our study we used human material and applied immunohistochemistry, Western blot analysis and real time/quantitative PCR (RT-qPCR). RESULTS: We have found that HSP90 mRNA level is higher in the temporal cortex of PD and in frontal cortex of DLB brains, even though level of protein does not change significantly. The mRNA level of SGT1 is higher in the frontal and temporal cortex of PD and in substantia nigra of DLB brains while no significant changes in the level of protein were noticed. Similarly, the mRNA level of CHP-1 was found to be higher in the frontal and temporal cortex of PD and in all examined regions i.e. substantia nigra, frontal and temporal cortex of DLB brains. In the case of CHP-1 the protein level was found to be higher in frontal cortex of PD and in all examined areas of DLB patients. CONCLUSIONS: Our data indicate that the level of HSP90, SGT1 and CHP-1 is upregulated in the majority of cases of PD and DLB, which suggests that the examined proteins might be involved in these pathologies.

Ordo ab Chao: framework for an integrated disease report.

CONTEXT: The volume of information that must be assimilated to appropriately manage patients with complex or chronic disease can make this task difficult because of the number of data points, their variable temporal availability, and the fact that they may reside in different systems or even institutions. OBJECTIVE .- To outline a framework for building an integrated disease report (IDR) that takes advantage of the capabilities of electronic reporting to create a single, succinct, interpretative report comprising all disease pertinent data. DESIGN: Disease pertinent data of an IDR include pathology results, laboratory and radiology data, pathologic correlations, risk profiles, and therapeutic implications. We used cancer herein as a representative process for proposing what is, to our knowledge, the first example of standardized guidelines for such a report. The IDR was defined as a modular, dynamic, electronic summary of the most current state of a patient in regard to a particular illness such as lung cancer or diabetes, which includes all information relevant for patient management. RESULTS: We propose the following 11 core data concepts that an IDR should include: patient identification; patient demographics; disease, diagnosis, and prognosis; tumor board dispositions and decisions; graphic timeline; preresection workup and therapy; resection workup; interpretative comment summarizing pertinent findings; biobanking data; postresection workup; and disease and patient status at follow-up. CONCLUSIONS: A well-executed IDR should improve patient care and efficiency for health care team members. It would demonstrate the added value of pathology interpretation and likely contribute to a reduction in errors and improved patient safety by decreasing the risk that important data will be overlooked.

Assessment of a direct acoustic cochlear stimulator.

This study aimed to assess the functional results of a new, active, acoustic-mechanical hearing implant, the Direct Acoustic Cochlear Stimulation Partial Implant (DACS PI), in a preclinical study. The DACS PI is an electromagnetic device fixed to the mastoid by screws and coupled to a standard stapes prosthesis by an artificial incus (AI). The function of the DACS PI-aided reconstruction was assessed by determining: (1) the maximum equivalent sound pressure level (SPL) of the implant, which was obtained from measurements of the volume displacement at the round window in normal and implanted ears, and (2) the quality at the coupling interface between the AI of the DACS and the stapes prosthesis, which was quantified from measurements of relative motions between the AI and the prosthesis. Both measurements were performed with fresh temporal bones using a scanning laser Doppler interferometry system. The expected maximum equivalent SPL with a typical driving voltage of 0.3 V was about 115-125 dB SPL up to 1.5 kHz in reconstruction with the DACS PI, and decreased with a roll-off slope of about 65 dB/decade, reaching 90 dB SPL at 8 kHz. The large roll-off relative to a normal ear was presumed to be a relatively high inductive impedance of the coil of the DACS PI actuator at higher frequencies. Good coupling quality between the AI and the prosthesis was achieved below the resonance (∼1.5 kHz) of the DACS PI for all tested stapes prostheses. Above the resonance, the SMart Piston, which is composed of a shape-memory alloy, had the best coupling quality.

Providing trust and interoperability to federate distributed biobanks.

