Melanoma is one of the most aggressive forms of skin cancer, with limited therapeutic options. Since its incidence has been rapidly rising in recent years, the study of new targeted therapeutic strategies has increased. The implication of nanoscience in the development of alternative targeted therapies for melanoma has multiple benefits and could significantly improve the outcome of melanoma patients. In this paper, we review the most recent progress in the field of targeted therapies, emphasizing the impact of nanoscale materials on the targeting and controlled release of anti-tumor drugs. The applications of nanomedicine in the management of melanoma are extensive and refer to sentinel lymph node mapping, chemotherapy, and RNA interference; each of these applications harboring the potential to develop efficient and personalized diagnostic techniques and therapies. Further research, especially in clinical trials, is needed to establish whether fighting melanoma on the nanoscale level represents the key to reaching a critical inflection point in mankind’s battle with metastatic melanoma. The incidence of melanoma, the most aggressive form of skin cancer, is alarmingly rising on a global scale. The American Cancer Society suggests a worrying estimate for 2019 in the U.S. of 96,480 newly diagnosed cases of invasive melanoma and 7230 deaths from advanced melanoma [The treatment of melanoma has evolved significantly over time, from surgery to chemotherapy, to radiation therapy, and nowadays to modern targeted therapy. When melanoma is diagnosed at an early stage, surgical excision may be curative. However, in the setting of advanced melanoma complex surgical and medical treatment is necessary and the clinician is often faced with difficult therapeutic challenges, long-term responses being difficult to achieve. However, recent advances in the modulation of immune responses against cancer have changed paradigms, and with the first regulatory approval of a novel immunotherapeutic agent targeting an immune checkpoint, cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), ipilimumab, the future of cancer care was changed forever. This very moment has subsequently led to the approval of over ten different immunotherapeutic agents, including other checkpoint inhibitors.checkpoint inhibition immunotherapies: programmed cell death 1 (PD-1) or programmed cell death ligand 1 (PD-L1) inhibitors  alone or in combination with anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) antibodies (ipilimumab);molecularly targeted therapies used in patients with driver mutations, against the mitogen-activated protein kinase (MAPK) pathway, with BRAF  and MEK (mitogen-activated protein kinase) inhibitors  in patients harboring a V600 mutation in the BRAF gene, and tyrosine-protein kinase (KIT) inhibitors (imatinib);tumor vaccines;gene therapy, such as locoregional immunotherapy with oncolytic viruses ;T cell directed therapies;other non-specific approaches  indicated only in a few subsets of patients due to their questionable benefits for the patient’s outcome .The main available medical therapies of melanoma are:Despite the continuous research and development of novel antineoplastic therapies, melanoma remains a disease that continues to outpace the solutions. The efficacy of the currently available melanoma therapies is limited by multiple drawbacks. The drugs’ interaction with biological fluids and membranes, as well as the heterogeneous tumor immunology and vasculature, are possible explanations for drug resistance ,4.Leading scientists focus their research on developing innovative nano-technologies that have the potential to increase therapeutic accumulation in targeted malignant melanocytes and enhance drug efficiency, while lowering systemic toxicity. The properties of nanomaterials  can be optimized in order to overcome all these barriers and bridge this gap. This has led to the development of ultra-small drug delivery platforms driven by nano-technology.clinicaltrials.gov database there are 2415 clinical trials on melanoma worldwide, but only ten investigating the use of nanotechnology in this type of cancer.Most of the research on the subject is currently preclinical. Officially, on the In this article, we aim to review the intensive research carried out at the interface between nanotechnology, medicine, biology, and chemistry, in the effort to enable nanosystem-based treatments for melanoma patients.Nanostructured materials have been comprehensively studied, with the purpose of enabling precision targeted drug delivery for melanoma treatment. They have the ability to navigate the body’s maze and to deliver drugs directly into a diseased cell only, without affecting healthy cells. This concept of drug targeting through nanosystems results in higher efficacy and fewer side effects. The results can be achieved through both passive and active targeting ,6.The EPR (enhanced permeability and retention) effect is a characteristic feature of tumor neovascularization and it represents the basis of nanotechnology-based drug delivery systems . The tumThese findings gave birth to an emerging field investigating different types of nanotechnologies capable of actually accumulating a medicine in a diseased area rather than affecting the healthy areas of the body.Active targeting  represents a similar strategy but with increased specificity in binding to certain tissues. This is due to the attachment of targeting ligands (such as antibodies) to the nano-carriers. Ligands are able to recognize surface receptors on the specific targeted cells and effectively bind to the receptor site, increasing the drug’s specificity.The most investigated nanoparticles effective in melanoma therapy are: polymeric nanoparticles, liposomes, inorganic nanoparticles, carbon nanotubes, and dendrimers.Polymeric nanoparticles are being researched extensively as drug delivery vehicles due to a number of factors, which include: (1) flexibility; (2) controlled release properties; (3) good biocompatibility and biodegradability; and (4) their ability to be used as multifunctional carriers .Various types of polymeric nanoparticles can be synthesized depending on the polymer’s properties and its applications, namely nanospheres and nanocapsules, polymeric micelles, polymersomes, dendrimer-based micelles, and polymer–drug conjugates .Liposomes are specialized lipid vesicles that possess a central aqueous compartment surrounded by a bilayer hydrophobic structure of phospholipids, similar to cell membranes . NanostrThey are able to increase drug circulation time through various mechanisms, such as size adjustment (under 100 nm) or functionalization  . In thisNoble metals such as gold, silver, and platinum biocompatible nanomaterials have applications in melanoma diagnosis and treatment.Gold nanoparticles (AuNPs) present with major biomedical advantages, such as: synthetic accessibility, versatility in size, shape or surface chemistry, valuable biocompatibility and stability, surface plasmon resonance .Due to the above properties, AuNPs have been extensively investigated for various functions, such as: (1) delivery vehicles for drugs and small interfering RNA (siRNA); (2) photodynamic and photothermal therapeutic utility.AuNPs are also suitable for an active delivery model, as they can be conjugated with antibodies against cancer markers. In an experiment on human melanoma cells, Choi B.B.R. et al. (2017) conjugated AuNPs with phosphorylated focal adhesion kinase (p-FAK), a protein highly expressed in melanoma cells, and combined the treatment with cold plasma treatment . G361 huLomelí-Marroquín D. et al. (2019) achieved the synthesis of bimetallic silver/gold nanoparticles, with the advantages of an increased biocompatibility, usually associated with AuNPs, and of a decreased toxicity of silver nanoparticles (AgNPs) . The proPlatinum nanoparticles (PtNPs) have also been used in preclinical trials. They showed good biocompatibility, as well as anti-inflammatory effects due to their enzymatic properties against reactive oxygen species ,16.Carbon nanotubes (CNTs) are molecular tubes formed from one or more rolled-up sheets of graphene . They are classified as single-walled (SWNT) and multi-walled nanotubes (MWNT) . CNTs arDendrimers are three-dimensional synthetic polymers with a branching tree-like structure . The worStructurally, dendrimers are constructed by the stepwise connection of a central core, multiple branched arms, and peripheral groups. They can be synthesized through two approaches: (i) a convergent approach  or (ii) the divergent approach  .Their main characteristics  can be easily optimized in the engineering process . Thus, dIn recent years nanotechnology has developed considerably, with extensive biomedical applications, also allowing the engineering of better therapies in melanoma. Multiple in vivo and in vitro studies showed significant improvements of almost each standard melanoma therapy by the association of nanoparticle delivery systems. In this subchapter we aim to review the multiple applications of nanotechnology in the treatment of melanoma, starting from the advantages and drawbacks of available therapies towards their improvement by the use of nanosystems.Cancer immunotherapy (passive or active) can specifically and persistently enhance the immune response to fight tumor cells. Over the years, promising results have been achieved in preclinical and clinical studies with several types of immunotherapeutic agents: monoclonal antibodies, immune checkpoint inhibitors, non-specific immunomodulatory agents, whole tumor cancer vaccines, and others ,21.Immunological drugs have changed the therapeutic landscape of advanced melanoma.The most significant advantages of checkpoint inhibition immunotherapy are the long-term response, the prolongation of overall survival, and progression-free survival.Dual therapy with nivolumab plus ipilimumab seems to achieve higher response rates and prolong survival more than nivolumab monotherapy in metastatic melanoma patients, regardless of the BRAF status ,23. AccoNivolumab is indicated as an adjuvant treatment in all patients diagnosed with stage IIIA-IIID melanoma or totally resected stage IV melanoma. Nivolumab and ipilimumab combined treatment also shows promising results as neoadjuvant therapy in patients with macroscopic stage III melanoma .Despite these encouraging results, the recent CheckMate 067 study showed that 52% of patients reached a five-year overall survival with immunotherapy combination strategy . One in Another major aspect to be considered is that responses to immunotherapy typically develop slowly and patients may experience transitory exacerbations of disease before the objective therapeutic response is achieved.Moreover, patients undergoing immunotherapy often experience an array of immune-related adverse events (irAEs) that range from mild to serious, or even potentially fatal reactions. The most frequently encountered irAEs are dermatological , diarrhea/colitis, hepatitis, pneumonitis, and endocrine toxicities . In the KEYNOTE-006 trial, patients treated with pembrolizumab, severe grade 3–4 treatment-related adverse events occurred in 96 (17%) of 555 patients in the combined pembrolizumab groups and in 50 (20%) of 256 patients in the ipilimumab group .Moreover, with more effective combined scheme therapy (nivolumab and ipilimumab), the incidence and severity of adverse effects is further increased. Mason R. et al. (2019) reported that 67% of patients developed treatment-related adverse events and 38% of individuals suffered from severe, grade 3–5 reactions .The treatment of moderate–severe irAEs requires discontinuation of the