DATE: February 09, 2021 TIME: 08:00am PST Flow cytometry immunophenotyping has become one of the mainstream applications for classification of several hematologic neoplasms. This technology is indispensable for detection of leukemic blasts at a single cell level, clonal lineage assignment, identification of aberrant expression of antigens, and detection of abnormal rare populations of blasts from normal progenitors, with extreme importance in tailoring decision-making. Moreover, functional flow cytometry testing can effectively provide new insights for research and evaluation of disease, and a more complete understanding of the complexities and challenges in the analysis of leukemic stem cells. In this webinar we will discuss simpler, faster and more affordable methods minimizing the effect of sample preparation on the sample biology. White blood cells in peripheral whole blood, among the most commonly analyzed cells in flow cytometry, are fundamentally rare in their native sample matrix, where red blood cells outnumber white cells by up to three orders of magnitude and platelets by nearly two orders. The challenges this rarity creates for no lyse no wash assays are the primary reason this technique does not enjoy wider use. Specifically, we will discuss how to detect candidate malignant primitive progenitor populations, using the alkaline phosphatase stem cell detection method in combination with flow cytometry immunophenotyping. Over a period of five years, we have been using this technique to study its activity in patients with leukemia and lymphoma, showing that changes in the alkaline phosphatase levels can be used to detect rare populations of highly refractory malignant cells. By screening different blood cancers, we have observed that this activity is not always restricted to CD34+ leukemic cells and can be overexpressed in CD34 negative leukemia. Moreover, we will discuss how to use these methods minimizing the effect of sample preparation on the PD-L1 expression at the single cell level in myeloid derived suppressor cells using a stimulatory functional assay to evaluate immunotherapy strategies in human cancer. Learning Objectives: Provide a short introduction and discuss the methods needed for minimal sample perturbation, cell damage and the number of variables affecting the cancer cell biology. Discuss flow cytometric significance of cellular alkaline phosphatase activity in acute myeloid leukemia PD-L1 expression at the single cell level in Myeloid Derived Suppressor Cells using a stimulatory functional assay to evaluate immunotherapy strategies Webinars will be available for unlimited on-demand viewing after live event. LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.

The fast spread and deadliness of SARS-CoV-2 has sparked much interest in understanding the underlying genomics and evolutionary patterns of this 30,000bp Coronavirus. Since the first reported case on January 19th, 2020, over 200,000 genomes are now available. Within this large set of SARS-CoV-2 genomic data, thousands of mutations have been, including the D614G mutation that now is found in nearly all genomes and mink-associated variants. In this talk, I will take a deep dive into the intrahost and interhost diversity of the virus responsible for the COVID-19 pandemic. Specifically, I will discuss within-host minor variants and consensus-level variants that are found across hosts and describe population-wide structural variations that highlight this underexplored intrahost diversity. Implications of these findings include improved transmission cluster identification and better understanding of how SARS-CoV-2 is changing over time. Learning Objectives: 1. Differentiating consensus-level vs within-host variation of SARS-CoV-2 2. Discovering the utility of genomic variants on tracking SARS-CoV-2 transmission 3. Understanding of how SARS-CoV-2 is changing over time

As part of a large collaborative effort, we have used systems biology approaches to identify host factors that participate in SARS-CoV-2 replication. Protein-protein interactions maps between viral and host proteins and global landscapes on protein phosphorylation changes taking place in SARS-CoV-2 infected cells were used as guides for the identification of host processes that could be targeted for antiviral therapeutic intervention. Drugs known to interact with these host processes and with known clinical profiles were tested for their ability to inhibit SARS-CV-2 replication in tissue culture. As a result, we found several promising therapeutic candidates to be evaluated in vivo for the treatment of COVID-19. Learning Objectives: 1. Understand the concept of drug re-purposing 2. Gain insights on the use of system biology approaches for the discovery of antivirals 3. Gain insights on the host-virus protein interactions contributing to SARS-CoV-2 replication

