Cardiovascular disease (CVD) complications have contributed significantly toward poor survival of cancer patients worldwide. These complications that result in myocardial and vascular damage lead to long-term multisystemic disorders. In some patient cohorts, the progression from acute to symptomatic CVD state may be accelerated due to exacerbation of underlying comorbidities such as obesity, diabetes and hypertension. In such situations, cardio-oncologists are often left with a clinical predicament in finding the optimal therapeutic balance to minimize cardiovascular risks and maximize the benefits in treating cancer. Hence, prognostically there is an urgent need for cost-effective, rapid, sensitive and patient-specific screening platform to allow risk-adapted decision making to prevent cancer therapy related cardiotoxicity. In recent years, momentous progress has been made toward the successful derivation of human cardiovascular cells from induced pluripotent stem cells (iPSCs). This technology has not only provided deeper mechanistic insights into basic cardiovascular biology but has also seamlessly integrated within the drug screening and discovery programs for early efficacy and safety evaluation. In this review, we discuss how iPSC-derived cardiovascular cells have been utilized for testing oncotherapeutics to pre-determine patient predisposition to cardiovascular toxicity. Lastly, we highlight the convergence of tissue engineering technologies and precision medicine that can enable patient-specific cardiotoxicity prognosis and treatment on a multi-organ level. The discovery of human induced pluripotent stem (iPSC) technology has made it possible to regulate organ-specific switches in stem cells to generate any cell type outside the body in a highly controlled artificial environment  and cancer. A 2018 statistics reported ∼17 million deaths globally from CVD and ∼9 million deaths from cancer . By 2040ironment . In the ironment . In the ironment . iPSC teefficacy . In thisAssessment of cardiovascular risks using conventional approaches such as non-clinical animal models have been challenging due to striking differences in both biochemical regulation and functional parameters such as beat rate and calcium handling . In contin vitro Proarrhythmia Assay (CiPA) initiative. CiPA aims to evaluate the proarrhythmic risk of new drugs through a comprehensive mechanistic assessment and validation on human iPSC-CMs. These drug assessments occur in four distinct stages: 1) Characterization of the drug effects on human cardiac currents; 2) In silico reconstruction of the ventricular electrophysiology; 3) modeling the effects on iPSC or embryonic (ES)- derived ventricular cardiomyocytes; and 4) clinical evaluation of cardiac risk. The primary endpoint assay is detection of electrophysiological abnormalities that could be due to changes in repolarising or depolarising ionic currents . Electrophysiological abnormalities are often underreported in several drug induced toxicities but are key early indicators of potential drug-induced adverse effects  anthracyclines; 2) tyrosine kinase inhibitors; and 3) checkpoint inhibitors. The co-incidence of cardiotoxicity associated with chemotherapeutic drugs was first identified and reported in 1970s . Since tin vitro , in particular have been shown to elicit a dose-dependent effect both in an acute or chronic setting  Sorense. Indeed,in vitro . These iin vitro . In anotin vitro . Indeed,in vitro . Such paHuman iPSC-CMs have been utilized to screen cardiovascular toxicities associated with tyrosine kinase inhibitors (TKIs) to mirror clinical phenotypes. TKI-associated cardiotoxicities in cancer patients include arrhythmias, myocardial infarction and reduced left ventricular ejection fraction (LVEF) . While TOne of the first large-scale studies looking at TKI-induced cardiotoxicity tested 21 FDA approved TKIs on iPSC-CMs derived from 11 healthy and two cancer patients on a high-throughput platform . Six TKICancer immunotherapy drugs are targeted biologics aimed to reinvigorate the immune cells’ capacity to target and eliminate cancer cells. In chimeric antigen receptor (CAR) T-cell therapy, patient’s T cells are isolated and engineered to express receptors, which upon binding to cancer cells lead to cell death. Sporadic clinical cardiotoxicity has been reported mainly arising from co-expression of tumor targets or antigen cross-reactivity with cardiac proteins . One of hiPSC-CMs have emerged as one of the important predictive tools is due to its ability to retain genetic identity of the patient for disease modeling and discovery of personalized medicine. However, somatic cell sources for pluripotency induction, differences in cardiomyocyte differentiation protocols and heterogeneity in myocyte composition can lead to inconsistencies in the outcomes of pharmacological studies. Several studies have indicated that differences in genetic background and iPSC-CM derivation methodologies can influence epigenetic landscape and consequently gene expression. For example, iPSCs derived from cardiac progenitor cells are shown to have higher cardiac differentiation potential compared to non-cardiac sources . This suTo date several key optimizations in cardiac differentiation have been explored to derive iPSC-CMs using simple, cost-effective and xeno-free methods . DespiteWith regard to function, in adult cardiomyocytes mature structural features are intertwined with functional regulation. Unlike adult cardiomyocytes, neonatal or iPSC cardiomyocytes do not show rectangular morphology, exhibit spontaneous generation of AP, have lower density of mitochondria and higher dependence on glycolytic metabolism over fatty acid oxidation. Several recently developed techniques help iPSC-CM maturation through metabolic supplementation , incremeCurrent drug discoveries and therapeutic testing takes place on conventional 2D tissue culture platforms. However, there are several limitations with using platforms that are non-physiological in a reductionist manner. 2D planar culture platforms offer flexibility and ease of use, however, fail to capture the structural complexities that are seen in 3D tissues and organ systems. These 3D tissues develop through gradients of biochemical and mechanical signaling, and multicellular crosstalk that contribute to the maintenance of tissue homeostasis. In a biological context, 2D platform offers the least resistance at a single-cell level wherein cellular turnover, metabolism, and protein synthesis occur in an accelerated manner. In contrast, in a 3D tissue-like architecture these processes occur in concert with changes in the local extracellular environment, receiving feedback responses from a multitude of cells that are tightly packed in a small volume . With thiPSC-CMs have been used to generate 3D myocardium as self-assembled, scaffold-free, spontaneously beating clusters referred as cardiac spheroids. Alternatively, iPSC-CMs can also be embedded in natural or synthetic extracellular matrix (ECM) in form of engineered heart tissues (EHTs) that allow anisotropic tissue-like orientation and cellular alignment. iPSC-CMs in both scaffold-free cardiac spheroids and matrix assembled tissues have shown to exhibit more mature characteristics when compared to 2D cultures . Maturitin vivo, unlike 2D culture systems. Furthermore, for the assessment of potential drug-induced toxicity, model systems that are relevant to human cardiac physiology and diseases may be more suitable over other species due to inherent differences in function and drug sensitivity. An overview of 2D and 3D stem cell derived cardiomyocyte platforms used for chemotherapeutic testing on cardiovascular cells is summarized in As a platform technology for high-throughput screening, iPSC-CMs have been assembled into EHTs on a 24-well  and 96-wAt a cellular level, decreased contractile function due to mutational cardiac disorder or drug-induced toxicity is often associated with cytoskeletal disarray and poor structural integrity. These changes influence electromechanical coupling and biomechanical properties both at the cellular as well as tissue level. A combination of atomic force microscopy (AFM) coupled with MEA platform could be used to measure sensitive biological changes in topography and force .in vitro cardiac models although offer very valuable insights in mimicking tissue complexities and multicellular interaction, they do not fully capture the dynamic pharmacokinetics and pharmacodynamics in volved in drug metabolism and absorption. Hence scaled-down models of interconnected multi-organ systems may provide additional insights that are not currently offered by static 3D in vitro models.In an adult human heart, over 60% of the tissue comprises of non-myocytes, namely endothelial cells, vascular stromal cells, cardiac fibroblasts and a small fraction of immune cells . Crosstain vitro models. Furthermore, integrated platforms such as these combined with biosensors can enable continuous monitoring over long period of time. A fully integrated multi-organ platform developed by Zhang et al. demonstrated drug-induced organ toxicity using a