Fluorinated(19F) probes have been increasingly showing promising potentialitiesfor in vivo cell tracking by 19F-MRI.Here we present the unique features of a bioorthogonal 19F-probe that enables direct signal correlation of MRI with Ramanimaging. In particular, we reveal the ability of PERFECTA, a superfluorinatedmolecule, to exhibit a remarkable intense Raman signal distinct fromcell and tissue fingerprints. Therefore, PERFECTA combines in a singlemolecule excellent characteristics for both macroscopic invivo19F-MRI, across the whole body, and microscopicimaging at tissue and cellular levels by Raman imaging.Molecular imagingtechniques are essential tools for better investigatingbiological processes and detecting disease biomarkers with improvementof both diagnosis and therapy monitoring. Often, a single imagingtechnique is not sufficient to obtain comprehensive information atdifferent levels. Multimodal diagnostic probes are key tools to enableimaging across multiple scales. The direct registration of  With the increasingrequest of correlation and integration of various imaging informationat different scales, the development of multimodal probes is highlyneeded.2A pressing need inmodern medicine is the development of ever moresensitive imaging tools to improve early diagnosis and support personalizedmedicine and precision surgery.4 Even if Raman microscopy is largely used as a label-free technique,enabling distinction between healthy and pathological cells or tissuesbypassing the use of fluorescent markers, specific Raman probes mayfurther extend sensitivity and specificity. These probes are typicallybased on small chemical groups with diagnostic vibrational features,such as alkynes5 and deuterated compounds.6 Alternatively, plasmonic nanoparticles (NPs)were studied as SERS (surface-enhanced Raman scattering) tags.7 All these probes have been widely used for imagingbiological targets in cells and tissues.9 While Ramanprobes paved the way toward super-multiplexed optical imaging10 and bioorthogonal cell- or organelle-targetedimaging, in most cases they are not directly detectable by clinicalimaging modalities . An exception is represented by someSERS tags, based on the synthesis of multimetal NPs, which have beenproposed as bimodal agents, but are often limited by toxicity andstability issues.12 For this reason, new biocompatibleand bimodal probes are essential to allow the integration of imaginganalysis of structural and chemical biology with in vivo functional imaging. Here we demonstrate, for the first time, theability of fluorinated molecules to work as bimodal probes (in vivo19fluorine-magnetic resonanceimaging (19F-MRI), used to explore whole body or organs, with Ramanmicroscopy, enabling tissue and cell imaging with subcellular resolution(∼500 nm) and high biomolecular specificity.3Raman spectroscopy (RS)is a direct and nondestructive approachbased on the highly specific recognition of chemical bonds presentin the analyzed sample. RS has been proved to be crucial in studyingbiological samples and has major potential for clinical translation.l probes 1, allowi19F-MRI isindeed emerging as a powerful noninvasiveimaging technique, easily translatable for clinical use,14 as it permits in-depth in vivo quantification withoutthe use of radioactive agents and can be combined with standard 1H-MRI protocols. Moreover, lack of endogenous organic 19F atoms in living tissues permits unambiguous invivo visualization of exogenous fluorinated probes as colored“hot spots” on the anatomical images obtained by multiparametric 1H-MRI.15 Until now, 19F-MRI has been combined withfluorescence imaging,16 enabling analysisat the cellular level. However, this approach has often been hamperedby the use of two distinct chemical agents, which are commonly co-incorporatedin nanoformulations.18 Moreover, the fluorescent signalis unstable over time, is easily quenched, and could be dissociatedfrom the fluorinated probe as the nanoformulation is degraded.19 Instead, direct covalent functionalization ofthe fluorinated probe with a fluorescent tag may alter its physicochemicaland pharmacokinetic properties.20 Comparedto fluorescence imaging, Raman probes are generally smaller, stable,and unaffected by quenching processes and may guarantee higher multiplexingcapabilities due to very narrow vibrational signals, permitting over20-color channels with Raman imaging.21 On the otherhand, Raman is mainly limited by the intrinsically low cross-sectionof the Raman effect, resulting in weak signals and low imaging speed,if compared to fluorescence-based approaches. In this context, recentlydeveloped imaging enhancing strategies, such as stimulated Raman scattering(SRS), permitted to accelerate imaging performance close to a fluorescent-basedone.22 The strength of our approach resides inthe combination of Raman microscopy and 19F-MRI for bioimagingat multiscales with high versatility and specificity by using a singlechemical imaging agent .These measurements were performed in water at room temperature (25°C). Further studies were also assessed in biological