Long gone is the time when tumors were thought to be insular masses of cells, residing independently at specific sites in an organ. Now, researchers gradually realize that tumors interact with the extracellular matrix (ECM), blood vessels, connective tissues, and immune cells in their environment, which is now known as the tumor microenvironment (TME). It has been found that the interactions between tumors and their surrounds promote tumor growth, invasion, and metastasis. The dynamics and diversity of TME cause the tumors to be heterogeneous and thus pose a challenge for cancer diagnosis, drug design, and therapy. As TME is significant in enhancing tumor progression, it is vital to identify the different components in the TME such as tumor vasculature, ECM, stromal cells, and the lymphatic system. This review explores how these significant factors in the TME, supply tumors with the required growth factors and signaling molecules to proliferate, invade, and metastasize. We also examine the development of TME-targeted nanotheranostics over the recent years for cancer therapy, diagnosis, and anticancer drug delivery systems. This review further discusses the limitations and future perspective of nanoparticle based theranostics when used in combination with current imaging modalities like Optical Imaging, Magnetic Resonance Imaging (MRI) and Nuclear Imaging (Positron Emission Tomography (PET) and Single Photon Emission Computer Tomography (SPECT)). Cancer is no longer considered as just a defect in the cell cycle control as a result of factors such as external radiation exposure, hereditary gene, or combinations thereof. Recent advancements in determining the factors that influence cancer development have indicated that even the immediate environment of the cancer cells influences their proliferation . The canOne of the predominantly preferred methods to study and understand TME is molecular or metabolic imaging. Recent improvements in the imaging field have greatly impacted on the way in which TME is deciphered and has also improved the efficiency in monitoring dynamic interactions between tumor cells and their surrounding factors . The patAnother way of monitoring a probe is using nanotheranostic techniques whereby probes in nanoscale are injected into the body for diagnostics and therapeutics or sometimes for both . These pOne of the most widely used tools in nanotheranostics is use of nanoparticles (NPs). They come in various types, shapes and sizes according to target site, pathway and application. Each target has a few specific requirements that needs to be satisfied by the particular NP, so that the theranostic drug delivery system is efficient. Three main aspects need to be considered before choosing a type of NP interaction with the cells. First is the knowledge of the target cells and the biomarkers present in that target. Route of administration of the probe, that is, whether an oral administration or an intra venous (IV) injection, is the next thing to be considered. Finally, we have to consider the in vivo stability of the NP for the chosen purpose. For example, we need to consider whether the IV administered nanoprobe will be stable and can resist attack from the immune cells in the blood circulation until it is absorbed by the targeted cells. By using these guidelines to choose nanoprobes, many nanomaterials like nanodots, quantum dots, nanotubes, and nanowells have been designed.Shape of the nano-probe is also an important factor to be considered. Although spherical nanoparticles are predominantly used in cancer theranostics, several non-spherical NPs like filamentous or worm like micelles, ellipsoid, nanorod, nanodisk etc. do have a number of favorable features including high drug loading capacity, long circulation time, better cellular uptake, longer tumor inhibition time etc. . BanerjeNPs have been proven to be a useful means for both early detection and drug delivery for cancer cells. Matrix metalloproteinases (MMPs) are hydrolases which are responsible for tumor growth, invasion, and metastasis. They also play a role in formation of tumor blood vessels. However, these have emerged as probable targets for early cancer detection. For example, matrix metalloproteinase (MMP-2) responsive nanoprobe is a type of NP developed to target breast, colon, and prostate cancer cells. MMP-2 plays an important role in the early detection of cancer due to its over expression on cancer cells compared to healthy cells . AmongstMost NPs either target the receptors on the cell, the intracellular target elements or the environmental factors specific to the cellular microenvironment . Thus, tThe physical factors which greatly influence the proliferation of cancer cells are hypoxia, pH, and the interstitial pressure in the surrounding environment. These factors also serve as targets for the development of specific theranostic nanoprobes, which will be further discussed in this section.Hypoxia is a state of oxygen deprivation in the tissues. Thomlinson and Gray observed necrotic cells at a diffusion distance around 150 μm off the closest capillaries in human lung cancer cells . They alAfter four decades, it was discovered that the hypoxia-inducible factors (HIF) consisting of HIF-1 and HIF-2 activated gene transcription for the expression of the phenotypes were associated with cancer cells . The hypSeveral molecular imaging modalities targeting tumor hypoxia such as Positron Emission Tomography (PET), Single Photon Emission Computer Tomography (SPECT), Magnetic Resonance Imaging (MRI), and Optical Imaging have been developed. In the upcoming section, nanotheranostic probes targeting tumor hypoxia and their corresponding imaging modalities will be discussed.18F-fluoromisonidazole (18F-FMISO), 61Cu-diacetyl-bis(N4-methylsemicarbazone) (61Cu-ATSM), 18F-flortanidazole (18F-HX4) etc. The FMISO PET determined the severity of tumors in a case study conducted on 22 patients with glioblastoma multiforme (GBM). It was found that the signal intensity of the image correlated with the severity of hypoxia in cancer cells -imidazole-1-ethanol (TF-MISO), that undergoes reduction when it binds to the endogenous molecules of hypoxic cells. This enables it to determine the severity of hypoxia in vivo [19F MRS, the tumors must be less than 3 cm in diameter and less than 4 cm in depth. To improve the imaging modality, positive contrast enhancement is achieved using gadolinium-tetraazacyclododecane tetraacetic acid monoamide conjugate of 2-nitro-imidazole (GdDO3NI). Apart from measuring the oxygen deficiency in tumors, it was possible to image the area with low blood vessel perfusion in a xenograft rat model of prostate cancer [Magnetic Resonance Spectroscopy (MRS), has evolved with probes for determination of the level of deoxygenation using the relaxation of Hexamethyldisoloxane  of carcinomatous cells . It was 19F MRS. This method can be applied using NPs for the delivery of molecular reagents such as siRNA, cDNA, and mRNA in clinical trials [Photoacoustic imaging is a hybrid technique which works on the principle of optical absorption and ultrasonic wave propagation. Electromagnetic radiation in the form of pulsed laser is irradiated on the sample which usually contains endogenous chromophores such as haemoglobin or deoxy haemoglobin or organic dyes and nanoparticles. This causes rapid heating and thermal expansion that leads to generation of acoustic waves which are detected by ultrasonic transducers. Since scattering of ultrasound is much less compared to light in human tissues, this leads to higher resolution images. Along with this, a higher contrast is obtained due to the use of the optical imaging technique. Hence, photoacoustic imaging generates high resolution and high contrast images . An appll trials .+/H+ pumps and carbonic anhydrases (CAs) I-XIII [The balance between the extracellular and intracellular pH is achieved by a group of transporters such as Na) I-XIII . They ca) I-XIII . With th31P MRS was used for characterization of extracellular pH (pHe). Unfortunately, the sensitivity of 31P MRS is low which limits the spatial resolution [1H MRS namely (imidazol-1-yl)3-ethyoxycarbonylpropionic acid. Later, an improved spatial resolution with enhanced contrast for imaging the pHe was achieved using another 1H MRS probe namely, 2-(imidazol-1-yl) succinic acid with increased specificity and sensitivity [Firstly, MRS can monitor the chemical shift of the mild signals from inorganic phosphate to characterize the variation of the intracellular pH (pHi) using pH marker such as 3-aminopropylphosphonate. solution . Thus, tsitivity .Kato and Artemony developed a novel dual-tracer MRI-based theranostic technique to track the drugs released from nanocarriers using the SPION and gadolinium diethylenetriamine-penta-acetic acid bismethylamide (GdDTPA-BMA) . The utiOptical probes have been designed to image the acidification in tumors using a pH-activated near infrared fluorescent (NIRF) probe with an elevated sensitivity and quicker data acquisition. Optical imaging using NIRF probe enhances the sensitivity and spatial resolution because it reduces the absorbance and auto-fluorescence of endogenous molecules . One of 13CO3 with prior intravenous injection in in vivo detection [However, the available probes have not been used in clinical trials so far. Furthermore, other pH probes like the chemical Exchange Saturation Transfer (CEST) has been used to image breast cancer in a xenograft model using contrast agents of CT such as iopromide . It can etection . The calBesides the use of imaging probes to track the fatality of cancer, there is an urgent need for the design of pH-sensitive carriers for a more specific and effective targeting. Polymeric micelles are well-known pH-sensitive hydrophobic carriers used to target tumors with great specificity because they contain ionizable groups . The hyd2S in HCT116 mice cell. This probe had a local concentration of Butylated, Octylated Diphenylamine (BODPA) within the NPs which is activated at pH 7.4 [Adding to the list, photoacoustic (PA) imaging is one of the emerging imaging modalities utilized in the field of theranostics for targeting tumor microenvironment. In recent times, Ying et al. have developed peptide functionalized gold nanostars (GNS) for photoacoustic tomography. This desirable targeting efficiency has improved the effect of Photothermal Theraphy (PTT) without damaging the adjacent tissues . In the t pH 7.4 . AlthougImbalance in the angiogenic factors like vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP) and angiopoietins leads to the production of abnormal vasculature. Sometimes, the hyperpermeability allows exchange of molecules in the tumor vasculature without any maintenance of gradient. This gives rise to an abnormality in IFP .Non-invasive imaging methods like dynamic contrast enhanced magnetic resonance imaging using Gd-DTPA (gadolinium diethylene-triamine penta-acetic acid) are helpful to analyze IFP, and pre-clinical study using xenograft mouse models of many human cancers was performed. It was concluded that, the velocity with which fluid flows correlates well with the magnitude of IFP . Liu et 15O-water PET/CT helped to analyze the outcome of Imatinib on the IFP in colorectal cancer [Nanoprobes designed for targeted therapy towards this abnormal IFP would help to suppress the rapid angiogenic process. A pre-clinical study of a probe designed to target and inhibit IFP on a mouse B16 melanoma model was performed using sterically stabilized lysosomes filled with Imatinib, a drug to reduce IFP by inhibiting platelet-derived growth factor receptor (PDGF-R) β. These lipid vehicles were distributed in the tumor. A reduction of tumor size was observed under fluorescence imaging . Developl cancer . TME is a key player in the development and progression of cancer. It can be targeted to suppress the severity of cancer. The need for nanoprobes has become important in current research because of the limitations of conventional therapies for cancer treatment. The following section will give an insight into the various therapeutic and imaging probes developed so far to target various components of TME.When a tumor is developing, it needs to be supplied with new blood vessels. The vessel formation (angiogenesis) does