liver-and-heart-on-a-chip comprising of human iPSC-derived cardiac spheroids and human primary hepatocyte-derived liver organoids  for metabolite analyses. This on-line rapid detection of drug metabolites and inhibitors can be utilized to identify new biomarkers that can predict cardiotoxicities (High-throughput quantitative multiplex assays provide a comprehensive understanding of signal transduction networks and molecular signatures that emanate from oncogenes. Similarly, they can detect potential pathways that are activated in other somatic cells that contribute toward organ-specific or systemic toxicity. xicities . A key aAdvances in the latest single-cell analyses tools allow profiling of transcriptomic changes at a higher resolution . The tecin vitro disease modeling approach that ensures reproducibility whilst capturing both acute and chronic effects will offer a boost in the predictive power and development of effective personalized cancer therapies.Cardiovascular diseases and several types of cancers have common risk factors and share intricately related pathogenesis that are linked via common cellular pathways. Therefore, the negative consequences of cancer drugs on the cardiovascular system is inevitable. Although conventional toxicology studies in animal models have translational value, they can be cost ineffective and time intensive. As an alternative, iPSC technology can be harnessed to provide a comprehensive phenotypic and genotypic trait that can be used clinically to make therapeutic decisions. Derivation of cardiovascular cells from cancer patients can be utilized to characterize unique functional and genomic signatures that may influence potential adverse reactions to clinically approved drugs. However, with regard to cell maturity there are several milestones that still needs to be reached for minimizing the batch-to-batch variability and the time taken to generate cell or tissue models for a timely and accurate pharmacological prediction. Creation of multiple tissue types using iPSCs can be tested alongside tumors from the same patient in a microphysiological tissue chip devices to tease the mechanistic effects of onocotherapeutics on the body. Such “patient-on-a-chip” models will aid in stratification of cancer patients based on the sensitivity and efficacy of oncotherapeutics . Current


Low-complexity nonlinear equalization is critical for reliable high-speed short-reach optical interconnects. In this paper, we compare the complexity, efficiency and stability performance of pruned Volterra series-based equalization (VE) and neural network-based equalization (NNE) for 112 Gbps vertical cavity surface emitting laser (VCSEL) enabled optical interconnects. The design space of nonlinear equalizers and their pruning algorithms are carefully investigated to reveal fundamental reasons of powerful nonlinear compensation capability and restriction factors of efficiency and stability. The experimental results show that NNE has more than one order of magnitude bit error rate (BER) advantage over VE at the same computation complexity and pruned NNE has around 50% lower computation complexity compared to VE at the same BER level. Moreover, VE shows serious performance instability due to its intricate structure when communication channel conditions become tough. Moreover, pruned VE presents more consistent equalization performance within varying bias values than NNE. Recent decades have witnessed the explosion of data traffic especially in regional spaces like data centers and supercomputers. The hardware infrastructure for supporting such massive connectivity has been turning to optical components and fibers even over a distance less than 100 m. The data rate of optical interconnects have exceeded 100 Gbps per lambda in commercialized products and will soon upgrade to 200 Gbps per lambda for 800 Gbps or 1 Tbps optical module , carrierAdvanced digital signal processing (DSP) technologies-based nonlinear equalization, such as Volterra series-based equalization (VE) ,16,17,18formance . More imIn this paper, we compare the efficiency and stability of a pruned three-layer neural network-based equalizer and a pruned three-order Volterra series-based equalizer for VCSEL enabled 112 Gbps optical interconnects. A threshold-based pruning and retraining algorithm, which is proposed in our previous publications ,12, is uP-order VE with memory length of The mathematical expression of Equation ,1)y(k)y(k)P-ordP and memory length P-order VE with memory length of It can be seen from Equation , the numgorithms . Therefoerations  and exprAccording to ,26, a thA basic Artificial neural networks have been widely deployed in various areas such as image classification and natural language processing, showing much better performance than traditional algorithms. Recently, it has been intensively researched in the field of optical communication, in which nonlinear equalization is one of hottest topic . To equax(k) is a sampled signal sequence, f means activation function of hidden layer. Rectified linear unit (ReLU), tangent hyperbolic (Tanh) and sigmoid functions are considered in this work. Function y is final result indicating a specific symbol of information. Loss functions of ANN, including mean square error (MSE) loss function, logarithmic loss function and cross-entropy loss function, are important for training model and final performance. The MSE loss function usually shows slow convergence and is seldom used in classification task. In this work, back propagation (BP) algorithm combined with cross-entropy loss function are used to train NNE to achieve better classification performance.The mathematical formula of NNE process can be expressed as Equation ,(4)y=arThe structure of the 3-layer NNE is shown in Equation ,(5)CompT is threshold. With weight value setting to zero, connections relating to this value is cut in these complex networks. Due to different features of two architectures, there also needs special design considerations for each equalizer. For VE, second- and third-order coefficients are pruned because we find linear terms only occupy a small amount of computation complexity while contribute most on the performance gain due to bandwidth limitation of the experimental system. For NNE, all weights both from input layer to hidden layer and hidden layer to output layer are run for pruning based on the threshold. An iterative pruning process is developed to achieve a more efficient network structure.It has been proved that most of multiplication operations in the nonlinear equalizers are redundant  as eitheTo avoid the risk of performance over-estimation from nonlinear equalization, we randomly disrupt the signals to create a sequence without periodicity before the received signals are fed into equalizer and conducted mini-batch training . After sFirst, memory length should be carefully tuned for VE and NNE to better understand the efficiency of pruning algorithms. If these parameters are intentionally set with large numbers, complexity reduction ratio will be exaggerated through pruning due to expanded redundancy, which is not fair for comparison. Therefore, we have tuned memory length of first order in VE by setting high-order memory length to zeros. Second, 100 Gbps PAM4 signaling at a typical 1-dBm received optical power is used for comparing efficiency of VE and NNE and their pruning version. Third, in order to better understand behavior of VE and NNE, we increase the data rate to 112 Gbps and eliminate the pre-distortion process for pressuring more on the bandwidth limitation as indicated in the inset of Performance stability for equalization is of great significance in the real application where output optical power or 3-dB bandwidth of critical devices in the optical interconnect will change within a life-cycle because of component aging or ambient temperature fluctuation. It is likely that performance stability will become worse with pruning algorithms since that the redundancy of connections in the equalization will be useful in changing scenarios. To verify the stability of pruning algorithm with changing link conditions, we choose DC bias of VCSEL as a key parameter for further evaluation because DC bias variation of VCSEL will not only change the output power but also bandwidth of directly modulated laser. The transmission experiment is run with a 100 Gbps PAM4 pre-distorted link. We set In this paper, we compare the efficiency and stability performance of pruned Volterra series-based equalization and neural network-based equalization for 112 Gbps VCSEL enabled optical interconnects. From the 112 Gbps PAM4 transmission experiment, we can conclude that NNE has more than one order of magnitude BER advantage over VE at the same computation complexity. By using pruning algorithms, NNE has around 50% lower computation complexity compared to VE at the same BER level. Moreover, when communication channel condition become tough, VE shows serious performance instability due to its intricate structure. Through performance stability evaluation by changing bias current of a VCSEL, pruned VE shows more consistent equalization performance than NNE.