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 and has spread globally, causing a pandemic of respiratory illness designated coronavirus disease 2019 (COVID-19). Two studies on the immunology of COVID-19 are included in this webinar: First, Miguel Muñoz-Ruiz, PhD from the Francis Crick Institute and his colleagues have identified several traits, including cytokine and chemokine profiles, and the status of defined immune cell subsets that may facilitate the early identification of COVID-19 patients at greatest risk for requiring prolonged hospital treatment. Second, Arutha Kulasinghe, PhD and his colleagues at Queensland University of Technology used spatial transcriptomics to generate an in-depth picture of the pulmonary transcriptional landscape of patients who died of COVID-19, and compared it to those who died of pandemic H1N1 (pH1N1) influenza and uninfected control patients. Host transcriptomics showed a significant upregulation of genes associated with inflammation, type I interferon production, coagulation and angiogenesis in the lungs of COVID-19 patients compared to non-infected controls. Learning Objectives: 1. Understand the current knowledge on the dynamics of the immune response to COVID-19 2. Learn what immune signatures are associated with a poor COVID-19 prognosis 3. Understand the differences between transcriptional profiles of lungs from COVID-19 patients versus those infected with influenza and non-infected controls 4. Learn how NanoString nCounter® technology and the GeoMx® Digital Spatial Profiler can be used to study COVID-19

DATE: November 5, 2020 TIME: 7am PST, 10am EST, 4pm CET Exosomes are a population of naturally occurring mobile, membrane-limited, 30 – 100 nm in diameter, extracellular vesicles containing a large number of proteins, lipids, messenger, and micro-RNAs. It was shown that they play a role in the mediation of intercellular communication, the modulation of immune-regulatory processes, tumor metabolism, and regenerative as well as degenerative processes. In recent years, there is increasing interest in the therapeutic potential of exosomes produced by Mesenchymal Stem Cells (MSCs). Stirred-tank bioreactors offer the possibility to tightly control and monitor the production of exosomes, as well as the scalability, to produce increasing amounts. However, the cultivation of stem cells in stirred-tank bioreactors requires profound knowledge and precise control of the process. In our talk, we will highlight the potential and benefits of stirred-tank bioreactors in the cultivation of stem cells for exosome production. We will share our knowledge on how we developed the process in our DASbox® Mini Parallel Bioreactor System and scaled-up the process to 1 L controlled by our newest parallel bioreactor control system, the SciVario® twin. Target groups: · Scientists new to the cultivation of stem cells in bioreactors · Experienced scientists looking for tips and tricks to improve their stem cell culture and EV production processes Learning Objectives: How to cultivate stem cells in a stirred-tank bioreactor: Learn how to identify the right cultivation method, vessel type, vessel material, and instrument configuration for your specific needs Learn how to define and optimize important parameters to improve culture results – incl. Tips & Tricks Live “Ask the Expert” session – Deepen your knowledge and optimize your process This webinar is part of a virtual event. With attending the live webinar you will also have access to the Eppendorf Planet of Knowledge Platform on which more engaging and educational content about stem cell bioprocessing will be provided. Webinars will be available for unlimited on-demand viewing after live event. LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.

The fast spread of SARS-Cov-2 sparked much interest in understanding the underlying genomics of this 30,000bp Coronavirus. Thus far, thousands of genome assemblies are available, yet they feature only a relatively small number of single nucleotide polymorphisms (SNPs) that differentiate the main SARS-CoV-2 clades that have evolved while rapidly spreading throughout the world. In this talk, I will describe the mutational landscape of thousands of SARS-Cov-2 genomes and take a deep dive into the intrahost diversity of this devastating virus. Specifically, I will discuss within-host minor variants that are found across hosts and describe population-wide structural variations that highlight this underexplored intrahost diversity. Implications of these findings include both high false-negative rates of qPCR-based tests and as a key tool in transmission analyses. Learning Objectives: 1. Differentiating consensus-level vs within-host variation of SARS-CoV-2 2. Discovering the potential impact of single nucleotide variation on COVID-19 diagnostics 3. Understanding the utility of minor variants on tracking SARS-CoV-2 transmission