checkpoint inhibitor and the administration of intravenous corticosteroids (1–2 mg/kg dose). The treatment can be resumed afterwards, but data on the results of this approach are limited .Benson Z. et al. (2017) proposed the concept that tumor response to immunotherapy may be increased by the pre-exposure of malignant tissues to nanotechnology or other compounds able to trigger immunogenicity, to achieve epigenetic modulation or inhibit cell cycle progression . IntensiAnother application of nanotechnology in immunotherapy was highlighted by Iwamoto N. et al. (2016), who developed a method of pharmacokinetic study using nano-surface and molecular-orientation limited (nSMOL) proteolysis to quantify nivolumab in human plasma, probably useful for the quantification of other checkpoint inhibitor agents .IL-2 was the first immunotherapy to influence the outcome of advanced melanoma patients, but its harsh toxicity with cardiovascular, respiratory, and infectious complications restrained its use. It is now considered an option in only a few selected patients .A very interesting ongoing phase 1/2 clinical trial is investigating the efficiency of an IL-2 pegylated molecule  in combination with nivolumab for the treatment of melanoma, with promising primary results .Wu T. et al. (2019) ingeniously developed a combinatorial drug delivery system by absorbing interleukin-2 (IL-2) in doxorubicin (DOX)-loaded vesicles . The nanIn order to reduce the severity of adverse effects, Xie Y.Q. et al. (2019) developed a complex delivery system, binding redox-responsive IL-2/Fc nanogels to the surface of adoptively transferred T cells . InteresImmunoliposome delivery systems of IL-2/anti-CD137 can achieve equivalent anti-tumor activity compared to the free molecular agents, with low or even absent systemic toxicity . Zhang YCancer immunotherapy can specifically and persistently enhance the immune response. A series of preclinical and clinical studies have explored the benefits of whole tumor cancer vaccines ,21. TheyAlthough they are currently under intensive investigation, most cancer vaccines have low to moderate clinical effects and are unsuccessful in eliciting relevant results , probablA current point of interest is the design of personalized micro- and polymeric nanoparticle cancer vaccines, to co-deliver melanoma antigens, as well as therapeutic adjuvants.Tumor membrane-coated nanoparticles share some similarities with whole tumor vaccines, since they induce immune responses against multiple tumor antigens, without the need to select specific peptides .Fontana F. et al. (2019) designed a cancer nanovaccine based on biohybrid (TOPSi@AcDEX) nanoparticles coated with membranes derived from tumors cells that promote antigen cross-presentation, enabling the maturation of immature monocyte-derived dendritic cells . The nanThe activated antigen-presenting cells contribute to the stimulation of Th1 lymphocytes, followed by CD8 T cells activation, and the secretion of interferon-gamma (IFN-γ) . MoreoveLiu W.L. et al. (2019) proposed a cytomembrane nanovaccine derived from dendritic cells and cancer cells, allowing the nanoparticles to exert an effect similar to the physiological antigenic presentation and lead to an efficient T cell activation .Conniot J. et al. (2019) from Tel Aviv University showed that nanoparticles could potentially be used in the treatment of melanoma, similar to the use of vaccines for viral diseases . The resFurther research is mandatory to establish whether it is safe to power-up melanoma approach with nanovaccines in humans.Nowadays, combined BRAF and MEK inhibition therapy has replaced BRAF inhibition alone. The available combinations include dabrafenib plus trametinib, vemurafenib plus cobimetinib, and encorafenib plus binimetinib ,44.The major advantage of such combined treatments is the capacity to induce an accelerated tumor regression in most of the patients with BRAF V600 mutation-positive melanoma. This is why they are now recommended as first line therapy, especially in patients in whom a quick objective response to therapy is crucial, such as symptomatic patients with high tumor load and poor prognosis factors . Patients who do not fit these criteria of severity should benefit from immunotherapy as first line treatment, regardless of the BRAF status .Moreover, when compared to the administration of single-agent BRAF inhibitor therapy, response rates and survival periods are improved, while resistance and toxicity are lowered .Acquired resistance mechanisms still represent the major drawback, since they are only delayed by the combined therapy . StudiesAnother very important problem is the unclear sequencing of treatments in BRAF-positive patients. A current major controversy is related to the initial decision between immunotherapy and molecularly targeted therapy, as well as the benefit of switching to immunotherapy after BRAF/MEK inhibitors as first line therapy. Recent data showed that the action on symptom-free melanoma brain metastases of different immunotherapies is lower in patients previously treated with BRAF/MEK inhibitors . MoreoveStarting from the rationale that BRAF and MEK inhibitors have the ability to increase T cell responses through a variety of immunological changes, researchers combined BRAF/MEK inhibitors and anti-PD1 antibodies to achieve a synergistic effect in the treatment of metastatic melanoma. The study is named COMBI-I and preliminary data have shown that more than 40% of patients had a confirmed complete response, while a large proportion, 78% of patients, expressed adverse events, grade 3 or higher .Ramesh A. et al. (2019) went even further and studied an improved nano-model of combined therapy . They enThe same study group showed that nanotechnology can also be used to modulate tumor microenvironment together with kinase inhibition . SupramoThe use of cytotoxic chemotherapy in the treatment of advanced melanoma is extremely limited.Melanoma is known to be chemotherapy-resistant, with a median duration of response of approximately four to six months . In consFunctionalized CNTs have been proven to boost the effectiveness of chemotherapy in melanoma cells. Chaudhuri P. et al. (2010) showed that a SWNT conjugated with doxorubicin (DOX) reduced the side-effects of chemotherapy drugs while increasing their bioavailability, thus generating better drug efficacy in a B16-F10 laboratory melanoma model .CNTs were also used for transdermal delivery of DOX. Back in 2010, scientists used multi-walled and double-walled functionalized nanotubes (through PEGylation—with Polyethylene glycol) for the transdermal delivery of indomethacin  and doxorubicin  . ResultsIn a recent study on melanoma mice models, doxorubicin-loaded liposomes were grafted with a T cell receptor (TCR)-like antibody targeted to melanoma antigen A1 (MAGE-A1) . These iThe efficacy of liposomes in anti-cancer therapies can also be increased by using pH as a trigger for drug release, considering the 6.5 pH of tumor tissues. In melanoma, pH-sensitive liposomes were used to co-deliver paclitaxel and Bcl-2 siRNA in acidic conditions, with significant inhibition of tumor proliferation .AuNPs were used for carrier-based chemotherapy in melanoma. Ultra-small gold nanoparticles conjugated to doxorubicin (Au-Dox) were investigated after intratumoural injection  and in cWhile cancer gene therapy represents a promising approach in the management of melanoma, one of the greatest challenges is represented by the development of high-performance delivery systems characterized by low toxicity and immunogenicity, as well as a high transfection efficiency. While viral carriers have also been approved for the treatment of melanoma, non-viral carriers using nanoparticles are under study.Talimogene laherparepvec (T-VEC) is an attenuated oncolytic herpes simplex virus that contains the granulocyte macrophage colony stimulating factor gene. In the randomized phase III trial OPTiM, which included patients with unresectable stage IIIB–IVM1c melanoma, Andtbacka R.H.I. et al. (2019) revealed the superiority of T-VEC compared to subcutaneous recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) regarding long-term efficacy and patient tolerance . T-VEC wSun Z. et al. (2018) tried to enhance the antigenic immune response by the co-encapsulation of GM-CSF with ovalbumin nanoparticles in a thermosensitive hydrogel . They prZhang Q. et al. (2019) studied the in vivo effects in a B16-F10 mouse melanoma tumor model of the tumor suppressor phospholysine phosphohistidine inorganic pyrophosphate phosphatase delivering its plasmid using a nanoparticle gene delivery system . The nanSTAT3-siRNA for melanoma treatment [Labala S. et al. (2016) investigated the efficiency of layer-by-layer chitosan-coated AuNPs as nano-carriers for the iontophoretic delivery of reatment . The forreatment . The samreatment .CNTs may also play a pivotal role in overcoming the biological barriers in siRNA delivery. Siu K.S. et al. (2014) developed a nanotube-based siRNA  topical delivery system . SiRNA iThe role of radiation therapy in melanoma is mainly palliative, as it is recommended as the primary treatment for inoperable tumors and as adjuvant therapy in patients with desmoplastic melanoma . AdjuvanRadiotherapy alone has not been shown to improve patient overall survival . HoweverInadequate tumoral vascularization, hypoxia, and deficiencies of radiation absorption may limit the effect of radiotherapy . Metal n131I by the conjugation of the radioisotope with polymer-grafted AuNPs [Le Goas M. et al. (2019) improved internal radiotherapy with ed AuNPs . The resed AuNPs .Daneshvar F. et al. (2019) combined X-ray radiotherapy with 808 nm diode laser photothermal therapy of melanoma B16/F10cells after their sensitization with PtNPs . They obPhotothermal therapy (PTT) has recently emerged as a promising alternative for tumor targeting therapy. Nanoparticles have the ability to absorb long-wavelength light  and convert its electromagnetic energy into heat. After the bio-accumulation of nanoparticles into the tumor, the external irradiation with a laser light source will induce a destructive heating of the cancer cells .Due to their capacity to effectively absorb near-infra-red (NIR) light and transform it into heat, AuNPs are extremely helpful in the photothermal therapy (PTT) of melanoma and other cancers . InfrareA gold–ferrite nano-composite (GFNC) proved to be suitable in melanoma PTT ablation. After the injection of the gold nano-shell as a photothermal agent and upon laser irradiation in melanoma mice models, the tumor volume tumor size was substantially diminished when compared to the control groups .In addition, CNTs are a promising approach in cancer hyperthermia. In a recent study, upon targeting melanoma cells in conjunction with photothermal therapy, the rate of cell death caused by CNT-mediated PTT ablation was explored on melanoma mice models . After tGorgizadeh M. et al. (2019) used carbon xerogel nanoparticles as a photoabsorber of 808 nm laser light in vitro and in vivo in a melanoma tumor-bearing mouse model with good biocompatibility and tumor control .Photodynamic therapy (PDT) is another alternative, two-step therapy that comprises the administration of a photosensitive drug to targeted cancer cells, followed by laser light application at a specific wavelength that activates the photosensitizer . As a coGold nano-shells can also be used as photosensitizers in PDT. Due to their capacity to produce major reactive oxygen species for the photo-dynamic technique under NIR light, they consequently proved to suppress the proliferation of melanoma cells ,78.In a recent study by Mohammadi Z. et al. (2017), 