Glioblastoma is the most common and aggressive adult brain tumour, with poor median survival and limited treatment options. Following surgical resection and chemotherapy, recurrence of the disease is inevitable. Genomic studies have identified key drivers of glioblastoma development, including amplifications of receptor tyrosine kinases, which drive tumour growth. To improve treatment, it is crucial to understand survival response processes in glioblastoma that fuel cell proliferation and promote resistance to treatment. One such process is autophagy, a catabolic pathway that delivers cellular components sequestered into vesicles for lysosomal degradation. Autophagy plays an important role in maintaining cellular homeostasis and is upregulated during stress conditions, such as limited nutrient and oxygen availability, and in response to anti-cancer therapy. Autophagy can also regulate pro-growth signalling and metabolic rewiring of cancer cells in order to support tumour growth. In this review, we will discuss our current understanding of how autophagy is implicated in glioblastoma development and survival. When appropriate, we will refer to findings derived from the role of autophagy in other cancer models and predict the outcome of manipulating autophagy during glioblastoma treatment. Gene expression analyses of patient-derived tumour cells revealed three distinct glioblastoma subtypes, classical, proneural and mesenchymal, which are classified based on their molecular genotypes . RecentlPTEN, TP53, NF1 and CDKN2a) [EGFRvIII, a truncated mutant that lacks the extracellular ligand-binding domain and signals constitutively in the absence of growth factors [PDGFRA is associated with the proneural subtype, whereas the loss of NF1 is linked to the mesenchymal subtype [All molecular subtypes of glioblastoma are driven by the gain-of-function of receptor tyrosine kinases (RTKs) and/or the loss of tumour suppressor activities  are a potential causative source . GlioblaThe current standard of care for glioblastoma patients is surgical resection followed by treatment with the chemotherapeutic agent, temozolomide ,10. HoweOne potential explanation for the molecular subtype-dependent variation in the survival of glioblastoma patients is the differences in the tumour immune microenvironment. Approximately, one-third of the glioblastoma tumour mass is composed of innate immune tumour-associated macrophages (TAMs) that have mostly infiltrated from the peripheral immune system ,12. ThesIrrespective of the underlying molecular mechanisms contributing to enhanced patient survival, glioblastoma remains a cancer of a high mortality rate . The chaThe aggressive nature of glioblastoma, including unconstrained growth, invasion into the normal brain parenchyma and resistance to therapy, indicates that glioblastoma cells have developed mechanisms to survive cell stress and proliferate under restrictive conditions. It is therefore important to understand cellular survival mechanisms used by glioblastoma cells in order to develop new treatments that can effectively target the tumour. Autophagy is one such pathway that is upregulated in cancer cells in response to stress. This review aims to explore what is known in the literature regarding the role of autophagy during the development and survival of glioblastoma cells. When appropriate, we will discuss the role of autophagy in other cancers and the potential applications to glioblastoma. However, we will first introduce the current models used to study glioblastoma and their drawbacks in order to better grasp the limitations of studying the role of autophagy in this aggressive cancer.2.The ideal model of glioblastoma should mimic various aspects seen in patients including molecular heterogeneity, interaction with stromal cells, invasion and exposure to growth restrictive conditions. To date, no individual model can recapitulate all these aspects. Here, we will briefly outline the advantages and limitations of the currently available models to study glioblastoma (recently reviewed in ).2.1.Cell culture models provide the flexibility to enable in-depth examination of molecular details and imaging analyses in a simplified and controlled manner. This comes at a cost of excluding the contribution of the tumour microenvironment and the underlying molecular complexity . In addiCulture conditions are likely to influence the maintenance of original cell identity. Recent studies have shown that culturing glioblastoma patient-derived stem cells under serum-free stem cell conditions in monolayers is more likely to preserve long-term stem cell properties in comparison to cells grown in neurospheres ,21–23. I2.2.Three-dimensional organoids are an emerging novel model system to study glioblastoma. Organoids were initially grown from dissociated patient-derived GSCs that can infiltrate and proliferate in human cerebral organoids . However2.3.Mouse models of glioblastoma provide an important tool to investigate the role of the tumour microenvironment, the interaction of tumour cells with stromal cells and the vasculature, and tumour cell invasion into the brain parenchyma. Brain tumours that resemble glioblastomas can be induced in mice using genetically engineered mouse models (GEMMs) or by injecting animals with the tumour-initiating mouse-or patient-derived cells. The different glioblastoma mouse models have been comprehensively reviewed in .A well-known GEMM of glioblastoma is the replication-competent avian sarcoma-leukosis virus LTR splice acceptor/tumour virus A (RCAS/TVA) model. This model has been engineered to express the avian virus receptor TVA in glial progenitor cells in order to allow the delivery of transgenes upon viral injection . Particu3.3.1.Autophagy is a catabolic process that involves the engulfment of cytoplasmic components in double-membrane vesicles, called autophagosomes, which can fuse with lysosomes for the degradation and recycling of their contents to provide metabolic substrates. Autophagy occurs constitutively in cells at basal levels to maintain homeostasis through the turnover of damaged organelles and unwanted cellular material . In periSeveral autophagy-related (ATG) protein complexes are required for autophagosome biogenesis. The activation of the Unc-51-like kinase 1 (ULK1) complex can initiate autophagy during periods of nutrient and energy depletion. This can occur upon the inactivation of the mammalian target of rapamycin (mTOR) complex 1 (mTORC1) in response to low nutrient levels or activation of the AMP-activated protein kinase (AMPK) in response to energy depletion . The ULKAtg genes in mice results in the loss of neonatal survival thereby revealing a vital role for autophagy during mammalian development [Atg5-deficient mice is rescued by re-expressing Atg5 in neurons, indicating that autophagy plays an essential role in brain development [Atg5 or Atg7 also resulted in neurodegeneration phenotypes in mice, further highlighting the role of autophagy in maintaining neuronal homeostasis [Knockout of core elopment . Intereselopment . Whole-beostasis –39. The eostasis ,40. WhilAtg genes in mice, the knockout of some autophagy players results in embryonic lethality [Unlike the neonatal lethality phenotype observed upon the knockout of core ethality –43. Thisethality ,45. Beclethality ,46. HoweSeveral ATG proteins, including ATG3, ATG5 and ATG7, are also required for lipidation of ATG8 family members on single-membrane vesicles, such as phagosomes and perturbed endosomes. The functional relevance of this non-canonical ATG8 lipidation on single-membrane vesicles may be distinct from its role during autophagosome biogenesis and can be triggered by various stimuli, including pathogen infection and treatment with lysosomotropic agents –49. This3.2.Assays to monitor autophagy mainly rely on measuring protein levels and modifications as well as cellular localization . These aTo accurately measure autophagic flux, careful assays are required to induce autophagy and concurrently block the degradation step. Autophagy can be stimulated following mTORC1 inhibition by nutrient starvation or with chemical agents such as rapamycin, while blocking the lysosomal degradation of autophagosome contents is achieved using inhibitors of lysosomal acidification, such as bafilomycin A1  and chloin vivo and in human patients. RNA expression of ATG genes has also been extensively used as a readout for autophagy, but changes in transcription or protein levels do not necessarily reflect alterations in ATG protein activity. As a result, additional readouts for autophagy, such as post-translational modifications of ATG proteins [Given the dynamic nature of autophagosome biogenesis, it is challenging to measure autophagy proteins , are nee4.Beclin 1, Atg7 and Atg5 [Atg5 expression showed that, while extended autophagy inhibition can accelerate ageing, reactivating autophagy after a period of inhibition can cause increased frequency of spontaneous tumour development [Autophagy has fundamental roles in maintaining cell health. In terms of oncogenic transformation, autophagy can play a role in preventing oxidative stress and subsequently DNA damage and cell transformation. This tumour-suppressive role of autophagy is supported by early studies that showed increased development of spontaneous, potentially benign, lung and liver tumours and lymphomas in mice deleted of autophagy genes and Atg5 ,46,56. Ielopment . This suelopment . Indeed,elopment . How autKRAS- and BRAF-driven lung tumours [Many solid tumours undergo metabolic rewiring to enable their unconstrained growth in nutrient-deprived microenvironments . Recycli tumours –62. In a tumours . Mouse m tumours ,64.The role of autophagy during cancer treatment has been extensively studied using both animal models and cultured cells. Autophagy can be induced by a wide range of anti-cancer drug treatments. This can occur directly, for example, following mTORC1 inhibition by rapamycin , or releV600EBRAF kinase inhibitor (vemurafenib) and restoring its efficacy in treating V600EBRAF-driven brain tumours [KRAS-driven PDAC to pharmacological suppression of the RAS/RAF/MEK/ERK signalling pathways [Based on promising findings from pre-clinical studies, several clinical trials have been initiated to investigate the effect of concurrently inhibiting autophagy during cancer treatment . Blockin tumours ,72. Morepathways ,73 and hpathways .G12DKRAS-driven PDAC and non-small cell lung carcinoma (NSCLC) models, whereby autophagy loss reduced tumour growth but not when p53 was additionally deleted [Overall, the role of autophagy in cancer is multifarious, varying from being a tumour suppressing mechanism to supporting tumour growth and survival. This may reflect tumour type- and stage-specific functions of autophagy or diverse effects of inhibiting early stages versus late stages of the pathway . Interestingly, an oncogene-dependent effect of autophagy during tumour growth has been described in  deleted ,75. Alto5.The relevance of autophagy in glioblastoma is not fully uncovered. The properties of this invasive and resistant solid tumour could suggest a likely role for autophagy in promoting its growth . Here, w5.1.ATG gene transcription, including ATG7, LC3B, LC3C, ATG16L1 and SQSTM1 (which encodes p62) [DRAM), were also increased [ATG gene expression signatures and reduced overall survival predominantly in patients with the mesenchymal subtype [ATG gene expression and a worse outcome for glioblastoma patients, thereby suggesting a potential role for autophagy in promoting glioblastoma growth.Assessing autophagy levels is one approach to examine its involvement in glioblastoma development. Enhanced autophagic activity evaluated by immunohistochemistry analyses of resected tumours has been shown to correlate with reduced glioblastoma patient survival ,77. Simides p62) . In addincreased . Interes subtype ,80. This5.2.G12DKRAS in a Cdkn2a−/− background where autophagy was disrupted by genetic knockdown of Atg7, Atg13 or Ulk1 in tumour cells [G12DKRAS-expressing Cdkn2a−/− glial cells in culture revealed that autophagy inhibition impaired anchorage-independent cell growth and induced senescence [RAS oncogene are only rarely detected in glioblastoma. Further GEMM studies using established glioblastoma oncogenic drivers are thus required to confirm the role of autophagy in initiating tumour development.As discussed previously, autophagy activation can impact tumour cells at various stages, including initiation, proliferation and response to treatment. A limited number of studies have examined how autophagy influences the formation of glioblastoma . An inveur cells . The molnescence . This moATG4B impeded the growth of GSCs expressing high but not low MST4 levels [MST4 and ATG4B transcripts correlated with GSCs encompassing the mesenchymal-like molecular subtype and reduced patient survival [Autophagy can also implicate tumour cell proliferation. Orthotopic xenograft experiments of patient-derived GSCs in immunocompromised mice suggest a link between autophagy and levels of MST4 , a serin4 levels . It may 4 levels . Interessurvival ,82, suggAdditionally, EGFRvIII-expressing glioblastoma cells, which are enriched in the classical subtype, have a greater dependency on autophagy for growth and survival during times of metabolic stress induced by nutrient-deprivation and hypoxia . Non-spe6.+ T cell infiltration [The interactions between glioblastoma and immune cells in the tumour microenvironment may have profound effects in determining treatment outcome. Manipulating autophagy has been described to be favourable in the context of immunotherapy, which includes dendritic cell (DC) vaccines, chimeric antigen receptor (CAR)-T cells and immune checkpoint inhibitors . Howeverltration . This imltration . Since altration , it woul+ T cell recognition, as seen in PDAC [By contrast, autophagy may facilitate immune evasion, for example by degrading major histocompatibility complex (MHC)-I in tumour cells thereby reducing antigen presentation and CD8 in PDAC . Additio in PDAC . This en in PDAC . It is t in PDAC –94.Autophagy proteins not only play a role in the tumour cells themselves, but their function in immune cells can also contribute to the outcome of anti-tumour immune responses. The non-canonical lipidation of ATG8 proteins on single membranes is required in Lewis lung carcinoma myeloid cells to impair the local anti-tumour T cell activation in response to the engulfment of dying cells . MHC-II Further investigation and pre-clinical models are necessary to discover the role of autophagy in immune responses associated with glioblastoma and to distinguish the contribution of canonical versus non-canonical lipidation of ATG8 proteins. Particular care should be taken when considering if autophagy inhibition or activation will potentiate the immune response to target glioblastoma since the outcome may vary depending on if autophagy is manipulated in the tumour and/or immune cells.7.The invasive nature of glioblastoma cells permits their infiltration to the surrounding normal brain tissue and hinders complete surgical removal. Tumour cells can acquire invasive properties by undergoing an epithelial-to-mesenchymal transition (EMT), a reversible process whereby cells lose epithelial cell properties and gain mesenchymal migratory characteristics. This process involves cytoskeletal remodelling, the loss of cell-cell adhesions and the degradation of the basement membrane and ECM . EMT is Autophagy has been shown to facilitate the degradation of SNAIL and SLUG in glioblastoma cells thereby consequently upregulating N-cadherin . This suDRAM1, SQSTM1 and ATG7 impaired the capacity of patient-derived, mesenchymal-like GSCs to invade using a transwell cell culture system, supporting a role for autophagy in glioblastoma invasion [α stabilization can reduce LC3C expression [Potential contradicting evidence exists that suggests the role of autophagy in supporting tumour cell motility and invasion . Downreginvasion . In addiinvasion  which isinvasion  and is ainvasion . In a xeinvasion . Activatinvasion . Interesinvasion . On the invasion . HIF2α spression ; therefopression . Given tOverall, these studies suggest a role of autophagy in regulating glioblastoma invasion. However, it is important to take into account the effects of the tumour–stroma interactions and the tumour microenvironment (such as oxygen and nutrient availability) which can affect cell invasion . More in8.α/β, have failed to significantly improve patient survival compared to temozolomide or irradiation alone [Inhibiting the RTK signal transduction pathways, which drive glioblastoma progression, is an attractive therapeutic strategy. Nevertheless, phase II clinical trials targeting RTKs, for example, tyrosine kinase inhibition of EGFR/EGFRvIII or PDGFRon alone ,114. Theon alone . TherefoAccumulating evidence suggest an interaction between RTK signalling and autophagy . AutophaRTKs, such as EGFR, are activated upon binding to their ligands when located on the plasma membrane and continue to signal following endocytosis. Signalling from endocytosed RTKs can be either terminated upon targeting to lysosomes or maintained upon recycling back to the plasma membrane. Autophagy can regulate RTK signalling through various mechanisms. The loss of autophagy in immortalized glial cells has been shown to disrupt endosomal homeostasis, consequently perturbing EGFR trafficking and signalling in response to growth factor stimulation thereby reducing cell survival . This waβ signalling has been shown to enhance autophagy via stabilization of HIF1α during hypoxia [α amplification is associated with 13.1% of glioblastomas [While trafficking and signalling of EGFR is well studied thereby facilitating our understanding of its interplay with autophagy, PDGFR activation  is less well understood. It remains unknown whether autophagy proteins play a regulatory role in PDGFR signalling and trafficking. Unlike autophagy inhibition by EGFR activation, autocrine PDGFR hypoxia , demonst9.Glioblastoma tumours inevitably relapse and are resistant to temozolomide and radiotherapy treatment. To date, alternative strategies designed to suppress glioblastoma tumour cell growth have been unsuccessful. The reasons behind this include failure to target quiescent tumour-initiating GSCs, inherent plasticity of tumour cells, reduced drug delivery across the blood–brain barrier (BBB), and the lack of pre-clinical studies that model the complex interactions of glioblastoma tumour cells with the tumour microenvironment.Quiescent stem cells are resistant to anti-cancer treatment that target actively proliferating cells by triggering cell death or inhibiting cell cycle progression. GSC heterogeneity and plasticity have contributed to the difficulty in translating drug responses observed in pre-clinical studies ,8, whichα/β (by imatinib treatment) have been found to upregulate autophagy in glioblastoma cell lines [In addition to supporting the development of glioblastoma, it is recognized that autophagy is upregulated in response to the first-in-line treatment, temozolomide plus radiotherapy . Autophall lines –124. Thell lines .10.Accumulating pre-clinical studies suggest that autophagy inhibition enhances the sensitivity of glioblastoma cells to radiation  and chemAlthough there is pre-clinical evidence that suppressing autophagy may be a promising mechanism to thwart resistance and enhance sensitivity to anti-cancer treatment, results from clinical trials have provided conflicting evidence . Early tα/β cytotoxicity in cell culture [Targeting HDAC activity is an emerging anti-cancer approach likely to exert its effects through the regulation of gene expression and chromatin remodelling . Yet, a  culture .Inhibiting mTOR kinase activity has been used to treat various cancers. The inactivation of mTORC1 can directly induce autophagy by releasing the ULK1 complex from its inhibitory phosphorylation. In the case of glioblastoma, treatment with rapamycin, which inhibits mTORC1 by limiting its access to substrates, augments the cytotoxic response of irradiated primary glioblastoma neurospheres  and glioThe disparity in the role of autophagy during glioblastoma response to therapy could indicate that the upregulation of autophagy beyond a specific threshold, for an example during rapamycin treatment, may cause intracellular stress that sensitizes cells to death-stimulating signals. Therefore, targeting autophagy as a means to treat glioblastoma requires careful investigation and modelling prior to clinical application.11.Numerous findings have described the relevance of autophagy in tumourigenesis. Autophagy plays a role in maintaining cellular homeostasis and thereby prevents oncogenic transformation and tumour growth in certain tissues. On the contrary, it is also appreciated that cell stress-induced autophagy is used by many established tumours to enable their unconstrained growth in the restrictive tumour environment. Furthermore, autophagy upregulation in response to chemotherapy and radiotherapy can lead to treatment resistance by aiding tumour cell survival. To date, evidence from glioblastoma models indicates that autophagy is involved in tumour initiation, development and response to treatment. Yet manipulating autophagy in patients has had limited success in improving their survival indicating that understanding the relevance of autophagy in various aspect of glioblastoma requires in-depth investigation.Currently, targeting autophagy in the clinical setting is limited to a few non-specific inducers and suppressors. For that reason, mechanistic understanding of autophagy requires additional research in order to develop novel and specific modulators. Additionally, it will be key to evaluate if autophagy dependency of glioblastoma tumour cells is influenced by molecular subtypes. Determining if a subset of patients is more likely to benefit from autophagy modulation could lead to personalized treatment and potentially better outcomes. Finally, given the impact of autophagy in both tumour and stromal cells, it would be important to employ whole-body genetic modulation of autophagy in pre-clinical studies in order to closely mimic its therapeutic targeting. It is possible that modulating autophagy in tumour and stromal cells may give opposing impact on improving survival.