The spread of the SARS-COV-2 (COVID-19) pandemic is driving an urgent need for extensive testing, including patients and health workers as a priority. The most sensitive and reliable method remains the RT-PCR molecular swab test but one challenge that still remains is to increase the throughput of the RNA extraction step in the process. Cytiva (formerly GE Healthcare LifeSciences) has developed Sera-Xtracta™Viral/Pathogen Kit which provides a highly automatable and sensitive extraction protocol, compatible with open system platforms and downstream RT-PCR processes. Learning Objectives: 1. Current COVID-19 diagnostic technologies 2. Effective RNA extraction through magnetic bead technology 3. Relevant performance data of magnetic bead extraction

DATE: July 29, 2020 TIME: 9:00am PDT, 12:009m EDT, 6:00pm CEST Hepatitis C infection, once a formidable enemy, is now a curable disease. Dramatic advances in antiviral therapies have launched a new era in how hepatitis C is treated and in turn have significantly improved the prognosis once an infection is identified. Unfortunately, many individuals today who are infected with hepatitis C are completely unaware of infection status; it is estimated that only 20% of cases worldwide are diagnosed. Only when individuals are tested and diagnosed with a hepatitis C virus (HCV) infection can they be treated and reap the benefits of these therapeutic innovations. Bridging the gap between unaware and diagnosed can make huge differences in a patient’s long-term prognosis. Learning Objectives: Underatand HCV disease state overview Discuss the Role of Dried Blood Spots (DBS) in management of HCV infection Discuss the Performance of m2000 RealTime HCV DBS Discuss Implementation strategies for HCV elimination Webinars will be available for unlimited on-demand viewing after live event. LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.

Brain function is remarkably reliable despite the imprecise performance of neurons and the continuous perturbations caused by aging, disease or injury. How does the brain succeed in producing stereotypic behaviors over long periods of time despite these perturbations? We are interested in studying the cellular and system mechanisms by which neuronal circuits are able to "self-tune" and adapt to perturbations. We are using genetic manipulations to perturb brain circuits to ask two types of questions: (i) how do brain circuits adapt when a large percentage of their neurons are deleted or silenced?, (ii) how do brain circuits adapt when electrical noise is injected into the system?. To study these questions we are focusing on the song circuits of birds because song is an extremely stereotypical behavior that can be rigorously measured. Learning Objectives: 1. Define the concepts of brain resilience and behavioral stereotypy 2. Explain the experimental methods that can be used to investigate brain resilience 3. Explain how investigating mechanisms of brain resilience in animals could be useful to improve outcomes in humans after diseases such as stroke

Since the establishment of reverse genetics techniques to manipulate the influenza virus genome, it has been possible to study viral molecular signatures responsible for virulence, and the generation of novel and improved vaccine strains. Moreover, high throughput screen technologies combined with functional genomics, transcriptomics and proteomics are now been used to identify novel mechanisms associated with virus replication and pathogenicity, as well as novel antivirals. Finally recombinant technologies can now be applied for the generation of broadly neutralizing influenza virus vaccines that could eliminate the need for annual vaccination and prevent pandemics by inducing protective immunity against conserved epitope shared by all influenza virus strains. Learning objectives: 1. Current influenza vaccines and antivirals are useful, but not optimal 2. New technologies allow the discovery of improved influenza antivirals and vaccines