5-aminolevulinic acid (5ALA), a photosensitizer limited by its low uptake by tumor cells, was conjugated with AuNPs and proved to be twice as efficient as 5ALA alone in promoting melanoma cell death .Our literature research with the terms “nanoparticle” and “melanoma” led to the identification of 10 clinical trials, most of them on nanoparticle bound-paclitaxel. This compound is better tolerated because it eliminates the hypersensitivity reaction caused by the Cremaphor solvent . The maiHersh E.M. et al. (2010) aimed to determine the antitumor activity of the formulation ABI-007  in patients with inoperable locally recurrent or metastatic melanoma and to determine the safety and tolerability of this drug . The stuOlencki T.E. et al. aimed to evaluate the response of patients with unresectable, metastatic uveal melanoma treated with paclitaxel albumin-stabilized nanoparticle formulation . Only foKotschade L.A. et al. (2011) published the results of a phase II clinical trial (NCT00404235) that included patients diagnosed with unresectable stage IV melanoma who received a combined treatment with nanoparticle albumin-bound–paclitaxel and carboplatin . The 76 Kottschade L.A. et al. (2013) studied the effectiveness of chemotherapy  in combination with vascular endothelial growth factor (VEGF) inhibition (bevacizumab), in patients with unresectable stage IV melanoma . The resIn another clinical trial (NCT01300533) the highest safe dose of BIND-014 (docetaxel nanoparticles for injectable suspension) was investigated in 52 patients with advanced or metastatic cancer ,86. OnlyThe results of a series of other completed clinical trials have not yet been published.Ribas A. et al. conducted a phase I clinical trial (NCT00689065) that included 24 adult patients with solid tumors (including melanoma) refractory to standard care who were treated with targeted nanocomplex containing anti-R2 siRNA administered intravenously . The resInteresting results are expected to be published from the clinical trial coordinated by Markovic S.N. et al. (NCT02158520), which compared the response to the combined regimen of nab-paclitaxel and bevacizumab with immunotherapy (ipilimumab) . The stuStambuk H. et al. from the Memorial Sloan Kettering Cancer Center are currently investigating a new experimental dye-labeled particle (targeted silica nanoparticles of fluorescent cRGDY-PEG-Cy5.5-C dots) for sentinel lymph node mapping prior to surgery in 105 patients with head and neck melanoma, breast cancer, or colorectal cancer (NCT02106598) .Svetomir M. et al. aim to include (NCT02020707) 36 patients diagnosed with stage IV melanoma that cannot be removed by surgery or with other types of gynecological cancers  in a phase 1 clinical trial in order to determine the maximally tolerated dose of a regimen with nanoparticle albumin-bound–paclitaxel and bevacizumab .Drug delivery platforms driven by nano-technology represent a standing example of precision medicine. As recent research has yielded promising results, we are confident that nano-structured materials will help empower the engineering of an end to melanoma mortality. By using nanotechnology, we are on the cusp of enabling personalized and targeted drug delivery in a way that could catalyze the next generation of therapeutics in melanoma.The pre-exposure of melanoma tumors to nanotechnology or other compounds in order to trigger immunogenicity, followed by immunotherapy, might represent the focus of research in the coming years. Further studies, especially clinical trials, are needed to establish whether fighting melanoma on the nanoscale level represents the key of reaching a critical inflection point in mankind’s battle with metastatic melanoma.


Significance: Differential phase contrast (DPC) is a well-known imaging technique for phase imaging. However, simultaneously acquiring multidepth DPC images is a non-trivial task. We propose simultaneous multiplane DPC imaging using volume holographic microscopy (VHM).Aim: To design and implement a new configuration of DPC-VHM for multiplane imaging.Approach: The angularly multiplexed volume holographic gratings (AMVHGs) and the wavelength-coded volume holographic gratings (WC-VHGs) are used for this purpose. To obtain asymmetric illumination for DPC images, a dynamic illumination system is designed by modifying the regular Köhler illumination using a thin film transistor panel (TFT-panel).Results: Multidepth DPC images of standard resolution chart and biosamples were used to compare imaging performance with the corresponding bright-field images. An average contrast enhancement of around three times is observed for target resolution chart by DPC-VHM. Imaging performance of our system is studied by modulation transfer function analysis, which suggests that DPC-VHM not only suppresses the DC component but also enhances high-frequency information.Conclusions: Proposed DPC-VHM can acquire multidepth-resolved DPC images without axial scanning. The illumination part of the system is adjustable so that the system can be adapted to bright-field mode, phase contrast mode, and DPC mode by controlling the pattern on the TFT-panel. The main idea in various volume holographic imaging systems is to reduce the requirements of axial scanning and to acquire multidepth images directly.,In this paper, we propose and experimentally demonstrate the DPC imaging technique in VHM using asymmetric illumination. We present two new configurations for DPC-VHM systems. Unlike previously mentioned volume holographic microscope systems that use uniform illumination of object and hence cannot provide the DPC images,22.1,In our imaging systems, DPC images are obtained by the method proposed by Dekkers and de Lang.If the pair of the asymmetric illumination images are added together, then the amplitude information can be obtained as By Eqs. (2) and (3), differential of the phase can be obtained as The differential of phase is sensitive to the phase variations, which results in contrast enhancement of high spatial frequencies as well as suppression of low frequencies.,Here, we are following the DPC method based on the asymmetric illumination.Next if we make above two equations to divide with each other, we can obtain 2.2The first configuration of DPC-VHM has two main parts, asymmetric illumination system and an angularly multiplexed volume holographic gratings (AMVHGs) imaging system as shown in ,Grating vector of each gratings in AMVHGs, ,The recording setup for AMVHGs and wavelength-coded volume holographic gratings (WC-VHGs) was built up following our previous work.,,In imaging system, a AMVHG, under Bragg-match condition,2.3To further reduce the image acquisition time, a second configuration based on WC-VHGs is presented that helps in simultaneously acquiring two different direction asymmetric illumination images with two different colors in single shot.Under Bragg-match condition, WC-VHGs display different wavelength images with different diffracted angles to the corresponding laterally separated position onto the CCD. Therefore, the DPC-VHM using WC-VHGs simultaneously acquires two different direction asymmetric illumination images with two different colors in single shot.33.1In addition, we use onion skin samples to test the ability of the DPC-VHM system to resolve weak phase features. Compared to To precisely compare the resolution and performance of our systems, modulation transfer function (MTF) analysis was performed. The 1951 AFRC was introduced as an object. A comparison of MTF for the proposed DPC-VHM and bright-field volume holographic microscope is shown in 3.2In ,The 4Two new DPC-VHM configurations are experimentally demonstrated. Two different kinds of volume holographic gratings are adopted for this purpose. AMVHGs, in DPC microscopy, help in acquiring multidepth-resolved non-axial scanning DPC images. MTF analysis of system shows that the low spatial frequency term component is reduced, whereas the contrast is enhanced for most of the frequencies. Image contrast can be significantly enhanced using our DPC-VHM as compared to the bright-field volume holographic microscope. We found that the DPC-VHM can simultaneously enhance contrast of fine features within a volumetric sample, such as onion skin, at three depths. In addition, we demonstrate that single-shot DPC microscopy is possible with the help of WC-VHGs. Our illumination system is dynamic and multipurpose, and offers many advantages. It can work in different microscopic modalities under different illumination conditions. A bright-field mode, phase contrast mode, and DPC mode can be adapted by just manipulating the mask on the TFT-panel. The presented configurations may be used for quantitative DPC measurements


There is a paucity of research on patterns of cyber-victimization in minority groups, including immigrants. This study aimed to identify individual, interpersonal and contextual characteristics associated with cyber-victimization among immigrants and non-immigrants.We drew on nationally representative data from adolescents and adults in the Canadian General Social Survey on victimization (2014). We used multivariable logistic regression to identify potential factors associated with cyber-victimization in the last 12 months, stratified by immigrant status and sex.Among 27,425 survey respondents, the weighted prevalence of cyber-victimization in the last 12 months was 2.1% among immigrants and 2.3% among non-immigrants. Cyber-victimization rates differed significantly by sex among immigrants , but not among non-immigrants . While most other factors associated with cyber-victimization were similar for immigrants and non-immigrants, there were pronounced associations of past child maltreatment  and residence in an unwelcoming neighbourhood  with cyber-victimization among immigrants that were diminished or absent among non-immigrants. Additionally, sex-stratified analyses among immigrants showed cyber-victimization to be strongly associated with having a mental health condition  among immigrant males only, and with perceived discrimination , as well as being under 24 years old  among immigrant females.Immigration status and sex were differentially associated with cyber-victimization. Findings support the salience of a social-ecological perspective and gender-stratified analyses to better elucidate complex pathways linking cyber-victimization to potential gender-based health inequities among immigrants. Cyberbullying is the use of computers, cell phones or other technological devices to deliberately threaten, abuse, or intimidate an individual or a group of individuals , 2. ThisAs individuals increasingly spend more time on-line, considerable attention has been placed on understanding and mitigating risks associated with cyberbullying. In two meta-analyses focused on young people, those who were cyber-bullied were identified as more likely to engage in frequent internet activities, experience off-line bullying, have low self-esteem, report depression and anxiety, and use drugs and alcohol , 8. ThisThere is a paucity of literature on cyberbullying among immigrants and it remains unclear what factors contribute to prevalence in this population. In one of the only studies comparing cyber-bullying rates of immigrant and US-born youth, immigrants were 2.27 times  more likely to report cyber-victimization than non-immigrants . SimilarGender is a prominent factor among socio-demographic variables examined in studies of cyber-victimization in immigrant populations. Since negative health outcomes  associated with resettlement stress are shown to differ by immigrant group, the context of migration, and further, within-group, by age and gender , 24, it