Solid dispersion is the preferred technology to prepare efficacious forms of BCS class-II/IV APIs. To prepare solid dispersions, there exist a wide variety of polymeric carriers with interesting physicochemical and thermochemical characteristics available at the disposal of a formulation scientist. Since the advent of the solid dispersion technology in the early 1960s, there have been more than 5000 scientific papers published in the subject area. This review discusses the polymeric carrier properties of most extensively used polymers PVP, Copovidone, PEG, HPMC, HPMCAS, and Soluplus® in the solid dispersion technology. The literature trends about preparation techniques, dissolution, and stability improvement are analyzed from the Scopus® database to enable a formulator to make an informed choice of polymeric carrier. The stability and extent of dissolution improvement are largely dependent upon the type of polymeric carrier employed to formulate solid dispersions. With the increasing acceptance of transfer dissolution setup in the research community, it is required to evaluate the crystallization/precipitation inhibition potential of polymers under dynamic pH shift conditions. Further, there is a need to develop a regulatory framework which provides definition and complete classification along with necessarily recommended studies to characterize and evaluate solid dispersions.The online version contains supplementary material available at 10.1208/s12249-020-01849-z. In the quest to access the complex targets, most of the New Chemical Entities (NCEs) in the development pipeline are becoming increasingly lipophilic, thereby restricting their aqueous solubility. Therefore, a drug candidate with good permeability but limited aqueous solubility is not drug-like due to consequent lower bioavailability,2. For sThe first reported solid dispersion attempt was from Sekiguchi and Obi in 1961. Howeveri.e., Soluplus® and various grades of poloxamers in solid dispersion technology (Early attempts (1960’s decade) of formulating solid dispersions were focused mainly on using small molecular weight water-soluble crystalline carriers like urea and sugars. Many of these reported solid dispersions belonged to the category of Eutectics. They exhibited excellent thermodynamic stability due to their crystalline nature; nevertheless, the drug release from such formulations was found much slower than amorphous forms ,22. Theschnology ,22.TableSince 1961, there is prolific literature available on the topic of solid dispersion technology. This is evident by almost 5457 records in the Scopus® database on the topic “Solid Dispersion” OR “Solid Dispersions” as of March 2020. The vast number of polymeric carriers like poly vinyl pyrrolidone (PVP), poly ethylene glycol (PEG), and natural polymers like cellulose derivatives  have been actively researched. These polymeric carriers possess interesting physicochemical and thermodynamic properties, which enables their usage in formulating solid dispersions. Aplenty dissolution and stability improvement mechanisms were deduced in the literature for each polymer. Seldom, there exists a single mechanism responsible for dissolution improvement. The same is true for the stability of prepared dispersions. Such numerous and overlapping mechanisms may overwhelm the early carrier researcher and limit his/her comprehension to select an appropriate polymer. In this context, the review article attempts to compile all the existing common knowledge and literature trends of dissolution and stability improvement mechanisms of commonly used polymers in solid dispersion technology to help make an informed decision, in particular for early-career researchers in the field. This was achieved by searching through the Scopus® database. The review will cover polymeric carriers employed in binary solid dispersions; however, the discussion about small molecular weight carriers is out of the scope of this manuscript, as this space is assumed by so-called co-amorphous technology. The interested readers are advised to read many interesting reviews about co-amorphous technology ,27.There are multiple bases to classify the polymeric carriers employed in the solid dispersion technology, like the physical state, relative hydrophilicity, ionization potential, and source of the carrier ,16,22,23Furthermore, the polymeric carriers can be classified based on their relative hydrophilicity into the categories of hydrophilic, hydrophobic, and amphiphilic polymer-carriers Fig. –35. The There exist other miscellaneous grounds to classify polymeric carriers like ionization tendency and source of polymers. Unlike afore discussed classification, these categories are not functionally relevant and hence not discussed in detail in this review.i.e., safety, kinetic, and thermodynamic aspects  molecular weight ranging from 10,000 (10K) to 120,000 (120 K) Table  68,69).,69.68,69 formula . It is a formula ,47.Tableet al. (2018) attributed the extended supersaturation to the reduced crystal growth rates of Dypyridamol and Cinnarazine in the presence of PVP K-30 . Th. Thet alPVP K-30 . The parPVP K-30 –81. Howeet al., (2019) found that Rivaroxaban solid dispersion with PVP underwent amorphous-amorphous de-mixing and subsequent API recrystallization after significant moisture uptake by PVP  reported that though there is a significant risk of moisture-induced amorphous phase separation in PVP-based dispersions, such phase separation does not have a profound impact on drug dissolution of slow-crystallizer compounds  also showed that high relative humidity and an increase in the temperature did not result in the recrystallization of Zopiclone from PVP-based amorphous dispersion and Plasdone S-630 . This is amorphous, water-soluble polymer, which has traditionally been used as a binder and film-forming agent in the pharmaceutical industry . However., 230°C ,88,89. T., 230°C .Fig. 2Liet al., (2019) concluded that Copovidone accounts for the ideal release of Ledipasvir from the solid dispersion making the process polymer controlled. This generated the supersaturated state of Ledipasvir, even exceeding the amorphous solubility of the drug leading to the formation of a colloidal drug rich phase. Such drug rich phase acts as a reservoir which replenishes the absorbed drug fraction leading to the sustained supersaturation  demonstrated that Anacetrapib forms nanoparticles during the rapid dissolution of amorphous domains of solid dispersions driven by Copovidone. However, the study employed TPGS as a surfactant to prevent rapid, local drug domain aggregation events  reported that Copovidone has strong crystal nucleation and growth inhibition properties through polymer adsorption onto the drug crystals. Such mechanism demonstrated to prevent the de-supersaturation of the drug and translated into the sustained parachute in the dissolution profile analyzed the stability of MK-0364 in Copovidone- and PVP-based solid dispersions under varied temperature and humidity conditions. Unlike PVP-based solid dispersions, no degradation was observed in Copovidone-based solid dispersion. Furthermore, under stress conditions, relatively less crystallinity was observed in MK-0364-Copovidone dispersions  investigated polymeric carrier-based solid dispersions of Rivaroxaban under elevated relative humidity conditions, and found that Copovidone-based dispersion resulted in less physically stable dispersion than its HPMCAS and Soluplus®-based counterparts  contended that higher drug loading of Indomethacin, a good glass-forming agent, increases the amorphicity of the polymer and inhibiting the crystallization of PEG, leading to the higher dissolution rates  demonstrated that a significant fraction of PEG stabilized in the amorphous state post-fusion and cooling, albeit the amorphous form reverted to recrystallized state gradually upon storage reported that albeit the crystalline nature of PEG may restrict the water uptake, nevertheless, once the hydration shell is formed during the prolonged exposure to higher relative humidity, then the moisture uptake enhances exponentially  demonstrated that the deliquescent nature of PEG depends on the molecular weight of the polymer, temperature, and humidity. The moisture sorption ability of the PEG negatively correlates with its molecular weight due to the hydrophobicity of the polymer by an increase in the chain length. It positively correlates with the temperature  observed the dissolution performance of Nifedipine/PEG1500 solid dispersion system was decreased and phase-separated after 6 months of storage. It can be inferred that due to the hygroscopic nature of PEG, there is a high chance of phase separation, and it may affect the dissolution performance as well , ethylcellulose (EC), hydroxypropylcellulose (HPC), hydroxyethyl cellulose (HEC), and hydroxypropyl methylcellulose (HPMC) are commonly used ether derivatives of cellulose. Esterification of cellulose alkyl ethers involves the reaction of ω-carboxyalkanoyl groups with monobenzyl adipoyl, suberoyl, and sebacoyl chlorides, and subsequent benzyl ester hydrogenolysis, to avoid cross-linking. Cellulose derivatives are also used as bonding, coating, stabilizing and film-forming agents, plastic sheets, and emulsion stabilizers in formulations. Cellulose derivatives can be classified based on their water solubility, chemical substituents, and pH responsiveness . Th. Thi.e.,que Fig. .Fig. 4Liet al., (2016) investigated and found that the inhibitory potential of HPMC on precipitation and solution nucleation of Celecoxib. Nevertheless, the study cautioned that in the presence of seed crystals emanating from the undissolved crystalline drug, it is difficult to prevent crystal growth, particularly at high supersaturation levels  also found the additional contribution of hydrogen bonding between API and HPMC towards preventing the crystal nucleation and growth in supersaturated solutions  observed that upon dissolution of Nifedipine/HPMC solid dispersion, the Nifedipine undergoes phase separation and dissolves independent of HPMC from amorphous Nifedipine rich phase  demonstrated that dry glassy HPMC transforms into the wet rubbery state after influx of water. This creates a gel layer, and diffusion of drugs through the gel layer is considered as the rate-limiting step during the dissolution of solid dispersion showed that HPMC slowed down the de-vitrification of Irbesartan due to its anti-plasticization potential  implemented the fluorescent-based method to investigate the indomethacin-HPMC miscibility. The study reported that the miscibility is highly dependent on relative fraction of a drug within the dispersion. They observed that the dispersion samples with less than 50% drug loading were maintained in amorphous form, while the