With humans pushing to live further off Earth for longer periods of time, it is increasingly important to understand the changes that occur in biological systems during spaceflight whether these be astronauts, their microbial commensals, or their plant-based life support systems. In a three-part presentation, we discuss GeneLab and recent discoveries regarding the microbiota of spacecrafts and space-flown animals. Part 1: GeneLab: Open Science for Life in Space, Jonathan Galazka, NASA Ames Research Center Abstract: To accelerate the pace of discovery from precious spaceflight biological experiments, NASA as develop the GeneLab data system (genelab.nasa.gov), which allows unfettered access to omics data from spaceflight and spaceflight relevant experiments. GeneLab houses metagenomic datasets from spacecraft and relevant spacecraft models. Users can download this data and associated metadata to make new discoveries about how microbial communities may change and adapt to spaceflight. Part 2: Reproducible changes in the gut microbiome suggest a shift in microbial and host metabolism during spaceflight, Peng Jiang, Northwestern University Abstract: The gastrointestinal microbiota interacts with multiple aspects of mammalian physiological functions. Microbiome changes in response to the challenging space environmental factors, such as microgravity and radiation, are thus thought to be important for astronaut health during long-term missions. Spaceflight-associated changes in the gut microbiome include an elevated microbial diversity and an altered community structure, which appeared to be consistent across studies in both humans and mice. Ongoing-studies are aiming to understand microbiome-host interactions during spaceflight and the significance of such interactions impact mammalian functions such as metabolism, immune functions, and sleep. Part 3: Novel technologies to identify Antimicrobial resistances in hospitals, urban environments and on the NASA International Space Station Abstract: With the revolution of next-generation sequencing technologies the field of microbiome and metagenomics research continues to expand and transform several fields. The Extreme Microbiome Project (XMP) launched in 2014 characterizes the microbial communities of extreme sites on Earth. In 2015 we launched the International MetaSUB consortium with more than 200 members in 25 countries. We experimenting with new technologies to find better solutions to for the remote and rapid sequencing of infectious diseases in hospitals, on the International Space Station (ISS) and in NASA clean rooms to inform the spacecraft assembly engineers and biological scientists of any potential bacterial or human contamination. Furthermore, we have been testing a broad range of cutting-edge technologies in the NASA Twins study. To better understand the impact of spaceflight on the human body and to prepare for future exploration-class missions, a pair of identical twin astronauts was monitored before, during, and after a one-year mission resulting in one of the most comprehensive studies ever have been made on one individuum.

DATE: September 17, 2019 TIME: 9:00am PDT Spatial Transcriptomics (ST) technology reveals gene expression from up to one thousand spots across a tissue section. In this webinar, Dr. Hanna Eriksson and Kim Thrane from Karolinska Institute will present the workflow of ST and show how they adapted the protocol for lymph node metastasis of cutaneous malignant melanoma (CMM). By exploring the spatial gene expression data from four lymph node metastasis, they confirmed the anticipated inter- and intratumoral heterogeneity as well as identify a shared gene expression profile in lymphoid tissue. ST is believed to be a valuable tool for a better understanding of tumor progression and responses to treatment. Kim will discuss: Spatial Transcriptomics - Bridging histology and RNA-seq The Spatial Transcriptomics (ST) technology provides transcriptome-wide RNA-seq data from hundreds of 100µm-sized spots within a tissue section. In this presentation I will go through the principles of the method, describe the tissue optimization procedure and show examples on data visualization. I will also talk about how Spatial Transcriptomics has been further developed by 10x genomics to become Visium, with increased sensitivity and higher spatial resolution. Hanna will discuss: Exploring genetic heterogeneity in Stage III Cutaneous Malignant Melanoma The prognosis in melanoma differ markedly within stage groups and melanoma is one of the most immunogenic and genetically heterogenous of all cancers. The ST technology based on melanoma lymph node biopsies has successfully generated sequencing data of transcriptomes from over 2,200 tissue domains. By using a strategy for deconvolution along with traditional approaches for dimensionality reduction of transcriptome-wide data, the transcriptional landscape within the tissue was visualized and gene expression profiles linked to histological entities were identified. Our unsupervised data demonstrated a more complex spatial intratumoral composition. By applying the ST technology to generate gene expression profiles, a new landscape of the melanoma metastases becomes evident. This should inspire further analyses to understand the multiple components of tumor progression and therapy outcome in a spatial context. Learning Objectives: Learn about the concepts and workflow of Spatial Transcriptomics. Learn about the limitations and needs in CMM diagnosis and treatment. Learn how spatially resolved transcriptomics can reveal insights into the tumor microenvironment of CMM. Webinars will be available for unlimited on-demand viewing after live event. LabRoots is approved as a provider of continuing education programs in the clinical laboratory sciences by the ASCLS P.A.C.E. ® Program. By attending this webinar, you can earn 1 Continuing Education credit once you have viewed the webinar in its entirety.