Considerable gaps in the literature are also evident in understanding the relationship between age and cyberbullying in immigrants, and no prior studies have yet examined age-related patterns. Looking more broadly at the general population, results have shown no association between age and cyber-victimization among youth of different age groups, while pointing to a weak positive association of older youth with cyberbullying perpetration . On thisWith the growing ubiquity of internet use, the unique challenges of the resettlement process, and rising anti-immigrant policies , 36, it In the present study, we used data from a nationally-representative sample of Canada, a country with a high proportion of immigrants 21%) that is second only to Australia (27%) . Adoptin% that isThis study used data collected from January to December 2014 by Statistics Canada’s Canadian General Social Survey (GSS), Cycle 28 on Victimization . The maiSurvey responses were obtained by computer-assisted telephone interviews conducted in the Canadian official languages (English or French) of the respondent’s choice. Households without telephones (approximately 1% of the target population) were not captured. The response rate was 53%. Total non-response was handled by adjusting the weight of households who responded to the survey to compensate for those who did not respond. Non-respondents included people who refused to participate, could not be reached, or could not speak English or French. The sample size was 33,127 respondents. Since cyberbullying was the focus of this study, we then excluded about 16% of respondents who reported either not using the internet in the past 5 years, did not know, or refused to answer, as well as < 1% of respondents who refused to answer questions related to cyberbullying and those for whom immigrant status could not be determined. The final analytic sample included 27,425 individuals.Cyber-victimization in the last 12 months: Cyber-victimization was assessed according to responses to a series of 5 questions asking whether in the last 5 years a respondent ever had any threatening or aggressive emails/messages; threatening or aggressive comments directed at them via group emails/messages or internet postings; embarrassing/threatening pictures posted of them; embarrassing/threatening information posted by someone pretending to be them; and any other type of cyber stalking/bullying. If a respondent answered affirmatively to any of these questions they were asked a follow-up question about whether any of these experiences occurred in the past 12 months. Cyber-victimization was thus constructed as a dichotomous variable specified as any cyber-victimization reported in the last 12 months versus none.Immigrant status: The key independent variable of interest was immigrant status. Respondents were classified as immigrants if they reported being born outside Canada and obtained legal permanent residency/citizenship. Respondents who did not report permanent residency, but reported being born outside of Canada and provided a year for when they first came to live in Canada, were also considered immigrants, assessed as likely to have arrived in Canada as asylum seekers, students or temporary workers.Explanatory factors were identified based on the literature and available data. Since a majority of previous research on cyberbullying involvement has been limited to individual-level factors, this study incorporated a broader social-ecological emphasis, including attention to how individual, interpersonal and neighbourhood factors may explain patterns of cyber-victimization. Individual characteristics included sex , age , education (high school completion vs. incomplete), visible minority status (visible minority vs. non-visible minority) based on the definition in the Canadian Census, excludes Indigenous peoples and consists mainly of the following groups: South Asian, Chinese, Black, Fillipino, Latin American, Arab, Southeast Asian, West Asian, Korean and Japanese, annual household income , disability status , mental health , and alcohol use . Interpersonal characteristics included history of childhood victimization , number of close friends or relatives , intimate partner violence (IPV) in last 5 years , and any discrimination experienced in last 5 years . Neighbourhood-level characteristics included respondents’ perspectives of how welcoming their neighbourhood is (residing in an unwelcoming community vs. residing in a welcoming community), neighbourhood discrimination , and neighbourhood trust  .Descriptive statistics were calculated for all variables and stratified separately by cyber-victimization and immigrant status. Subsequently, we estimated the odds of cyber-victimization with each covariate using unadjusted logistic regression models and applying bootstrapped sampling weights to account for complexity of the survey design and produce nationally representative estimates. To obtain estimates stratified by immigrant status and, further, by immigrant status and sex, six multivariable logistic models were constructed: Model 1: immigrants-only; Model 2: non-immigrants only; Model 3: immigrant females; Model 4: immigrant males; Model 5: non-immigrant females; and Model 6: non-immigrant males. Explanatory variables for final models were selected based on Hosmer et al.’s (2013) purposeful selection of covariates criteria . To avoip-value for interaction of immigrant status with sex: 0.008), to have experienced prior child maltreatment , and to have a larger network of close friends and relatives   among Canadian-born counterparts. Results also showed a strong association between cyber-victimization and having a history of child maltreatment, which was most notable among immigrants. Additionally, at the interpersonal-level, while we found that having a history of IPV was strongly associated with cyber-victimization among non-immigrants, there was no evidence of an association among immigrants. Further, at the neighbourhood-level, we found that perceiving to reside in an unwelcoming neighbourhood, rather than individual or interpersonal factors, was associated with the strongest association to cyber-victimization for immigrants, and did not find evidence of a similar pattern among non-immigrants.At the individual-level, among immigrant females, we found those who were cyber-victimized tended to have significantly higher odds of being younger, a pattern that was also observed, though to a lesser degree, among non-immigrant females. Further, results showed a positive association between cyber-victimization and having a mental health condition among immigrant males that was also evident among non-immigrant males and females, yet notably absent among immigrant females. Additionally at the interpersonal-level, we found evidence of a strong association between cyber-victimization and perceived discrimination among immigrant females, which was far more modest among non-immigrants groups, and absent among immigrant males.The link between cybervictimization and being male among immigrants suggests a differential vulnerability between immigrant men and women that is consistent with findings in one previous study of school-age youth that similarly showed immigrant males to have higher odds of both cyber and other forms of bullying than females . It is pThe high degree of overlap between cyber-victimization and other forms of interpersonal victimization is consistent with prior literature on young people in the general population , 46, whiOur results also point to an important role of perceived neighbourhood characteristics in explaining patterns of cyberbullying, and show these links to be more strongly and consistently observed among immigrants than non-immigrants. Most significantly, we found that the perception of residing in an unwelcoming neighbourhood was very strongly associated with cyber-victimization across all immigrants regardless of sex, but found no discernible association among non-immigrants, for whom perceived neighbourhood-level discrimination was more often associated with cyber-victimization. These results, and recent research , encouraOur findings extend prior literature in several ways. First, our stratified approach allowed for interpretation of differences by immigrant status and sex, making it possible to examine comparisons that were not explored in previous studies. Second, our leveraging of a demographically and geographically dispersed weighted sample improved generalizability of findings to the entire country-level population. Third, our analysis expands consideration of cyber-victimization beyond youth to include older adults, a population whose experiences of the internet and social media are often overlooked. Lastly, we incorporated a broad social-ecological analytic approach, including neighbourhood factors, which attempted to respond to cautions raised in recent studies about the tendency for an overly narrow focus on individual and family-level factors explaining patterns of cyber-victimization.Results, however, must also be interpreted within the limitations of our study. The study’s primary limitation is the cross-sectional study design and data, which implies the directionality of many associations found, are unknown. We were also not able to identify how associations may vary over time, especially for immigrants whose experiences and perspectives on what constitutes bullying, may change the longer they reside in the host country. Second, there was a lack of available variables related to our cyber-victimization outcome. For example, we could not know the types or sources (medium and perpetrators) of cyber-victimization in order to better understand this phenomenon across immigrant/non-immigrants and males/females. Third, we have only a crude measure of biological sex in available data and not a measure of gender, which precluded capturing various aspects of gender identity and how these may also intersect with our outcome. Additionally, English/French language ability for the GSS was an inclusion criterion, so the most marginalized immigrants, such as refugees, for whom forced migration has been associated with lower fluency of official language, are more likely to be excluded. In future, national-level data that are not limited by language ability are needed to better represent immigrants’ experiences.Our findings highlight several directions for future research in this area. Importantly, our main finding underscores the need to investigate the complex manner in which immigrant status and gender appear to place immigrant men at higher risk than women. Also, though we did not find an association between cyber-victimization and visible minority status, consistent with some prior research examining race and cyber-bullying , 26, we This study expands knowledge about factors associated with cyber-victimization among immigrants and non-immigrants using, for the first time, a large, nationally-representative sample. Our main findings show that immigrants and non-immigrants experienced similar rates of cyber-victimization and that there was a notable increased vulnerability of males among immigrants that was not detected among non-immigrants. Increased vulnerability to cyber-victimization was also more pronounced among immigrants with a history of child maltreatment and those residing in an unwelcoming neighbourhood. These findings suggest that gender-stratified analyses should be a key avenue for future research on cyber-victimization among immigrants, and also demonstrate strong support for incorporating a broad social-ecological perspective to better elucidate relationships between immigrant status, gender and cyber-victimization.