not happen sequentially and hence it has increased permeability and leaky membranes. Because of this malformed vasculature, it becomes difficult to have drug delivery. Hence, these days nano-therapeutic applications have emerged to tackle the drug delivery inefficiency . The nano-probe contains photofrin or photofrin and iron oxide NPs. Photofrin is a photodynamic agent which acts like a sensitizer that is taken up by the tumor and upon photo irradiation releases singlet oxygen . These N5β1 integrin and these vessels were visualized with 3D spatial resolution of the MR signals. They did another experiment with a theranostic nanoprobe targeted towards α5β1 (ανβ3) integrin along with a fumagillin drug. Results showed a significant reduction in tumor size compared to ανβ3 fumigillin which did not show much reduction [Schmider et al. later developed a probe to target neo-vasculature expressing αeduction . The adv2 in short [Increasing the specificity of a nanoprobe by adding a peptide can be seen from the research conducted by Grange et al. where neural cell adhesion molecule (NCAM) expressed on tumor endothelial cells or Kaposi Cells can be particularly targeted with a specific NCAM peptide conjugated with a nanoprobe. A comparison was made between a PEG liposome with no specific NCAM target and C3d liposome which had a specific peptide moiety. They concluded that C3d liposome was more efficient in delivering the doxorubicin drug into the tumor cell which resulted in a reduction in tumor size and also with less toxicity of the drug and this was observed on SCID male mice at a pre-clinical level. On the other hand, the PEG liposome was accumulated extracellularly. The doxorubicin drug release was observed through MRI by tagging the liposome with a gadolinium—1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)—monoamide or Gd DOTAMA (C18)in short . Gd DOTASimilarly, another study with Gd DOTAMA as a contrast agent in targeting tumor vasculature where a glucocorticoid drug prednisolone phosphate (PLP) was incorporated inside long circulating stealth liposomes (LCL) and this combination of LCL-PLP could permeate through the tumor vasculature easily compared to normal endothelial cells to achieve a targeted therapeutic effect. Two kinds of drug release study were conducted: one with only LCL-PLP and other with an MRI imaging agent Gd-PLP-LCL. The latter helped to understand the distribution of the drug delivered in vivo on B16.F10 melanoma cells injected into Male C57BI6 mice and concluded that this had a good accuracy in analyzing the therapeutic response at a pre-clinical level .124I-VG67e is a monoclonal antibody that binds to VEGF-A that can be imaged through PET and this was done on HT1080-26.6-bearing mice at a pre-clinical stage [64Cu-DOTA-VEGF121 are performed on mice bearing U87MG human glioblastomas where DOTA acts as a chelator [VEGF regulates tumor angiogenesis and is over-expressed in a tumor tissue. The usage of many other imaging modalities such as PET and SPECT in targeting the VEGF is evident from the following examples which were performed at a pre-clinical stage. al stage . To targchelator . 124-L19chelator   which used multifunctional mesoporous silica nanoparticles and 800CW which acts as the NIRF dye. This experiment was performed on the 4T1 murine breast tumor model and resulted in a 2-fold enhancement of tumor accumulation compared to targeting without TRC105(Fab) [124I MORAb-004 has shown to specifically target hTEM1 which is a tumor vascular marker and this was expressed by engineering immortalized murine endothelial cells (MS1) to express luciferase, enabling optical imaging in nude mice [Dual Modality imaging using both PET/NIRF was carried out using 105(Fab) . In vivoude mice .A multifunctional theranostic approach involving photoacoustic imaging was performed to image tumor vasculature associated with mouse bearing the 4T1 breast tumor model using (PEG) coated copper(II) sulfide NP. These NPs act as a contrast agent and also help to destroy the tumor by photothermolysis .89Zr bevacizumab, a tracer which helps to target VEGF-A in breast cancer patients. This study showed that the standardized uptake value of the tracer was higher (1.85 ± 1.22) in the tumor compared to normal tissues (0.59 ± 0.37) [123I-L19(scFv)2 targeted towards fibronectin of five male patients with head and neck cancer could be imaged by SPECT/CT modality [Clinical PET Imaging studies have also been done for example by using  ± 0.37) . 123I-L1modality .The lymphatic system plays an important role in both the immune system and the spreading of cancerous cells from the primary site to the secondary site in different organs, i.e., cancer metastasis. In many diseases including cancer, there is an excess growth of lymphatic vessels called lymphangiogenesis . LymphatLymphatics-homing peptide-1 (LyP-1) is a 9-amino-acid cyclic peptide that binds to its receptor (p32/gC1qR) in some type of tumor cells especially, in highly malignant breast tumor cells MDA-MB-231 and MDA-MB-435S . Luo et Changes in the rigidity of the ECM can be a trigger for tumor development. Therefore, when nanocarriers are targeted to this lineage of TME to stop these changes associated with ECM, the growth of the tumor can be suppressed. Hyaluronan (HA) metabolism is a predominant feature for breast cancer progression. During tumor progression, the amount of hyaluronidases is at increased levels. Peer and Margalit discussed HA tagged to gold NPs along with a fluorophore which helps to detect the cancerous tissue due to increased levels of hyaluronidases .d,l-lactide-co-glycolide)-block-PEG copolymer. This nanoprobe system was successfully implemented in vitro and in vivo and resulted in significant tumor suppression. The research team conducted a fluorescence spectroscopic analysis using Coumarin-6 dye that helped to analyze the nanoprobe accumulation inside the tumor [A nanotherapeutic probe targeting ECM was designed by attaching an antibody of lysyl oxidase to poly and NIR imaging .Matrix metalloproteinase (MMP-2) is usually over expressed in certain tumors. A probe that follows FRET principle was designed by attaching a low molecular weight heparin associated with quantum dot and this moiety was attached to the low molecular weight protamine which contains the MMP-2 substrate cleavable by MMP-2. This substrate acts as a spacer and also contains the fluorophore. MMP-2 overexpressed in the tumor tissue cleaves this spacer leaving the fluorophore free. To make this probe permeable through the blood brain barrier (BBB), a transferrin receptor T7 peptide was used for targeting the tumor correctly .A recent advancement in this field was the use of an external alternate magnetic field which makes the liposomes covered by heat sensitive magnetic NPs undergo hysteresis and this localized increase in temperature triggers the release of protease substrates embedded inside the liposome. MMPs excessively present on tumor tissues degrade the substrate and these short peptides are detected by diagnostic tests like enzyme-linked immunosorbent assay (ELISA) .9) and fluorescence revealed the accumulation of the triple helical structure in the tumor tissue [Collagen undergoes many changes in the TME. These structural proteins become denatured due to action by MMPs and several other proteases. Collagen mimetic peptides (CMP) are photo-triggered and have a tendency to form a triple helix with the denatured collagen. The CMPs were tagged with Near Infrared Fluorophore (NIRF) (IR-Ahx-(GPO)r tissue .64Cu-DOTA-antiperiostin-F(ab′)2 was developed by Heidari et al. which showed better binding affinity towards periostin expressed in esophageal squamous cell carcinoma mouse models in comparison with 18F FDG [An extracellular matrix protein named periostin has functions of cell adhesion and motility and is an important component of TME. This protein was imaged using PET/CT modality. A PET tracer  18F FDG .2-terminal [Tumor-associated stromal cells (TASCs) such as cancer-associated fibroblasts (CAFs) and tumor-associated macrophages (TAMs) have been found to play a key role in promoting cancer progression and metastasis ,89,90. Cterminal ,97. Hencterminal ,98,99. Jterminal . Ji et aterminal .Furthermore, cysteine cathepsins especially cathepsin B (CtsB) are excessively expressed in TME. Normally, CtsB interacts with lysosomes to perform functions such as autophagy and immune responses . The excBesides the role of TAMs in tumor progression, invasion and metastasis, they can also be a potent drug target. The over-expression of mannose receptor on the TAMs was used for targeting TAM positive carcinomas . Zhu et Efficiency of drug delivery is the key element in improving the present status for both diagnosis and therapy of cancer. Multifunctional upconversion NPs (UCNPs) are used for effective cancer therapy. These NPs are used in nuclear targeting therapy where they deliver the anti-cancer drug into the cancer cell nucleus and also monitor the process using imaging modalities such as MRI, with the help of trans activator transcription (TAT) peptide. This is utilized as a cell penetrating peptide (CPP) derived from human immunodeficiency virus 1 (HIV-1) and thus, helps efficiently in the uptake of the NP . The mulNanoparticles that are used particularly for targeting tumors have a drug releasing system which releases the drug when exposed to a particular pH. This system is based on the fact that the pH level of the tumor cells is much lower compared to that of normal cells. Griset et al. developed a paclitaxel-loaded polymeric nanoparticle that targets tumor cells and releases the drug when exposed to mild acidic condition. It was demonstrated that even at a low dose, the drug delivery of this nanoparticle was found to be highly efficient . Further2+ released in this process enables imaging using MRI thus providing a theranostic approach [Recent advancements of this pH based drug delivery system include pH responsive hyaluronic acid functionalized manganese dioxide nanosheets. These nanosheets can absorb cisplatin, a therapeutic drug which is delivered at the tumor target on acquisition of lower pH and reduced glutathione. The Mnapproach . Aggregaapproach .The mesoporous silica nanoparticle (MSN) is another type of efficient drug delivery system that acts as a novel vehicle for the transport of anti-cancer drugs, which provides a large structural central cavity for drug encapsulation which in turn improves the rate of accumulation of the drug . This siwt/wt) ratio of 0.46 showed effective drug release [Liposome mediated nanoparticle drug delivery is non-toxic and non-immunogenic. It is mostly encapsulated in order to protect it from degradation. Liposome formulated nanostructures are the most widely used method for treatment of various types of cancer . The use release . Doxorub release . Chang e release . Ze Lie  release . Another release . dl-Lactic acid), poly(dl-glycolic acid) which are used for polymer based nanotheranostics. Along with this, an imaging agent is also incorporated such as an MRI contrast agent, radionuclide, fluorophore etc. This can further be enhanced by tagging a targeting ligand on the NP that specifically targets the tumor site [dl-lactide)). This new copolymer nano-particle (377 nm size) carries and effectively (approx. 