In recent years, many countries have passed legislation regulating the use of cannabis for medical purposes. This has granted patients and clinicians access to both herbal material and cannabis-based products, generally bypassing the strict regulatory procedures that normally apply to pharmaceutical development. Further, legalization of cannabis for recreational purposes in several states of the US, as well as in other American countries, such as Uruguay and Canada, confers cannabis an intriguing dual status, pharmaceutical substance and mere commodity, which creates a complex regulatory scenario. Irrespective of their purpose, cannabis products are limited by the lipophilic nature of cannabinoids, the main active substances in Cannabis sativa L., which difficults their formulation and makes their oral absorption erratic. Also, the psychoactive effects induced by Δ9-tetrahydrocannabinol (THC), the main cannabinoid, limit the therapeutic index of cannabis medications. Current efforts to achieve efficient, reliable dosing of cannabinoids aim at exploring different routes of administration (e.g., pulmonary, oral, mucosal, transdermal) and delivery vectors (e.g., inhaling powders, nanovehicles) to optimize their pharmacokinetic profile. This presentation will take a glance at the status of cannabis therapeutics, summarizing the ongoing efforts and strategies to develop new cannabis-based medical products. We will focus on indications for which cannabis and cannabinoids’ efficacy is either under clinical evaluation or has already been stablished. Also, we will explore differences in pharmacokinetic profiles among various routes of administration and novel pharmaceutical strategies to optimize both ADME properties (administration, distribution, metabolism and excretion) and therapeutic index of cannabis and cannabinoids. Learning Objectives: 1. Understand the state of the art of cannabis therapeutics, reviewing approved medications that target the endocannabinoid system and summarizing ongoing efforts and strategies to develop new cannabis-based medical products. 2. Learn about the differences in pharmacokinetic profiles of cannabinoids among various routes of administration and explore novel pharmaceutical strategies to exploit them.

Analgesics are commonly employed drugs for perioperative procedures and are required for painful procedures. They include not only non-steroidal anti-inflammatory drugs (NSAIDs) but also opioids and local anesthetics. Although NSAIDs may be adequate for minor procedures, where light pain would be expected, opioids should be administered for more painful procedures since they may provide the highest analgesic efficacy. Among them, potent full-agonist opioids should be considered for severe pain. Local anesthetics can be also provided to block pain when administered locally, for example at the wound site. To improve analgesic efficacy NSAIDs and opioids, but also local anesthetics, are usually combined, thus reducing their doses a related side effects. Unfortunately, analgesics are not devoid of side effects. NSAIDs block the inflammatory response and may not only interfere with research results when this response is evaluated, but also promote a reduction in renal and gastrointestinal blood flow, which may become dangerous when hypotension is favored by the use of inhalation anesthetics. Local anesthetics should be administered considering the maximum dose since their nerve blocking actions may not only affect nociceptive stimulation but also other vital organs such as the heart or the brain. Opioids have been found to produce not only the expected analgesic (hypoalgesic) action but also a delayed opposite action (hyperalgesia) which may appear in the postoperative period. Other analgesics may share this dual and opposite action. It remains unclear whether this hyperalgesic effect has clinical relevance in the painful animal or should be considered only in the healthy non-painful animal suggesting indiscrimate use of opioids should be avoided.