fluidsat 37 °C, and results confirmed the NPs stability at least upto 72 h in all investigated media . All these nanoformulations, which proved successful as labelingagents for cell tracking and in vivo imaging by 19F-MRI,28 thanks to their distinctive 19F-NMR signals, are also characterized by important Ramanfeatures related to their molecular structures, although with substantialdifferences  . Similarly, the Raman marker of PFOB (724cm–1) is assigned to collective symmetric stretchingand bending vibrations, exhibiting a good intensity, whereas the markerof PFCE (820 cm–1) is assigned to collective symmetricbending , which explainsits reduced intensity compared to those obtained with the other fluorinatedprobes. Since the molecular structures of both PFOB and PFCE are conformationallymore flexible than PERFECTA, their Raman bands not only are less intensebut also have a broad line width. Importantly, despite that the mostintense Raman peaks of all tested fluorinated probes fall in a relativelysilent biological region, between 500 and 800 cm–1, it also includes the typical DNA vibrational modes (around 790cm–1). However, DNA detection is not compromisedin biological samples, as we show in detail in Figure S2).Regarding olecule) 2d,e. Simresponse2d,e. Conresponse2f, PERFEntration 2g. The min vitro experimentsin immune effector cells of the brain were performed to screen andevaluate the three fluorinated probes. Thanks to their phagocyticcapacity, murine microglial cells were easily labeled with each fluorinatedprobe separately, at a fixed fluorine dose, and then imaged. As shownon 19F-MR images acquired at the NMR frequency of eachfluorinated probe  , but Raman microscopy allowed the directintracellular visualization of the same fluorinated molecule detectedby MRI. Moreover, as suggested by spectral analysis, we confirm thatthe Raman signal from each fluorinated probe does not prevent thedetection of label-free intrinsic chemical contrast of key cellularcomponents such as the nuclear DNA. Remarkably, signals of both PERFECTAand PFOB were strong and clearly distributed in the cytoplasm, whereasPFCE was more difficult to detect due to its broader and less intenseRaman signal. Of note, similarly to the use of fluorophores in fluorescencemicroscopy, tricolored Raman was obtained on a mixture of cells treatedseparately with each fluorinated probe  3c–h. Allctively) S6a–c, buin vitro labeled cells by MRI,fluorinated probes have been widely used for imaging inflammatoryactivity by in vivo labeling following systemic administration.Indeed, nanoformulations of fluorinated probes are avidly taken upby phagocytes as leukocytes35 that are highly present in injured tissues or organs with an immunereaction.Besides tracking in vivo in the well-establishedautoimmune encephalitis (EAE) mouse model that develops multifocalneuroinflammation along the spinal cord,36 similarly to patients affected by multiple sclerosis. From in vivo19F-MRI of mice treated with the PERFECTAnanoformulation, most of the 19F signal from PERFECTA wasfound in the liver . Following in vivo MRI, sections of spinal cord were collected anddirectly analyzed by Raman microscopy without further labeling orstaining , as shown by immunofluorescence microscopyin adjacent tissue slices .Interestingly, Raman images were also obtained in an EAE mouse witha reduced concentration of PERFECTA (estimated by 19F-MRIabout 0.2 mM) in the spinal cord at the limit of detection for bothimaging modalities .For this reason, the bimodal properties of the PERFECTAnanoformulation were tested he liver 5a, but ahe liver 5b, whereph nodes 5b, whichstaining 5c–f.Thi signals 5d,e were signals 5e. Indee signals 5e. These19F-MRI probe PERFECTAhas also a strong, specific, and stable over time Raman signal thanksto its characteristic C–F bonds, thus allowing a direct andreliable correlation between the two imaging modalities, which iscrucial for the validation and development of imaging probes. Thissignal falls in a relatively silent biological region, making it biorthogonalto the biological matter. Intracellular uptake of PERFECTA is demonstratedwithout use of additional fluorescent dyes. Thus, this enables theuse of PERFECTA as a bright bimodal probe for both macroscopic in vivo imaging across the entire body and organs by 19F-MRI and microscopic imaging at histological and cellularlevels by Raman microscopy. Of note PERFECTA shows unparalleled performancesin both imaging modalities when compared to other fluorinated probes,such as PFCE and PFOB. On the other hand, the three probes show separatedsignals, and simultaneous intracellular detection of the three fluorinatedprobes has been achieved, indicating the possibility to use theseprobes for multiplexing imaging.We showed that the superfluorinated ex vivo Raman imaging.38 Importantly, the use of Raman microscopy allows localization ofthe tracking agent at the tissue level and also permits obtainingunique additional information on the biomolecular composition of thebiological environment, distinguishing healthy tissues from diseasedregions. PERFECTA