Breast cancer with various biological diversity known as the common reason of death in the world and despite progress in novel therapeutic approaches, it faced with failure and recurrence in general. Recent clinical and preclinical statistics support cancer stem cells (CSCs) hypothesis and its similarities with normal stem cells. Evaluation of related paper conclude in significance finding in the further characterization of CSCs biology such as surface biomarkers, microenvironment regulatory molecules, cell signaling pathways, cell to cell transition and drug efflux pumps to overcome multidrug resistance and effective therapy. Emerging novel data indicate biological concepts in the base of unsuccessful treatment. A powerful understanding of the cell signaling pathways in cancer and CSCs topics can be led us to define and control treatment problems in cancer. More recently nano medicine based on drug delivery system modification and new implications on combinatorial therapy have been used to treat breast cancer effectively. The aim of this review is focus on CSCs as a potential target of cancer therapy, to overcome the limitation and problems of current therapeutic strategies in cancer. Breast cancer is a disease with histological, molecular and epidemiological heterogeneity which this heterogeneity creates big challenges to the development of effective cancer treatment.3 There is a lot of biological diversity in the breast cancers that occur because of variations in transcriptional programs. To distinguish patients with a high risk of progression, breast cancers are classified into subtypes according to gene expression profiles:Breast cancer is the most common malignancy and the 5th cause of cancer related death among the US women.1) Luminal A2) Luminal B,3) Human growth factor receptor 2 overexpressing (HER2-OE), and64) Basal-like tumors as shown in 7 Despite progress in common treatment strategies of cancer like chemotherapy, radiotherapy, and surgery, an untreatable population of tumors remains that metastasize to distant organs. These population displays stem cell properties that we focused more in next part.8In the breast cancer treatment, the recurrence and failure cause a big obstacle in treatment of the cancer. To reduce the cancer-related mortality rate, studying the biological properties of cancer and discovering new therapeutic approaches can be helpful. Several mechanisms cause to chemoresistance like alterations in apoptotic signaling pathways, metabolic enzymes defection, mutations in tumor suppressor genes, increased drug efflux pumps, reduced drug uptake and tumor microenvironmental changes in response to therapy.9 Stem cells in the normal breast tissue produce early and late progenitors, that these progenitors finally differentiate into (Ι) the luminal or alveolar epithelial cells; (II) the ductal epithelial cells and (III) the myoepithelial cells .al cells .8,10,1112 The aldehyde dehydrogenase (ALDH) enzyme is another normal breast stem cell marker that plays a functional role in stem cell differentiation. Studies of the normal breast tissue show that about 6% of the epithelial cells in the TDLU were ALDH+ and these cells can generate mammospheres in the suspended culture condition.13 Another subset of normal breast stem cell was found that are keratin K19 negative and the part of these cells increased under proliferative conditions such as epithelial hyperplasia; so, they can be the origin of breast cancers.14The normal breast stem cells are CD49f +/EpCAM- that are capable of self-renewal and differentiation to various types of breast tissue cells. According to recent studies, the breast stem cells can be enriched within a CD49f +/EpCAM- population with a basal cell specification.15 So, the origin of these cells is important for the prevention, early detection, and breast cancer therapy.16 CSCs have the similarities with normal stem cells like being quiescent, multipotency and self-renewal capacity that these specifications helps preserve the tumor.17 CD44+/CD24low/- and ALDH are common CSC markers that are the same with normal stem cells. Common markers of BCSCs have been briefed in 19 During carcinogenesis, these factors reprogram differentiated tumor cells into undifferentiated stem-like cells.14 Breast cancer CD44+ cells are like to normal stem cells and they show basal-like cells properties. But CD24+ cells have shown similarity with luminal cells. In the breast cancer cell lines, we can find these two cell types which demonstrating a similar hierarchy to tumor cells.20 BCSCs can survive in the therapeutic condition in comparison with the more differentiated tumor cells.21Cancer stem cells (CSCs) firstly were discovered in acute myeloid leukemia and they have become an important part of research as a potential target for cancer therapy. The origin of breast CSCs is the mammary multipotent stem cells with genetic defects that affect pathways related to self-renewal and differentiation.22 CSCs may play a central role in contributing to this circumstance and understanding critical molecular components of cell signaling pathways is the priority of defining new therapeutic strategies. Here some important molecules participated in the regulation of network signaling concept that have been shown in 23 Numerous studies consider the roles of mentioned molecules and their signal transduction, but it seems there is a much more complicated context to reach an exact treatment without gross side effects.Breast cancer-related deaths in the United States make a dramatic number (40 000) every year, and involved with the metastatic condition generally.24 Previous studies showed Wnt also involved in chemo- and radio-resistance of breast CSCs.11 The subpopulation of CSCs is maintained by the employment of Wnt ligands by periostin, a component of the fibroblast’s extracellular matrix. Wnt signaling in tumor cells induces its expression and colonization in the context of lung tissue. Despite inhibition of the Wnt function show poor progression and metastasis in some cancer, but we need more information on the Wnt signaling pathway to finalize the therapeutic approach.25One of the key regulatory and embryonic originated molecules in the cancer formation and progression is Wnt and its ectopic expression which increased the breast stem cells that occurred by alteration of the epithelial hierarchy. Besides of embryological approach for Wnt function, it is involved in the function of some somatic cells such as the brain, blood, lung and mammary gland. The abnormal function of Wnt can lead the cell to tumor formation and expands breast stem cells.26 Inhibition of Notch receptors signaling pathways affected mentioned items as a novel therapeutic approach. Generally, Notch plays so much role in the developmental biology of breast tissue.29 Various subtypes of Notch such as Notch1 and Notch4 have a different function in cleaved/activated form. For example, upregulation of the Notch 4 induced metastatic conditions in the breast epithelial tissue.30 The differential expression of Notch4 versus Notch1 receptors in breast CSCs can suggest different characters for each receptor and so our therapeutic strategies.Like Wnt, deregulation of Notch as a very important embryonic molecule involved in tumor progression and self-renewal of CSCs in adult which conclude in mammosphere formation.31 But more recently, it has been shown not only downstream effectors of HH signaling such as polycomb group involved as a central regular in self-renewal of adult stem cells but also participated in CSCs pluripotency.33 Epithelial-mesenchymal transition (EMT) procedure as a key factor in tumor progression has been affected by the HH signaling pathway and could be defined as a therapeutic approach in pancreatic cancer.34 Until now we numbered characteristics of some of the most important molecules involved in developmental biology and abnormal issue in the body. But in continue, we showed another essential molecule that affected the growth of tumor cells passively or negatively.The mentioned molecules have principle roles in the embryonic period, but more important functions can be found for some of them in abnormal and pathologic conditions. HH pathway implicated in the viability of adult tissues. Regards knowledge about the role of deregulation of HH in cancer progression, there are very few studies links this role in CSCs too.35 TGF-β signaling synchronization with the WNT pathway in breast cancer, induce EMT and cause more progression of the tumor. Several molecules have been involved in cancer metastasis but the role of TGF-β is essential because of the interaction of other immune cells. Indeed, the dysregulation TGF-β detected in several types of cancers especially having a role in progression to malignancy of breast cancer. But it seems a careful clinical study needs to approve this function completely.36TGF-β is a multifunctional cytokine, produced by white blood cells and affect stemness properties through the expansion of CSCs in breast cancer. More recently a study declared that chemotherapy decline the growth of tumoral cells in breast cancer probably via CSCs expansion modification through the alteration of IL8 and TGF-β signaling pathways.25 It was possible to assume that activation of JAK/STAT signaling pathway is associated directly with tumor fate in term of therapeutic approach.Some of the proposed molecules having a different role in animals and humans. The Jak/Stat in Mouse embryonic stem cells promote selfrenewal and pluripotency but in human leukemia inhibitory factor have not this function. One of the known activated molecules of this pathway, named STAT3 promotes tumorigenesis and increases the survival/proliferation of breast CSCs.38 The similarity between CSCs and stem cells shows a lot of cue to clinic and experiment aims. EMT normally occurred during the development of a fetus and indeed it is one of the most important events, which has been detected in CSCs growth and tumorigenic process.39 Based on embryological understanding facts and similarity we can use those molecular approaches and signaling transduction in defining of novel therapeutic strategy. There are several known cells signaling such as Wnt, Hedgehog and Notch which involved in both CSCs and normal stem cell viability and renewal. In contrast expression of genes related to inducing epithelial differentiation, can target as a good alternative in treatment  in cancer treatment produced therapeutic limitations based on CSCs renewal and progression after the onset of the therapeutic procedures. Thus, focus on CSCs is so critical to improving the quality of treatments in the clinic. In this review, several strategies have been designed to gain final goal in CSCs biology such as developmental anatomy principles, EMT, surface biomarkers recognition/targeting, oncogenes profile association, cell signaling in CSCs, the role of pumps in drugs efflux and resistance, and microenvironment and immune cell behavioral. Apart from these