DATE: November 15, 2018 TIME: 06:00am PST, 09:00am EST, 3:00pm CET Part 1 - Presented by Valentina Svicher Hepatitis Delta Virus: insights and emerging concepts into a peculiar pathogen Hepatitis D virus (HDV) is a unique RNA virus that requires the hepatitis B surface antigen (HBsAg) to infect the hepatocytes. Chronic Hepatitis D is recognized as the most severe form of viral hepatitis, leading to accelerated progression of cirrhosis and hepatocellular carcinoma and a high mortality rate. HDV infection affects an estimated 15–20 million individuals, is spread worldwide and is endemic in some regions. Moreover, HDV prevalence is increasing in many countries as a consequence of immigration from highly endemic geographic areas. Despite the severity of hepatitis D, screening and treatment has been often neglected in part due to the lack of an effective therapy. Indeed, interferon alpha is the only anti-HDV drug approved for the treatment of chronic hepatitis D, even if its efficacy is limited and side effects can be severe. Future therapeutic options are under investigation targeting HDV entry, HBsAg secretion, and viral assembly. This webinar will summarize current knowledge on virological and epidemiological aspects of HDV infection with particular attention of categories of individuals in which HDV testing should be prioritized. It also provides an overview on the upcoming treatment options against this virus. Learning Objectives: Discuss epidemiological and virological aspects of HDV infection Identify categories of individuals at risk of acquiring HDV infection Describe current and future HDV treatments Part 2 - Presented by Ana Avellon HDV infection: how and when to diagnose a forgotten disease Although the most aggressive infectious liver disease, hepatitis delta virus (HDV) infection may be underdiagnosed both for the unawareness or the unavailability of accurate diagnostic methods. Any hepatitis B virus (HBV) infected patient is susceptible to HDV infection for life and consequently searching for HDV should be included in HBV infected patient monitoring guidelines. Periodically detection of HDV antibody and viral RNA in HBV infected patients can early diagnose HDV and may impact the progress of the liver disease. Learning Objectives: How to diagnose HDV: methods and interpretation of results When to ask for HDV diagnosis: guidelines review

Historically single cell isolation techniques suffered from poor efficiency, low recovery and difficult automation, among other limitations. This presentation will highlight cellenONE, a piezo-acoustic and visual feedback technology to isolate and dispense single cells. Case studies have shown outstanding accuracy, high viability; both crucial parameters for cell line development.

DATE: January 26, 2017 TIME: 8:00am PT, 11:00am ET The demand for low-cost, disposable devices with short response times capable of performing routine electrochemical biosensing has increased dramatically. Primarily driven by the medical diagnosis and food fields, disposable screen-printed electrodes provide fast, sensitive detection and quantification of various analytes in complex samples. Screen-printed electrodes (SPEs) are preferable over other types of electrodes for these applications as they can be fabricated from several different materials, made in diverse geometries and multiplexed formats and offer the possibility for mass production at a low fabrication cost. In addition, the small size of these electrodes enables a very small volume of sample to be used for analysis. The planar shape of SPEs facilitates the incorporation of magnetic bioconjugates by simple attraction using a magnet positioned under the electrode. In this webinar Dr. Susana Campuzano will demonstrate SPEs-based electrochemical biosensors for the rapid, sensitive and selective biosensing of analytes in food and clinical samples. Learn about the simultaneous detection of multiple components such as adulterants and allergens in food samples as well as the analysis of cancer reporters in the clinical lab. Key Learning Objectives: Learn tips and tricks from Dr. Susana Campuzano’s case studies in clinical and food applications Understand the advantage and utility of screen-printed electrodes in drug discovery research and other applications Learn how to perform high throughput SPE applications using 96 well plate and multichannel potentiostats Learn how to choose the right SPE electrodes and potentiostats for your application needs Who should attend: Anyone performing decentralized assays to develop specific sensors and performing other electrochemical studies Anyone performing clinical and life science application looking for high throughput solutions Biopharmaceutical and food scientists involved in assay and new sensor development Research lab manager and scientists involved in new biosensor development and miniaturized electrochemical studies