Biomedical research requires large numbers of well annotated, quality-assessed samples which often cannot be provided by a single biobank. Connecting biobanks, researchers and service providers raises numerous challenges including trust among partners and towards the infrastructure as well as interoperability problems. Therefore we develop a holistic, open-source and easy-to-use IT infrastructure. Our federated approach allows partners to reflect their organizational structures and protect their data sovereignty. The search service and the contact arrangement processes increase data sovereignty without stigmatizing for rejecting a specific cooperation. The infrastructure supports daily processes with an integrated basic sample manager and user-definable electronic case report forms. Interfaces for existing IT systems avoid re-entering of data. Moreover, resource virtualization is supported to make underutilized resources of some partners accessible to those with insufficient equipment for mutual benefit. The functionality of the resulting infrastructure is outlined in a use-case to demonstrate collaboration within a translational research network. Compared to other existing or upcoming infrastructures, our approach has ultimately the same goals, but relies on gentle incentives rather than top-down imposed progress.

Postmortem and imaging based analyses reveal CNS decreased myelination in restless legs syndrome.

BACKGROUND: Restless legs syndrome (RLS) is a neurological disorder characterized by a strong urge to move the legs and has been shown in many studies with abnormally low brain iron. Iron deficiency is associated with hypomyelination in brains of animals. Therefore we hypothesized that a myelin deficit should be present in the brains of patients with RLS. METHODS: We performed Western blot analysis on myelin isolated from RLS (n=11) and control (n=11) brain tissue obtained at autopsy for the expression of the integral myelin proteins, myelin basic protein (MBP), and proteolipid protein (PLP) and the oligodendrocyte specific enzyme 3'5'-cyclic nucleotide phosphohydrolase (CNPase). To expand the postmortem findings to in vivo, we analyzed the brains of RLS patients (n=23) and controls (n=23) using voxel-based morphometry (VBM). RESULTS: The expression of MBP, PLP and CNPase in the myelin from RLS was decreased by approximately 25% (p<0.05) compared to controls. The amounts of transferrin (Tf) and H-ferritin (H-Frt) in the myelin fraction were also significantly decreased in RLS compared to controls. The imaging analysis revealed significant small decreases in white matter volume in RLS patients compared to controls in the corpus callosum, anterior cingulum and precentral gyrus. CONCLUSION: A decrease in myelin similar to that reported in animal models of iron deficiency was found in the brains of individuals with RLS. The evidence for less myelin and loss of myelin integrity in RLS brains, coupled with decreased ferritin and transferrin in the myelin fractions, is a compelling argument for brain iron insufficiency in RLS. These data also indicate the need to look beyond the sensorimotor symptoms that typically define the syndrome and its assumed relation to the dopaminergic system. Understanding the full range of RLS pathology may help us better understand the complex, intermittent nature and diversity of the clinical features of RLS and expand our consideration of treatment options for RLS.

Efficacy and safety of new medicines: a human focus.

The introduction of safe and effective new medicines is proving ever more difficult, a problem arguably due at least in part to over-reliance on experimental animal-based test systems. In light of the increasing awareness of the lack of predictiveness of such non-human approaches, the necessity to focus on human-based test methods is clear. There has been considerable progress in human in vivo (microdosing) and in silico approaches, primarily to identify ADMET issues, however, in vitro functional studies using human tissues are receiving inadequate attention. The potential scope of human tissue-based research is considerable, but much methodological development is required, which necessitates an increased willingness on the part of the Pharma industry to support it. This approach also requires considerably improved access to the cells and tissues themselves. While current acquisition is almost exclusively from surgery and post mortem, the range of tissue types, the quantity, quality and frequency of supply will remain inadequate to support human tissue as a key component of pre-clinical efficacy and safety testing. Additional routine access to non-transplantable tissues from organ donors for research purposes would be of inestimable value, but in order to realise this, true collaboration will be required between NHS, the Pharma and biotech industries, and the general public.

[Embryonic stem cell research].

Research using human embryonic stem cell (hESC) lines has expanded dramatically because of two attractive capacity; self-renewal and differentiation into almost all cell types. For therapeutic purposes, many researchers are trying to establish methods for maintaining pluripotency in defined xeno-free conditions and scalable culture systems. Banking of hESC lines is important for the wide spread of personalized cell therapy and transplantation. We introduced the ongoing clinical trials using hESC-derived cells in patients with subacute spinal cord injury and Stargardt's macular dystrophy. We also discussed opportunities and an example for the use of hESC in drug discovery. Finally, we introduced transgenic hESC as a disease model.