items, nanotechnology and its recent powerful advance also use in combinatorial therapy ) as a specific direct ALDH inhibitor and ATRA ) indirectly target the activity of ALDH by the retinoic acid (RA) pathway.43 Mentioned signaling items directly involved in the developmental of the body during intra uterine life and similarity of normal stem cells to CSCs reveal more knowledge in both of them mutually.52 The notch signaling pathway and its cascade in multiple biological aspects finally affected CSCs stemness, MDR, and hypoxia inducible factor (HIF) pathway.53 On the intermediated modalities in this cascade such as γ-secretase inhibitors and antibodies directed showed accepted resulted in a clinical trial in various types of cancers.54 It is critical to know about the diversity of Notch signaling to manipulee of the final events in special cancer like breast cancers. Utilizing of Notch signaling inhibitors in oncology related to many alternatives such as type of cancer, exact pharmacodynamic effects identification and mechanistic understanding of it signaling. For example, CD44 as a biomarker in CSCs is Wnt signaling target and knockdown of CD44 diminished tumor formation.55 In another study administration of an antagonist of Wnt decreased mammosphere formation.56Signaling pathways and deregulation of its modalities such as Stat3/mTor, Hedgehog, Notch and Wnt/-catenin cascade defined as a new therapeutic strategy in the prevention of cancer extension, especially CSCs growth behaviors modification . Multipl58 Molecular modification of the tumor microenvironment can block the growth of CSCs. Repertaxin as an inhibitor of IL-8 increase chemotherapy efficiency and diminishing tumor size.59 Here also combinatorial therapy makes the curing process more effective than before multiple biological approaches by using IL-6 and IL-8 inhibitors or HIF pathway blocking.60 It seems there is more detail information about signaling pathways needed to overcome CSCs microenvironment suitability.Preservation of CSCs in the tumor, depends on the microenvironment of cancer and recently it is defined as a novel pharmaceutical target. Indeed, a suitable niche has been provided by (a) immune cell secretion, conclude in chronic inflammation, (b) perivascular space involved in proliferation and differentiation and, (c) finally routine hypoxia which is known as a critical issue in the growth of CSCs . The Sta61 Combinatorial therapy with cell death agents like engineering in mesenchymal stem cells to express TRAIL based viral vectors62 and anticancer drugs like cisplatin, daunorubicin and cytarabine, inhibition on transcription factor of NF-kB by proteasome inhibitor is critical in tumoral cells survival.63 Suppress growth of CSCs, trigger and induce apoptosis through inhibition of the Wnt signaling pathway in various types of cancers.64One of the most critical issues during primary stages of cancer, development and metastasis is the impairment of apoptosis procedures. Indeed, the cells in the normal tissues of the body without the apoptosis process be cancerous definitely. Thus, internally or externally induction of this onset of the apoptotic signaling pathway can be defined as a reliable method in novel cancer therapeutic studies. Modification in apoptotic genes and receptors by synthetic or natural materials have been used in several studies. P53, bcl-2, ras and c-myc cell cycle regulating genes and death related receptors like CD95, the tumor necrosis factor (TNF) receptor, trimeric human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) are the part of apoptotic induction targets which finally activated caspase 8 and then caspase 3 which finally conclude in permeabilization of mitochondrial membrane and cell death.Modification in apoptosis mechanism beside anti-cancer drugs provide a good preclinical study goal, but there is some issue in specific effects, side effects problems and toxicity of the mentioned approach most be solved to allow utilization in the clinic.65 Another alternative to the silence of P-gp is the use of RNAi such as siRNA66 alone or in the combination with nanoparticles application can simulate the effects of inhibitors of P-gp. For example, utilizing conjugated copolymers with ritonavir (ABC transporter inhibitor) increase the sensitivity and mortality of drug-resistant tumoral cells through enhancement in the cytotoxicity of doxorubicin in leukemia. Combinatorial therapy by application of nanotechnology in this field provides promise to overcome CSCs resistance to routine drug and classic treatments.67MDR in cancer treatment, especially chemoresistance highly related to overexpression of ABC transporter as drug efflux pumps (P-glycoprotein (P-gp)). Thus, inhibition of the function of transporter simply can be recognized as an anti-MDR procedure. But low specificity and high side effect defined as an obstacle in this alternative. Although some of these inhibitors like Vardenfl showed the accepted result in P-gp blocking and increasing anti-cancer drugs like (paclitaxel and vincristine) intracellular concentration.68 Among them, circulating miRNA as a valuable molecular tool with its stability and approximately noninvasive isolation procedure, make it a valuable alternative in diagnosis, prognosis and even a good target for monitoring of anti-tumor interventions response.69 During the onset and progress of cancer, various types of miRNA can be upregulated or downregulated in different stages of the diseases. Deliberate dysregulation of mentioned oncogenic miRNA with mimic miRNA leads to alteration in tumor-suppressive genes and trigged or blocking a signal transduction pathway conclude in defining a new therapeutic strategy with knowledge about the complication of this procedure. Previous studies with sequencing and PCR techniques showed some miRNAs expression are modified in a lot of the diseases including breast cancer and profiling of miRNA has been considered to determination and evaluation of disease progress.70 MiRNA based treatment goes to establish a feasible approach to enhance current anti-cancer therapeutic strategies. But there are several obstacles to reach the final aim before clinical utilization such recognition of differential expression pattern of miRNA that involved in particular cancer on experimental animal model, degradation and instability problems solving in in-vivo, target base function to avoid side effects on normal tissue.68 For example, Yu et al. showed that in SKBR3 cells, expression of some miRNA like let-7, miRNA-16, 107, 128 significantly changed compared with differentiated cells in breast tissue context.71To good monitoring and diminishing the risk of cancer progression, scientists attempt to define a new powerful method in the cellular and molecular levels. Early detection by sensitive and specific biomarkers placed on top of the list. Body fluids like peripheral blood contain molecular targets called nucleic acids such as DNA, RNA and miRNA. Thus genetic, epigenetic modification and microenvironmental alteration as a very novel approach has been recognized in this field.72 retinoic acid  and even signaling pathways modification (Akt/mTOR and P53).75 Knockdown of special genes or proteins such as pancreatic differentiation 2 (PD2) protein in CSCs affected cell viability, trigged apoptosis process76 and decrease expression of stem cell markers. Combinatorial therapy with a focus on differentiated cells and CSCs by various alternatives can be so useful to define a new therapeutic strategy in the cancer treatment field.In general, the target of cancer therapy is the differentiated cells but recently induction of CSCs differentiation is established as a novel method to decreasing tumoral cells population. These approach handle by some differentiative inductive agents like: histone deacetylase inhibitors .79 The gist of these studies can highlight the fact that that lower chronic inflammation equals a lower risk of cancers. Epidemiological research on nonsteroidal anti-inflammatory drugs (NSAIDs) administration provides more good results in cancer treatment,80 albeit side effects of NSAIDs not be ignored on the cardiovascular system and so need to develop of a new therapeutic approach. Apart from inflammation factors, proinflammatory factors, TNF-α,81 IκB kinase, the upstream NF-κB activator kinase, interleukins 1, 6, 8 and certain chemokines and their receptors inhibitors82 can be defined as potential targets for cancer therapy. Another strategic alternative is protumorigenic factors to recognize and modify them to reach the final goal of treatment in cancers.83 The future direction seems must be the focus on immune cell’s genetic or epigenetic modification to gain an accepted therapeutic approach is the change of tumor-infiltrating immune cells gene expression through the cytokine cells balance alternation. Immune tolerance can be changed by cytokines effectors administration to reduce immune system response enhancement, but there is a very important part and it is side-effects of utilizing these methods. Therefor final strategy must be localized to decrease the toxicity of the mentioned therapeutic alternatives. Recently studies focus on the modulation of immune cells functions by CSCs microenvironment. Chronic inflammatory condition is a key factor in the tumor microenvironment that leads to proliferation and metastasis of the cancerous cells through the immunosuppressing of immune cells like T cytotoxic and Natural killer cells.86 In some cases, dendritic cells (DCs) anti-tumor antigens are masked from tumor cells by tolerizing antigens presenting cells. Indeed, DCs stimulate T cytotoxic.87 Immune cells (tumor-associated macrophages (TAMs), tumor-associated neutrophils (TANs), and myeloid-derived suppressor cells (MDSCs)) recruited by tumor cells through the cytokines and chemokine secretion. TAMs secrete TGF-β with a role in immunomodulation and immunosuppression, recruits T regulatory cells (Tregs).88 MDSCs are bone marrow derived origin, also involved in cytokines secretion like  and also recruit proinflammatory secreted cells such as T helper 17 cells which improve immunosuppression.86 TANs and TAMs cause cancer progression with the breakdown of ECM lead to metastasis and tumor angiogenesis.86 These cell’s exosomes can be internalized mRNAs and microRNAs into different cell types89 for tumor cell proliferation, metastasis and chemoresistance through the drug efflux pumps transporter.90 In the inflammatory TME, secretion of TNF-α by TAMs and CD4+ T cells, involved in upregulates of NF-kB signaling pathways. Downstream cell signaling continuously caused in self-renewal and finally promote migration and invasion of CSCs. The conversion of non-CSCs to CSCs may be induced by stemness and EMT correlation.91The correlation between inflammation and neoplasia has been considered by several studies, which showed that chronic inflammation of pancreatic to helicobacter pylori infection associated with gastric, colorectal, bladder, pancreatic, liver, lung and even