Quality animal research is based on quality animal care and use. This is achieved by coordinating several key personnel and activities, and implementing appropriate animal care and use procedures. Areas to be covered include institutional responsibilities in terms of personnel (resources and training); occupational health and safety; oversight and ethical review process; animal environment, housing and management; veterinary care; and physical plant. All these components together will be the institutional animal care and use program. The first thing to do to implement the right animal program is to review in a systematic manner all components of the program currently in place and identify potential gaps. The AAALAC International Program Description template, freely available from the AAALAC website (http://www.aaalac.org/programdesc/index.cfm) provides a very simple and useful self-evaluation tool to do this, as it encompasses all necessary animal program areas. Once gaps are identified, a scheduled plan for corrective actions can be established. But achieving the right animal program cannot be a single person task. Cooperation from and coordination with several others in the institution is needed: management; animal care personnel; IACUC or equivalent oversight body (ethics committee, animal welfare body…); research team; health and safety personnel; maintenance service; or quality assurance unit (if existing). That will be the key to success. Examples of typical findings on all different program areas identified during AAALAC site visits will be described, along with the expectations AAALAC has for those same areas. Quality animal studies depend on quality animal care and use, which can be only achieved by a systematic self-evaluation and implementation process.

Increasing prevalence and severity of multi-drug-resistant (MDR) bacterial infections has necessitated novel antibacterial strategies. Ideally, new approaches would target bacterial pathogens while exerting selection for reduced pathogenesis when these bacteria inevitably evolve resistance to therapeutic intervention. As an example of such a management strategy, we isolated a lytic bacteriophage, OMKO1, (family Myoviridae) of Pseudomonas aeruginosa that utilizes the outer membrane porin M (OprM) of the multidrug efflux systems MexAB and MexXY as a receptor-binding site. Results show that phage selection produces an evolutionary trade-off in MDR P. aeruginosa, whereby the evolution of bacterial resistance to phage attack changes the efflux pump mechanism, causing increased sensitivity to drugs from several antibiotic classes. Although modern phage therapy is still in its infancy, we conclude that phages, such as OMKO1, represent a new approach to phage therapy where bacteriophages exert selection for MDR bacteria to become increasingly sensitive to traditional antibiotics. This approach, using phages as targeted antibacterials, could extend the lifetime of our current antibiotics and potentially reduce the incidence of antibiotic resistant infections.

During my presentation, we will discuss: - Normal patterns of behavior (repertoire): social, food intake, territorial, agonistic, anti-predation, sleep and rest, play behavior, grooming, sexual(courtship and maternal), disposal, thermoregulation, exploratory, kinetic. - Characteristics and species - specific differences in the behavioral repertoires of rodents - Animal Welfare and its relationship to the behavioral repertoire of the species - Evaluation of the behavior pattern expression in animals bred and maintained in an animal facility - Applied Ethology in laboratory animals - Environmental Enrichment: concepts and examples of its application - Learning Objectives: - To understand normal behavior patterns in animal species frequently used in biomedical experimentation - To evaluate the proper development and expression of these patterns of behavior in animals kept in captivity in an animal facility and to relate it to Animal Welfare

perioperative analgesia aims to minimize, or even eliminate pain during a surgical procedure. Currently this approach is rarely performed using a single analgesic drug and more commonly different types of analgesic drugs, such as opioids and non-steroidal anti-inflammatory drugs (NSAIDs), are combined. This approach is known as multimodal or polymodal analgesia and also includes other analgesic drugs and techniques such as alpha-2 adrenergic receptor agonists, ketamine, or loco-regional analgesic techniques. Another approach is the anticipation to perioperative pain by administering analgesic drugs before surgical injury occurs and thus preventing the animal suffering from pain. This approach is known as preemptive analgesia and may reduce pain not only intraoperatively but also postoperatively. Other relevant aspects include the expected pain intensity, which allows us to select the appropriate drugs or drug combinations. The duration of analgesic therapy should also conform to the expected duration of pain so as to provide the necessary coverage postoperatively. Fortunately, there are on the market analgesic formulations with an effect typically lasting 24 hours with some of them providing analgesia up to 4 days following a single drug administration. Despite these advances, a good postoperative analgesia can only be obtained with an appropriate pain assessment system that would allow changes to the analgesic regimen in order to limit or prevent pain effectively.