Banco de tejido ovárico para jóvenes con cáncer de mama: informe preliminar de los primeros casos / Ovarian tissue bank for young women with breast cancer

Introducción: el número de pacientes jóvenes con cáncer de mama parece ser mayor en Uruguay que en los promedios internacionales. Muchas de estas jóvenes no tienen hijos y la consejería en reproducción es actualmente recomendada por las sociedades científicas de referencia. En nuestro país funciona desde 2005 un programa de Preservación de la Función Ovárica ante el Cáncer, en el ámbito del Centro Hemato-Oncológico Pediátrico del CentroHospitalario Pereira Rossell, que realiza consejería y técnicas de riopreservación de tejido ovárico destinadas a pacientes de todos los ámbitos y en forma gratuita. Objetivos: el objetivo principal de este trabajo es presentar los primeros casos de cáncer de mama en pacientes jóvenes que ingresaron en el protocolo de criopreservación de tejido ovárico. El objetivo secundario es difundir y lograr incorporar en la práctica clínica la derivación de las pacientes jóvenes sin hijos que reciban este diagnóstico a una consejería en reproducción. Material y método: en el marco del protocolo previamente aprobado por la Facultad de Medicina se tomaron los primeros cinco casos de pacientes que ingresaron con diagnóstico de cáncer de mama, sin hijos jóvenes y en estadios iniciales. La extracción de tejido ovárico se realizó luego de la cirugía oncológica y antes del tratamiento complementario mediante laparoscopía. El tejido se almacenó en el Instituto Nacional de Donación y Trasplantes de Células, Tejidos y Órganos (INDYT). Resultados: cinco pacientes de entre 26 y 35 años ingresaron al protocolo en el período mayo de 2006 a enero de 2009. Una de las pacientes falleció, comprobándose que era un estadio superior al planteado inicialmente. Las cuatro pacientes restantes están en tratamiento complementario con buena evolución y en amenorrea. Por lo tanto, no están planteados procedimientos de reimplante al momento.

Introduction: the number of young patients with breast cancer seems to be higher in Uruguay than international averages. Many of these young women have no children and reproductive counseling is currently recommendedby the reference scientific societies. In Uruguay, an ovarian function preservation program for cancer patients has been operative since 2005, at the Pereira Rossell Hosital Pediatrics Hemato-Oncologic Center. The program provides for counseling and ovarian tissue cryopreservation techniques for all patients, for free. Objectives: the main objective of this study is to present the first breast cancer cases in young patients who entered into the ovarian tissue cryopreservation protocol. The secondary objective is to disseminate the reference of young patients with no children who are diagnosed with breast cancer to reproductive counseling, and to include this into clinical practice. Method: within the framework of the protocol previouslyapproved by the School of Medicine, the first five cases of patients admitted with a diagnosis of early stage breast cancer who had no children were included in the study. Extraction of ovarian tissue was performed after the oncologic surgery and before the laparoscopic complementary treatment. Ovarian tissue was stored in the National Institute of Donation and Cell, Tissue and Organ Transplant. Results: five patients between 26 and 35 years old entered into the protocol from May 2006 through January 2009. One of the patients died and thus, it was proved that the disesae stage was more advanced than initially diagnosed. The four remaining patients are undergoing complementary treatment and evidence a good evolution and amenorrhea. Therefore, no reimplantation procedures have been defined so far.

Introdução: o número de pacientes jovens com câncer de mama parece ser maior no Uruguai do que as médias internacionais. Muitas dessas jovens não têm filhos e atualmente o aconselhamento em reprodução é recomendado pelas sociedades científicas de referência. Desde 2005 funciona no Uruguai um programa de Preservação da Função Ovárica frente ao Câncer no Centro Hemato-Oncológico Pediátrico do Hospital Pereira Rossell que realiza aconselhamento e técnicas de criopreservação de tecido de ovário, destinadas a pacientes de todos os âmbitos e gratuitamente. Objetivos: o objetivo principal deste trabalho é apresentar os primeiros casos de câncer de mama em paciente jovens incluídas no protocolo de criopreservação de tecido de ovário. O objetivo secundário é difundir e incorporar à prática clínica a derivação das pacientes jovens, sem filhos, que recebam esse diagnóstico ao aconselhamento em reprodução. Material e método: de acordo com um protocolo aprovado previamente pela Faculdade de Medicina-UdelaR ingressaram ao programa as cinco primeiras pacientes jovens, sem filhos com diagnóstico de câncer de mama em fase inicial. A extração do tecido ovárico foi realizada por laparoscopia depois da cirurgia oncológica e antes do tratamento complementar. O tecido foi armazenado no Instituto Nacional de Doação e Transplantes de Células, Tecidos e Órgãos (INDYT). Resultados: cinco pacientes, com idades entre 26 e 35 anos, foram ingressadas ao protocolo no período maio de 2006 - janeiro de 2009. Uma das pacientes faleceu comprovando que o estadio da neoplasia era superior ao inicialmente considerado. As demais pacientes estão recebendo tratamento complementar com boa evolução e amenorréia; por essa razão ainda não foram realizados procedimentos de reimplantaçao.