71.6%) releases the BSA  which was also confirmed using H NMR, SDS-PAGE and circular dichroism spectra [Polymeric based nanospheres, nanocapsules, and nanoparticles are made up of a biodegradable and biocompatible polymeric compound which encapsulates the drug inside the cavity. In the field of cancer theranostics, PNPs (Polymeric Nano Particles) serve as a promising vehicle for drug delivery because they carry the drug effectively to the specific organ. The nanometer size of the PNPs facilitate effective and easy transportation through the cell membrane  The polymor site . A studymor site . Incorpomor site . Another spectra .One of the major challenges faced in nanotheranostics is the stability of the functionalization between the nanoprobe and the particular drug. Though this challenge can be overcome by modifying the surface of the nanoprobe, it gives rise to another problem—the control over the size of the nanostructure. This is because some applications, such as the probes for brain or liver cancers require a much smaller nanocarrier in order to efficiently penetrate the cells . The effNanotoxicity is an important factor in appraising the safety of the probe’s use in the future. It also proves to be a major concern in therapeutic techniques because negative interaction between NP and the drug might produce free radicals. This may in turn target the healthy neighboring cells instead of the cancer cells . This phIn recent years, attaching immunosuppressors to the nanoprobes seems to have been highly welcomed as it eliminates the risk of triggering the immune system thus, possibly leading to autophagy . AutophaRadioactive imaging probes have an attribute called half-life, which denotes the amount of time they remain active. Usually, the probes have low half-time denoting their poor-viability and thus demanding a faster imaging process . Hence, It has been a milestone in cancer research to observe the paradigm shift of cancer cells from an isolated mass of cells displaying uncontrolled division to one that undergoes complicated communication with the TME. Since then, many studies have expanded our knowledge on the different components of TME and their roles in tumor development. The rapid advancements in nanotechnology allow nanotheranostics to be implemented in the combat against cancer.The abnormal pH, IFP, and oxygen concentration in the TME serve as a suitable target for nanoprobes for cancer detection. Moreover, the difference in the environment of healthy cells and TME allows researchers to design drug carriers that release anticancer drugs in the TME. However, many of these drug carriers and molecular probes are still confined to the research domain and have yet to be transferred to a clinical setting. This review discussed the methods with which different elements in TME are targeted using nanotheranostics for detection, therapy and drug design in cancer, for example, the tumor-associated stromal cells (TASCs) that consist of tumor-associated macrophages and cancer-associated fibroblast. The overexpression of genes in TASCs poses a potential nanotheranostic target. Abnormally high level of lymphangiogenesis with overexpression of antigens is also another target for nanotheranostics. Apart from that, scientists have developed different nanoprobes to detect angiogenesis and the proteins attached to the newly made blood vessels using modalities like PET and MR imaging. Besides that, the signaling molecules and growth factors supplied by the ECM for tumor progression have provided potent cancer-specific target nanoprobe development. Lastly, we highlighted different drug delivery systems designed based on pH, enzyme, liposome, and topology. The shortcomings of these drug delivery systems, such as cytotoxicity and instability of functionalization between the drug and the NPs were discussed.In summary, the rapid knowledge advancement in tumor microenvironment and its role in tumor development proffer researchers a better foundation to conduct cancer research. The development of TME-specific nanotheranostics will provide a faster and more accurate cancer diagnosis and therapy in the future, if it is translated in the clinical setting.As the role of TME in cancer progression becomes established, the future direction is to develop TME-targeted therapies and drug delivery systems to halt or slow down cancer malignancy. Although numerous studies have highlighted TME-specific cancer treatment, there are challenges that are still unresolved. These include the dynamic changes in TME, heterogeneity of TME, and the complex nature of different molecular factors in TME. In addition, many studies discuss the interplay between a TME element and cancer growth. It is important to look into the network of all TME elements to elucidate the crosstalk between cancer growth and TME as a whole. Another area of interest is to establish the TME network based on specific types of cancer. For example, the TME of hepatocellular carcinoma and prostate cancer is different. Based on this difference, more specific and effective diagnostic or therapeutic methods can be developed.In the near future, once the TME of a particular type of cancer has been established, personalized and targeted treatments can be tailored to replace non-specific chemotherapy or radiotherapy. Besides, understanding the cancer-TME interplay will help us look into therapies to reduce the recurrence of cancer. By doing so, there will be less societal and governmental burden imposed. Cancer progression can be controlled and monitored using TME-targeted therapies and diagnostics.


Beta-blockers are commonly prescribed for patients with cardiovascular disease. Providers have been wary of treating chronic obstructive pulmonary disease (COPD) patients with beta-blockers due to concern for bronchospasm, but retrospective studies have shown that cardio-selective beta-blockers are safe in COPD and possibly beneficial. However, these benefits may reflect symptom improvements due to the cardiac effects of the medication. The purpose of this study is to evaluate associations between beta-blocker use and both exacerbation rates and longitudinal measures of lung function in two well-characterized COPD cohorts.We retrospectively analyzed 1219 participants with over 180 days of follow up from the STATCOPE trial, which excluded most cardiac comorbidities, and from the placebo arm of the MACRO trial. Primary endpoints were exacerbation rates per person-year and change in spirometry over time in association with beta blocker use.Overall 13.9% (170/1219) of participants reported taking beta-blockers at enrollment. We found no statistically significant differences in exacerbation rates with respect to beta-blocker use regardless of the prevalence of cardiac comorbidities. In the MACRO cohort, patients taking beta-blockers had an exacerbation rate of 1.72/person-year versus a rate of 1.71/person-year in patients not taking beta-blockers. In the STATCOPE cohort, patients taking beta-blockers had an exacerbation rate of 1.14/person-year. Patients without beta-blockers had an exacerbation rate of 1.34/person-year. We found no detrimental effect of beta blockers with respect to change in lung function over time.We found no evidence that beta-blocker use was unsafe or associated with worse pulmonary outcomes in study participants with moderate to severe COPD. Chronic Obstructive Pulmonary Disease (COPD) is a progressive and debilitating disease that burdens the healthcare system with frequent office visits and hospitalizations. In recent years, studies have examined both traditional treatments for COPD  , 2 as we1, respiratory symptoms, or responsiveness to beta-agonist inhaled therapy. Sub-group analysis extended this to patients with severe obstruction as well as those with bronchodilator reversibility demonstrated on spirometry [Beta-blockers are regularly prescribed in patients with cardiovascular disease, a common comorbidity in patients with COPD. Providers have historically been reluctant to treat COPD patients with beta-blockers due to a concern for precipitating bronchospasm. These concerns have been expressed in review articles and practice guidelines that cited case studies of acute bronchospasm in patients treated with non-selective beta blockers , 6. Cardirometry .Multiple retrospective studies have suggested that beta-blockers may reduce the mortality of patients with COPD as well as the risk of AECOPD , 10. MorThe COPD Clinical Research Network has conducted several randomized, placebo-controlled prospective trials in study participants with COPD assessing rates of AECOPD.The STATCOPE study showed no benefit attributable to daily simvastatin, whereas theMACRO trial demonstrated reduced rates of AECOPD with azithromycin treatment , 4. The 1/FVC) < 70% and being at high risk for experiencing an AECOPD as a result of using supplemental oxygen, or being treated with oral glucocorticoids, antibiotics or being hospitalized in the previous year for an AECOPD. Patients also had no exacerbation within 4 weeks of enrollment 4. Patients in the azithromycin treatment arm of the MACRO study were excluded due to the significant treatment effect of azithromycin on reducing exacerbation rate. The STATCOPE trial had similar inclusion criteria, but excluded patients who were taking a statin, had contraindication to the use of statins or who were found to have an indication for statin therapy [We performed a retrospective review of 1219 study participants who had at least 180 days of follow up from the STATCOPE trial or the placebo arm of the MACRO trial. Entry criteria for the MACRO trial included having a ratio of forced expiratory volume in one second to forced vital capacity  reported taking beta-blockers at enrollment. In the STATCOPE cohort 63 of 688 (9.2%) participants were taking beta-blockers along with 107 of 531 (20.2%) in the MACRO placebo arm. The majority of participants in the study had severe or very severe airflow obstruction classified as GOLD stage III or IV. Those taking beta-blockers tended to have a slightly higher FEVPatients in the STATCOPE cohort taking beta-blockers had the lowest rate of AECOPD , followed by patients in STATCOPE not taking beta-blockers . In the MACRO cohort the rate of AECOPD was higher when compared with STATCOPE, but nearly identical with respect to beta-blocker usage within the cohort  who had follow-up spirometry. The data shows that the presence of beta-blocker medications had no clinically or statistically significant effect on the change in airflow limitation in this cohort.1 over time and no statistically significant difference in the rate of acute exacerbation of COPD in an at-risk population, regardless of the presence of cardiac comorbidity.We found no harmful effect of beta-blockers with respect to change in FEVOther retrospective studies have shown beta-blockers to be beneficial in patients with COPD but those observations may represent cardiovascular benefits of beta-blockers rather than pulmonary specific improvements in COPD symptoms or severity.One recent Swedish nationwide observational study concluded that patients with COPD discharged on a beta-blocker after an MI had a lower all-cause mortality compared with those not discharged on a beta-blocker . HoweverProposed mechanisms by which beta-blockers may have an effect on the COPD process and potentially decrease exacerbation frequency include reduction of ischemic burden and tempering the sympathetic nervous system. COPD has been associated with systemic inflammation, and it has been proposed that the negative effects of neurohumoral activation  can contribute to the cycle of COPD exacerbation and pathophysiology. Beta blockade theoretically could have an impact on neuro-humoral activation and COPD.  In additOther observational studies have shown conflicting results. For instance, Bhatt and colleagues evaluated a cohort of over 3000 patients from the COPDGene cohort and found that patients taking beta-blockers had a decreased incidence on AECOPD. This association held true for severe exacerbations as well as mild to moderate exacerbations . Short eAdditionally, the expected outcome of beta-blockers having a greater effect on the cohort with cardiac comorbidity (MACRO) may not have been present due to baseline differences in the two groups. The MACRO cohort on beta-blockers trended to be an older population, with an increased number of pack-years which may be reflective of a sicker patient population when compared with the STATCOPE cohort on beta-blockers Table .The relatively low number of patients taking beta-blockers at enrollment in these studies limited our ability to show a statistically significant difference between the groups with respect to demographic and baseline spirometry Table . MedicatWe found no evidence that beta-blockers significantly affected the rate of AECOPD regardless of whether the patients did or did not report cardiac comorbidities. We did find that beta-blockers had no detrimental effect on lung function in a population that was at increased risk for AECOPD. Because previous observational studies have reported conflicting data, the question will require a large, prospective and randomized trial to determine the benefits of beta-blocker therapy in COPD patients.