cervical cancers92Besides of very clear role of CSCs in proliferation, metastasis and therapeutic resistance of tumor cells, more recently the role of CSCs in extent of the tumor through diminishing immune cells surveillance intra tumor microenvironment has been investigated. Additionally, CSCs are potent to escape from immune cells by alteration of immunogenicity. Furthermore, an experimental model showed CSCs directly related to the progression of tumor cells and can be useful if we destroyed or ablated those.93 A very complicated response and organized gene expression and secretion of particular effectors (TGF-β) of CSCs conclude in attenuating MHCII and depleting intratumor immune cells finally even through the expansion of T regulatory cells.96 In contrast, not only tumor supportive phenotypes cells (TAMs) recruited through periostin secretion to the microenvironment of the tumor but also macrophage migratory factors (colony-stimulating factor-1 (CSF-1), macrophage inhibitory cytokine 1 (MIC-1) and C-C motif ligand 2 (CCL2) induced by CSCs make the condition worse than before. These factors lead to a monocyte context to the immune suppressive M2 phenotype.97 As mentioned above macrophage is critical for maintains stemness in CSCs in breast via interactions of CD90 (Thy1) and EphA4 and induction of downstream signaling such as Src and NF-κB. Indeed NF-κB is so important for the recruitment of macrophage in breast cancers. The self- renewal of CSCs in breast tumor increased by IL6 that activated signaling pathway of STAT3. It seems more studies are required about the reciprocal interaction of CSCs and immune cells in micromovement of the breast cancer tumor and its progression behaviors.98 insert clinical point, mentioned future studies suggestion and point your limitation of your study.CSCs delayed or cut the response of immune cells by secretion of immunosuppressive factors, mimicking of antigen-presenting cells (expression of MHC I) and the co stimulator molecule  inhibition or lack of co-stimulatory molecule  activation which lead T cells to abnormal immune response to a specific antigen. The secretion of galectin-3 by CSCs triggered an apoptotic process in T cell.Breast cancer is a disease with histopathological and epidemiological heterogeneity. Here we discussed about various alternatives and biological characteristic concept of CSCs include targeting surface biomarkers, targeting cell signaling, microenvironmental modalities alteration, apoptosis and differentiation pathways, efflux-mediated drug pumps in MDR, manipulation of miRNA expression, differentiation inductive agents, inflammation, immune modulation of microenvironment, etc in CSCs and breast cancer treatment. For example, exact information about biomarkers means the intervention of certain signaling transduction, which makes CSCs good candidates in cancer treatment or it seems more detail information is need about signaling pathways to overcome CSCs microenvironment suitability. Modification in apoptosis mechanism beside anti-cancer drugs provide a good preclinical study goal, but there is some issue in specific effects, side effects problems and toxicity of the mentioned approach most be solved to allow utilization in the clinic.Numerous experiments and clinical studies consider the roles of mentioned pattern, but it seems there is a much more complicated context to reach an exact treatment without gross side effects. It seems so much careful clinical study needs to approve this function completely. Indeed, knowledge about stem cells and particularly CSCs can be led us to define a novel therapeutic strategy based on new technology (nano biomedicine) and CSCs interventions. In fact, nanotechnology and its recent powerful advance also use in combinatorial therapy. Finally, emerging novel data indicate biological concepts in the base of unsuccessful treatment. A powerful understanding of the cell signaling pathways in cancer and CSCs topics and their interaction with other biological modalities can be led us to define and control treatment problems such as metastasis and chemoresistance. It will be critical for the developing of effective anti-tumor drugs, utilizing nanomedicine.Not applicable.The authors declare that they have no relevant affiliations or financial involvement with any organization.


Segregation of homologous chromosomes is the defining event of meiosis and is central to sexual reproduction. To ensure regular segregation, homologs must establish strong but reversible physical links capable of providing the resistance to poleward spindle forces needed for stable biorientation of homologous centromeres. What form do such linkages take? In the most widespread mechanism, the linkers are chiasmata, in which crossovers between homologous chromatids are anchored by sister chromatid cohesion distal to the crossover see  1]. Thu. Thu1]. PLOS Genetics [stromalin in meiosis (snm), mod(mdg4) in meiosis (mnm), and teflon (tef) [snm or mnm do not affect homolog pairing in early prophase I but disrupt all 4 bivalents beginning in late prophase I [tef cause the same phenotypes but affect only the 3 autosomal bivalents [This question has led some researchers, including the authors of a paper in the current issue of Genetics , to inveGenetics . In the Genetics . Althougon (tef) ,7. Null ophase I . Up to 8ivalents . SNM andivalents . These fivalents .CG8712 ), an uncharacterized gene expressed mainly in testes. UNO lacks conserved domains, but extensive genetic and cytological characterization revealed it to be a new conjunction protein remarkably similar in function to MNM and SNM [uno mutations were found to cause the same primary phenotype as snm and mnm mutations, premature dissolution of all 4 bivalents and random assortment of the resulting univalents. The spatial–temporal localization pattern of UNO protein recapitulated that of SNM and MNM, and the chromosomal foci formed by all 3 proteins were observed to be fully codependent (and dependent on tef on autosomes). These features strongly suggest that SNM, MNM, and UNO form a complex, but, until now, there has been no direct evidence. In support of this idea, the native MNM and SNM proteins were among the strongest interactors with both baits. Moreover, TEF copurified with MNM-EGFP, consistent with it being a component of autosomal AHC complexes [Although existing data indicated roles of SNM and MNM in homolog linkage, both the mechanism of conjunction and the means by which it is removed were unclear. With the goal of identifying additional conjunction proteins, the authors of the current paper conducted a proteomic screen for testis proteins that interacted with either SNM-enhanced green fluorescent protein (EGFP) or MNM-EGFP . Among t and SNM ,7. uno mThe finding that Separase is required for release of conjunction at anaphase I  was surp3 volume of an amorphous “chromosome territory” and after chromosome condensation when they are compacted into dense “blobs” and subjected to spindle forces [As a result of recent progress exemplified by the featured paper , severale forces ,15. (3)


The Australasian Virology Society (AVS) aims to promote, support and advocate for the discipline of virology in the Australasian region. The society was incorporated in 2011 after 10 years operating as the Australian Virology Group (AVG) founded in 2001, coinciding with the inaugural biennial scientific meeting. AVS conferences aim to provide a forum for the dissemination of all aspects of virology, foster collaboration, and encourage participation by students and post-doctoral researchers. The tenth Australasian Virology Society (AVS10) scientific meeting was held on 2–5 December 2019 in Queenstown, New Zealand. This report highlights the latest research presented at the meeting, which included cutting-edge virology presented by our international plenary speakers Ana Fernandez-Sesma and Benjamin tenOever, and keynote Richard Kuhn. AVS10 honoured female pioneers in Australian virology, Lorena Brown and Barbara Coulson. We report outcomes from the AVS10 career development session on “Successfully transitioning from post-doc to lab head”, winners of best presentation awards, and the AVS gender equity policy, initiated in 2013. Plans for the 2021 meeting are underway which will celebrate the 20th anniversary of AVS where it all began, in Fraser Island, Queensland, Australia. The 10th Australasian Virology Society (AVS10) meeting was held on 2–5 December 2019 in stunning Queenstown, New Zealand. The inaugural AVS meeting, then known as the Australian Virology Group, was held on Fraser Island, Queensland, in 2001. The Australian Virology Group (AVG) was founded by Professor Paul Young, who led the group for 10 years, holding meetings every two years. The AVG became an incorporated Society (AVS) in 2010 and Professor Young was our first president before passing on the baton to Professor Damian Purcell (AVS President 2011–2015), followed by Professor Nigel McMillan (AVS President 2015–2017) and now Professor Gilda Tachedjian (AVS President 2017–current). Eighteen years later, we marked our 10th AVS scientific meeting by returning to Queenstown, the site of the 2013 (AVS7) meeting. The AVS biennial meeting is the premier virology conference in Australasia, providing a forum to disseminate the latest research on the biology of human, animal, plant or prokaryotic viruses in a relaxed yet engaging environment. AVS aims to foster collaboration between different fields of virology, to strengthen multidisciplinary virus-related research, and to encourage the participation of students and post-doctoral researchers. We were delighted to welcome our plenary speaker Dr. Ana Fernandez-Sesma, who delivered the Ruth Bishop Oration on dengue viruses and innate immunity, and Professor Benjamin tenOever, the Robert Webster Orator, who presented on the evolution of antiviral defenses. We were thrilled that Professor Richard Kuhn, our third international speaker, presented a keynote on structural studies of flaviviruses. Professor Kuhn was the plenary speaker at our meeting in 2007 (AVG4) and he was keen to return to the meeting. Professor Lorena Brown and Professor Barbara Coulson, who have made outstanding contributions in virology throughout their distinguished careers in Australia, were honoured at AVS10 as lead invited session speakers to present their research on influenza and rotavirus, respectively. AVS10 featured an additional 12 invited speakers from Australasia, 41 speakers selected from proffered abstracts, 16 early career and student rapid-fire three-minute talks, and over 100 poster presentations. We introduced several new initiatives to promote and support the goals of the society including a session on indigenous research that highlights the disparate burden of virus borne by indigenous communities in Australasia. For the first time at an AVS conference, we used the “Catchbox”, an alternative to a roving microphone that requires a good throwing arm and catching ability of chair and