The Organization of European Cancer Institute Pathobiology Working Group and its support of European biobanking infrastructures for translational cancer research.

BACKGROUND: Today's translational cancer research increasingly depends on international multi-center studies. Biobanking infrastructure or comprehensive sample exchange platforms to enable networking of clinical cancer biobanks are instrumental to facilitate communication, uniform sample quality, and rules for exchange. METHODS: The Organization of European Cancer Institutes (OECI) Pathobiology Working Group supports European biobanking infrastructure by maintaining the OECI-TuBaFrost exchange platform and organizing regular meetings. This platform originated from a European Commission project and is updated with knowledge from ongoing and new biobanking projects. This overview describes how European biobanking projects that have a large impact on clinical biobanking, including EuroBoNeT, SPIDIA, and BBMRI, contribute to the update of the OECI-TuBaFrost exchange platform. RESULTS: Combining the results of these European projects enabled the creation of an open (upon valid registration only) catalogue view of cancer biobanks and their available samples to initiate research projects. In addition, closed environments supporting active projects could be developed together with the latest views on quality, access rules, ethics, and law. CONCLUSIONS: With these contributions, the OECI Pathobiology Working Group contributes to and stimulates a professional attitude within biobanks at the European comprehensive cancer centers. IMPACT: Improving the fundamentals of cancer sample exchange in Europe stimulates the performance of large multi-center studies, resulting in experiments with the desired statistical significance outcome. With this approach, future innovation in cancer patient care can be realized faster and more reliably.

Preparing for growth: current capacity and challenges in hematopoietic stem cell transplantation programs.

During the past decade, the demand for hematopoietic stem cell transplantation has grown dramatically, and there are expectations that this will continue or even accelerate over the next decade. This prompts a variety of questions about the ability of the health care system to accommodate the increased demands on transplantation centers; for example, what is the current patient capacity of transplantation programs, and how much elasticity do they have to accept a larger volume of patients? An informal survey of a sample of medical directors of transplantation programs found that existing facilities might be able to increase their patient volume by about 7%. Expanding much beyond that limit will require an infusion of resources to enlarge current programs and/or establish new programs.

[Ethical aspects of human embryonic stem cell use and commercial umbilical cord blood stem cell banking. Ethical reflections on the occasion of the regulation of the European Council and Parliament on advanced therapy medicinal products]. / Bioethische und sozialethische Probleme bei der Verwendung von Geweben aus humanen embryonalen Stammzellen sowie der Aufbewahrung von Stammzellen aus dem Nabelschnurblut durch private Firmen zum autologen Gebrauch. Ethische Uberlegungen anlässlich der Verordnung (EG) Nr. 1394/2007 des europäischen Parlaments und des Rates über Arzneimittel für Neuartige Therapien.

The regulation of the European Council and Parliament on advanced therapy medicinal products also includes therapies with human embryonic stem cells. The use of these stem cells is controversially and heavily discussed. Contrary to the use of adult stem cells, medical and ethical problems concerning the use of human embryonic stem cells persists, because this use is based on the destruction of human life at the very beginning. The regulation foresees, therefore, subsidiarity within the European Member States. Although there are no ethical problems in principle with the use of stem cells from the umbilical cord blood, there are social ethical doubts with the banking of these stem cells for autologous use without any currently foreseeable medical advantage by commercial blood banks. Also in this case subsidiarity is valid.