Aims. This study evaluated the effects of L-PRF presence and implant surface texture on bone healing around immediately placed implants. Methods. The first mandibular molars of 8 beagle dogs were bilaterally extracted, and implants  were placed in the mesial or distal extraction sockets in an interpolated fashion per animal. Two implant surfaces were distributed per sockets: (1) dual acid-etched  and (2) micrometer/nanometer scale textured (Ossean™ surface). L-PRF  was placed in a split-mouth design to fill the macrogap between implant and socket walls on one side of the mandible. The contralateral side received implants without L-PRF. A mixed-model ANOVA (at α = 0.05) evaluated the effect of implant surface, presence of L-PRF, and socket position , individually or in combination on bone area fraction occupancy (BAFO). Results. BAFO values were significantly higher for the Ossean relative to the DAE surface on the larger mesial socket. The presence of L-PRF resulted in higher BAFO. The Ossean surface and L-PRF presence resulted in significantly higher BAFO. Conclusion. L-PRF and the micro-/nanometer scale textured surface resulted in increased bone formation around immediately placed implants. The phenomenon of osseointegration has allowed multiple specialties in dentistry and medicine to better rehabilitate form and function of a multitude of clinical scenarios that include tissue loss due to pathology or trauma. In dentistry, one of the most commonly encountered clinical situations is progressive alveolar bone loss that occurs after tooth extraction. It has been well characterized that this progressive loss may substantially reduce the ridge dimensions prior to implant placement resulting in less favorable clinical conditions both functionally and aesthetically, before an implant is placed on the healed ridge –3.In an attempt to improve the distribution and quantity of bone present around implants, along with decreasing treatment time for final prosthetic rehabilitation, minimally traumatic extraction techniques followed by immediate implant placement have been widely utilized and investigated –6. GivenRegardless of the surgical technique utilized, it is unequivocal that the extraction socket cervical dimensions are larger than the implant diameter and that a gap is going to be present between the implant and extraction socket walls . Hence, While this gap may be bridged with new bone forming within the extraction socket, several reports indicate that grafting procedures may be indicated to avoid soft tissue downgrowth between implant and the socket walls –11. The Autologous tissue engineering approaches such as platelet-rich plasma (PRP), fibrin glues, and platelet-rich fibrin have been utilized in both orthopedic and oral and maxillofacial surgery in an attempt to hasten both bone and soft tissue healing , 14. A rThis study utilized screw root-form Blossom implants  of 3.75 mm in diameter and 13 mm in length. Two different implant surfaces were utilized, namely, a dual acid-etched (DAE) surface and the commercially available Ossean surface . Both surfaces have previously been characterized, where the DAE surface presents textured micrometer scale and smooth nanometer scale whereas the Ossean surface presents textured nanometer scale within the textured micrometer scale . A totalFollowing approval of the bioethics committee for animal experimentation at the Ecole Nationale Veterinaire D'Alfort, France, 8 beagle dogs (approximately 2-year-old dogs) were acquired for the study and allowed to acclimate for 2 weeks prior to surgery. All surgical procedures were performed under general anesthesia. The preanesthetic procedure comprised an intramuscular (IM) administration of acepromazine maleate (0.2 mg/kg), diazepam (0.5 mg/kg), and fentanil (4 mg/kg). Anesthetic induction was then achieved through ketamine (3 mg/kg), and general anesthesia was then obtained and maintained by 1 to 2% halothane.Bilateral extractions of the mandibular first molars were performed. Mucoperiosteal flaps were elevated and teeth were sectioned in the buccolingual direction to allow nontraumatic individual root extraction by means of root elevators and forceps. One implant of each surface was placed in a split-mouth design at the mesial or distal extraction sockets in an interpolated fashion per animal so that the number of DAE and Ossean implant surfaces was equally distributed per mesial and distal sockets. On one side of the mandible, the implants were placed to the level of the buccal bone plate and soft tissue closure was achieved through standard suture procedures. On the contralateral side, L-PRF was prepared from each individual animal, by drawing venous blood using proprietary tubes and spinning them through a proprietary centrifuge at 2700 rpm for 12 minutes, or 400 RCF  . Both meμm by means of a series of diamond blade sectioning and SiC abrasive papers  in a grinding/polishing machine  under water irrigation [At necropsy, the mandibles were retrieved by sharp dissection. The implants in bone were then separated from the mandible allowing blocks containing the implants in the mesial and distal sockets. The bone blocks were kept in 10% buffered formalin solution for 24 h and gradually dehydrated in a series of alcohol solutions ranging from 70 to 100% ethanol. Following dehydration, the samples were embedded in a methacrylate-based resin  according to the manufacturer's instructions. The sections, performed in a buccal-lingual direction, were then reduced to a final thickness of ~30 rigation . The secα = 0.05). The statistical unit utilized for evaluation was the number of animals. The independent variables considered  were implant surface, socket position , and the presence of L-PRF. The dependent variable considered was BAFO.Statistical analysis was performed by mixed model ANOVA , no complications were observed during animal surgical procedures or follow-up assessments, including postoperative infection or any other clinical concern.General histologic analysis depicted the implant placed in the center of the socket at a distance from the buccal plate in proximity with the inferior alveolar nerve . The gapp = 0.47) were observed between groups despite lower mean values for dependent variables observed for the mesial socket relative to the distal socket (p = 0.11) were observed despite higher mean values for both dependent variables observed for the Ossean surface relative to the DAE surface  than the DAE surface in the mesial socket. A significant difference was observed between the mesial socket without L-PRF and the distal socket with L-PRF (p = 0.02). Finally, the combination of the Ossean surface and L-PRF presence resulted in significantly higher BAFO level relative to its no L-PRF counterparts and DAE surfaces with and without L-PRF (p = 0.012) .In general, the histomorphologic results obtained from implants placed in fresh extraction sockets are in direct agreement with previous studies , 30, 32,When the presence or absence of L-PRF was evaluated as a function of socket position, substantially higher degrees of BAFO were observed for sockets filled with L-PRF. The substantial increase in BAFO observed for both sockets due to the presence of L-PRF is likely accounted by improved cell migration through the stable L-PRF scaffold present between implant and socket walls.Our histometric results for BAFO further highlight the biologic and scaffolding properties of L-PRF when placed in combination with the Ossean implant surface. This result unequivocally demonstrates the synergistic effect that exists between a micrometer and nanometer length scale and the L-PRF structure that allows significantly higher amounts (~90%) of bone formation between implant threads than the combination of the DAE surface and L-PRF. A potential physical explanation for the BAFO discrepancy observed between DAE and Ossean surfaces placed with L-PRF lies in their roughness pattern differences , where lGeneral histomorphologic observations for the implants placed along with L-PRF in fresh extraction sockets differed from those where no L-PRF was utilized primarily due to the lack of soft tissue migration through the gap formed between implant and socket wall, suggesting L-PRF's efficiency as a barrier during healing. The osteogenic potential and adequacy of L-PRF substituting the blood clot during socket healing were confirmed since bone growth occurred from the socket walls towards the implant leading to substantial bone formation around implants irrespective of implant surface group.It should be acknowledged that this study is an in vivo preclinical model on beagle dogs, suggesting that the synergistic effect of leukocyte platelet-rich fibrin (L-PRF) and micrometer/nanometer surface texturing on bone healing around immediately placed implants should be evaluated in well-designed clinical trials in humans. Further studies are therefore needed to confirm the evidence emerging from the present research.The postulated hypothesis that the combination of L-PRF with an implant surface presenting micrometer/nanometer scale texturing would result in higher degrees of osseointegration of immediately placed implants was accepted. Further studies are warranted to shed light on possible physical and molecular mechanisms that result in the substantial increase in the amount of bone in proximity with Ossean surface implant placed along with L-PRF.