Background and Objectives: Epilepsy patients have a higher risk of sudden unexplained death compared to the rest of the population. Cardiac repolarization abnormalities might be seen in epilepsy during interictal periods. We aimed to evaluate the changes in electrocardiography (ECG) parameters in generalized tonic-clonic seizure patients treated with carbamazepine or valproic acid (VPA) drug. Materials and Methods: A totally of 129 subjects  were enrolled in the study. Of the patients, 36 were on carbamazepine and 30 were on VPA. There were 12-lead ECGs obtained from all participants. RR interval (time between consecutive R peaks), QT interval (defines the period of ventricular repolarization), corrected QT , QTc-maximum (QTc-max), QTc-minimum (QTc-min), QTc dispersion (QTcd), P -maximum (P-max), P-minimum (P-min) and P dispersion (Pd) were measured. Results: QTd (QT dispersion), QTcd, and Pd values were significantly higher in the patients compared to the controls (p < 0.01). QTcd, Pd, and P-max values were statistically higher in male patients compared to healthy male controls. QTcd values were significantly higher in female patients using carbamazepine compared to the female patients on VPA and healthy controls (p = 0.01). Male patients using VPA had significantly higher QTcd values against the male population in carbamazepine and control groups. Conclusions: This study demonstrated that QTd, QTcd, and Pd values were significantly higher in epilepsy patients than in healthy controls. In addition, female patients using carbamazepine and male patients using VPA were prone to ventricular arrhythmia compared to the control group. Epilepsy, a chronic disorder of the central nervous system, includes recurrent seizures or unusual behavior, and sometimes loss of consciousness . The risIt is not uncommon to observe cardiac rhythm disorders during ictal, interictal, and postictal phases in patients with epilepsy. Cardiac repolarization abnormalities , increased QT dispersion (QTd) might be seen in epilepsy during interictal periods . ShorterAntiepileptic drugs (AEDs) have different effects on the cardiovascular conduction system. Use of carbamazepine may cause atrioventricular blocks, bradycardia, and arrhythmias . MoreoveThis study was planned as a cross-sectional study and it was conducted between May 2014 and June 2015 in the neurology and cardiology departments of Van Training and Research Hospital. The study population included 66 patients and 63 healthy controls. Sixty-six patients diagnosed with GTCS and epilepsy according to the International League Against Epilepsy (ILAE) criteria [All patients underwent detailed history taking, cardiac and neurological examinations, routine biochemistry, electrocardiography (ECG), and transthoracic echocardiography (TTE). The inclusion criteria were: Being diagnosed with GTCS and to be aged between 18–65 years old. The exclusion criteria were: Being on medications other than carbamazepine and VPA that could affect ECG intervals  stimulants, and lithium), using a cardiac pacemaker, having cardiac rhythm disorders, diabetes, bundle branch block, or known cardiac and psychiatric disease, liver failure, substance abuse, acute or chronic infection, and serum electrolyte abnormalities.The patients were rested for 15 minutes to stabilize the heart rate and the traces of 12-lead ECGs were recorded. The Nihon Kohden Cardiofax GEN ® device  was used with a rate of 25 mm/s in 12-channeled form and with a standard of 10 mm/mV. A magnifying glass was used for measurements. The ECGs were examined by two different cardiologists, who did not have any information about the study, and they did not have any contact during the interpretation of the results. The RR interval was defined as the time intervals between two consecutive heartbeats (R-waves). QT interval (distance between the start of QRS (ventricular depolarization) and the end of the T-wave (ventricular repolarization)) was measured and minimum (QT-min) and maximum QT (QT-max) values were recorded. The end of the T-wave was defined as the intersection of the terminal limb of the T-wave with the isoelectric baseline. The Bazett formula (QTc = QT/√RR)  was used to assess the corrected QT (QTc); hence, QTc-minimum (QTc-min), the QTc-maximum (QTc-max), and QTc dispersion (QTcd) values were recorded [A complete TTE was performed in all participants following the American Society of Echocardiography guidelines. M-mode from the parasternal long-axis view was used to determine the left ventricular end-systolic and end-diastolic diameters . Pulse wave Doppler echocardiography measured the early and late diastolic mitral flow velocities . The thicknesses of posterior wall (PW), left atrium diameter, inter-ventricular septum (IVS), ejection fraction (EF), and aorta systolic and diastolic diameters were measured. All participants gave their written informed consent before the study. This study was performed under the principles stated in the Declaration of Helsinki. The local Ethics Committee of the hospital approved the study (086 – 26 September 2014).p < 0.05 was taken as statistical significance.SPSS  was used for data analysis. The Kolmogorov–Smirnov test was used assess the distribution of the variables. Normally distributed variables were expressed as mean ± standard deviation (SD). The categorical variables are given as percentages. Student’s unpaired t-test or the Mann–Whitney U test were used to assess differences between two groups for parameteric and nonparametric variables, respectively. Fisher’s exact or chi-square tests were used to calculate the frequencies of nominal variables. For multiple comparisons, one-way analysis of variance (ANOVA) test followed by the Tukey post hoc test was used, and The cardiologic assessment results and demographic data of the participants are presented in p > 0.05). Heart rate, RR interval, P-min, QT-min, QT-max, QTc-min, and QTc-max were similar between patient and control groups (p > 0.05). QTd, QTcd, and Pd values were significantly higher in patients than in the healthy individuals (p < 0.01). There was no significant difference between groups in terms of age, body mass index, gender, blood pressures, biochemical and hematological analyzes, and echocardiographic evaluation . QTcd and Pd values were similar in patients using carbamazepine and VPA (p > 0.05). The clinical and demographic data were similar in study groups except for ECG analysis. QTd, QTcd, and Pd values were significantly higher in patients using carbamazepine and VPA than in control subjects . QTcd, Pd, and P-max (P-maximum) values were statistically higher in male patients compared to healthy male controls . You can find the result of ECG analysis on carbamazepine, VPA, and control groups in terms of gender in Female patients had significantly higher QTcd and Pd values compared to healthy female subjects . QTcd values were significantly higher in female patients using carbamazepine compared to female population in VPA and control groups (p = 0.01). P-min, P-max, QTc-min, and QTc-max values were similar in female subjects in carbamazepine, VPA, and control groups (p > 0.05). Pd values were significantly higher in male patients using carbamazepine compared to male epilepsy patients on VPA and healthy controls (p = 0.02). Male patients using VPA had significantly higher QTcd values compared to male epilepsy patients on carbamazepine and controls (p = 0.01). In the male population, P-min, QTc-min, and QTc-max values were similar in carbamazepine, VPA, and control groups (In our study, we demonstrated five main results in patients with GTCS. Our first result was significantly higher QTd, QTcd, and Pd values in epilepsy patients compared to healthy controls. Our second result was that female patients using carbamazepine had higher QTcd values compared to female patients on VPA and control groups. Third, male epilepsy patients using carbamazepine had higher Pd values compared to male epilepsy patients on VPA and control groups. Four, female patients had higher QTcd and QTd values compared to male patients. The last but not least finding was the significantly higher QTcd values in male patients on VPA group compared to male epilepsy patients on carbamazepine and control groups. Dogan et al. have evaluated depolarization indices in patients with partial epilepsy and found significantly longer QTc-max and QTcd intervals in patients with epilepsy compared to controls . MoreoveQT interval is an indicator of the period of ventricular depolarization and repolarization. A prolonged QT interval and QTd are defined as cardiac repolarization abnormalities and they are relate to sudden cardiac death and ventricular tachyarrhythmia . The P-wAEDs can facilitate the development of ventricular tachyarrhythmias by acting on ionic channels. AEDs may pose a pro-arrhythmic effect if used in combination . CarbamaIn addition, we evaluated epilepsy patients in terms of gender. We found that female patients with epilepsy were more prone to ventricular arrhythmias compared to healthy female individuals. Male epilepsy patients were prone to ventricular repolarization and atrial depolarization abnormalities compared to healthy male controls. Also, we evaluated repolarization indices in epilepsy patients in terms of carbamazepine and VPA usage. QTcd values in female patients using carbamazepine were significantly higher compared to the female epilepsy patients on VPA and control groups. Male patients using VPA had significantly higher QTcd values than the male epilepsy patients on carbamazepine and control groups. We concluded that female patients using carbamazepine and male patients using VPA were prone to ventricular arrhythmia compared to the control group. The possible influences of VPA use on cardiac conduction, duration of the epilepsy, VPA dosage, and the influences of the epilepsy disorder itself are also important factors, which would be responsible for some higher repolarization parameters in VPA group. Studies with more patients and multicenters are necessary to confirm these results. This study has some limitations. It was a single-center study enrolling a relatively small group of patients, and our results cannot be generalized to all health centers. Another study limitation was the single measurement of repolarization indices, and these parameters might change in the course of time, especially with AEDs. One other limitation of this study was the lack of information on carbamazepine and VPA dosages. Moreover, patients with other AEDs were not investigated in the study. In addition, we did not evaluate peri-ictal ECG parameters and different epilepsy types in the study. Despite these limitations, our study provides important information on the repolarization indices in epilepsy patients.In conclusion, we determined that QTd, QTcd, and Pd values were significantly higher in epileptic patients than in healthy controls. Also, we showed that female patients using carbamazepine and male patients using VPA were prone to ventricular arrhythmia compared to the control group. More studies are necessary to confirm these results.