participants, respectively . www.avs.org.au). We are pleased that the percentage of women on our AVS committee and AVS10 local organising committee (LOC), as well as lead invited speakers, chairs and proffered oral abstracts, has increased to or close to 50%, reflecting the composition of our AVS membership  to present on the “big stage” in front of national and international virology experts. AVS10 also introduced the “Poster Video Awards”, where delegates could prepare a three-minute video describing their poster. This educational initiative, championed by Professor Paul Young, aimed to promote skills in science communication, with first prize being awarded to Wilson Nguyen . The vidAs part of our commitment to career development, we organised a breakfast session on “Successfully transitioning from post-doc to lab head” that included a panel of early, mid and experienced researchers who provided their pearls of wisdom to post-docs, moderated by Professor Heidi Drummer . We congratulate and thank the AVS10 2019 organising committee for their efforts in delivering a highly successful meeting led by AVS10 Meeting Convenor, A/Prof Merilyn Hibma and co-Convenor Dr. John Taylor (AVS10 co-Convenor), both based in New Zealand. Members of the organising committee  and theiAna Fernández-Sesma  (The conference organisers were extremely fortunate in attracting  States)  to preseBenjamin tenOever  ssRNA viruses, where the target exists in incoming viral genomes, viral replication was slowed, but not eliminated, by the miRNA pathway acting for RNAi. Replication delay resulted from the short lag used precisely to excise the target miRNA sequences from the viral genome, probably due to homologous recombination. Targeting RNAi to nucleoprotein-coding RNA of (−)ssRNA viruses additionally leads to an induction of interferon response, whereas targeting to other regions, such as RNA-dependent polymerase, did not. These experiments of curiosity demonstrate that fully functional RNA defense pathways exist in vertebrates that may be limited by the lack of sufficiently active RNA-dependent RNA polymerases needed to produce the quantity of RNA effectors, especially for larger animals, or a mutually exclusive incompatibility between RNAi and interferon pathways. These findings have implications for new antiviral strategies in humans.A thought-provoking keynote presentation was delivered by invited international speaker, NY, USA) , who exaRichard Kuhn  presented a keynote lecture detailing his group’s recent re-examination of flavivirus structures solved by cryo-electron microscopy [croscopy . Kuhn hyAllison Imrie  spoke of the emerging threat of dengue virus in Australia driven by increasing international travel to and from endemic countries, mainly Indonesia. The number of infections detected in Western Australia, mainly involving DENV serotypes 1 and 2, has been on the rise over the past decade, except for 2017/2018, when an unexplained decline occurred. The numbers have surged again in 2019. A prospective follow up of infected patients in Western Australia has been established to observe neutralization antibody responses against homologous and heterologous viruses from one-week post-infection to up to seven years. Data shows that although antibody-mediated neutralization persists against the autologous virus, it wanes over time for within-serotype homologous viruses, questioning the dogma of lifelong immunity against the infecting serotype.In this session, Narayan Gyawali  presented work on a novel scaled-down version of the standard plaque reduction neutralization test (PRNT), re-named as micro-PRNT. He demonstrated its potential utility to sample blood from mosquitoes as an indirect measurement of the prevalence of immunity against Ross River virus (RRV) infection in vertebrate hosts. This assay circumvents the need to directly sample from animals to establish changes in Ross River virus reservoirs in the wild.Alice Michie  presented the most comprehensive phylogenetic analysis of RRV in Australia to date, generating and analyzing 94 new full-genome sequences (and 12 sequences from public databases). The analysis dated the most recent common ancestor of currently circulating variants at approximately 94 years prior and challenged the traditional geographical classification of RRV lineages. The work demonstrated periodic bursts of genetic diversity in RRV phylogenies with four distinct genotypes within Australia, with genotype 4 being the contemporary circulating variant.Nasir Riaz and Chaturaka Rodrigo  introduced a novel wet and dry lab (bioinformatics) workflow to optimise nanopore sequencing to identify within-host variants of hepatitis C virus (HCV). They demonstrated that nanopore sequencing is as accurate as state of art Illumina sequencing for viral consensus sequence generation, providing a coverage of at least 300 reads. Furthermore, nanopore sequencing can reduce sequencing costs to $24–$40 AUD per sample if multiple samples are pooled (multiplexing up to 96 samples) in a single run. The novel bioinformatics pipeline was introduced to differentiate within host variants of HCV by phylogenetically arranging closely related subject specific nanopore reads into clusters and generating a mini-consensus per cluster.Grace Yan  presented work on the utility of wastewater analysis of norovirus and adenovirus sequences and compared the findings with those from clinical samples during the same period in 2018/2019. She argued that data from clinical samples may be skewed towards more severe disease as only symptomatic patients seek treatment, while sampling from wastewater allows a better representation of circulating strains. The genotypes and strains identified from clinical samples demonstrated similar patterns but were not identical in abundance to genotypes and strains identified from wastewater samples. Sampling both sources for viral variants can provide better data for understanding the true prevalence of circulating enteric viruses.Barbara Coulson , who gave an overview of her career in Rotavirus research. This was tightly woven with discovery of rotavirus by Ruth Bishop, Ian Holmes and others, and an outline of her role in the development of an antibody-based diagnostic test for rotavirus. Professor Coulson’s additional advances in rotavirus research included identifying that IgA in stool is protective against reinfection in children, as well as identification of the host receptors for rotavirus entry. The remainder of the talk focused on her research in mice, which has detailed the immunopathological process that leads to rotavirus induced diabetes. Her mouse model has shown that during the initial phase of rotavirus infection, rotavirus induces bystander activation, via TLR7, resulting in strong activation of plasmacytoid dendritic cells and strong upregulation of interferon-dependent gene expression with a skewed Th1-specific response, resulting in activation of pre-existing islet autoreactive lymphocytes.The invited speaker for this session was T. H. Oanh Nguyen  presented work detailing the adaptive immune response to the inactivated influenza vaccine (IIV) [ne (IIV) . Using aCarolyn Samer  presented evidence that the herpes simplex virus 1 (HSV-1) rapidly downregulates both cellular and surface MHC I-related gene protein, MR1 [ein, MR1 , the antAndreas Suhrbier  presented his novel findings surrounding the counter intuitive role of a high-fibre diet in the exacerbation of Chikungunya viral infection [nfection . High-fiAlexander Underwood . He described a human cohort of hepatitis C virus-infected individuals and that antigenic sin, the recall of previous immune responses rather than new immune responses, was commonly observed in reinfection of hepatitis C virus. This may be problematic for hepatitis C virus, which consists of a highly genetically diverse genera. His work has demonstrated that broad neutralizing antibodies on their own were not associated with protection, but instead the quality of the memory B cells and their ability to proliferate, which may be a better marker of protection against reinfection.The final speaker for this session was PhD student, Ed Gane  entitled “Eradication of chronic hepatitis B virus (HBV) infection—translating clinical virology into practice.” Global elimination of HBV has become a reality due to the availability of a safe, effective and inexpensive vaccine but 300 million adults currently live with chronic HBV infection. Although nucleoside polymerase inhibitors can maintain viral suppression and reduce complications, life-long therapy is associated with high cost, risk of breakthrough and potential toxicity. Other steps of the HBV lifecycle can be targeted by small molecules including inhibitors of entry, capsid and virion assembly/release. Capsid assembly modulators (i) prevent encapsidation of pregenomic RNA (pgRNA) and block HBV replication; and (ii) inhibit de-novo formation of covalently closed circular DNA (cccDNA) by interfering with disassembly of the capsid. These inhibitors have recently been demonstrated to be synergistic with nucleos(t)ide analogues (NUCs) in reducing HBV DNA, pgRNA and HBsAg. Translation inhibitors such as small interfering RNAs and antisense oligonucleotides can silence multiple HBV transcripts, potentially blocking the production of the core, polymerase, surface, X-protein. The first clinical studies show potent suppression of HBsAg with only three doses. Chronic HBV infection is characterised by high viral load and antigen burden and inadequate host immune responses. Innovative approaches to restore innate and adaptive immune responses against HBV are currently in clinical development and include therapeutic vaccines, RIG-I agonists, and TLR-7 and TLR-8 agonists. Pilot studies with checkpoint inhibitors, which reverse T cell exhaustion, have also produced promising results. It is likely that HBV Cure will require combinations of novel immunomodulatory, antiviral and cccDNA silencing strategies. Efficacy, safety, route of administration and cost will ultimately determine the impact of these new regimens on the burden of HBV.This session opened with a keynote lecture from Dongsheng Li  discussed research on a novel antiviral to combat dengue virus (DENV) infection, disease and transmission. There is currently no specific DENV antiviral available and the first dengue vaccine licensed has limited data on use and effectiveness. The group’s early research has identified DENV defective interfering particles (DIPs) from all 4 serotype DENV-infected patients. DIPs are virus-like particles that contain the same viral proteins as wild-type virion, and a defective, largely internal deleted viral defective genome (DI RNA). DI RNA can replicate with the help of wild-type virus, while inhibiting wild-type virus replication. DIPs can co-transmit and co-evolve with wild-type virus to produce a durable and resistance-proof antiviral. The group’s early results indicated that DIPs were transmissible and strongly inhibited DENV replication in vitro