This data article contains analyzed data for the article “Continuous Production of Stretchable Conductive Multifilaments in Kilometer Scale Enables Facile Knitting of Wearable Strain Sensing Textiles”  [1]. Details of wet-spinning conditions to achieve scaled-up production of stretchable and conducting polyurethane/poly:poly(styrenesulfonate) (PU/PEDOT:PSS) multifilaments are provided. The stress-strain curves for tensile and stretch-relaxation tests on the multifilament and different knitted textile structures  are presented. It is shown that the PU/PEDOT:PSS multifilaments can also be knitted into fabrics that when worn on various body parts, such as knee, elbow, and finger, can monitor their various movements. Specifications tableValue of the data•The data presented here shows how we can solve some of the challenges in transitioning from lab-scale wet-spinning of functional monofilaments to scaled-up fabrication of multifilaments which are suitable for textile processing.•The data in this article provide details of the experimental conditions to achieve stretchable and conductive PU/PEDOT:PSS multifilaments.•The tensile and stretch-relaxation data of multifilament and textile prototypes may be used to elucidate the effects of structure on strain sensing behavior of the knitted textiles.•The data explain in details how to use the strain sensing textile prototypes for wireless monitoring of movements of various body parts.1This Data in Brief article describes the fabrication of stretchable and conductive PU/PEDOT:PSS multifilaments via a scaled-up wet-spinning process and the mechanical and strain sensing properties of the multifilaments when used individually and within various knitted textile structures as shown in Ref. −1) in several solvents including dimethyl sulfoxide (DMSO), dimethyl formamide (DMF), and DMSO/DMF (50/50 V/V). We used a 20-hole spinneret  for spinning PU multifilaments. We observed that when we used water as the coagulating solvent, sever filament sticking occurred and there was no clear boundary between the filaments. We then replaced water in the coagulation bath with isopropanol (IPA). While PU multifilaments were achieved from the DMF solution, wet-spinning of PU solutions in DMSO and DMSO/DMF using IPA in the coagulation bath led to droplets or short fiber fabrication. The low coagulation rate in these solvent/non-solvent systems could be the reason for the unsuccessful fiber spinning. We found that it was possible to achieve spinnability form all solutions using an IPA/water (50/50 V/V) coagulation bath with the presence of noticeable filament sticking from DMSO and DMSO/DMF formulations. Interestingly, further increasing the IPA content to IPA/water 90/10 V/V resulted in PU multifilaments with distinct separation of filaments from all formulations  in DMSO as it resulted in frequent breakage of the filaments within the coagulation bath. By slightly decreasing the IPA content of the coagulation bath to IPA/water 80/20 V/V, we achieved continuous spinnability of the PU/PEDOT:PSS formulation in DMSO, albeit with a noticeable filament sticking c. By cha−3, and elastic recovery of ~70% at the applied strain of 100%. The excellent mechanical properties of the PU/PEDOT:PSS multifilaments enabled their knitting into various structures such as plain-knit, co-knit, co-knit-alternate, co-knit with conductive stitch, and plain with non-conductive stitch , we were able to monitor the sensing response of the knitted prototypes on a personal computer using the Bluetooth® connection. TM with the knitted prototypes. The knitted sensors responded to diverse bending deformations of knee, elbow, and finger including very fast actions such as kicking . In all cases, the voltage signal of the sensor decreased when the limb was bent and then increased upon straightening similar to the wired connection , and testing procedures  used to generate the data reported here, are described in Ref. TM sensor. The AnEx board allows the ShimmerTM platform to connect to a third party analog devices (the textile strain sensor in this case) TM  board of the Shimmercase) TM  and the


Nonparametric Spearman correlation analysis was used to study associations between ECog and z-transformed total CERAD-NAB scores in both groups. The results revealed a significant correlation between total ECog and CERAD-NAB scores , and category verbal fluency test with the executive domain of the ECog scale . The CERAD-NAB scores were also significantly correlated with AD-related hypometabolism . The memory domain of the ECog scale was significantly correlated with FDG uptake in the angular gyrus and posterior cingulum gyrus . In conclusion, both total and memory domain ECog scores were correlated with the neuropsychological tests and neuroimaging biomarkers.We applied the Everyday Cognition (ECog) scale in normal aging adults and patients with Alzheimer’s disease (AD) to investigate associations between neuropsychological tests and neuroimaging markers. A total of 160 normal aging adults and 40 patients with the early stage of probable AD were included. Neuropsychological performance was assessed using the Consortium to Establish a Registry for Alzheimer’s Disease Neuropsychological Assessment Battery (CERAD-NAB).  The most prominent cognitive deficit in early stage AD is episodic memory loss, which can be detected by performance-based neuropsychological tests3. Functional impairments or a decline from previous levels in performance are also included in the core diagnostic criteria1, and can be assessed by informant-rated instruments such as the Clinical Dementia Rating (CDR) scale4, basic activities of daily living (ADLs) or “instrumental” activities of daily living (IADL)6. The early detection of this functional decline has been shown to have prognostic value in predicting future disease progression or conversion to dementia8. In addition, the functional decline in AD has been linked to objective biomarkers such as cortical gray matter loss9, small hippocampus volume10, and increased amyloid burden on positron emission tomography (PET) scans11. However, the relationship between functional activity and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) has yet to be fully elucidated. The Everyday Cognition (ECog) scale is a validated informant-rated questionnaire that includes one global factor and six domain-specific factors12. The psychometric properties in the ECog scale focus on mild problems in everyday function and cognition that may complement and expand those included in older informant-rated instruments.Of the dementia syndromes, Alzheimer’s disease (AD) is the most common degenerative form. Its diagnosis is based on core clinical features and supported by neuroimaging studies or cerebrospinal fluid markers14. It has been translated into several languages including German, Finnish, and simplified and traditional Chinese for use in Hong Kong, China and Taiwan20. The total score of the CERAD-NAB has been shown to have good accuracy in diagnosing the early stage of AD in several multi-center studies22. Moreover, in a brain-behavior study, the CERAD-NAB was significantly associated with regional cerebral metabolism as measured by FDG-PET23. However, associations between the ECog scale and neuropsychological measurements have not been extensively studied. Previous studies have shown that the ECog scale is significantly correlated with global cognition and individual domains of cognitive tests24. Furthermore, other functional tools such as the AD8 have been shown to be superior to performance-based cognitive measurements in identifying the underlying pathology in the early stage of AD25. Although one neuroimaging study demonstrated that episodic memory and hippocampal volume were associated with each domain of the ECog scale in patients with AD24, the association between ECog and neuronal metabolism measured using FDG-PET is unknown. According to Jack’s hypothetical model of dynamic biomarkers in AD, abnormalities in FDG-PET can be detected earlier than changes in volumetric magnetic resonance imaging (MRI)26.The Consortium to Establish a Registry for Alzheimer’s Disease Neuropsychological Assessment Battery (CERAD-NAB) is a well-validated, widely used tool to study cognitive deficits in patients with dementiaIn the current study, we investigated associations among the ECog scale, CERAD-NAB and FDG-PET scans in the early stage of probable AD. We hypothesized that the ECog score may be correlated with neuropsychological results and FDG-PET abnormalities. In addition, we compared the sensitivity and specificity between the ECog scale and CERAD-NAB in the diagnosis of AD.P < 0.01) and total CERAD-NAB scores  between the normally aging and probable AD groups. In the early stage of probable AD group, 38 patients had a CDR score of 0.5 and two had a CDR score of 1. In the normally aging group, the regression analysis of total CERAD-NAB score showed a significant association with age and years of education (P < 0.01), but not with gender (P = 0.58). The R-square of regression was 0.42 (P < 0.01).A total of 160 normally aging adults and 40 patients with the early stage of probable AD were included for analysis. There were no significant differences in mean age, gender and years of education between the two groups. The years of education ranged from 0 to over 20  and AD group = 9.2 (4.6)), with 25% of the participants having less than 6 years of education. Table P < 0.01). The mean ECog scores in each domain were all significantly lower in the normally aging group than in the early stage of probable AD group  . Pairwise correlation analysis revealed a significant correlation between the total ECog scores and total CERAD-NAB scores in all participants . The word-list learning test with delayed recall and recognition from the CERAD-NAB test showed a significant correlation with all domains of the ECog scale . The category verbal fluency test also showed a significant correlation with planning and organization domains of the ECog scale  . The average total score of the ECog scale was not significantly correlated with the AD t-sum scores. Only the average score of the memory domain of the ECog scale demonstrated a trend of correlation with the AD t-sum scores . In addition, the executive domain of the CERAD-NAB score was significantly correlated with the SUVr of the posterior cingulum gyrus . The memory domain of the ECog scale also demonstrated a significant inverse association with the SUVr of the angular gyrus and posterior cingulum gyrus  in the ECog scale were significantly correlated with executive CERAD-NAB subtest score. Using a functional tool, we demonstrated that the ECog scale could differentiate patients with the early stage of probable AD from normally aging adults. In the AUC analysis conducted to discriminate both groups, the average total score of the ECog scale was associated with a significantly higher AUC value compared with the z-transformed total CERAD-NAB score. Furthermore, the memory domain of the ECog scale and CERAD-NAB score were both significantly associated with the regional SUVr in the angular gyrus and posterior cingulum gyrus, suggesting their diagnostic potential for the early stage of AD.12. Moreover, this functional score was significantly negatively correlated with the CERAD-NAB score, suggesting that functional impairment is related to cognitive dysfunction12. In a regression study, the average total ECog score was significantly negatively correlated with z-transformed total CERAD-NAB scores in patients with the early stage of AD (P < 0.01) but not in normally aging adults, which may be due to the flooring effect of the ECog scale. In the current study, the average domain-specific scores were significantly correlated with memory subtest scores of the CERAD-NAB test. The results indicated that memory performance may be associated with memory and other cognitive domains of the ECog scale. Executive function as assessed using the category verbal fluency test was also significantly correlation with the executive domain of the ECog scale, which is compatible with the dysfunction of executive ability in the early stage of AD28. However, the cognitive deficits revealed in the CERAD-NAB test but not in the ECog scale were associated with AD t-sum scores, as observed in FDG-PET, demonstrating that cognitive dysfunction may not be completely equal to functional impairment. In confirmatory factor analysis of the ECog scale, one global factor and six domain-specific factors have been reported12. Other factors such as neuropsychiatric symptoms  have also been reported to independently contribute to the ECog score29. These results support that functional impairment and cognitive dysfunction share a common factor but are also independent.The ECog scale is a useful tool for measuring general and domain-specific everyday functions