shows a peculiar advantage over other Raman probesdescribed in the scientific literature. SERS tags are, in fact, highlybright and can be detected at a single-probe level.10 However, they usually have a complex spectrum characterizedby multiple peaks and fluorescence background hiding signals fromthe surrounding tissue. Alkyne and deuterium tags have instead simplerspectra and can be easily incorporated into the sample, but theircross-sections are very small, and they are challenging to detectby Raman microscopy. In this regard, PERFECTA, with its 36 carbon–fluorinebonds, displays a single Raman peak at 770 cm–1 witha quite high cross-section. Furthermore, besides its unique spectroscopicand magnetic properties, PERFECTA has also an intrinsic chemical versatilityfor further functionalization, differently from other fluorinatedprobes. Its molecular design allows substitution of one of the foursymmetrical arms with an alkyl chain bearing different functionalgroups such as thiol,39 carboxylic acid,alcohol, azide, and amine, still maintaining 27 magnetically equivalent 19F in the molecule. This versatility paves the way for thedevelopment of a new set of dual-imaging functional tags, based ona library of PERFECTA derivatives, for detecting biological targetsby 19F-MRI and Raman microscopy. Finally, given the strongstretching vibrations of the C–F bond in the 1000–1400cm–1 region, these tags may also be foreseen asactive IR markers suitable for the emerging mid-infrared (MIR) microscopy40 technology, which will be studied in the nearfuture.Commonly used bimodal probeshave to overcome the complex synthesisneeded to combine two different molecular moieties in the same structure and their potential differences inpharmacokinetics. The use of a unique molecule as bimodal probe notonly permits reliable correlations but also avoids misinterpretationsand facilitates the translation for clinical purposes. For instance,a single bimodal probe could be used to identify by MRI the tumorfor surgical extraction or biopsy and to further characterize thetissue by intraoperative or


Results at room temperature and pressure showed that the blending of 0.26 wt.% MoS2 increased the lubricity of thermally stable base fluid by 27% and enhanced the thermal and electrical conductivities by 7.2% and 8.8%, respectively.Bentonite-based drilling fluids are used for drilling, where inhibitive fluids are not required. The rheological and the density properties of the drilling fluids are highly affected by high temperature and pressure. Due to high temperature, the clay particles stick together, and the fluid system becomes more flocculated. Poorly designed drilling fluid may cause undesired operational issues such as poor hole cleaning, drill strings sticking, high torque and drag. In this study, the 80 °C thermally stable Herschel Bulkley’s and Bingham plastic yield stresses drilling fluids were formulated based on lignosulfonate-treated bentonite drilling fluid. Further, the impact of a MoS Global energy demand is increasing rapidly due to population growth and industrial activities. The petroleum industry is expanding exploration activities in the deep-water and arctic regions. Moreover, green energy exploration such as geothermal energy is receiving lots of interest, where one can extract thermal energy from the high-temperature formation. However, it is evident that inventions of improved technologies are required to handle high pressure and high temperature (HPHT) conditions.Drilling fluids are an essential part of drilling operations, which have several functions such as to maintain well pressure, cool and lubricate the well, and bring cuttings up to the surface. The most important considerations for the selection of drilling fluids are their performance, cost, and environmental impact ,2. Oil- Drilling fluids are formulated with density, viscosity, and filtrate loss control additives to achieve the desired drilling fluid’s physical and rheological properties. Bentonite is the common additive used to provide viscosity and gel strength of the water-based drilling fluid. In the presence of electrolyte, or other flocculating compounds, bentonite particles may flocculate due to the net force between the negative charge on the surface of the plates and the positive charge of the edge of the bentonite or ions in the systems . Additioquebracho, tannate, Q-Broxin, lignosulfonate, and lignite  . In deepHigh temperature and high pressure influence the rheological and physical properties of the drilling fluids. As a results, the well pressure changes. In order to maintain the well pressure within the desired allowable drilling operational window, it is vital to design of the thermal and pressure resistance fluid. 