samples with drug loading higher than 50% crystallized within 15 days  attributed the stability of Nimodipine/HPMC dispersion to disruption of intramolecular hydrogen bonding within the drug molecules  put forth an alternate narrative about crystallization inhibition potential of HPMCAS. The group demonstrated that HPMCAS suppresses crystallization of Carbamazepine, Nifedipine, Mefenamic acid, and Dexamethasone through molecular level hydrophobic interactions between the drug and polymer demonstrated that during crystal growth of Rivaroxaban, small API crystals adhere to the polymer surface due to hydrogen bonding interactions, consequently inhibiting the crystal growth and translating into the improved dissolution . However, the presence of lipophilic methoxy and acetate substituents limits the solubility of the polymer at pH values above 5.0. Hence, once the solubility limit is exceeded, HPMCAS forms the colloidal phase of Nano-dimension in the intestinal pH conditions, and the negatively charged succinate group keeps the e stable . Such nae stable . Much lie stable . The drue stable , Carbamae stable , therebyd polymer. Kapourasolution .et al., (2020) evaluated the intermolecular interactions between Carbamazepine and HPMCAS by employing 1D -1H NMR spectroscopy and found that the mobility of Carbamazepine was strongly suppressed in the presence of HPMCAS due to intermolecular interactions  applied coarse grain simulation approach to study the interactions between Phenytoin and HPMCAS and demonstrated that the drug and polymer form a complex due to strong intermolecular interactions. Interestingly though, the protonated polymer chains were found more effective than deprotonated ones in inhibiting the recrystallization of the drug , polyvinyl caprolactam (57%), and polyvinyl acetate (30%). It is an amphiphilic polymer, wherein PEG provides hydrophilicity, while vinyl caprolactam and vinyl acetate domains are lipophilic within the polymer matrix. The molecular weight of Soluplus® usually ranges from 90,000 to 1,40,000 g/mol. It is an amorphous polymer with a relatively low glass transition temperature (Tg) 70°C ,150. Solet al., (2019) demonstrated that hydrogen bonding interactions between Andrographolide and Soluplus® lead to enhanced interface wetting, consequently leading to improved dissolution rate  reported that Soluplus® synergistically inhibited crystal nucleation and growth of Lacidipine, leading to the prolonged supersaturation  reported that Tadalafil release from Soluplus® dispersion retarded due to swelling of the polymer during dissolution  demonstrated that Scopoletin/Soluplus® dispersion formed drug entrapped Soluplus® micelles with an average size of 59.4 nm after dissolution  found that the solubilization capacity of Candesartan Cilexetil from Soluplus®-based dispersion reduced in biorelevant media  found that the dissolution advantage of API from Dolutegravir/Soluplus® dispersion in USP phosphate buffer pH 6.8 was found bridged in biorelevant media. The bridging in the dissolution extent was due to the improvement of dissolution of pure Dolutegravir in biorelevant media which is attributed to micellar solubilization of Dolutegravir in the bile and lecithin micelles, and much less due to the retardation of dissolution from solid dispersion  demonstrated the physical stability of ABT-102/Soluplus® dispersion as a consequence of strong hydrogen bonding between –C=O function of Soluplus® and –N-H moiety of the drug prepared Itraconazole/Soluplus® solid dispersion through hot-melt extrusion. The dispersion was found stabilized due to hydrogen bonding interactions between API and polymer. However, while attempting to formulate the tablets from prepared dispersion, Soluplus®-rich regions were formed during compression. It was indicated that the disruption of hydrogen bonding leads to phase separation . Therefore, seldom, high Tg is attributed to the physical stability of Soluplus®-based dispersions Fig. . One of  the drug. Singh eparation . Oftentii.e., spray drying and HME.Solid dispersions are usually prepared by solvent, fusion, or solvent-fusion methods . At the et al. reported the highest residual solvent content in Naproxen-PVP solid dispersion at lower inlet temperature and airflow, while phase separation was observed in response to increased inlet temperature and airflow rate


Time-resolved contrast enhanced magnetic resonance angiography, if fast enough, can not only provide arterial anatomical and pathological detail, but can also follow the first pass of contrast through visceral parenchymal tissue in order to evaluate vascular flow dynamics or perfusion.To evaluate the feasibility of a new magnetic resonance imaging (MRI) technique that would provide unified anatomic and functional evaluation of the kidneys in a single scan with a single dose of Gadolinium contrast. We hypothesize that dynamic mask-mode subtraction yields perfusion weighted images of the renal parenchyma.2) on a 1.5 T Siemens MRI scanner. A single dose of Magnevist  was administered intravenously. Images were processed with a dynamic mask-mode subtraction which has been shown to be beneficial in artery-vein separation. The raw data was reconstructed offline with a sliding window and a sliding mask subtraction technique generating sequential angiographic images at a rate of 3 frames per second. Perfusion analysis was also performed offline by two experienced diagnostic radiologists, implementing the upslope and deconvolution methods, as well as using the maximum value subtracted image. All reconstructions and analyses were performed using Matlab software. The maximum value subtraction image was compared on a pixel-by-pixel basis to the upslope and deconvolution methods using correlation and Bland-Altman plots.A new magnetic resonance angiography (MRA) pulse sequence, with a high frame rate, capable of simultaneous determination of renal arterial anatomy and pathology, and renal perfusion, was developed. Eleven healthy volunteers and one renal transplant patient were recruited to undergo MRI examination using a radial three-dimensional FLASH acquisition with sliding window view-share reconstruction  and Bland-Altman plots .This technique provides anatomic and functional evaluation of the kidneys with a single scan and single dose of contrast that would currently require multiple scans or examinations that require ionizing radiation. The maximum value subtracted image provides an accurate estimate of renal perfusion compared to the established upslope and deconvolution methods. This technique could be applied clinically to a population of potential renal transplant donors, providing arterial anatomic information for surgical planning as well as pre-transplant functional evaluation.


Humans are the only primate that walk bipedally with adducted hips, valgus knees, and swing-side pelvic drop. These characteristic frontal-plane aspects of bipedalism likely play a role in balance and energy minimization during walking. Understanding when and why these aspects of bipedalism evolved also requires an understanding of how each of these features are interrelated during walking. Here we investigated the relationship between step width, hip adduction, and pelvic list during bipedalism by altering step widths and pelvic motions in humans in ways that both mimic chimpanzee gait as well as an exaggerated human gait. Our results show that altering either step width or pelvic list to mimic those of chimpanzees affects hip adduction, but neither of these gait parameters dramatically affects the other in ways that lead to a chimpanzee-like gait. These results suggest that the evolution of valgus knees and narrow steps in humans may be decoupled from the evolution of the human-like pattern of pelvic list. While the origin of narrow steps in hominins may be linked to minimizing energetic cost of locomotion, the origin of the human-like pattern of pelvic list remains unresolved. By virtue of adducted hips and valgus knees  humans also display step widths that are approximately 2.5–3.3 times narrower than those measured for non-human apes4.As the only habitually bipedal primates, humans display a suite of unique gait attributes not seen during facultative bipedalism in other primates. Several of these traits relate to the need to maintain balance and stability in the frontal plane Fig. . NotablyAustralopithecus afarensis (KSD-VP-1/1)5. Direct morphological evidence of human-like swing-side pelvic drop however, is less clear. Many have argued based on aspects of iliac blade and/or femoral head and neck morphology that human-like pelvic and hip kinematics may have been present as early as Au. afarensis9, Ardipithecus ramidus11, or perhaps even earlier13. These arguments, however, have been vigorously debated19.When these characteristic aspects of human-like bipedalism emerged in the hominin fossil record has long been a matter of debate. Valgus knees and femoral bicondylar angles, indicative of narrow steps and some level of hip adduction during bipedalism, are the clearest morphological trait indicative of these locomotor characteristics. Human-like bicondylar angles are present as early as ~3.6 million years ago (mya) from 20, and thereby reduce metabolic energy expenditure during walking20. The selective advantage of swing-side pelvic drop is unresolved. Classically, it has been suggested that pelvic drop serves to reduce vertical oscillations of the CoM21. More recent data refutes this idea as pelvic list has a minimal effect on vertical CoM oscillations24. Alternatively, swing-side pelvic drop may be a consequence of changes to other biomechanical variables during bipedalism. Pelvic list, hip abduction, and step width are all part of body coordination in the frontal plane26. Thus the selective pressure to change one feature, for instance step width, could be expected to cause correlated change in hip abduction and pelvic list or vice versa. In order words, the origin of narrow steps in hominins may have driven changes toward the human-like pattern of pelvic list.Understanding when and why these locomotor characteristics evolved also requires knowledge about their function during bipedalism. Bicondylar angles, and the narrow steps resulting therefrom, have been shown to reduce mediolateral oscillations of the center of mass (CoM)4. If step width and pelvic motion are linked, we expected that wide steps should cause humans to adopt a pattern of chimpanzee-like swing-side pelvic elevation and hip abduction. Second, we had subjects walk with altered pelvic kinematics, including bipedal-primate-like swing-side pelvic elevation3. We expected that, if pelvic motion and step width are linked, pelvic elevation should increase step widths to a value similar to those seen in bipedal primates.Here