in elderly people, and in patients with mild cognitive impairment (MCI) and dementia. There were significant differences in total score and the six average domain-specific scores between the normally aging and early stage of probable AD groups, which is consistent with the findings of a previous study12. In addition, an informant-based assessment tool provides an opportunity for clinicians to assess changes in a patient’s functioning level and determine the interference in their daily tasks, whereas a neuropsychological battery is a performance-based measurement tool that requires a group of age- and education-matched normal participants for comparison to demonstrate the deficits. Theoretically, compared with a measurement tool for inter-participant comparisons, a tool for intra-participant comparisons could be more sensitive in detecting the subtle changes in the early stages of AD30. Our results demonstrated that the memory domain score of the ECog scale was associated with regional FDG uptake in the bilateral angular gyrus and posterior cingulum gyrus. These findings suggest that the ECog scale may reflect cortical regional metabolism status. Furthermore, the ECog scale includes numerous everyday cognitive domains other than memory function, such as language, visual–spatial, and executive function. Various dementia syndromes such as behavioral variants of frontotemporal dementia, posterior cortical atrophy, and primary progressive aphasia may initially present as distinct everyday cognitive deficits rather than memory problems, which could possibly be identified using the ECog scale31. The ECog scale may therefore potentially help clinicians to more efficiently differentiate among degenerative dementia syndromes.The discriminative ability of the average total ECog score between the two groups was associated with a significantly higher AUC value compared with the z-transformed total CERAD-NAB score. Most cognitive tests are performance-based measurement tools that require a group of age- and education-matched normal participants for comparison to demonstrate deficits. However, the ECog scale is weakly correlated only with age and years of education, which could be beneficial when studying a group with a wide range of years of education as in the current studyet al. reported that the hippocampal volume and total brain volume were associated with most ECog domains, whereas the frontal lobe volume was independently associated with two everyday executive domains in normal participants and in patients with MCI and AD24. According to a review of the relevant literature, no previous study has investigated associations between the ECog scale and FDG-PET scans in relation to dementia. We found that the memory domains of the ECog were significantly correlated with FDG uptake in the angular gyrus and posterior cingulum gyrus in the patients with the early stage of probable AD (P < 0.01), whereas other non-memory domains of the ECog scale were neither correlated with AD t-sum scores nor regional SUVr from FDG-PET scans. The reason for this may be because the disease severity of our patients with the early stage of probable AD was very mild (mean CDR-SB = 2), and because most abnormal hypometabolism regions in the mild stage of AD are related to the memory domain. Of all CERAD-NAB test items, the memory-related domain contributed 50% , whereas of all ECog scale items, the memory domain contributed only 23.6% . Another possibility is that the ECog scale is composed of six cognitive domains, and that the association between average total ECog scores and AD t-sum scores may decrease in patients with the early stage of probable AD. On the basis of this finding and because all six ECog domains exhibited a significant difference between the normally aging adults and early stage of probable AD groups, we suggest that in patients with the early stage of probable AD, the functional impairment measurements from the ECog scale can provide complementary information in addition to that from the performance-based cognitive tests.In a neuroimaging study, Farias 32. We will consider this aspect in our next MCI study. Caregiver distress has been shown to affect the patient’s behavior rating in previous studies, which may in turn affect the ECog scores33. Other limitation including incorporation bias in the ECog scale and CERAD-NAB test may also have confounded our findings. The application of these approaches in a large cohort of participants with MCI in the future is warranted.There are several limitations to the present study. First, due to the low number of patients with the early stage of probable AD and a lack of patients with MCI, we could not generalize our results to clinical practice. The low number of normally aging adults may also have affected the z-transformed total CERAD-NAB score. In addition, we did not perform further studies in our normally aging adult group which may have included patients with MCI or even the early stage of AD. These factors may have decreased the correlation between ECog scores and CERAD-NAB scores. We used the CERAD-NAB test scores and MRI scans in the diagnostic process, however not the FDG-PET scans or ECog scores. According to the hypothetical model of dynamic biomarkers in AD, this process may have limited the enrollment of patients with probable AD at an earlier stage, and may have led to more significant differences in ECog scores between the normally aging adults and patients. It would be interesting to investigate whether both ECog scale and FDG-PET scans can improve the diagnostic repertoire in future studies. Second, the diagnosis of probable AD in this study was based on clinical criteria rather than neuropathological evidence. The FDG-PET scans only represented the neuronal injury markers in the diagnostic criteria. As the number of patients with amyloid biomarkers in this study was insufficient to perform subgroup analysis, further studies are needed to investigate the relationships between ECog scores and FDG-PET scans in patients with AD who are positive for amyloid markers. Third, we did not apply the ECog scale using self-rated participant tests and did not assess the mood characteristics or caregiver burden of the informants. A recent study revealed that self-reported complaints as measured by self-rated ECog scale were not correlated with cognitive dysfunction but with depressive symptoms, and that this may confound the misclassification of MCIIn summary, the present study is the first to provide the results of applying the ECog scale to normally aging adults and patients with the early stage of probable AD. The ECog scores exhibited significant correlations with standard neuropsychological tests and could also discriminate between both groups. The memory domain of this functional ability scale and CERAD-NAB scores were associated with AD-related hypometabolism derived from FDG-PET scans.We recruited 160 normally aging adults aged 50 years or older from several senior citizen community centers whose first language was Chinese or Taiwanese. Registered nurses, social workers, and occupational therapists reviewed the recent medical records of all participants. No memory or other cognitive complaints were confirmed in the participants before they were enrolled into the study, and they all underwent detailed physical and neurological examinations. Those with a history of major neurological, psychiatric, or severe cardiovascular diseases were excluded.1. Cognitive dysfunction was assessed using the CERAD-NAB test, and brain MRI scans were used to exclude the possibility of structural brain lesions such as hydrocephalus or brain tumors. The severity of dementia measured by the Clinical Dementia Rating (CDR) scale score was limited to 0.5 to 14. All procedures were approved by the Institutional Review Board (IRB) of Chang Gung Memorial Hospital, and all participants signed informed consent to participate in the study (IRB number:103-1994B). The study was performed in accordance with the approved guidelines and regulations.In addition, we recruited 40 patients aged older than 50 years from the outpatient clinics of our hospital as the disease group. In this study, the early stage of probable AD was diagnosed according to the criteria of the National Institute on Aging-Alzheimer’s AssociationThe informants of the normally aging adults and the patients with the early stage of probable AD completed the ECog scale. The ECog scale is comprised of 39 items covering six domains , and is scored based on a four-point scale: 1 = better or no change compared to 10 years earlier, 2 = questionable/occasionally worse, 3 = consistently a little worse, 4 = consistently much worse. An “I don’t know” response option is also included. The total score was calculated as the sum of all 39 items, and the average score was derived from the mean average of all responses. If the response was “I don’t know”, then the item was not included in the calculation. The response rate for each item was calculated as the total number of responses from 1–4 (not including “I don’t know” responses) divided by the total number of participants. The translation and adaptation of the ECog scale involved a bilingual member of the medical staff translating the original ECog version to the Chinese version, followed by a neurologist with a subspecialty in dementia reviewing the translation and applying the translated version to a small group of early stage dementia caregivers.34. The CERAD-NAB is comprised of five subtests derived from previously established cognitive tests: an executive domain of the category verbal fluency test35, a language domain of the Modified Boston Naming Test (BNT)36, a memory domain of the Word-List-Learning test with delayed recall and recognition37, and a visuospatial domain of the visual construction test38. The total score of the CERAD-NAB was calculated by summing the six subtest scores . The CDR sum of box (CDR-SB) was calculated as the sum of all six items in the CDR scale.The neuropsychological battery included the CERAD-NAB, Mini Mental State Examination (MMSE) and CDR scales, which were evaluated by a single rater for all participantshttps://www.pmod.com/web/?portfolio=32-alzheimers-discrimination-for-fdg-palz) was used for image analysis. The AD t-sum score was calculated using this software as the global indices of AD-related hypometabolism39. We further used individual FDG-PET scans to perform spatial normalization, and then the global mean intensity in the gray matter mask was used as the reference to calculate the standard uptake value ratio (SUVr) in each region according to the AAL template40. The mean SUVr values from both hemispheres in AD-related regions including the middle frontal gyrus, angular gyrus, calcarine gyrus, middle temporal gyrus and posterior cingulum were calculated for further analysis41.All of the patients underwent brain MRI and FDG-PET scans on an integrated PET-MR system (Siemens Biograph mMR scanner), which could simultaneously acquire PET and MRI data using a vendor-supplied 12-channel phase-array head coil according to standard procedures. In brief, the participants were required to fast for 4 h before imaging. FDG data were collected 30 min after the injection of a dose of 185 MBq FDG. This 30-min PET data reflected the distribution of FDG tracer after homogeneous and complete distribution in the brain. A high-resolution magnetization-prepared rapid acquisition gradient-echo T1-weighted image (voxel size 1 × 1 × 1.1 mm) and a FLAIR image (1 × 1 × 1 mm) were obtained simultaneously using the integrated PET-MR system. PMOD Alzheimer’s discrimination tool . Continuous and dichotomous variables are expressed as mean and standard deviation and number and percentage, respectively. Ordinal variables are presented as median and interquartile range. Socio-demographic variables included age, gender, and years of education, and the results of the neuropsychological tests including the MMSE, CDR, CDR-SB, and ECog were compared between the normally aging and early stage of probable AD groups using the chi-square test and Mann–Whitney U test. Nonparametric Spearman pairwise correlation analysis was used to study associations between the total ECog scores, total CERAD-NAB scores and their subtest scores in both groups. The raw CERAD-NAB score may have been confounded by age, gender, and years of education. Therefore, we used regression analysis to adjust for any confounding effects. The raw total CERAD-NAB scores in the normally aging group were entered into the regression analysis model using age, years of education, and gender as independent variables to generate the regression coefficients of the total equation. The variables with significant regression coefficients were used to generate the adjusted CERAD-NAB score in the whole sample. The raw total CERAD-NAB scores of all participants were converted into standardized z-scores according to the means and standard deviations of the adjusted CERAD-NAB score of the normally aging group. Receiver operating characteristic (ROC) curves were generated to investigate the accuracy of the average total ECog scores and z-transformed total CERAD-NAB scores for all participants. Moreover, areas under the curve (AUCs) with 95% confidence intervals (CIs) were calculated for each test. To compare the discriminative abilities of these tests in both groups, the AUCs were assessed using the Wald test. In addition, the cutoff points were determined according to the sensitivity and specificity. To study the association between the AD t-sum scores, regional SUVr and total and domain-specific scores of the ECog scale and z-transformed total CERAD-NAB scores, we used Spearman pairwise correlation analysis. The level of significance was set at