in serotype-cross and dose-dependent manners, suggesting DENV DIPs may represent a paradigm-shift in strategy of DENV treatment and prevention. The preclinical trials for using DENV DIPs to treat DENV infection and to attenuate DENV transmission in vivo are scheduled.Kanta Subbarao  posed the question: what do post-vaccination antibody kinetics suggest about the timing of the seasonal influenza vaccine? The aim of this study was to estimate the optimal timing of influenza vaccination using antibody decay curves in different age groups, relative to the start, peak, and end of previous influenza epidemics. The final sample consisted of 71 adults (staff aged 18–50 year), 15 ambulatory elderly  and 16 frail elderly (aged care residents aged 65 year+). Estimated geometric mean titres (GMTs) increased for all viruses, peaking 1 month post-vaccination in all groups, before the peak of the season. Titres declined at 3 months, thereafter remaining steady but above baseline, with 6 month post-vaccination GMTs 1.4-, 1.9-, 1.9- and 1.9-fold above baseline for A(H1N1)pdm09, A(H3N2), B/Yamagata and B/Victoria, respectively. The pattern of antibody decay was not significantly different between age groups. The data suggest that subtype-specific antibody-mediated protection following vaccination persists throughout the typical influenza season.Kirsty Short  presented research demonstrating that a history of obesity reduces the immune response to influenza virus in an NLRP3-dependent manner. Obesity significantly increases the risk of death following an influenza virus infection. A novel mouse model was developed to study the long-term effects of obesity on antiviral immunity. Upon infection with influenza virus (A/Auckland/09(H1N1)), previously obese (PO) mice displayed increased viral replication, inflammation, body weight loss and pulmonary dysfunction compared to lean-fed mice. Cells in the lung lumen of PO mice also had an altered metabolic state compared to those of lean fed mice. Importantly, in mice deficient in the NLRP3 inflammasome, obesity had no long-term effect on susceptibility to influenza virus infection. The group propose that obesity can have long-term, NLRP3-dependent, effects on the metabolism of innate inflammatory cells such that they are impaired in their antiviral response.Michelle Tate  posed the question: could a World War II drug be repurposed to limit viral hyperinflammation? Severe influenza A virus (IAV) infections are associated with damaging hyperinflammation that can lead to mortality. The group have identified two existing drugs; Probenecid (used to extend the life of penicillin and is currently used to treat Gout) and AZ11645373 (used for treating inflammatory diseases such as arthritis) which target P2X7 receptor signaling and dampen NLRP3 inflammasome responses during severe IAV infection. Intranasal therapeutic treatment of mice displaying severe influenza disease reduced proinflammatory cytokine production, cellular infiltrates in the lung and provided protection against disease. Importantly, Probenecid and AZ11645737 could be administered at either early or late stage of disease and provide therapeutic efficacy. The group propose these drugs are effective as they dampen NLRP3 responses to a level that is not detrimental or damaging, but sufficient to provide protection.Damian Purcell and his research team, Samantha Grimley and Ashley Hirons   and DNA were isolated and the HTLV-1c genomes were sequenced. Newly identified spliced mRNA variants of the viral HBZ regulatory and p16 accessory genes were discovered, which may contribute to disease severity (later presented in detail by his PhD student Ashley Hirons). Furthermore, Purcell described how HTLV-1c proviral load (PVL) was quantified in PBMCs and T cells using a novel digital droplet PCR (ddPCR) assay developed by his research team. High PVL in T cells was associated with chronic inflammatory diseases of the lung and potentially linked to the novel mRNA spliced proteins. Dr. Samantha Grimley’s research focused on the seroprevalence of neutralizing antibodies measured in the plasma of 21 patients from the HTLV-1c cohort. Over 70% of these patients displayed neutralizing antibodies to the HTLV-1 envelope protein, assessed in a reporter infection assay, which correlated with patients of higher PVL and disease outcomes. This study was the first to describe HTLV-1c seroprevalence in a unique cohort of Indigenous Australians and is an important step toward future vaccine development. Lastly, PhD student Ashley Hirons presented on the novel truncated HBZ mRNA transcript that was discovered from the genome sequencing data. She presented that this novel variant may affect the regulation of important down-stream transcriptional processes. Overall these novel findings will provide and enhance the understanding of the pathogenesis of HTLV-1c infection and inform therapeutic and prevention strategies. It was also a terrific example of how Australasian virologists can contribute to the awareness and prevention of significant viral infections in our Indigenous communities.The Inaugural Indigenous Virology Session was held to highlight the important research undertaken by our Indigenous researchers and/or on viruses affecting our Indigenous communities. In this first iteration, the session was spearheaded by stralia) , who prePeter Waterhouse  spoke on the interaction between Tomato yellow leaf curl virus (TYLCV) and the model plant Nicotiana benthamiana including its wild relatives. The virus TYLCV has a DNA genome and belongs to the genus Begomovirus of the family Geminiviridae. It has a wide host range with a major economic impact on tomato production. Peter and team have utilised this virus to understand more deeply the differences between the laboratory strain of N. benthamiana and its relatives collected recently from its endemic centre of origin, Australia. The team identified RDR3 encoded by the lab strain of the host plant as carrying a premature stop codon that conferred hyper-sensitivity to the TYLCV. By contrast, a Queensland ecotype of the plant that did not bear the mutation was highly resistant. Furthermore, RDR amplifies short interfering RNAs and adds a new player in RNAi mechanism. Using this approach, that maximises the useful differences between the N. benthamiana ecotypes, Peter’s team is exploring further mechanistic aspects of RNA interference (RNAi) and its impacts on virus–host plant interactions.Invited speaker, Sassan Asgari  described the discovery of a new negev-like virus, Aedes albopictus negev-like virus (AaINLV). The negeviruses are insect only viruses, with AaINLV isolated from Aedes albopictus Aa23 cell line and in wild-caught mosquitoes. Sequencing, showed the AaINLV viral genome is a positive sense RNA molecule, containing 3 ORF’s, flanked by 5′ and 3′ untranslated regions (UTR’s). A report showing Wolbachia pipientis is able to limit arbovirus  infection in insects presented an opportunity to investigate Wolbachia (wAlbB) as an agent to limit or suppress infection of the newly discovered AaINLV. Surprisingly, initial investigation by real-time PCR found AaINLV to be present in high copy numbers in Aa23 cells in the presence of wAlbB, suggesting that no suppression of the virus is occurring. Further investigation added weight to the hypothesis that the longevity of association between AaINLV and wAlbB may be a factor in that both wAlbB and AaINLV have effectively learned to “live” with each other due to a long association, whereas, if introduced to each other in cell culture , the virus and bacterium have to go through a period of “adjustment”. Callum Le Lay  presented his PhD research to date using metagenomics to investigate RNA virus diversity in termites and their bacterial symbionts. From this presentation, it was evident that our current knowledge grossly underestimates the diversity of RNA viruses in the environment, with our understanding limited to only a fraction of medically and agriculturally significant members. Focusing on six Australian termite species, Le Lay discovered 98 novel RNA viruses along with 18 novel single-stranded DNA viruses. Subsequent annotation and phylogenetic tree construction established the majority of the viruses occupied basal positions, a testament to the longevity of the association between the RNA viruses and termites. The substance of this “fishing expedition” showed the power of viral metagenomic analysis with a tantalising glimpse into what will turn out to be just the tip of the RNA “virusburg”.Tanja Strive  on the biological control of rabbits in Australia—“an ongoing co-evolutionary arms race”. Two self-disseminating viral biocontrol agents of European rabbits were discussed; myxoma virus (MYXV), which attenuated shortly after its release; and the calicivirus, rabbit haemorrhagic disease virus (RHDV), which appears to be evolving towards high levels of virulence to facilitate insect transmission from virus-laden carcasses. Dr. Strive highlighted that long-term management of rabbit populations will stem from increased research into rabbit caliciviruses, particularly in the context of recent increases in the genetic diversity caliciviruses in Australia and New Zealand.The session opened with an invited presentation by Janine Duckworth , which detailed the effects of RHDV1 K5 release in New Zealand to control wild rabbits. In this study, it was shown that the Czech strain of RHDV1 (released in 1997) was active at all study sites prior to RHDV1 K5 release and that both virus strains were present post-release of RHDV1 K5. The average rate of spread of RHDV1 K5 was 3.5 km/month, with 64%–67% of rabbit carcasses testing positive to RHDV1 K5. This resulted in a 30%–40% reduction in rabbit numbers in the 6–8 weeks following RHDV1 K5 release.The theme of biological control of rabbits was continued with a presentation by Joshua A. Hayward  presented the discovery and characterisation of a group of gammaretroviruses that were found in Australian and Asian bats and are closely related to the koala retrovirus (KoRV) [s (KoRV) . Using aKarla Helbig  spoke on the differential immune responses of New Zealand paua and Australian hybrid abalone to Haliotid herpesvirus 1 (HaHV-1), which causes abalone viral ganglioneuritis (AVG). The research showed that the HaHV-1-resistant päua, upregulated differentially expressed genes in the haemocytes that are involved in extracellular matrix remodelling upon viral challenge. In contrast, these transcripts were absent in the HaHV-1-susceptible abalone.Alice Russo  presented data on comparisons of the viral prevalence and diversity of cane toads (Rhinella marina) found in their native range (French Guiana) and two introduced ranges . In the native range toads, multiple phylogenetically distinct viruses were detected that were not present in the Australian cane toads. These data provide a foundation for studies looking at the influence of viruses on cade toad invasion.The first of two sessions on virus–host interactions