we investigated the hypothesis that step width is linked to the unique pattern of human hip adduction and pelvic motion during bipedalism. We tested this hypothesis in two ways; first by having humans walk with altered step widths, including chimpanzee-like wide steps20). The distance between the strings was calculated based on subject’s normal step width and set to achieve a step width of approximately three times the normal width. This represents the approximate difference in relative step widths between humans and chimpanzees4. Subjects were given ample time to practice walking with the various gait conditions prior to data recording. Ten strides were analyzed for each condition for each subject.All experimental protocols were approved by the New York Institute of Technology’s Internal Review Board and were performed in accordance with all relevant guidelines and regulations and with the principles outlined in the Declaration of Helsinki. Subjects provided informed written consent prior to experiments. Three-dimensional kinematic data were recorded on ten human subjects  and compiled in Polygon and ProCalc (Vicon Motion Systems) for further analysis. For plots of pelvic list over the stride, the mean pelvic list for two chimpanzee subjects, previously published and recorded using similar methods4, was also included for reference  at 150 frames-per-second and using the full body Plug-In Gait marker set (based on the Newington/Helen Hayes gait model)Step widths were calculated by establishing a line between the positions of the left heel marker at two consecutive ipsilateral heel strikes, and subtracting the mediolateral distance of the right heel marker at contralateral heel strike from this line. Step width was reported both absolutely (in cm) and relatively, by dividing step width by standing lower limb length (from greater trochanter to the ground).30 for R 3.6.131. Two classes of LMMs were conducted based on the fixed effect of either step width condition or pelvic condition. Subject was included as a random factor with subject-specific slopes and intercepts. Each kinematic variable  in the package lme4p < 0.001; Fig. 4) as intended. Narrow steps decreased step widths from 9.2 ± 2.9 to 1.9 ± 3.4 centimeters .For pelvic conditions, both target conditions achieved the intended results during stance phase, while swing phase kinematics were largely unaltered Fig. . The pelWalking bipedally with chimpanzee-like wide steps did not cause human pelvic kinematics to drastically vary from the unique human pattern. It did reduce the normal range of motion by 22%, or about 2°, however the pattern of motion remained distinctly human-like Fig. . AlterinAu. afarensis5 though perhaps earlier. Narrow, human-like, step widths engender a clear energetic advantage compared with wide steps33 and femoral bicondylar angles in early hominins represent the presence of this energetic benefit.Taken together, our results show that experimentally altering either pelvic motions or step widths does have a strong effect on hip adduction. Yet, neither of these effects are so extreme that they cause major deviations in underlying step widths  nor the pattern of pelvic list . Therefore, the evolution of femoral bicondylar angles and valgus knees in hominins and the associated narrow step widths may not have evolved in concert with the appearance of human-like swing-side pelvic drop. These two characteristic features of human bipedalism may have evolved at different times in hominin evolution, and for different reasons. It is clear that valgus knees and thus narrow steps emerged fairly early in hominin evolution, at least by ~3.6 mya in 2, increase stride length2, or shift the center of mass towards the stance side foot4. Furthermore, during quadrupedalism, mammals utilize swing-side pelvic elevation . It is possible that the retention of similar pelvic kinematics during bipedalism in non-human primates points to some degree of neuromuscular conservatism retained from quadrupedal gait patterns (e.g.36).However, the biomechanical reason underlying the change from a primate-like pattern of pelvic list to a human-like pattern is still unresolved. Swing-side pelvic elevation during facultative bipedalism in primates may function to aid in foot clearance3. Classical descriptions of human bipedalism attribute stance-phase pelvic drop to minimizing center of mass vertical displacements, which could reduce energy requirements21. More recent studies have shown that pelvic list only has a small24, and perhaps negligible23 effect on vertical CoM displacements. Moreover, decreasing vertical CoM displacement likely increases, rather than decreases, the metabolic cost of locomotion38. Ultimately, the best way to understand when and why a human-like pattern of pelvic list evolved may be through forward dynamic modeling of fossil hominins. The only current forward dynamic model of fossil hominin bipedalism  suggests some slight degree of stance-phase hip abduction, but whether or not this was due to stance-phase pelvic elevation was not reported39.The current experimental evidence suggests that it is only within hominins that a bipedal gait entailing swing-side pelvic drop evolvedFuture forward dynamic modeling may also offer a resolution to the biggest limitation of this study, which is that we utilized human subjects walking in ways that targeted non-human primate-like gait parameters. We were fundamentally unable to experimentally alter underlying bicondylar angle and hip morphology within our subjects, and therefore could not directly investigate the effect of morphology. Thus, the earliest hominins may not have displayed the same relationship between variables that we measured here in modern humans. Forward dynamic simulations may allow for further investigation of step width, pelvic motion, and hip adduction, while simultaneously modeling non-human skeletal morphology.33. The specific reasons why humans, and likely some hominins, utilize swing-side pelvic drop remains an open question.The data herein suggest that human-like pelvic list and valgus knees (and corresponding narrow steps) represent two different biomechanical phenomenon which may not be directly coupled in human evolution. While this would not preclude the simultaneous appearance of both factors, it does open the possibility that narrow steps and valgus knees may have temporally preceded the human-like pelvic kinematics, or vice versa. It is well-established that relatively narrow, human-like steps offer an energetic advantage for bipedsSupplementary Information.Supplementary Information2.


To evaluate the effect of asphericity and blue light filter (BLF) of three different intraocular lenses (IOLs) on the visual performance, second- and third-order aberrations , and contrast sensitivity after uneventful cataract surgery.One hundred and twenty eyes of 60 patients with clinically significant cataract were randomly assigned to receive one of the three IOL types: Bioline Yellow Accurate , BioAcryl 60125 , and H65C/N . Each IOL was implanted in 40 eyes. Complete ophthalmologic examination, functional acuity contrast testing and wavefront analysis were performed 60 days postoperatively.P = 0.83). Mean defocus and coma values did not yield any statistically significant difference through the IOL groups varying from –0.784 to –0.614 μm and 0.129 to 0.198 μm . Bioline Yellow Accurate IOL presented less trefoil aberrations, 0.108 PP The mean postoperative best-corrected visual acuity (BCVA) was 0.95 Bioline Yellow IOL indicated lower contrast sensitivity under mesopic conditions when glare was applied but resulted in less trefoil aberrations after uneventful cataract surgery. No further differences were noted in postoperative visual performance among three IOL groups. Modern cataract surgery has recently evolved into a refractive procedure aimed at improving visual quality in addition to increasing the visual acuity. Therefore, it is routinely combined with the implantation of intraocular lenses (IOLs) of various materials and designs.,2,3,4,5 Ultraviolet (UV) light-filtering lenses have been the dominant IOLs used in the modern era since growing evidence indicated that UV light could result in photic retinopathy and other retinal pathologies. There has recently though been support for increasing the range of light absorption by IOLs. The rationale was that UV light-filtering IOLs cannot offer protection to the retina from phototoxic damage induced by the high-energy, short-wavelength blue light (400–480 nm), which is considered to contribute to the pathogenesis of age-related macular degeneration (AMD).,8 In response to this potential damage, blue light-filtering (BLF) IOLs were introduced in 1996 and have been thereafter widely used, especially in cataract surgery candidates with signs of AMD as a possible measure of preventing associated retinal pathology.,10,11,12 The yellow tint of BLF IOLs replicates the spectral transmission properties of the aged human crystalline lens in a much closer manner than the UV light-filtering IOLs do.Since the initial introduction of IOLs, there has been great debate over the importance of light filtration.While the possible visual benefits of BLF IOLs are still under debate, controversy has been raised whether a yellow-tinted IOL could modify the visual performance of patients, specifically regarding the postoperative best-corrected visual acuity (BCVA), contrast sensitivity, color vision, and glare. It is well-established that conventional-spherical IOLs degrade image quality by increasing the spherical higher-order aberrations (HOAs), and several authors have published studies indicating that aspheric IOLs may improve retinal image quality and mesopic contrast sensitivity at low spatial frequencies.,19,20,21In addition to BLF, another recently commercialized property of IOLs that has become increasingly popular is asphericity. Spherical aberration has a strong influence on image quality.However, the combination of BLF and asphericity in IOLs has not been clearly investigated regarding their effect on contrast sensitivity, aberrometry, and quality of vision. The purpose of this prospective randomized study was to evaluate the effect of blue light-filtering and aphericity of IOL on visual quality by comparing the three different IOL types: one aspherical IOL with BLF, one aspherical IOL without BLF, and one spherical IOL without BLF.This prospective, randomized clinical study comprised patients who underwent bilateral cataract surgery for visually significant cataract. Sixty patients were randomly assigned to receive one of the three IOL types. Group A received Bioline Yellow Accurate IOL , Group B had H65C/N IOL , and Group C had BioAcryl60125 IOL . Each IOL was implanted in 40 eyes of 20 randomly selected