The environment in which laboratory rodents are kept has an impact on experimental data and breeding performance. The quality of the environment is defined by both the quality of the primary enclosure and the secondary enclosure and their fluctuations. There are regulations and international recommendations for optimal values and ranges of most relevant environmental parameters, but it is actually the whole environment and animal care and use procedures what might (and certainly will) impact experimental results. Standardization of animal models is an issue of paramount relevance in biomedical research and this standardization must include procedures, genetics, health status, but also the environment animals are kept in. The presentation deals with environmental standardization and the impact that lack of control of these variables can have on experimental results.

I will present the most common hazards, as they are perceived by workers (traditionally zoonoses, bites and scratches, allergies). Then I will present real data on accidents in research facilities including our facility. This will show that generally we are not focused on risks that are medically or numerically important. Finally I will talk about ‘emergent risks' (psychological and emotional risks) and will present the results of a Spanish study and the proposal for a ‘Spanish-speakers' global survey focused on these risks.

The popularity reached by the genetic manipulation of laboratory animals to create new models for studying human diseases, produced in turn, that the techniques for assisted reproduction consolidate as routine in the laboratory of an animal facility. However, the costs of maintaining these mouse strains have increased deeply, and the physical space to housed these animals is practically the same one, for what we should admit that it is a concrete problem. In order to alleviate this problem is why embryo cryopreservation took importance in the early 90s. The cryopreservation technology has the advantage of being a tool to control the animal space at a low cost, but also has the benefits of being able to stop the risk of possible natural mutations, and genetic or infectious contamination (a tragic epidemic).Nowadays we can freeze embryos, sperm and ovaries to cover any situation of protecting the genetics of a mouse line. At the same time, the progress in the manipulation of the sperm and in the methods of in vitro fertilization does this task a more efficient process. With the sperm collected from a mouse there is enough material to fertilize approximately thousand of oocytes, which facilitates the task to preserve, to rederivate, and to expand or to distribute a mouse strain. Another application of this technology is to recover strains that otherwise it could be lost or by negligence, lack of breeding or by death. Today it is also possible to recover sperm from a death mouse and fertilize in-vitro up to 4 days after his dead.

It is increasingly recognized that the genetic background (i.e., all genomic sequences other than the gene(s) of interest) can have profound influences on the phenotype of an animal model. It has been shown that mutations (spontaneous and induced), transgenes, and targeted alleles (knockouts and knockins) that are moved onto a different background can show a change in phenotype. One of the first cases involved the classical diabetes (Leprdb) mutation that presented transient diabetes on a C57BL/6 background but overt diabetes on C57BLKS. Other examples include background effects on survival rate in Egfr (epidermal growth factor receptor) KO mice and effects on tumor incidence and spectrum in Trp53 and Pten (tumor suppressors) KO mice. In order to highlight the importance of this issue, I will present a selection of published articles showing the influence of genetic background on different mouse and rat models. I will also discuss some of the problems arising from the use of undefined substrains, the use of genetically engineered mice with mixed backgrounds (e.g., after breeding chimeras), as well as the flanking genes and passenger mutations concerns. Finally, I will present different ways to avoid or resolve these drawbacks, including the development of congenic strains by marker-assisted backcrossing and the use of newly available ES cells from strains other than 129. In order to stay away from confounding or unreliable experimental results, particularly with the increasing number of mouse and rat strains, attention to the genetic background is crucial.