highlighted strategies evolved by viruses to exploit proteins and small molecules co-opted from the host cell to facilitate steps in the viral lifecycle or molecular defense mechanisms mounted by the host cell to interfere with viral replication. Alexander Khromykh  presented two high-throughput methodologies used in elucidating determinants of flavivirus pathogenesis and evasion of host response. In the first approach, he used deep mutational scanning to identify viral protein function in replication and interactions with host factors . In the Agathe Colmant  presented a new host restriction mechanism of Bamaga virus, a newly discovered flavivirus that replicates in mosquitoes. Interestingly, replication in vertebrate cells was attenuated at 37 °C but restored at 34 °C, whereby the temperature-dependent restriction was observed after cell entry. Adaptive mutations that allowed replication at the higher temperature were mapped to residues close to a viral protease site suggesting that suppression of proteolytic processing prevents replication of Bamaga virus in vertebrate cells.K M Rifat Faysal  presented new single-molecule fluorescence imaging approaches to investigate the uncoating kinetics of HIV-1 capsids. Analysis of capsid mutations in this assay allowed identification of residues involved in selectively binding the host molecule inositol hexakisphosphate (IP6) as assembly factors. IP6 stabilises the immature virus during assembly in the producer cell and is then recycled to catalyse assembly of the mature capsid inside the viral particle, revealing a role of this host molecule at multiple stages of the viral lifecycle.Jade Forwood  and his team determined how Hendra and Nipah viruses hijack cellular machinery for import of viral W proteins into the nucleus. Nuclear localisation of W proteins is required to counteract innate immune responses that would otherwise prevent the virus from replicating. X-ray crystal structures of complexes between nuclear import machinery and the nuclear localisation sequence of W proteins revealed the residues involved in the high-affinity interaction with the transport receptor importin α3, adding to our understanding of virus–host interactions and nuclear transport adaptor cargo specificity.Botrytis gemydayivirus 1, which represents a new genus and species. The virus is mechanically transmitted and infects the Botrytis cinerea, a fungus that leads to disease in plant species including grapes. The new virus restricted the growth of its fungal host.Robin Macdiarmid  presented on the reciprocal control of replication between a new ssDNA mycovirus and its host. The team identified and purified infectious viral particles of an exotic virus, named Jeremy Barr , emphasised that bacteriophages are one of the dominant groups of biological entities in our body, outnumbering both bacterial and human cells. Despite this observation, very little sequence data is publicly available for bacteriophages and how homologous bacteriophage species are between people. To remedy this knowledge gap, Dr. Barr has employed the bioinformatics tool, VirSorter, to identify new bacteriophages. He has also studied bacteriophage populations (virome) in humans and identified that these populations are very unique between individuals. This is a new and exciting area of research and, given that bacteria are their natural hosts, phages are an integral member of the human microbiome, and so their potential association with disease is of great interest.Invited speaker, Jessamine Hazlewood , reported on the use of the poxvirus-derived vector system, Sementis Copenhagen vector (SCV), to deliver arboviral structural proteins in mice. Their aim was to utilise RNA-Seq to understand successful signatures of protective immunity, and also, especially in the case of the chikungunya virus, to test for signatures of arthritis. Natural chikungunya virus and a previous chikungunya vaccine have been associated with the development of arthritis. Hazlewood’s aim is to understand RNA signatures associated with adverse vs. safe expression profiles and to use that information to help inform vaccine development. The data so far indicates that signatures of Toll-like receptor activity are associated with SCV immunogenicity.Andrii Slonchak  presented work on the importance of non-coding subgenomic flaviviral RNA (sfRNA) in successful replication and transmission of Zika virus, and its subsequent impact on disease pathogenesis. Dr. Slonchak made mutant Zika viruses unable to make sfRNA and then discovered that while they could replicate in vitro, their ability to spread to the salivary glands in vivo was attenuated. Further investigation suggested that while sfRNA had no effect on the RNAi pathway, it is important for preventing cell apoptosis by inhibiting Caspase 7 expression, and preventing innate immune functions by inhibiting phosphorylation of STAT1 [Karyn Johnson  started off the session by summarizing and sharing new data on the role of host micro RNAs (miRNAs), now commonly-known regulators of gene expression, during virus infections. Karyn emphasised that miRNAs could exert both proviral and antiviral functions during virus infection. By using the natural Drosophila melanogaster-Drosophila C virus (DCV) model system, she showed that miR-956, through its target Ect4, supports DCV replication [Invited speaker lication , whereasAlice McSweeney  presented her work on murine norovirus genome-linked protein VPg, which is encoded by all noroviruses and known to arrest host cell cycle at the G0/G1 interphase to benefit virus replication. Alice shared that the N-terminal region of VPg is essential but not sufficient on its own for cell-cycle arrest. Furthermore, the VPg N-terminal region possesses a functional positively-charged amino acid motif (KGKxKxGRG) and has the ability to non-specifically bind RNA at low affinity.Stephen Rawlinson  highlighted that despite carrying out key replication steps in the host cell cytoplasm, many RNA viruses target their proteins to the host nucleolus. He presented that Nipah virus and Hendra virus M proteins localise to a subnucleolar region and interacts with nucleolar protein, Treacle. This interaction results in the inhibition of Treacle-mediated host ribosomal RNA biogenesis and facilitates virus replication [lication . In addiByron Shue  presented data from a CRISPR/Cas9 genome-wide knockout screen that identified RACK1 as a proviral host factor for multiple flaviviruses including Zika, Dengue and West Nile. RACK1, a scaffold protein in host cell homeostatic processes, interacts with flavivirus NS1 protein and is associated with recruitment of NS1 to viral replication complex in endoplasmic reticulum.Thomas Tu  closed the session by reporting that the integration of hepatitis B virus (HBV) DNA into host cell genome, a step associated with liver cancer progression and viral persistence, is not driven by an active mechanism involving viral proteins. He found that the rate of integration is similar in cells infected with wild-type HBV or a replication-deficient HBV mutant or cells transfected with viral DNA. These data suggest that DNA integration occurs through a passive mechanism involving host cell DNA repair machinery.David Jacques  gave an elegant talk that described the molecular structure of the HIV capsid complex, where each capsid hexamer has a size-selective pore bounded by a ring of six arginine residues, and the role of a highly conserved arginine amino acid residue (R18). The HIV capsid is a fullerene cone comprising ~250 hexamers and exactly 12 pentamers of the capsid protein. R18 is located at the centre of each hexamer and pentamer. Despite exerting a strong electrostatic force at a critical junction of the capsid lattice, this residue is essential for the function of many retroviruses [oviruses . Using aSvenja Fritzlar  investigated the late stages of the human cytomegalovirus (HCMV) replication cycle, which is currently not well characterised. A library of >100 HCMV mutants was screened for viral mutants that showed typical viral assembly complex formation (a cytoplasmic megastructure associated with late stage HCMV infection), but which were unable to complete the viral lifecycle. In this way, she identified a subset of viral mutants and from this characterised a viral protein as crucial for HCMV egress. These proteins represent a set of novel antiviral targets for inhibiting HCMV spread.Daniel Watterson  described a novel chimeric virus platform which can be used to rapidly solve the structures of flaviviruses by cryo-electron microscopy [croscopy . Using aThao Huynh  described a model for investigating the interactions between hepatitis B virus (HBV) and host proteins. Using a plasmid-free monomeric HBV (“1-mer”) transfection model which expresses cccDNA-like molecules, the viral genomic reservoir involved in HBV persistence, the researchers asked whether expected viral replication intermediates could be produced following the co-transfection of host factors known to modulate HBV infection (DDBI proteins). By co-transfecting these plasmids into hepatoma cells, including 1-mers from different viral genotypes, Huynh confirmed that this system produced the expected HBV replicative intermediates observed using more conventional in vitro models, but with the advantage of being able to detect cccDNA-like molecules usually absent in equivalent systems. Given the role of cccDNA in driving HBV persistence, this model may present opportunities for future development of novel therapeutics to cure chronic HBV infections.Kate Stacey’s  first piece of data was a very early photo of the “Three Musketeers” of RNA virology in Queensland–Alex Khromykh, Paul Young and Roy Hall–who have all made significant contributions to virology at the national and international level. Following on from this, she discussed a role for dengue virus NS1 in activation and induction of TLR4 responses. TLR4 is the canonical receptor for bacterial LPS; however, recent studies have shown that viral and host factors during influenza, Ebola and respiratory virus infections can activate TLR4. Stacey presented data showing that insect- and mammalian cell-expressed DENV NS1 drives inflammatory cytokine expression, which could be blocked by competitive inhibitors for LPS and sepsis. She linked this to NS1 disruption of the endothelial barrier both in the gut and the liver, and proposed that TLR4 antagonists may have therapeutic potential against Dengue infections.Invited speaker, Matloob Husain  presented on the role of host acetylation machinery during influenza A virus (IAV) infection. Host acetylation is regulated by a delicate balance between histone acetyltransferases (HATs) and histone deacetylases (HDACs), and the presentation proposed a proviral role for HATs, and an antiviral role for HDACs during IAV replication. Focusing on HDAC4, he showed that HDAC4 knockdown enhanced IAV replication and reduced the production of interferon-stimulated genes (ISGs) such as Viperin, ISG15 and IFITM3. Furthermore, IAV