patients. All patients were recruited from the outpatient anterior segment unit of the Clinic Pallas Ophthalmology Department in Olten, Switzerland. The study was performed in adherence with the Declaration of Helsinki for research involving human subjects after approval was obtained from the Institutional Review Boards of Pallas Clinic.Patients with bilateral cataract with visual disturbance and no history of color vision deficiency were eligible for inclusion in the study. The exclusion criteria were ocular diseases such as corneal opacity or irregularity, astigmatism greater than 2.5 D, dry eye syndrome, inadequate visualization of the fundus, amblyopia, anisometropia, calculated IOL power less than 10.0 diopters (D) or more than 30.0 D, surgical complications, IOL tilt, previous or current use of medications known to cause color vision deficiencies, and incomplete follow-up. Also, patients with a history of uveitis and current intraocular inflammation, uncontrollable glaucoma, proliferative diabetic retinopathy, or retinal detachment were excluded from the study.One experienced surgeon (GP) performed all surgeries with standard small incision phacoemulsification and IOL implantation in the capsular bag. The time between first eye surgery and second eye surgery was six–eight days in all cases. All eyes were targeted for emmetropia. Table 1 shows the characteristics of the IOLs implanted during the study period. All patients were given combined antibiotic–steroid eye drops for four weeks postoperatively. Patients were examined before surgery and one, seven, and one to three months postoperatively. At all visits, the best corrected-distance visual acuity (BCDVA) and uncorrected distance visual acuity (UDVA) were measured. Contrast sensitivity assessment and aberrometry by means of wavefront analysis were evaluated at the baseline (preoperatively) and last follow-up visit which was performed one to three months postoperatively.Visual acuity was measured using Snellen chart under scotopic conditions (target luminance 1.5 candelas [cd]/m2). Contrast sensitivity was assessed using Functional Acuity Contrast Testing  with spectacle correction under photopic conditions  and mesopic conditions  was used for all aberrometry measurements. Zywave was used to assess and compensate for the refractive errors, and, eye fogging system was acquired before each wavefront measurement to avoid patient accommodation. Before use, the aberrometer was calibrated by an experienced Bausch & Lomb technician to ensure the accuracy. Five measurements were performed by a single experienced technician to avoid interobserver variability in the results; of these, two measurements with higher deviations from the mean were excluded and the three best measurements were averaged and used for statistical analyses. Patients were instructed to blink between measurements, and acquisition was obtained after a blink to ensure higher quality results by limiting tear film disruption. All results were exported as raw data so that individual Zernike terms could be analyzed independently. The details of the Zernike coefficients up to the third order were recorded and used for the statistical analysis. Zywave measurements were obtained without any pharmacologic mydriasis and dark adaptation. Nonetheless, all measurements were made in certain mesopic lighting conditions and it was confirmed that pupillary diameter was at least 6 mm in every case. Total, corneal, and internal components for each of the high-order aberrations were obtained and used for the analysis.t-test was used to compare the outcomes between two IOL groups. Categorical variables were compared using the Fisher's exact test. Nonparametric Kruskal–Wallis and Mann–Whitney tests were also used to examine the associations between categorical variables and continuous or ordered outcomes. A P-value of Statistical analysis was performed using the SPSS  and MedCalc statistical software . Normality was checked using the Kolmogorov–Smirnov test. Since data were not normally distributed in all cases, both parametric and nonparametric methods were used. For normally distributed data, Pearson correlation was used to evaluate the association between two continuous variables and the one-way analysis of variance (ANOVA) was applied to evaluate the influence of a qualitative factor on another continuous variable. The association of not normally distributed data was assessed using Rank correlation calculating Spearman's coefficient rho. When parametric analysis was possible, the Student's A total number of 120 eyes of 60 patients . Table 2 demonstrates the preoperative and postoperative refraction data in more details.There were no statistically significant differences among the study groups in preoperative clinical and refractive values. Preoperative total and internal components of aberrometry showed great deviations between cases, as patients with various degrees and types of cataracts were included. However, the preoperative corneal component of coma, defocus, and trefoil did not have any statistically significant difference among the three groups . Bioline Yellow Accurate presented less trefoil aberrations, 0.108 PThere was no statistically significant difference among the three IOL groups in contrast sensitivity at any spatial frequency under all three lighting conditions. Figure 1 (photopic 85 cd/m2), Figure 2 (mesopic 3 cd/m2), and Figure 3 (mesopic with glare) depict postoperative contrast sensitivity for all IOL groups. In a separate analysis, Bioline Yellow was found to have a statistically lower contrast sensitivity under glare conditions compared to the BioAcryl and PhysIOL in 12 and 3 cpd spatial frequencies, respectively , Kim et al,28,29 We compared in our study the outcome of the three different IOLs, two aspheric and one spherical and found no statistically significant difference in the second- and third-order aberrations other than trefoil aberrations that was lower in the eyes implanted with aspheric IOL with a BLF. Surprisingly, no difference was noted between the two non-tinted IOL groups, despite one of them having an aspherical design. One may hypothesize that BLF added on yellow-tinted IOLs could reduce some spherical aberrations along with the longitudinal chromatic ones; however, this theory needs to be examined by further prospective randomized studies with larger population sizes to compare aberrations between IOLs of identical design and material.On the contrary, several researchers have reported no statistically significant differences in visual acuity and contrast sensitivity between spherical IOLs and aspheric IOLs.,3 Violet and blue lights are much more important for vision in dim-light environments than in bright-light environments, providing 45% of rod-mediated aphakic scotopic sensitivity but only 7% of photopic sensitivity for an iso-illuminance light source.,2 However, the results reported in the literature are controversial regarding postoperative contrast sensitivity after the BLF-IOL implantation and do not indicate a significant decrease in the mesopic and scotopic visual function.In the past decade, several manufacturers and distributors have promoted commercially available IOLs with BLF properties. Theoretically, BLF IOLs may induce a reduction in mesopic and scotopic visual performance attributed to the Purkinje shift, where differing peaks of spectral sensitivity for scotopic and photopic vision are identified. investigated the long-term possible side effects after implantation of an IOL with a BLF. The authors found no significant differences in color perception, scotopic contrast sensitivity, or photopic contrast sensitivity between the BLF IOL and the IOL with a UV-light filter only. In another study, Greenstein et al investigated contrast sensitivity in nine patients implanted with a BLF IOL (AcrySof SN60AT) in one eye and a UV-only filtering IOL (AcrySof SA60AT) in the fellow eye. In addition, they compared the results with those obtained in nine young phakic patients and found no significant difference in hue discrimination or dark-adapted sensitivity between the two IOLs. These results were comparable to the outcome of a study by Muftuoglu et al who compared photopic and scotopic CS in eyes with an AcrySof SN60AT IOL (with BLF) and eyes with a conventional AcrySof SA60AT IOL (UV-only filtering) and reported no statistically significant differences between the two IOL types. Furthermore, Hayashi et al measured contrast visual acuity in 74 patients implanted bilaterally with either tinted IOL (HOYA YA60BB) or non-tinted IOLs (VA60BB) and reported no significant difference between the IOL groups.Kara-Junior et alIn concordance with these aforementioned studies, our results showed no statistically significant difference in contrast sensitivity between IOLs with and without BLF. In an additional analysis, we evaluated the loss of contrast sensitivity after glare was applied in mesopic conditions and found that the tinted IOL had a statistically greater loss of contrast sensitivity under glare compared to the non-tinted IOLs, but only in some spatial frequencies. Although this may be an accidental finding, it is noteworthy as most previous studies did not include contrast sensitivity measurement in mesopic conditions under glare, a situation that is rather common in real life, such as night driving, and can substantially affect the patient's quality of life after cataract surgery.A weakness worth mentioning of all studies reporting mesopic CS results after the implantation of IOLs with BLF is the fact that all have utilized measures that are a function of only central vision, where macular pigment is also acting as a BLF. Moreover, one should consider that mesopic vision is mediated, at least in part, by cones, and therefore it is less likely to be adversely influenced by the transmittance properties of such blue-blocking IOLs. Other limitations of our study include the relatively small sample size and the lack of a group with implantation of a spherical IOL with BLF; this type of IOL was not commercially available at the time the study was conducted.In summary, the present study compared three IOLs varying in terms of asphericity and BLF and showed only minimal differences in postoperative contrast sensitivity and aberrometry. All IOLs achieved comparable results in postoperative visual performance; an aspheric IOL with BLF, however, resulted in less trefoil aberrations and a greater loss of contrast sensitivity in mesopic conditions when glare was applied. Further randomized patient-centered studies are needed to evaluate the long-term results of aspheric IOL design and BLF and to investigate whether these features are desirable for the patients' quality of life.Nil.There are no conflicts of interest.