2 impact on the thermal stable drilling fluids enhances the lubricity, electrical and heat conductivity properties without impacting the thermal stability rheological behavior of the base fluids. In terms of filtrate losses, several investigators reported that different nanoparticle reduced the filtrate loss [2 used in this paper did not show any reduction impact. The observations indicate that the performance of nanoparticles in filtrate loss reduction depends on the chemistry of the nanoparticle’s interaction with the base fluid. In this paper, the lignosulfonate treated base fluid and the MoS2 are highly negatively charges and the system might have been deflocculated/dispersed well. Therefore, the performance of MoS2 with respect to filtrate loss is valid only in the base fluid formulated in this paper. However, MoS2 could reduce the filtrate loss in other base fluid. In the future, the effect of MoS2 will be evaluated in different commercial and in-house formulated base fluids. The effect of hot rolling will also be investigated.In this paper, at first, bentonite based thermal stable drilling fluid was formulated by varying the lignosulfonate concentration in bentonite drilling fluid. Lignosulfonate is an important additive when design bentonite base drilling fluid for geothermal and high temperature oil and gas wells to control undesired clay flocculation. The right concentration of the flocculant is determined through experimental works. Despite the thermal stability, water drilling fluids have a lower lubricity as compared with the Oil based drilling fluid. In order to improve the thermal stabile drilling fluid with other properties, nanoparticles have been treated. In literature as reviewed, nanoparticles have shown an improved performances on the drilling fluids. In this paper, MoSate loss ,18,19,202 on improving the lubricity and heat conductivity can be reflected in terms of allowing to drill longer offset by reducing the drag as well as cooling the bit in order to reduces early bit damage, respectively. The engineering implication of MoSThe addition of 0.17 wt.% lignosulfonate maintains thermally stable H-B and Bingham plastic yield stresses.The 0.26 wt.% molybdenum disulphide additive enhanced the lubricity of the thermally stable base fluid by 27%. The nanoparticles maintain the thermal stability of the fluid and did not show a significant impact on viscoelasticity.2 nanoparticle solution increased the thermal and electrical conductivity of the base fluid by about 7.2% and by 8.8%, respectively.Addition of 0.26 wt.% MoSResults from the experimental study showed that:2 reported in this paper is valid only on the considered base fluid, the measurement temperature up to the 80 °C and atmospheric pressure conditions. By changing any one of these conditions, one may achieve different results.To sum up, the impact of the MoS


Lupus is a complex autoimmune disease that is difficult to diagnose and treat. It is estimated that at least 5 million Americans have lupus, with more than 16,000 new cases of lupus being reported annually in the United States. Social media provides a platform for patients to find rheumatologists and peers and build awareness of the condition. Researchers have suggested that the social network Twitter may serve as a rich avenue for exploring how patients communicate about their health issues. However, there is a lack of research about the characteristics of lupus patients on Twitter and their attitudes toward using Twitter for engaging them with their health care.This study has two objectives: (1) to conduct a content analysis of Twitter data published by users (in English) in the United States between September 1, 2017 and October 31, 2018 to identify patients who publicly discuss their lupus condition and to assess their expressed health themes and (2) to conduct a cross-sectional survey among these lupus patients on Twitter to study their attitudes toward using Twitter for engaging them with their health care.This is a mixed methods study that analyzes retrospective Twitter data and conducts a cross-sectional survey among lupus patients on Twitter. We used Symplur Signals, a health care social media analytics platform, to access the Twitter data and analyze user-generated posts that include keywords related to lupus. We will use descriptive statistics to analyze the data and identify the most prevalent topics in the Twitter content among lupus patients. We will further conduct self-report surveys via Twitter by inviting all identified lupus patients who discuss their lupus condition on Twitter. The goal of the survey is to collect data about the characteristics of lupus patients  and their attitudes toward using Twitter for engaging them with their health care.This study has been funded by the National Center for Advancing Translational Science through a Clinical and Translational Science Award. The institutional review board at the University of Southern California (HS-19-00048) approved the study. Data extraction and cleaning are complete. We obtained 47,715 Twitter posts containing terms related to “lupus” from users in the United States published in English between September 1, 2017 and October 31, 2018. We included 40,885 posts in the analysis. Data analysis was completed in Fall 2020.The data obtained in this pilot study will shed light on whether Twitter provides a promising data source for garnering