Background: Altered white matter connectivity, as evidenced by pervasive microstructural changes in myelination and axonal integrity in neuroimaging studies, has been implicated in the development of autism spectrum disorder (ASD) and related neurodevelopmental conditions such as schizophrenia. Despite an increasing appreciation that such white matter disconnectivity is linked to social behavior deficits, virtually no etiologically meaningful myelin-related genes have been identified in oligodendrocytes, the key myelinating cells in the CNS, to furnish an account on the causes. The impact of neurodevelopmental perturbations during pregnancy such as maternal immune activation (MIA) on these genes in memory-related neural networks has not been experimentally scrutinized.Methods: In this study, a mouse model of MIA by the viral dsRNA analog poly(I:C) was employed to mimic the effects of inflammation during pregnancy. Transcriptional expression levels of selected myelin- or oligodendroglia-related genes implicated in schizophrenia or ASD development were analyzed by in situ hybridization (ISH) and quantitative real-time PCR (qRT-PCR) with brain samples from MIA and control groups. The analysis focused on SOX-10 (SRY-related HMG-box 10), MAG (myelin-associated glycoprotein), and Tf (transferrin) expression in the hippocampus and the surrounding memory-related cortical regions in either hemisphere.Results: Specifically, ISH reveals that in the brain of prenatal poly(I:C)-exposed mouse offspring in the MIA model (gestation day 9), mRNA expression of the genes SOX10, MAG and Tf were generally reduced in the limbic system including the hippocampus, retrosplenial cortex and parahippocampal gyrus on either side of the hemispheres. qRT-PCR further confirms the reduction of SOX10, MAG, and Tf expression in the medial prefrontal cortex, sensory cortex, amygdala, and hippocampus.Conclusions: Our present results provide direct evidence that prenatal exposure to poly(I:C) elicits profound and long-term changes in transcript level and spatial distribution of myelin-related genes in multiple neocortical and limbic regions, notably the hippocampus and its surrounding memory-related neural networks. Our work demonstrates the potential utility of oligodendroglia-related genes as biomarkers for modeling neurodevelopmental disorders, in agreement with the hypothesis that MIA during pregnancy could lead to compromised white matter connectivity in ASD. SOX10, Iwamoto et al., MAG, Fujita et al., Growing evidence suggests that functional disconnectivity within and across large-scale neural networks is a central hallmark of clinically overlapping neurodevelopmental disorders such as autism spectrum disorder (ASD) and schizophrenia ] induction in animals is a robust method for investigating neurodevelopmental disorders  in the brain of adult mouse offspring. Based on the literature on white matter disconnectivity associated with the hippocampus-amygdala complex or temporal lobes in schizophrenia and ASD , the University of Hong Kong. Timed-pregnant mice were kept in a 12:12-h reversed light-dark cycle (light onset at 19:00), and temperature and humidity-controlled  animal vivarium. Animals had access ad libitum to food and water supplied by the LAU. All experiments had been approved by the Committee on the Use of Live Animals in Teaching and Research at the University of Hong Kong, and every effort had been made to minimize the number of animals used and their suffering.via the tail vein under mild physical constraint. Control animals were similarly administered with 5 ml/kg saline via tail vein on GD9. The animals were returned to their home cages after injection. The resulting offspring from six litters  were weaned and sexed at postnatal day (PND) 21. Pups were weighed and littermates of the same sex were caged separately (3–4 per cage). Only adult male offspring were used in subsequent experiments.Prenatal treatment was performed as previously reported by our group , and Tf  and reverse-transcribed with SuperScript II (Invitrogen). Based on the cDNAs from reverse transcription, gene-specific primers for , and Tf  were useDouble-strand (ds) DIG (digoxygenin)-labeled DNA probes were prepared with a PCR DIG Probe Synthesis Kit (Roche) by using specific PCR primers  accordinIn situ hybridization was performed on several selected regions of the mouse brain, based on methods previously reported by our group  from adult offspring of different pregnant mice (n = 3 per group) either exposed to poly(I:C)- or saline at GD9 were collected. The brains were perfused under terminal anesthesia, fixed in 4% paraformaldehyde, and embedded in paraffin wax. Brain sections of 5 μm in thickness were prepared on a Leica RM2135 microtome  and mounted onto pre-coated Superfrost Plus glass slides . The tissue sections were stored at −80°C for subsequent experiments. For in situ hybridization, brain sections were dewaxed with xylene, rehydrated with decreasing levels of ethanol, and post-fixed briefly in 4% paraformaldehyde. After that, the sections were digested with proteinase K (Roche), washed, and incubated at 37°C for 10 min with hybridization solution . For in situ hybridization, DIG-labeled DNA probes for mouse SOX10, L-MAG, and Tf were added to brain sections, followed by incubation at 42°C overnight in a humidified chamber. On the following day, post-hybridization washing was performed in decreasing concentrations of SSC solution at 42°C. Signal development was conducted with an anti-DIG antibody  by using NBT and BCIP as the substrates. In these experiments, hybridization with coronal sections of the saline-challenged mouse brain was used as a control.n = 3 per group) brain regions including mPFC, hippocampus, amygdala, neocortex, and thalamus from an independent batch of the samples used by in situ experiments, was isolated with TRIzol reagent (Invitrogen). Next, 4 μg of each total RNA sample was digested with DNase I (Invitrogen) and reverse-transcribed into cDNA (RT) with SuperScript II (Invitrogen). Gene-specific primers for real-time PCR were designed based on the sequences of mouse SOX10, L-MAG, T-MAG , Tf, and GAPDH. The RT samples obtained were then subjected to quantitative PCR in a RotorGene 6,000 Real-time PCR System  and PCR reactions were conducted with a Light-cycler DNA Master SYBR Green I Kit (Roche). The primer sequences and real-time PCR reactions for gene expression studies are as shown in Total RNA from individual mouse .For quantitative real-time PCR for transcript expression of SOX10, L-MAG, and Tf transcript expression was generally reduced in several brain regions (both right and left sides) including the hippocampus, retrosplenial granular cortex (RSG), and retrosplenial dysgranular cortex (RSD), in the prenatal poly(I:C)-exposed group relative to saline-exposed control.SOX10 signals, while the CA1 and CA3 regions showed mild to moderate expression in saline control mice , left CPu, RSG and RSD, or SC, AC or PC  showed the highest density of rol mice . Besidesmen CPu; , but no e of CPu  in salin and RSG , the som and RSG . HoweverAC or PC  comparedAC or PC .L-MAG expression in saline-exposed control, the hippocampal regions  and the lateral amygdaloid nucleus dorsolateral part -exposed group was conspicuously absent or dramatically reduced in the same brain regions examined above when compared with that of the saline-exposed control - and saline-exposed groups -exposed group , d groups , S6A, whed group , S6B.GAPDH as a reference gene. Also, the qRT-PCR results reveal that there was no significant difference between the prenatal poly(I:C)- and saline-exposed groups in all the brain regions compared (in situ hybridization for these regions (To clarify whether poly(I:C) induced MIA would give rise to nonspecific modulatory effects on global gene expression in the mouse brain, we included the housekeeping gene compared , which i regions –S6.SOX10 in prenatal poly(I:C)-exposed group were significantly lower (by at least one-fold difference) than that of saline-exposed control - and saline-exposed groups -exposed group was significantly lower -exposed mice -exposed mice. Specifically, we found that several brain regions with prominent functions in learning, memory and social interaction  showed markedly reduced SOX10, MAG, and Tf mRNA expression, suggesting that they may be high-risk regions susceptible to the effects of poly(I:C)-induced MIA during pregnancy. This disruption of the normal course of development could result in a compromised capacity to maintain synchrony of widely distributed neural networks, which is ultimately manifested in the heterogeneity of symptoms and cognitive deficits of schizophrenia and autism -exposed brain, this interhemispheric difference was not observed in in situ hybridization. In qRT-PCR, SOX10, L-MAG/T-MAG, and Tf transcript levels were reduced by over three folds , whereas only about one-fold difference was observed for L-MAG/T-MAG mRNA in the right but not left amygdala. As a structural protein with cell signaling functions mediated by its additional cytoplasmic domain, L-MAG contributes to most functions of total T-MAG in the CNS. The proportionally more significant reduction in SOX10 mRNA expression could mean that SOX10 as a transcription factor contributes significant weight to altered myelination or oligodendrocyte function in the postnatal offspring.Apart from the hippocampus and entorhinal cortex, a memory-related brain region that showed clear transcriptional alterations was the retrosplenial cortex  early in brain development could well have a more profound effect than the case of later stage disturbances. Consistent with our findings, reduced SOX10 expression was found in the hippocampus and anterior cingulate cortex (ACC) of subjects with schizophrenia  and MAG in the cerebella of mouse offspring  exposure induced MIA during pregnancy and selected oligodendroglia-related genes involved in myelination are as delineated in Quarles, . Also, Min situ hybridization, we examined a typical coronal section (Bregma –1.70 mm) revealing the hippocampus but did not explore some other brain regions deemed important in canonical schizophrenia or autism pathophysiology such as the prefrontal cortex. Similarly, a sagittal section would reveal more information on oligodendroglial markers in anatomical structures relevant to autism, such as the cerebellum. In qRT-PCR, we chose to examine transcript expression patterns separately in the left and right brains, but our findings were restricted by relatively small sample sizes. Differences in tissue sampling methods used in in situ hybridization and qRT-PCR mean that their results are coupled to differential spatial resolution and variability. Therefore, qualitative differences seen in histology  may not always be reflected in quantitative differences in qRT-PCR .The prevalent nature of gene expression changes seen in many brain regions has unexpectedly stretched the scope of this study. As a result, we only chose to focus on describing the effects of poly(I:C)-induced MIA on a few ROIs known to be involved in learning, memory, and social interaction. For Overall, our work is relatively exploratory. It attempted to address the need for specific molecular biomarkers relevant to myelin- or oligodendroglia-related changes in murine MIA models, in particular, that of poly(I:C)-induced MIA in pregnant mice. While previously some of our works supported the utility of C57BL/6N mice in evaluating behavior on social-affective functions  exposure dramatically altered the spatial distribution and transcript level of oligodendroglial molecular signatures including The datasets analyzed in this article are not publicly available. Requests to access the datasets should be directed to wongnksz@163.com.The experiments had been reviewed and approved by the Committee on the Use of Live Animals in Teaching and Research at the University of Hong Kong, and every effort had been made to minimize the number of animals used and their suffering.N-KW, YZ, and AW conceived and designed the experiments. X-FZ, TC, AY, PC, and Y-LX performed the experiments. N-KW, Y-LX, JY, and JX analyzed the imaging data. X-FZ, YZ, and TC performed the statistical analysis. N-KW, X-FZ, YZ, and TC wrote the article. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Mosquito-borne viral infections have in recent years, become a public health threat globally. This review aimed to provide an overview of the ecological and epidemiological profiles of mosquito-borne viral infections in the Democratic Republic of the Congo (DRC).A search of literature was conducted using Google Scholar, PubMed and the WHO website using the following keywords: “Democratic Republic of the Congo”, “Zaire”, “Belgian Congo” and either of the following: “mosquito-borne virus”, “arbovirus”, “yellow fever”, “dengue”, “chikungunya”, “West Nile”, “Rift Valley fever”, “O’nyong’nyong”, “Zika”, “epidemiology”, “ecology”, “morbidity”, “mortality”. Published articles in English or French covering a period between 1912 and October 2018 were reviewed.Aedes aegypti and Ae. simpsoni were documented as the main vectors of most of the mosquito-borne viral infections. Heavy rains, human movements, forest encroachment and deforestation were identified as drivers of mosquito-borne viruses occurrence in DRC.A total of 37 articles were included in the review. The findings indicate that the burden of mosquito-borne viral infections in DRC is increasing over time and space. The north-western, north-eastern, western and central regions have the highest burden of mosquito-borne viral infections compared to south and eastern highland regions. Yellow fever, chikungunya, dengue, Zika, Rift Valley fever, West Nile and O’nyong’nyong have been reported in the country. These mosquito-borne viruses were found circulating in human, wildlife and domestic animals. Yellow fever and chikungunya outbreaks have been frequently reported. Mosquito-borne viral infections are becoming common and a serious public health problem in DRC. In the current context of climate change, there is urgent need to improve understanding on ecological and epidemiology of the diseases and strengthen surveillance systems for prompt response to epidemics in DRC. Aedes are the major vectors of Chikungunya (CHIKV), Dengue (DENV), Rift Valley fever (RVFV), Yellow fever (YFV) and Zika (ZIKV) , morphological investigation of affected organs, tissue culture cell lines  and molecular biological methods including reverse transcription polymerase chain reaction (RT-PCR) detection, genome sequencing and molecular genetics analysis. The majority of sample specimens analysed were from humans; few were from animals and mosquitoes. Some studies reported co-occurrence or co-infections of mosquito-borne viruses. These included CHIKV and YFV, CHIKV and DENV, YFV and DENV , 14–16.The first yellow fever outbreak in DRC was reported among ship passengers from Europe in 1912 at Matadi Beach . Later iThe Democratic Republic of the Congo belongs to an endemic geographical area at high risk for YFV transmission . YFV traThe two first reported chikungunya cases in the DRC occurred during a yellow fever outbreak in north-eastern region. The first was in 1958 in Haut-Uelé Province , and theThree serotypes of dengue virus  have been recorded in DRC, the most frequent being serotype DENV-1 , 10, 30.Aedes mosquitoes collected around Ndjili River in Kinshasa [Rift Valley Fever virus has been reported in humans and cattle in DRC. A low rate (4%) of IgG antibodies from human against RVFV was reported in 1998 from Kisangani in the north-eastern region . In 2009Kinshasa .Two studies on Zika were identified during the review. In the first study, of 453 archived sera collected country-wide from 2003–2011 that tested YFV negative were also negative for ZIKV virus using RT-PCR . The secIn the DRC, antibodies against WNV in humans have been detected by virus neutralization tests, first in 1942 in Buta, Ango and Bondo from Bas-Uélé Province located in the north-eastern part, and later in 1963 from Gemena in Sud-Ubangi in the north-western part , 37. TheAnopheles mosquito. Its RNA was detected by RT-PCR from a pool of Anopheles mosquito collected in Kinshasa [O’nyong’nyong virus (ONNV) is a closer CHIKV related alphavirus vectored mainly by the Kinshasa . SerologKinshasa . O’nyongKinshasa . HoweverSignificantly high rates of neutralizing antibodies against flaviviruses and alphaviruses have been reported in buffaloes, elephants, gorillas and duikers from the Congo River basin . In KinsAe. aegypti and Ae. simpsoni were the most recorded in the regions where mosquito-borne virus outbreaks occurred. Yet, in certain areas, the presence of these Aedes species were not significantly recorded during outbreaks [Ae. aegypti at that time [Ae. aegypti predominated during the second yellow fever outbreak in Matadi and neighbouring townships [Ae. simpsoni and Ae. aegypti were found in proportions of 90% and 10%, respectively during these outbreaks [Ae. aegypti predominated Ae. simpsoni [Ae. aegypti larvae were collected from various breeding habitats in both natural and artificial breeding sites, while most positive breeding habitats were artificial [Anopheles and unspecified flavivirus was detected in Culex mosquitoes (Table Aedes albopictus has been recently established in Kinshasa [Despite the medical and veterinary importance of mosquitoes, published reports on mosquito vectors of viruses are extremely rare in DRC. Entomological indices even during outbreaks within the regions with high mosquito-borne virus transmission risk are lacking in most cases. Moreover, utbreaks . The enthat time . Ten yeaownships , while iutbreaks . In the simpsoni . In the tificial –22. A ratificial . O’nyonges Table . Aedes aKinshasa , where tKinshasa , 28, 30;Numerous factors might have contributed to the emergence and re-emergence of mosquito-borne viral infections in DRC. These driving factors range from demographic, socioeconomic, environmental to climatic factors. Moreover, evidence of population movement as key driving factor of spreading yellow fever has been documented . High poAe. aegypti was rarely reported [The overall findings indicate that mosquito-borne viral infections are an emerging and re-emerging health concern in the DRC. However, little attention has been paid and many questions on its epidemiology, socio-ecology and anthropology need to be addressed. While mosquito-borne viruses are re-emerging or emerging across the whole country, the north-western, north-eastern and western parts of the DRC are the most affected areas, experiencing multiple outbreaks compared to other parts of the country , 19–22. reported  comparedreported , 44–48. reported , 50. Thireported . Most prAedes mosquitoes invaded the city and successfully established in their new habitats closer to humans, leading to a high human mosquito biting rate in a deforested area. Indeed, the increased carbon dioxide in the atmosphere due to deforestation warms up the climate and causes changes in rainfall patterns leading to conditions of flood or drought. The establishment of new suitable habitats is creating an opportunity for new mosquito species to proliferate [Aedes mosquitoes [This review indicates that there is a dearth of information as regards to vector and transmission indices of mosquito-borne viral infections in DRC. This underlines the importance of better identification of different local vectors in each region or specific area within regions, and to assess competence of the vectors to transmit mosquito-borne viruses. Intensive deforestation activities in Gemena city provided an opportunity for mosquito-borne viruses to emerge . It is lliferate . This isliferate . It was liferate . Similarsquitoes . A studysquitoes . This obClimate change influences the frequency of mosquito-borne disease occurrence by affecting host and vector diversity, abundance, genetics and the pathogen infection load in the host and vector. Climate affects growth, survival and abundance of the vectors and virus replication load in the vector. Temperature is the major climate component susceptible to accelerate or decelerate mosquito reproduction and virus replication in the host depending on low and high temperature thresholds. Increased levels of precipitation lead to multiple microhabitats for oviposition and an efficient larval development pattern . The higAedes mosquito productivity and might explain in part the frequent chikungunya and yellow fever outbreaks in the city.RVFV is remarkably distributed in the eastern region of the DRC starting from the South to the North extremity. This region constitutes long borders with Zambia, Tanzania, Burundi, Rwanda, Uganda and South Sudan. Most of these countries are known to be endemic of Rift Valley fever –60. It iAedes, which is known to be the main vector of major mosquito-borne viruses. The lack of information is a barrier to appropriate knowledge on the ecology and epidemiology of mosquito-borne viruses, which is crucial in the design of appropriate interventions. Evidence for circulation of mosquito-borne viruses in livestock and domestic animals from the DRC, and the increasing mosquito-borne virus distribution over time and space, call for robust holistic mosquito-borne virus surveillance systems, including One Health approaches with emphasis on integrated vector control.Despite the limited information on mosquito-borne viruses and their vectors in DRC, available evidence indicates the wide-range of mosquito-borne virus species, potential hosts and vector species occurring in the country. The diversity of mosquito-borne viruses is underestimated, as well the current mosquito-borne viral disease burden and its driving factors. The tendency is that the continuous geographical expansion of mosquito-borne viruses leads to fears that multiple epidemic outbreaks are likely to arise in the DRC. There is a dearth of information on mosquitoes, in particular


The Trial-Ready Cohort for Preclinical/Prodromal Alzheimer’s Disease (TRC-PAD) Informatics Platform (TRC-PAD IP) was developed to facilitate the efficient selection, recruitment, and assessment of study participants in support of the TRC-PAD program.Describe the innovative architecture, workflows, and components of the TRC-PAD IP.The TRC-PAD IP was conceived as a secure, scalable, multi-tiered information management platform designed to facilitate high-throughput, cost-effective selection, recruitment, and assessment of TRC-PAD study participants and to develop a learning algorithm to select amyloid-bearing participants to participate in trials of early-stage Alzheimer’s disease.TRC-PAD participants were evaluated using both web-based and in-person assessments to predict their risk of amyloid biomarker abnormalities and eligibility for preclinical and prodromal clinical trials. Participant data were integrated across multiple stages to inform the prediction of amyloid biomarker elevation.TRC-PAD participants were age 50 and above, with an interest in participating in Alzheimer’s research.TRC-PAD participants’ cognitive performance and subjective memory concerns were remotely assessed on a longitudinal basis to predict participant risk of biomarker abnormalities. Those participants determined to be at the highest risk were invited to an in-clinic screening visit for a full battery of clinical and cognitive assessments and amyloid biomarker confirmation using positron emission tomography (PET) or lumbar puncture (LP).The TRC-PAD IP supported growth in recruitment, screening, and enrollment of TRC-PAD participants by leveraging a secure, scalable, cost-effective cloud-based information technology architecture.The TRC-PAD program and its underlying information management infrastructure, TRC-PAD IP, have demonstrated feasibility concerning the program aims. The flexible and modular design of the TRC-PAD IP will accommodate the introduction of emerging diagnostic technologies. Alzheimer’s Disease (AD) has emerged as one of the most significant public health issues of the 21st century. In 2020, it was estimated that 5.8 million people in the United States (U.S.) were living with Alzheimer’s Disease; this number was expected to rise to 13.8 million by 2050 , 5. The The TRC-PAD Informatics Platform (TRC-PAD IP)  was concwww.aptwebstudy.org) is a public-facing online longitudinal study that invited members of the general population who were over 50 years of age and had sufficient interest in participating in AD research