degraded HDAC4 transcripts by its endonuclease PA-X, as well as targeting HDAC4 protein by cleavage through host caspase 3 [aspase 3 .Larisa Labzin  discussed the detection of viral infection, and how the location of pathogen sensing could determine the strength of the responses. Using an adenovirus model, she showed that infection of macrophages with antibody-coated adenovirus triggers strong inflammasome responses from damaged lysosomes. She also went on to describe that antibody-coated adenoviruses that escape into the macrophage cytosol could initiate a second wave of inflammasome signaling, and that this was mediated by the intracellular Fc receptor TRIM21 and cGAS.Ebony Monson  continued with the intracellular response theme, presenting data that supported an emerging role for lipid droplets in early antiviral responses. Lipid droplet size and number increased with virus infection , and with treatment of DNA/RNA ligands. This increase in lipid droplet production correlated with decreased ZIKV replication and enhanced expression of interferon. Interestingly, this was dependent on EGFR signaling, as EGFR inhibitors increased ZIKV production and limited lipid droplet and interferon production. Monson finished off her talk with some exciting work elucidating the proteome of purified lipid droplets and discussed the potential for localisation of antiviral factors to lipid droplets as an antiviral hub.Greg Moseley  who provided for the first time, structural data from NMR and super-resolution microscopy, on how the rabies P protein interacts with STAT1 to block the antiviral interferon response. NMR analysis of the P protein and STAT1 complex revealed multiple contact points between the proteins with mutational analysis at these sites preventing STAT1 binding and IFN antagonism. Importantly, insertion of these mutations in a pathogenic field stain of rabies revealed significant attenuation of disease, highlighting the importance of this interaction. This work furthers our understanding of host–virus interactions that block the innate response to infection and may pave the way for possible novel vaccine strategies.The final presentation was by Invited speaker Lorena Brown  discussed recent results from her laboratory, in collaboration with colleagues at Oxford University, relating to the molecular determinants of gene segment packaging in influenza virus . She desFlaviviridae family  continue to cause havoc worldwide. With close to 400 million cases of dengue infections per year and no dengue virus (DENV)-specific antiviral drug approved, vaccination is the most promising tool to control this arboviral disease. Although one tetravalent live-attenuated chimeric vaccine (CYD-TDV) is currently commercially available in some dengue endemic countries; poor efficacy in children and in seronegative individuals has been observed [Mosquito-borne flaviviruses cause a significant global health burden and have potential for pandemic emergence . During observed . Jovin Choo  and colleagues described the use of the BinJV platform described by Dr. Hobson-Peters to develop a new chimeric DENV vaccine. They exchanged the structural proteins of BinJV with two proteins from dengue virus 2  to construct the BinJV/DENV2-prME chimera. Using an AG129 infectious DENV mouse model, they demonstrated the protective efficacy of BinJV/DENV2-prME chimera by showing (i) high production of neutralizing antibodies and (ii) 80% to 100% survival after challenge with lethal doses of DENV-2. This study confirmed the potential use of BinJV chimera constructions to develop novel vaccines—in this case, against dengue virus.Anjali Gowripalan  described how the CRISPR/Cas9 system can be effectively harnessed and adapted into a powerful selection tool for recombinant poxviruses. In recent years, CRISPR/Cas9 has enabled the manufacture of recombinant large dsDNA viruses from various families enabling and accelerating mechanistic studies into viral pathogenesis, host–virus interactions and development of novel interventions. For these viruses, CRISPR/Cas9 is used to facilitate homologous recombination, in much the same way as has been widely adopted for engineering genomes in cells and animals. Dr. Gowripalan’s work, however, highlights the unreliability and poor efficiency of this approach for a vaccinia virus, which has a cytoplasmic site of replication. Moreover, she dissected the reasons why this approach does not simply translate to poxviruses. Using this knowledge, a highly efficient and innovative two-stage method for virus generation was developed and tested. This approach was able to reduce the time for virus generation from months to 1–2 weeks, which extends the utility of vaccinia virus as a vaccine vector to situations requiring a rapid response, such as emerging infections and personalized cancer immunotherapeutic strategies.With close to 37 million people living with HIV-1 worldwide , HIV/AIDNatalia Salazar-Quiroz  was the recipient of the Australian Society for Microbiology award for her three-minute oral presentation demonstrating the dual activity of bovine antibodies to broadly neutralize as well as elicit Fc-effector function to HIV-1 envelope glycoproteins, while Natalee Newton  received the Microorganisms MDPI award for her presentation of the first cryo-EM structure of the insect-specific flavivirus BinJV. Other presentations of note were those from Tasmin Gordon (Burnet Institute) and Fernando Villalon Letelier (The Peter Doherty Institute for Infection and Immunity at the University of Melbourne), who both presented novel work describing the activity of host antiviral proteins. Tamsin Gordin characterised the unique ability of Egyptian fruit bat tetherin to control both EBOV and MARV viral particle release, while Villalon Letelier described the ability of MARCH 8 to reduce the infectivity of IAV particles by reducing the incorporation of integral proteins into the virion. Overall, these sessions showcased the remarkable research conducted by early career researchers throughout Australia and New Zealand which helps further understand both animal and human viral diseases and their treatment.AVS10 provided the opportunity for numerous early career researcher—many of whom were current PhD students—to present their novel research findings during two sessions of three-minute oral presentations. These sessions encompassed a broad range of virological research, ranging from investigations into livestock viral pathogens, antiviral and vaccine trials, immune surveillance, to basic viral and host research. AVS held its second career development workshop over breakfast with a panel discussion led by AVS Vice President Professor Heidi Drummer (Burnet Institute) with panelists Professor Allison Abendroth (University of Sydney), Professor Vernon Ward (University of Otago), Dr. Michelle Tate (Hudson Institute) and Dr. Kirsty Short (University of Queensland). The experiences of the five panelists were diverse, and this was equally reflected in their diverse views on this important topic.Every panel member made a conscious decision to become a lab head and pursued funding opportunities relentlessly, showing a high degree of resilience to knock backs and obstacles. The time required post-PhD varied considerably, with some undertaking several post-doc stints, and others stepping straight out of their first post-doc into a lab head position with funded grants. Overseas post-doc experience was not essential amongst this panel, with several never having worked overseas. A university academic appointment can assist significantly, providing essential personal salary support, access to small grants and high-quality students. These opportunities are limited and highly competitive but should be considered even if relocation is required.Mentoring is often considered an essential component of training and professional development. On this panel, one member believed strongly that their mentor was essential to their development and future independence. However, this was not identified as being essential for the majority of the panelists. Instead, panelists identified a strong advocate as being perhaps more important; someone who actively pushes you onto review panels, into speaking positions, chairing sessions, and positions of influence. Advocates are often already successful, have nothing to lose by passing on an opportunity to you, and they believe in you and the calibre of your work.All panel members agreed that once funding has been secured, it is tricky to know how much time should be devoted to writing new grants, as this time commitment detracts from writing manuscripts and performing experiments. There was division amongst the panel, with some believing that every funding opportunity should be pursued, and others believing that a period of consolidation is necessary to ensure that funded work is initiated and providing an important respite from the application process. Small grants should not be overlooked, particularly early in your career, as they provide essential seed funds to generate preliminary data for bigger grants. Sometimes two or three small grants can equal the budget of an Australian Research Council (ARC) or Australia National Health and Medical Research Council (NHMRC) grant for one year, providing consumable funds to support students or short-term employees. Look at University and Institute web pages for lists of funding opportunities, track down people previously successful and ask for advice.An important consideration for all post-docs is the issue of family responsibilities, whether this is having children, looking after children or elderly parents. The importance of flexible work hours was identified by all panel members and participants as essential requirements to maintain a healthy and productive work–life balance. Time management and being efficient assist in making the most of the working day and many identified the importance of a good network of people to provide support as essential success factors.Overall, every individual must carve their own path to success; there is no golden ticket, there are no set rules and the path is unlikely to be linear. However, resilience was identified as the one key attribute that you must possess and actively display if you are to make the transition from post-doc to lab head. None of the panel members regretted their decision and medical research remains one of the most exciting and fulfilling careers available to those who are scientifically minded.Overall, the presentations at AVS10 showcase the breadth and quality of virology being undertaken in the Australasian region, spanning fundamental research, and translational, clinical and epidemiological studies. AVS10 consistently showcases the talent of our budding virologists. Our next meeting in December 2021 marks an auspicious occasion celebrating 20 years of AVG/AVS, and we are planning to hold the meeting in Fraser Island, Queensland, Australia, the location of the very first Australian virology meeting.