Both Raman and Fourier-transform infrared (FT-IR) imaging have been applied to obtain detailed information on the chemical composition of biological materials, ranging from single microbial cells to tissues. Due to its compatibility with methods such as stable isotope labeling for the monitoring of cellular activities, vibrational spectroscopy also holds considerable power as a tool in microbial ecology. Chemical imaging of undisturbed biological systems (such as live cells in their native habitats) presents unique challenges due to the physical and chemical complexity of the samples, potential for spectral interference, and frequent need for real-time measurements. This Mini Review provides a critical synthesis of recent applications of Raman and FT-IR spectroscopy for characterizing complex biological samples, with a focus on developments in single-cell imaging. We also discuss how new spectroscopic methods could be used to overcome current limitations of single-cell analyses. Given the inherent complementarity of Raman and FT-IR spectroscopic methods, we discuss how combining these approaches could enable us to obtain new insights into biological activities either  A full understanding of these topics also requires an ability to study these processes at the level of single cells . However, a small fraction is inelastically scattered, involving a decrease or an increase in energy compared with the excitation wavelength . By providing information on vibrational and other low-frequency transitions in a molecule, both types of inelastically scattered light can be used to determine and differentiate between the chemical composition of solids, liquids and gases. For a more detailed introduction to this technique (as well as infrared spectroscopy), the reader is referred to 13C labeling to identify cells that fixed carbon dioxide in culture and in field-collected seawater samples. By reducing spectral acquisition times to milliseconds, resonance Raman spectroscopy – a method in which the excitation wavelength matches the electronic transition of a selected molecule – was key to enabling the rapid imaging of these samples. It is also possible to visualize selected strains and their locations within habitats including human endothelial cells  and linear discriminant analysis (LDA)], Two features that make Raman microspectroscopy an ideal technique for single-cell analyses include its direct compatibility with aqueous samples (due to water exhibiting only weak Raman scattering) and its high spatial resolution . While aal cells , macrophal cells  and otheal cells . ThroughPantoea sp. YR343) on Arabidopsis thaliana root surfaces    as well ization)  and Ramaization)  are likeization) , spatialization) . This teization) . CARS haization) , and a sization) . Anotherization) . Howeverization) .While Raman spectroscopy relies on irradiating a sample with a monochromatic laser beam, Fourier-transform infrared (FT-IR) spectroscopy is based on measuring the absorption of polychromatic infrared light. The functional groups in a given molecule are identified according to their vibrational modes at different IR frequencies  and nitrogen (15N) compounds (2O), due to absorbance peaks corresponding to O–H and O–D bending modes occurring at different wavenumber regions , which led the authors to suggest that FT-IR analyses could be particularly useful for characterizing mixed cultures . The suggestions provided in Table 1 additionally highlight the promising role that live-cell FT-IR imaging could play in environmental microbiological research, further to Raman measurements which have traditionally been more common in this field. The future development of vibrational spectroscopy instrumentation and analytical methods may serve to further enhance the cross-compatibility of Raman and FT-IR techniques .The spectroscopic imaging of microbial cells in physically and chemically complex samples involves diverse analytical challenges. While addressing these will often require sample-specific optimization steps . Synchrotron-FT-IR microspectroscopy has been paired with synchrotron ultraviolet microspectroscopy and time-of-flight-secondary ion mass spectrometry (ToF-SIMS) for the analysis of human liver tissue, with each technique yielding unique information on the chemical composition of the sample , and that result using these methods may need to be verified using additional methods. For example, Raman-activated cell sorting has recently been combined with single-cell genomics to identify members of a novel cyanobacterial order within seawater samples . UltimatAll authors listed, have made substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Pseudomonas putida. Disruption of fleQ caused strong defects in flagellar motility, biofilm formation and surface attachment, and the ability of this mutation to suppress multiple biofilm-related phenotypes associated to cyclic diguanylate overproduction suggests that FleQ mediates cyclic diguanylate signaling critical to biofilm growth. We have constructed a library containing 94 promoters potentially involved in motility and biofilm development fused to gfp and lacZ, screened this library for FleQ and cyclic diguanylate regulation, and assessed the involvement of alternative σ factors σN and FliA in the transcription of FleQ-regulated promoters. Our results suggest a dual mode of action for FleQ. Low cyclic diguanylate levels favor FleQ interaction with σN-dependent promoters to activate the flagellar cascade, encompassing the flagellar cluster and additional genes involved in cyclic diguanylate metabolism, signal transduction and gene regulation. On the other hand, characterization of the FleQ-regulated σN- and FliA-independent PlapA and PbcsD promoters revealed two disparate regulatory mechanisms leading to a similar outcome: the synthesis of biofilm matrix components in response to increased cyclic diguanylate levels.Most bacteria alternate between a free living planktonic lifestyle and the formation of structured surface-associated communities named biofilms. The transition between these two lifestyles requires a precise and timely regulation of the factors involved in each of the stages that has been likened to a developmental process. Here we characterize the involvement of the transcriptional regulator FleQ and the second messenger cyclic diguanylate in the coordinate regulation of multiple functions related to motility and surface colonization in  The alternation between a free-swimming planktonic lifestyle and the formation of structured highly cooperative polymer-encased sessile communities, named biofilms, is a common feature of bacterial life in the environment . BiofilmThe nucleotide cyclic diguanylate (c-di-GMP) is a second messenger that is ubiquitously used in bacteria for signaling the transition between the planktonic and sessile lifestyles. c-di-GMP is synthesized from GTP by diguanylate cyclase (DGC) activities, associated to proteins with GGDEF domains, and degraded by specific phosphodiesterase (PDE) activities, associated to proteins with EAL or HD-GYP domains. Bacterial genomes often encode multiple proteins containing one or more of these domains, implying that the c-di-GMP levels are likely regulated in a complex fashion. Changes in c-di-GMP concentration are sensed by effectors, which in turn regulate a variety of processes, generally related to motility, biofilm development or virulence, acting at the transcriptional, translational or posttranslational levels. The biology of c-di-GMP signaling has been extensively reviewed in recent years –8.N-dependent promoter activators, and as the master regulator of flagellar biogenesis in Pseudomonas aeruginosa and other bacteria N-depende(PDF)Click here for additional data file.S1 Table1ORF identification code according to the Pseudomonas genome annotation. 2Operon structure, according to the DOOR algorythm prediction for the P. putida KT2440 genome. 3Description of the relevant gene products and the criteria used for selection of each promoter.(PDF)Click here for additional data file.S2 TableThe results shown are the averages and standard deviations of at least 3 independent experiments.(PDF)Click here for additional data file.S3 Table1Coordinates are relative to the start codons. 2A plus (+) sign designates the strand corresponding to the lapA or bcs transcript 3Positions matching the consensus are indicated in bold.Prediction was performed according to . 1Coordi(PDF)Click here for additional data file.


Conventional asphalt binder derived from the petroleum refining process is widely used in pavement engineering. However, asphalt binder is a non-renewable material. Therefore, the use of a co-production of renewable bio-oil as a modifier for petroleum asphalt has recently been getting more attention in the pavement field due to its renewability and its optimization for conventional petroleum-based asphalt binder. Significant research efforts have been done that mainly focus on the mechanical properties of bio-asphalt binder. However, there is still a lack of studies describing the effects of the co-production on performance of asphalt binders from a micro-scale perspective to better understand the fundamental modification mechanism. In this study, a reasonable molecular structure for the co-production of renewable bio-oils is created based on previous research findings and the observed functional groups from Fourier-transform infrared spectroscopy tests, which are fundamental and critical for establishing the molecular model of bio-asphalt binder with various biomaterials contents. Molecular simulation shows that the increase of biomaterial content causes the decrease of cohesion energy density, which can be related to the observed decrease of dynamic modulus. Additionally, a parameter of Flexibility Index is employed to characterize the ability of asphalt binder to resist deformation under oscillatory loading accurately. Conventional hot mix asphalt (HMA) is produced with asphalt binder that derives from the petroleum refining process. However, the total storage of petroleum is limited and petroleum is non-renewable. Thus, it is necessary to develop an alternative binder material to entirely or partly replace the petroleum asphalt with respect to sustainable asphalt pavements. In recent years, biomass materials that can be derived from various bio-resources such as corn stover, animal waste wood and waste cooking oil are gaining interest from the pavement industry ,2,3. SigThough the performance of WCO-based bio-asphalt has been comprehensively studied, there is still a lack of studies addressing the effects of WCO-based bio-oil on petroleum asphalt from a micro-scale perspective to understand the fundamental modification mechanism. Molecular Dynamics (MD) simulation is one of the effective analytical approaches to investigate the micro-structure and micro-properties of asphalt materials ,35,36. BThe objective of this paper is to quantify the WCO-based bio-oil modification effect on the performance of petroleum asphalt from the micro-scale perspective. The molecular model of WCO is first created to represent bio-asphalt binders with various bio-oil contents. The validation of these molecular models is conducted by means of thermodynamic properties including the density, cohesive energy density and surface free energy. Then the bio-asphalt binders are simulated by means of a MD simulation to investigate several possible relationships between micro-scale characteristics and laboratory-tested macro-scale performance, which effectively reveals the micro-scale modification mechanism for the WCO-based bio-oil modified asphalt binders.In this study, a 60/80 penetration-grade asphalt binder was used as the control asphalt binder. This petroleum asphalt source was obtained from the Hebei province and has been widely applied to produce the HMA in the Beijing area. The physical properties of this control asphalt are shown in The bio-oil used in this study was a