health-related attitudes among lupus patients. The data will also help to determine whether Twitter might serve as a potential outreach platform for raising awareness of lupus among patients and implementing related health education interventions.DERR1-10.2196/15716 Lupus is a chronic disease characterized by an autoimmune response that can range in its frequency and affect any part of the body . It is estimated that at least 5 million Americans have lupus, with more than 16,000 new cases of lupus being reported annually in the United States [Lupus is a difficult disease to diagnose as its symptoms can often mimic those of other diseases . SystemiAs most people with lupus develop the disease between the ages of 15 years and 44 years, we hypothesize that social media provides a potentially promising tool for raising awareness and supporting early diagnosis and management of lupus. This study aims to shed light on the use of Twitter among patients who publicly discuss their lupus condition on the platform and to assess their attitudes toward using Twitter to engage them with their health care.The term “social media” describes widely accessible web-based and mobile technologies that allow users to view, create, and share information online and to participate in social networking -6. SociaThe social network Twitter is used by 23% of American adults, and users are diverse, including Hispanics (25%), Blacks (24%), and Whites 21%) . Twitter% . TwittThe emergence of social media has created new sources of analyzable data  and led However, previous research examined user-generated content about lupus on Facebook . The autThis study has two objectives: (1) to conduct a content analysis of Twitter data published by users (in English) in the United States between September 1, 2017 and October 31, 2018 to identify patients who publicly discuss their lupus condition and to assess their expressed health themes and (2) to conduct a cross-sectional survey among the lupus patients on Twitter to study their attitudes toward using Twitter for engaging them with their health care.Our findings will shed light on whether Twitter provides a promising data source for garnering insights and attitudes about lupus expressed among patients. The findings will help to determine whether Twitter might serve as a potential outreach platform for raising awareness of lupus among patients and implementing related health education interventions.This is a mixed methods study that analyzes retrospective Twitter data and conducts a cross-sectional survey among lupus patients on Twitter.This study will analyze user-generated posts in English that include keywords related to “lupus”  from theTwitter posts containing terms related to “lupus”  were obtThe following types of irrelevant tweets were excluded: (1) non-English language tweets identified using the Liu method , (2) retAs we attempt to understand attitudes, we relied on machine learning to identify Twitter posts by social bots or marketing-oriented accounts that could possibly influence the results and introduce bias ,42. We uStudy data were collected using the system REDCap (Research Electronic Data Capture) at the University of Southern California (USC). REDCap is a secure, web-based application designed to support data capture for research studies . All anaAny identifying and personal health information was redacted from the dataset by the coders. Since the “Tweet ID,” “Tweet URL,” “Profile thumbnail URL,” “Username,” and “Display Name” in the dataset can potentially identify the person directly, we removed these from the initial data collection sheet and used a unique code identifier instead. We maintained the link between the unique code and the identifiable elements in a separate file. We retained the data only for use in this project and destroyed the identifiable  information prior to the data analysis as requested by the local IRB.The data will be retained in a secure database called REDCap at USC. The anonymous data will be kept for future research. Individuals are informed that they should not participate in the study if they do not want their data kept.The proposed survey study involves contacting lupus patients in the United States who discuss their health in English on Twitter. Eligible survey respondents will be patients with lupus 18 years of age and older. To focus on feasibility, we will limit this pilot to lupus patients who discuss their health on Twitter. Other individuals who talk about how the condition affects a family member or friend  will be excluded from this study.The goal of the survey is to collect data about the characteristics of lupus patients  and their attitudes toward using Twitter for health care engagement among lupus patients . The full survey is included in We will conduct self-report surveys via Twitter by inviting all identified lupus patients who discuss their health on Twitter. We will recruit via the project Twitter account using a personalized message package approach  and replEligible lupus patients who are 18 years and older will proceed to the information