[Tissue bank of the National Centre for Tumour Disease. An innovative platform for translational tumour]. / Gewebebank des Nationalen Zentrums für Tumorerkrankungen. Innovative Plattform für die translationale Tumorforschung.

The tissue bank of the National Centre for Tumour Diseases (NCT) in Heidelberg, Germany, was founded in 2005 by the University Hospital of Heidelberg and the German Cancer Research Centre as a section of the NCT. It is a nonprofit organization with a completely evaluated legal and ethical framework and supports the Comprehensive Cancer Centre concept. Its main aim is the acquisition and characterization of fresh-frozen and paraffin-embedded human tissues according to the standards of good scientific practice and the promotion of interdisciplinary tumour research of the comprehensive cancer centre and its cooperating partners. It also offers expert project assistance: a project leader can submit a short proposal, and the tissue collecting/preparing process will be performed in cooperation with a specialised pathologist and, if applicable, an experienced clinical researcher. The tissue bank is also a central platform for further developing of innovative technologies for tissue handling, e.g. multi-tissue-array and virtual microscopy, with links to digital image analysis and bioinformatics. Thus, the NCT tissue bank represents a model for innovative biobanking and for institutions with active interdisciplinary cancer research.

Tissue-based research in kidney cancer: current challenges and future directions.

The past several years have seen unprecedented advances in the application of various therapeutic strategies for the treatment of patients with renal cancer. The availability of active immunotherapy, antiangiogenic therapy, and targeted therapy for this disease has brought front and center issues related to choosing the appropriate treatment for particular patient populations. It is increasingly evident that the most promising treatment selection strategies will incorporate identifying specific features of the tumor itself. To facilitate this move toward personalized medicine, it is critically important to establish some standard principles for renal cancer tissue collection, preparation, and analysis for translational research studies. In this article, we identify and discuss some critical issues related to tissue-based kidney cancer research. We focus on five major areas as follows: (a) surgical and image-guided techniques for tissue collection; (b) quality control of specimen collection, processing, storage, and review; (c) issues related to analysis of paraffin embedded tissues; (d) genomic studies; and (e) assessment of reproducibility of assays across institutions. In addition, some practical implementation strategies are proposed. Although many of the topics discussed are specific for renal cancer, several are also relevant to tissue based biomarker investigations in a broad array of malignancies.

The Environmental Specimen Bank for Human Tissues as part of the German Environmental Specimen Bank.

The German Environmental Specimen Bank for Human Tissues (ESBHum) as part of the German Environmental Specimen Bank (ESB) focuses on documenting and assessing trends of human exposure via real-time monitoring of body burden and long-term storage of samples under stable deep freezing conditions (-150 degrees C) for later retrospective analyses. Real-time monitoring is performed after completing sampling processes of one year and covers actually 20 inorganic and 5 organic substances. While concentrations of several substances, e.g., arsenic, cadmium and mercury, are remained unchanged over time, other substances, e.g., lead and pentachlorophenol (PCP), show a clearly perceptible decrease. Substances which are not routinely analyzed in real-time monitoring are retrospectively measured by indication in the stored human specimens. Indications of retrospective monitoring are availability of valid analytical methods, e.g., in case of PCDF and PCDD, or assessment of concentration trends of substances with actual interest of toxicology and/or environmental medicine, e.g., polybrominated diphenyl ethers (PBDE), perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA). While over time the body burden of dioxins as well as PFOS and PFOA decreased, the PBDE concentrations in human blood increase. The observed decrease of blood lead and PCP levels over time is a consequence of legal prohibition and restriction. The time-dependent concentrations of the aforementioned substances agree with results of other national studies. So it can be concluded that the German ESBHum is an important instrument for health-related environmental observation and protection in Germany.

A Non-Frozen Living Tissue Bank for Allotransplantation Using Green Tea Polyphenols

Generally, mammalian cells and living tissues can be cryopreserved in a frozen state at very low temperatures over a long storage term. The survival rate of cell suspensions is often acceptable however, living tissues suffer a variety of injuries. In this paper, it was demonstrated that the addition of polyphenols extracted from green tea to conventional cell culture medium and tissue compatible liquid, can control cell proliferation and also preserve tissues for several months at ordinary room temperature, including such tissues as blood vessels, cartilage, islet cells and corneas. This protocol allows a non-frozen living tissue bank to be established using the preservation fluid described.