black oily liquid as shown in The control asphalt was first heated in the oven at a constant temperature of 150 °C for about one hour. Then 5%, 10%, and 15% bio-oil by weight of control asphalt binder was respectively added into the control asphalt when the temperature was stable at 140 °C. Afterwards a continuous 30 min of blending with a mixing speed of 4000 rounds per minute (rpm) using a high shear mixer followed to achieve a homogeneous mixing state. The sample IDs of the prepared asphalt binder materials are given in −1.The FTIR  test has been an effective means for analyzing the chemical properties of asphalt binders. The chemical functional groups of asphalt can be identified based on the principle that the molecular rotation or oscillation at specific frequencies would result in the absorption of infrared spectra. Measuring the FTIR absorbance, the change in chemical functional groups of asphalt binder due to the incorporation of bio-oil residues can be detected and compared. The binder samples for FTIR tests were prepared by mixing asphalt binder with a common asphalt solvent of toluene followed by dropping the solution onto a KBr table. The toluene was then evaporated and the prepared sample was then scanned with the test spectrum range of 400 to 2000 cmThe rheological tests in this study were completed with an Anton Paar MCR 302 dynamic shear rheometer . The asphalt binder was a well-known viscoelastic material. The complex shear modulus of the oscillation loading was a measurement of the undamaged binder stiffness within the linear viscoelastic range using small strain amplitude, which was also dependent on the loading frequency and temperature. In this study, the complex shear modulus tests were performed under 1.58 rad/s at the temperature of 20 °C with the 8-mm parallel plate and 2-mm gap geometry.Fatigue cracking due to repeated traffic loading is the main pavement distress that shortens the service life of asphalt pavements, which has been widely validated . Based oAt least two replicates were run for all of the chemical and rheological performance tests to get the average test results. Three or even more replicates were completed to ensure the coefficient of variation was within 10 percent.−1) and C=O stretch (1735–1750 cm−1) exhibits much higher values than those of control 70# binder, indicating the existence of oxygen-related functional groups within the bio-oil. More specifically, it is also observed that the amount of C=O stretch and C–O stretch increases when increasing the content of bio-oil, which is expected from the spectra property of the bio-oil. Meanwhile, the S=O stretch (1030 cm−1) is also an important functional group for asphalt binder; however, the sulfoxide functional group within the control 70# binder and three bio-binders show slight differences.A step of baseline correction is conducted for the raw FTIR spectra of each binder to perform a reliable spectra comparison. The corrected FTIR spectra of control asphalt and bio-binders are shown in To remove the variability of peak absorbance heights due to the various sample film thicknesses, the carbonyl index (C=O), carbon-oxygen index (C–O) and the sulfoxide index (S=O) developed by Lamontagne et al. were utilized to quantitatively describe the chemical structure change upon the bio-oil addition, as expressed in Equations (1)–(3) respectively .(1)IC=OC=O, IC-O and IS=O indices for the tested materials are given in C=O and IC-O indices of bio-oil are much higher than those of control 70# binder. When further adding the bio-oil to the 70# binder, the IC=O and IC-O are observed to be monotonically increased. However, the IS=O index of all materials almost stays around a constant level, indicating that the S=O content within bio-oil is similar to that of the control asphalt.The quantified IThe complex shear modulus at a temperature of 20 °C under a loading frequency of 1.58 rad/s for the control 70# binder and three bio-binders are presented in The control 70# binder and three WCO-based bio-binders were subjected to the LAS test procedure followed by the fatigue performance analysis according to the S-VECD modeling approach ,57. The Asphalt binder is well known as a typical organic material, which contains millions of complex chemical components . In ordeThe unit numbers of the above 12 chemical components in the molecular system of asphalt binder are listed in Normally the WCO contains lots of complex chemical components due to its wide range of sources, so a feasible typical component should be selected to represent the WCO. The main chemical component within the WCO is the triglyceride (TG), which is an ester derived from glycerol and three fatty acids ,74,75, aThe number of C=O and C–O within these created molecular structures for control 70# binder and three bio-binders are calculated to compare with the measured C=O and C–O by FTIR. The comparison results are shown in To establish the molecular models of control 70# binder and three bio-binders respectively with 5%, 10% and 15% WCO-based bio-oil, four molecular models of asphalt binders with 0, 2, 4 and 6 WCO molecules are constructed according to the molar mass of the WCO (993.6 g/mol), and the number of WCO is also adopted as an integer. These four molecular structure models with different bio-oil contents are created by means of the software Material Studio 6.0 . The force field used in this study is the condensed-phase optimized molecular potential for atomistic simulation studies (COMPASS) force field, which is the first parameterized and validated first-principles-based force field and has been proved to be accurate for covalent molecules including most common organics, small inorganic molecules and polymers . Then th3 [The density information is first investigated to estimate the properties of these created molecular models from the MD simulation with duration of 100 ps under NPT conditions. The computation process took about 12 h and the simulated density results are given in 3 , which ia is the SFE; Esurface and Ebulk represent the potential energy of the confined asphalt layer and bulk asphalt, respectively; and A is area of the new surface to be created.The parameter Surface Free Energy (SFE) measures the energy required to disrupt intermolecular chemical bonds when creating a new surface area . In orde2, as reported in the literature [As shown in terature .The Cohesive Energy Density (CED) is a mutual attraction between the molecules within the same material, which represents the manifestation of the molecular force between molecules of the same species and can be utilized to assess the intermolecular interaction inside an asphalt molecule model . A serieThe CED results of the control 70# binder and three bio-binders are compared in The above MD simulation of density, SFE and CED properties validates that these created molecular structure models for control 70# binder and three bio-binders with different bio-oil contents are reasonable since the simulation results of these models are identical to the previous experimental results and research findings ,79.The CED parameter characterizes the cohesion energy within a unit volume. Previous research  showed tThe complex shear modulus represents the typical linear viscoelastic (LVE) material property of a non-damaged body. Fatigue resistance of asphalt binder is important for fatigue performance of asphalt mixture and pavement. Currently main research efforts are focused on the fatigue performance evaluation and prediction based on laboratory testing. However, there are only limited studies addressing the fatigue damage and failure mechanisms from the micro-scale perspective. Essentially, the fatigue resistance of asphalt binder is mainly determined by various chemical properties such as the chemical composition, molecular weight, functional group and molecular structure. Therefore, investigating the micro-scale material property enables insightful understanding for comparing the distinguished fatigue resistance of different asphalt binders.1 index; the branching or relative spatial density of molecules is encoded into the K2 index. The K1 and K2 indices are respectively derived as Equations (5) and (6).In h of one .Then, the α-modified shape indices xth heavy atom and 3 carbon, which is assumed to be 0.77 Å [The modifier item of α is given by Equation (9).e 0.77 Å .Finally, the Flexibility Index (FI) can be calculated using Equation (10).The FI parameters of the fraction structure are calculated and shown in (1)From FTIR tests, it is demonstrated that the bio-oil modified binders displayed increased carbonyl index when increasing the bio-oil content, whereas the sulfoxide index almost exhibited the same level as that of the control asphalt. Further rheological performance tests indicated that the addition of bio-oil softened the stiffness and improved the fatigue resistance of the control asphalt.(2)A reasonable averaged molecular structure for bio-oil is created based on previous research findings and further validated with the FTIR results and the micro-scale properties in terms of density, SFE and CED from the MD simulation.(3)A larger CED at micro-scale level represents a stronger link between the molecules within asphalt binder to provide a better stability against shear deformation in the macro-scale. Therefore, the observed complex shear modulus decrease due to the bio-oil addition is well related to simulated CED properties for all tested binders. Meanwhile, a parameter of Flexibility Index (FI) is utilized to investigate the micro-scale fundamental mechanisms for the improved binder fatigue life from the addition of bio-oil. The more flexible the binder molecule is obtained, the better its fatigue resistance.This paper investigates the performance of WCO-based bio-asphalt binders from both rheological and molecular simulation approaches. The bio-oil effects on the chemical functional groups, complex shear modulus and fatigue performance of control asphalt is firstly characterized and then followed by the MD simulation to explain the modification mechanisms of bio-oil from a micro-scale perspective. The specific findings of this study are summarized as follows:In further research, more material properties of WCO-based bio-oil modified asphalt binder still need to be covered from both macro-scale and micro-scale. Also, the long-term aging effects on these material properties need to be addressed.


Ranolazine (RAN) is a novel late sodium current inhibitor, effective in treating angina pectoris in patients with chronic stable CAD. Its therapeutic effectiveness is not well understood. It is thought to reduce myocardial energy utilization by enhancing diastolic relaxation and by increasing myocardial blood flow. The purpose of the present study is to define the mechanism for the effectiveness of RAN. We applied myocardial phosphorus-31 magnetic resonance spectroscopy (P-31 MRS) at rest and during handgrip stress to demonstrate the relationship between RAN treatment and stress-induced energy utilization with myocardial ischemia.Patients with angina pectoris defined by the presence of sub sternal pain, associated with stress, and effectively reduced with rest (or nitroglycerine), and with evidence of "ischemia" by conventional imaging approaches including: stress SPECT, PET or echocardiographic studies; or the presence of >70% diameter stenosis on conventional or CT coronary angiography were enrolled. RAN was administered in doses of 500 mg twice daily for 2 weeks added to existing medical therapy, and titrated to 1000 mg twice daily for 2 weeks. All subjects were evaluated at the start and upon completion of the study for the presence and severity of angina pectoris using the Seattle Angina Questionnaire (SAQ).Prior to and after maximal RAN dose titration, a P-31 MRS study was performed at rest and during isometric hand-grip stress of moderate intensity. With P-31 MRS, myocardial metabolic ischemia was defined as an abnormal decrease of phosphocreatine:ATP ratio (PCr/ATP) during handgrip stress.Nine of 16 subjects were identified as having metabolic ischemia by P-31 MRS prior to RAN. At 4 week follow-up, five of the nine P-31 studies (56%) with metabolic ischemia at baseline demonstrated evidence of improvement in the myocardial HEP response (p = 0.005). Four subjects without improvement with their abnormal baseline P-31 study and seven subjects with a normal baseline P-31 showed no significant differences in HEP response at follow-up . There was a modest degree of agreement between improvements in HEP response identified by P-31 MRS, and angina frequency from the SAQ with 78% of subjects with ischemia at baseline similarly classified (κ=0.571).This preliminary study demonstrates that Ranolazine treatment is associated with a significant reduction in myocardial ischemia in the majority of patients with angina at baseline.