to register for a user account. Once registered, users were presented with a guided walkthrough during which they provided their contact information and preferences, current AD-related diagnosis (if known), and interest in participating in AD-related research. Subsequently, users were offered additional study information, asked to consider the benefits and risks of study participation, and allowed to consent and enroll in the APT Webstudy using an online consent process. Participants were subsequently asked to provide additional information regarding their demographics, family and medical history, lifestyle, current AD biomarker status (if know), and complete assessments of their subjective memory perceptions using the Cognitive Function Instrument (CFI), and cognitive performance using the Cogstate Brief Battery  using responsive web design principles .To improve the prediction of participant risk, the APT Webstudy was designed to incorporate unsupervised cognitive assessments as part of a longitudinal observational study. Thus, participants are invited to return to the website every 3 months to complete a new round of assessments and update their information as needed. These follow-up assessments are designed to take 10 to 15 minutes to complete. To encourage compliance, participants received quarterly electronic mail reminders to log-in to the APT Webstudy website to complete their web-based assessments.An important consideration in a public-facing website is the variability of website traffic patterns. Websites with highly volatile traffic patterns can exhibit occasional periods of poor performance or unavailability. This is especially true when an unexpected media event, such as a viral social media mention or a celebrity endorsement, draws a burst of traffic toward the website. To mitigate the effects of these unexpected traffic spikes, the APT Webstudy has an architecture that dynamically adjusts its capacity to respond to incoming requests as changes in traffic patterns emerge. This architecture combines a cloud-based fleet of webservers, a multi-region database cluster, and a dynamic networking configuration to allow the website to automatically activate and deactivate computational resources on-demand to maintain performance during high-traffic periods in a cost-effective manner.brainhealthregistry.org), the Alzheimer’s Prevention Registry , Trial Match , and Healthy Brains , partnered with the TRC-PAD program to refer participants from their cohorts. These registries, collectively known as the “feeder” registries, provided the APT Webstudy with access to a large, engaged group of potential participants. The performance of these recruitment efforts was tracked by using Urchin Tracking Module (UTM) codes which associated participants with the campaign that referred them to the APT Webstudy  used to assess and manage the performance of the support team.APT Webstudy participant data were regularly evaluated using an adaptive algorithm that assessed each participant’s risk of AD biomarker positivity and ranked participants based on predicted risk . ParticiA key concept guiding the design of the SRS was the idea that site referral queues would be managed collaboratively among multiple site personnel. To support collaboration, the SRS website incorporated several features that optimized team-based management and disposition of participant referrals. For example, as new referrals became available in the referral queue, site personnel received notifications via electronic mail. The status of every referral in the queue was summarized in a dashboard view facilitating management and reporting. Each referral provided site personnel with access to a summary of participant-reported demographic, medical history, lifestyle, and contact information, as well as status changes. As a final step, referrals were assigned an outcome code and marked complete.The TRC was conceived as the final stage in the TRC-PAD process. The initial target for the TRC was to enroll a standing longitudinal cohort of 2,000 biomarker-confirmed participants  . An ElecThe TRC EDC was built to provide centralized management of all critical cohort study dataflows and workflows. This approach provided study teams with broad transparency and management capabilities over the study’s complex dataflows and processes. Implementing this approach, however, has proven challenging for traditional systems, which struggle to scale up as larger and more complex data types are introduced. Historically, a solution to this problem has been the implementation of interconnected purpose-specific systems. While feasible, this solution suffers from several shortcomings: 1) supporting evolving study requirements requires complex multi-system impact analysis, 2) training on multiple systems increases the burden on study teams, and 3) data integrations require ongoing maintenance as systems are updated. The TRC EDC was designed to avoid these issues by implementing a cloud-native architecture, which allowed it to harness the full range of capabilities of the underlying cloud platform on which it was hosted. In this architecture, the TRC EDC served as an orchestration engine that coordinated and delegated computational workloads, such as uploading and processing large binary objects , to the underlying cloud-based service best-suited for the required function, without impacting the study team’s interaction with the system.An initial step toward the establishment of the TRC was the creation of the TRC-PAD site network, a set of 35 academic clinical sites distributed across the large population centers in the contiguous United States. Sites were selected based on multiple criteria including study team research experience and expertise, amyloid positron emission tomography (PET) imaging and radiotracer availability, and track record in AD/AD and related disorders (ADRD) clinical trials. Site selection and activation were conducted in two stages, the “vanguard” or initial phase, which included 8 pilot sites, and the broad activation phase. The vanguard phase was used as a learning exercise to test and refine TRC-PAD data management tools and processes. These learnings were applied during the broad activation phase to ensure the full site network was optimized.The TRC-PAD selection and recruitment processes were managed by a series of learning algorithms that assessed participant data at multiple points. Newly acquired data were used to update participant risk predictions and rankings . These rTRC participants who meet eligibility criteria will be referred for screening and potential enrollment to downstream clinical trials, temporarily suspending additional longitudinal assessments and data collection activities in the TRC EDC. In these cases, a link between a participant’s TRC data record and their downstream clinical trial data record will be established via the use of the National Institute on Aging’s (NIA) Global Unique Identifier (GUID) , 15. ThiTRC-PAD IP supported a single Analytical Platform (AP) that aggregated data from multiple sources in a single semi-structured repository, also known as a «data lake» . This apThe TRC-PAD IP was constructed using open source web development and scientific computing tools hosted on Amazon Web Services (AWS), a public cloud computing platform . The TRCThe TRC-PAD IP was built on cloud-based computational infrastructure that was optimized to support each component system. The infrastructure supporting the APT Webstudy, which was subject to temporary bursts of traffic due to recruitment campaigns or unexpected media events, was designed to dynamically adapt to changing web traffic patterns by increasing or decreasing its fleet of webservers, within set cost parameters. Likewise, the TRC EDC, which managed a large, regulatory-compliant cohort database, used a cost-effective, highly durable storage strategy. In this architecture, computational resources were used on-demand, reducing idle capacity and providing flexibility as the TRC-PAD program’s requirements evolved.Ensuring participant confidentiality and data security were the primary requirements for the TRC-PAD IP. Thus, the TRC-PAD Informatics team worked with AWS ProServ to build a Health Insurance Portability and Accountability Act (HIPAA)-eligible architecture and implement AWS’s best practices . This arRegulatory oversight of the APT Webstudy was provided by the University of Southern California Institutional Review Board (USC IRB). Regulatory oversight for the TRC was provided by Advarra, Inc., under a single IRB (sIRB) model. The sIRB model was established as a National Institute of Health (NIH) requirement for multi-site studies starting in 2018 to streamline the review of research that involves human subjects  enrolled in the study via online consent and had completed more than 280,000 remote assessments . RecruitTRC sites began in-person screening of participant referrals in August 2019. As of July 6, 2020, 27 of 35 TRC sites were activated and had received 1,675 risk-ranked participant referrals via the SRS. Of these, 246 (14.7%) participants were referred to the TRC for initial screening, 123 (50.0%) participants completed the initial screening visit, 99 (80.5%) participants were authorized for amyloid testing, 55 (55.6%) participants were biomarker-confirmed using amyloid PET or CSF assessment, 26 (47.3%) participants were found to be amyloid elevated, and 23 (88.5%) participants were enrolled into the TRC . The demOur early experiences with the TRC-PAD program and its underlying information management infrastructure, TRC-PAD IP, have demonstrated feasibility concerning the program aims. The TRC-PAD IP supported recruitment, screening, and enrollment of TRC-PAD participants by leveraging a secure, scalable, cost-effective cloud-based information technology (IT) architecture. The APT Webstudy has demonstrated effectiveness in terms of selecting and recruiting individuals from the general population who have an elevated risk of amyloid positivity. The APT Webstudy has proven effective in remotely assessing participants on a longitudinal basis. The SRS has demonstrated effectiveness in terms of allowing TRC sites to manage site referrals promptly. The TRC EDC has supported the coordination of the multi-stage risk-based screening and enrollment process into the standing TRC. Much work remains to demonstrate the effectiveness of the TRC-PAD program in accelerating recruitment into downstream AD clinical trials.Several limitations should be considered. First, the TRC-PAD program has struggled to select and recruit a representative sample of participants across multiple sociodemographic dimensions . EffortsIn May 2019, the TRC-PAD principal investigators selected the first set of downstream clinical trials slated to utilize the standing TRC to recruit participants. These clinical trials are scheduled to begin recruiting participants in North America (U.S. and Canada) in May 2020. The efficiencies that will accrue to these clinical trials are two-fold: 1) by drawing participants from the TRC, the expectation is that recruitment into these clinical trials will be accelerated by reducing the number of screen failures; and 2) the TRC clinical, neuropsychological, biofluid, and imaging data will be available to use as high-quality run-in data for downstream clinical trials . These cAs the TRC-PAD program has been established over the past few years, promising new AD biomarkers have been developed. Plasma-based AD biomarkers, for example, have been shown to effectively predict amyloid positivity . When fuFinally, as news of the TRC-PAD program’s progress has spread, a global network of programs modeled on its approach has begun to take shape. Investigators in several countries have been collaborating with the TRC-PAD program leadership to establish similar programs in their home regions. By using global cloud computing infrastructure and the TRC-PAD IP as a model architecture, these programs have been able to rapidly establish similar platforms in their local jurisdictions. Once fully operational, the global TRC-PAD network aims to provide AD clinical trials with a steady stream of well-characterized, biomarker-confirmed participants yielding savings in time, effort, and expense.