The biomarkers commonly utilized in diagnostic evaluations of kidney disease suffer from low sensitivity, especially in the early stages of renal damage. On the other hand, obtaining a renal biopsy to augment clinical decision making can lead to potentially serious complications. In order to overcome the shortcomings of currently available diagnostic tools, recent studies suggest that exosomes, cell-secreted extracellular vesicles containing a large array of active molecules to facilitate cell-to-cell communication, may represent a rich source of novel disease biomarkers. Because of their endocytic origin, exosomes carry markers typical for their parent cells, which could permit the localization of biochemical cellular alterations in specific kidney compartments. Different types of exosomes can be isolated from noninvasively obtained biofluids; however, in the context of kidney disease, evidence has emerged on the role of urinary exosomes in the diagnostic and predictive modeling of renal pathology. The current review summarizes the potential application of exosomes in the detection of acute and chronic inflammatory, metabolic, degenerative, and genetic renal diseases. Kidney disease, which encompasses various acute, chronic, or end-stage conditions, incurs a considerable health burden due to high prevalence and costly management . While mThe conventional biomarkers or renal disease in clinical practice are suboptimal: serum creatinine is limited by poor sensitivity in mild-to-moderate kidney failure and eGFR by its dependence on creatinine measurement in the early stage of renal dysfunction ; in addiExosomes are bilipid membrane-bound vesicles measuring 40-120 nm in diameter; they are distinct from other extracellular vesicles (EVs) such as microvesicles and apoptotic bodies because their biogenesis is linked to the endosomal pathway. They are generated by an inward blebbing of the endosomal membrane that produces multivesicular bodies (MVBs), which are then fused with the plasma membrane and released via exocytosis . Those miRNAs can be analyzed by RNA sequencing. There are currently bedside RNA sequencing techniques that give results within a few hours , 20.Human models of renal disease have demonstrated that kidney damage is primarily driven by immune dysregulation and alterations in hemostasis, vascular integrity, and matrix modulation that are regulated by exosomes . CirculaThe potential of exosomes to serve as therapeutic agents or drug delivery vehicles in chronic kidney disease  to allevMicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression mainly through RNA silencing . ExosomeExosomal cargo which determines cell-targeting can give us a wealth of information about the original cytosolic environment and relevant biochemical changes and also serves as a potential source of biomarkers. In this review, we aim to synthesize published data from human studies to date. The candidate biomarkers are presented in Tables 1We searched the online MEDLINE® database of the U.S. National Library of Medicine with the complex term (exosomes OR “extracellular vesicles”) AND  which produced 393 results . Articles referring to “extracellular vesicles” were included as long as the described experimental method included exosomal isolation. Ultimately, the literature cited herein includes 95 peer-reviewed, original articles of studies in humans published in English. We used simple narrative analysis to summarize the data from the studies selected for review.α [Chronic kidney disease (CKD), which is characterized by the gradual irreversible deterioration of kidney function, is a multifactorial condition caused mainly by metabolic and inflammatory changes and is typically diagnosed and staged based on the estimated glomerular filtration rate (eGFR). In CKD, urinary exosomal cargo is characterized by higher levels of ceruloplasmin  and the α  and is iα ; miR-451α . These bα . It is sα , or contα . Renal fα  as well α .Hyperglycemia secondary to diabetic nephropathy gradually damages all compartments of the kidney, beginning with glomerular capillary dysfunction with hyperfiltration and microalbuminuria and ultimately leading to interstitial fibrosis, tubular atrophy, and interstitial inflammation in advanced disease. Evidence of exosome-mediated podocytal injury is evident by either increased Elf3 protein  or WT1, Hypertensive nephropathy is the result of either long-standing essential hypertension causing vascular-glomerular damage and remodeling or a primary renovascular lesion leading to renal hypoperfusion and secondary hypertension. The level of urinary exosomal plasmalemma vesicle-associated protein (PLVAP), a protein expressed in the peritubular capillaries, is associated with clinical measurements such as blood pressure and eGFR, and also the histological count of peritubular capillaries and degree of fibrosis in renal patients with essential or renovascular hypertension. This association makes PLVAP a potentially specific biomarker of microcirculation injury . UrinaryPreliminary data from small clinical studies in critical care medicine have identified two urinary exosomal proteins as candidate biomarkers of acute kidney injury (AKI): (i) activating transcriptional factor 3 (ATF3), which is activated in models of ischemic reperfusion injury , and , a cytosolic protein expressed by increased number of renal cells in kidneys exhibiting ongoing injury . In idioNephritic syndrome, defined by the presence of hematuria in association with hypertension, oliguria, fluid retention, and a decline in renal function, is an inflammatory process with a histological picture of glomerular basement membrane ruptures and usually diagnosed by renal biopsy. Common causes of nephritic syndrome include anti-GMB disease, IgA nephropathy, and lupus nephritis.α-1-antitrypsin, and ceruloplasmin have been used to distinguish between IgA and thin basement membrane nephropathy which is another common cause of glomerular hematuria [In IgA nephropathy, increased expression of urinary exosomal miR-215-5p and miR-378i and decreased expression of miR-29c and miR-205-5p have been described compared to healthy individuals . Higher ematuria . A decreematuria , whereasematuria .Lupus nephritis is characterized by downregulation of urinary exosomal miR-29c  and upreCystic kidney diseases (CKD) are heterogeneous in origin, distribution, and pathogenesis; many are related to genetic defects. Autosomal dominant polycystic kidney disease (ADPKD), the most common inherited CKD, mainly results from PKD1 mutations. Hogan et al. described lower PKD1 and higher transmembrane protein 2 (TMEM2) urinary exosomal protein secretion in ADPKD, suggesting that the PKD1/TMEM2 ratio may have some diagnostic utility . Thirty Exosome isolation and characterization have assisted in the diagnostic challenge to differentiate between medullary sponge kidney (MSK), a cause of medullary nephrocalcinosis, and idiopathic calcium nephrolithiasis. Bruschi et al. found that higher FCN1 and C4PBP, as well as lower MASP2 serum exosome protein levels, were positively associated with MSK . FurtherA pilot study with patients with cystinuria highlighted that 165 urinary exosomal proteins, analyzed by mass spectrometry, could be utilized to identify patients and also determine the severity of disease . In two Treatment optimization of renal disease depends on the availability of diagnostic and prognostic biomarkers. The use of renal biopsy, which remains the gold standard in the diagnosis of kidney disease, is also affected by potentially serious postoperative complications and the possibility of improper or nonrepresentative sampling.+/H+ exchange regulatory cofactor 2, and angiotensin-converting enzyme [It is evident that exosomes may be the solution to finding accurate renal disease biomarkers without the need for invasive procedures. Proteomic profiling of urinary exosomes by mass spectroscopy and subsequent computational analysis, reveals an abundance of proteins implicated in pathophysiologic processes such as sodium ion transport, immune activation, and epithelial cell differentiation . Additiog enzyme . Productg enzyme  or totalg enzyme .Another advantage of exosomes as potential biomarkers is the expression of markers that are specific for their cell of origin that allows the tracking of alterations in specific cellular compartments within a tissue . Hogan eDespite all the theoretical advantages of exosomal biomarkers, there are many challenges, both technical and translational that need to be addressed before the routine application in the clinical practice.The main challenge in exosome isolation is to differentiate exosomes from other EVs. Unfortunately, to date, there is no single isolation technique that guarantees purity, speed, cost-effectiveness, and ability to process large sample volumes at the same time. Ultracentrifugation-based techniques are inexpensive with a low contamination risk and are suitable for large volume preparation; however, they require expensive nonportable equipment and are labor-consuming, making them unsuitable for small volume sampling. Additionally, high centrifugation speeds may mechanically damage EVs . UltrafiThe exosomal cargo is speculated to be reflective of complex intracellular changes. However, it is unclear and dependent on the condition whether exosomal biomarkers can be more sensitive for the detection of a pathologic process than nonexosomal biomarkers. In general, exosomal and nonexosomal EV cargoes can overlap considerably but are not identical. For example, analysis of the proteomic composition of urinary EVs has revealed that some proteins are detected exclusively either in microvesicles or exosomes . In lupuAnother consideration is which biofluid is optimal for exosomal biomarkers in renal disease diagnostics. The vast majority of biomarkers examined in this review were identified in urinary exosomes and very few in serum. Other biofluids such as peritoneal dialysis aspirate contain exosomes which could carry biomarkers associated with membrane failure . It has The diurnal variations of urine consistency should also be accounted for in the evaluation of urinary biomarkers. Urine creatinine is commonly used to normalize the values of soluble urinary biomarkers, but its relevance to exosomal biomarkers remains unknown . As far Lastly, comorbidities should also be taken into account. Changes in circulating and urinary exosomal contents have been reported in patients treated with antihypertensive agents  or cycloExosomes represent a valuable source of candidate diagnostic and/or prognostic biomarkers for a variety of renal conditions. Their potential to reflect changes in specific cellular compartments of the nephron is of particular interest. Exosomes, particularly urinary ones, may provide a dynamic image of the processes taking place in the affected renal tissue. Exosomal biomarkers unlike renal biopsies are not limited by the possibility of obtaining unrepresentative sampling. Exosomal purification and analysis require minimally to noninvasive techniques depending on the biofluid of interest, and exosomal isolation technology is constantly improving. This allows for serial analyses in follow-up clinical visits for comparison. Moreover, exosomal miRNAs have a potential diagnostic and therapeutic potential mainly to their active role in disease pathophysiology. In many cases, exosomal biomarkers may complement renal biopsy in risk stratification and prognostic evaluation. Clinical correlations of currently available data on exosomes in kidney disease are difficult to make currently because of a considerably low to moderate sample size in most research efforts found in the literature and the lack of a standardized methodology of exosome isolation that prevents the direct comparison of study results. Further work is warranted in order to identify accurate and reliable exosomal biomarkers that could complement or replace currently available diagnostic tools. Although great progress has been achieved in exosome research so far, further work is warranted in order to identify accurate and reliable exosomal biomarkers that could complement or replace currently available diagnostic tools.