sheet form and access the survey once they give consent via a check box in the survey form.Survey participants will be able to enter a raffle to win one of three US $100 gift cards after they complete the survey. We will use a standard coding approach for characterizing the Twitter messages and users. Two independent team members will use a range of text classifiers  to identThe analyses will rely on public, anonymized data, adhered to the terms and conditions, terms of use, and privacy policies of Twitter. This study was performed under IRB approval from the authors’ university. No Twitter posts will be reported verbatim in the report of the findings to protect the privacy of the users. Representative examples of tweets within each category will be selected to illustrate additional themes and will be shown as paraphrased quotes.P=.05 for statistical tests.We will use descriptive statistics to analyze the data and identify the most prevalent topics in the Twitter content. Units of analysis will be unique terms in posts as well as the number of Twitter messages and users. For each analysis, we will present findings in a confusion matrix where the diagonal line indicates the prevalence of a topic and the off-diagonal lines indicate topic overlap. The number of posts containing 2 or more topics is found at the intersection of the matrix for these topics. We will further describe the patient characteristics such as age, gender, race/ethnicity, and other characteristics and survey responses. We will use multiple regression to assess which variables  are significantly associated with acceptance of using Twitter for health care engagement. Analyses will be performed in SPSS (v.24), using The sample size estimate (and survey protocol) is based on previous similar research that demonstrated the usefulness of user data to identify and engage cancer patients on Twitter .Twitter data from users in Los Angeles County posted over the course of 12 months were used to identify 134 cancer patients who had discussed their cancer condition on Twitter. Nearly one-quarter  of them responded positively to the outreach on Twitter that was focused on clinical trial recruitment. As the prevalence of SLE is lower in the United States, with 20-150 reported cases per 100,000 , compareThis study presents minimal-risk research. We will use public data from the social network Twitter. We will de-identify any subject’s names or Twitter handles, and they will not appear in the analysis dataset. We have implemented a number of measures to ensure data security and confidentiality . We will further abide by USC IRB regulations and the USC Privacy of Personal Information policy. In general, all data will be entered into a computer and database that are password protected. The data will be stored using appropriate, secure computer software and encrypted computers.The study authors plan to publish the study findings in a peer-reviewed journal and at topic-related conferences (to be determined at a later date). All listed authors or contributors are compliant with guidelines outlined by the International Committee of Medical Journal Editors for author inclusion in a published work. Furthermore, to support research transparency and reproducibility, we will share the de-identified research data after publication of the study results. We will share the de-identified data on Figshare, a repository where users can make all of their research outputs available in a citable, shareable, and discoverable manner.This study was approved by the IRB at USC , college graduates (32%), and located in urban areas (26%) . NonetheFinally, we will take several steps to reduce the chance of fraudulent survey responses on Twitter, including sharing the survey link only via private messages on Twitter once a user has followed the project account on Twitter. In the case that the majority of users seems reluctant to follow the Twitter account, we will send reminder messages with the personalized link to the survey via a public reply message on Twitter to increase the survey response rate.This pilot project will provide preliminary data and practical insight into the application of publicly available Twitter data to gain a better understanding of lupus patients who publicly discuss their condition on Twitter and their attitudes toward using the platform to engage them with their health care. The data will also help to determine whether Twitter might serve as a potential outreach platform for raising awareness of lupus and implementing related health interventions.


Osteopathic principles and philosophy suggest the use of osteopathic manipulative treatment (OMT) to restore, augment, or facilitate lymphatic fluid flow to maintain body fluid balance, and/or to stimulate immune system responses to aid in the recovery from illness and maintain normal body defense mechanisms. This review provides an osteopathic view of the role of the lymphatic system in health and disease, with an emphasis on the use of OMT to alleviate somatic dysfunctions (SD) that inhibit the optimum function of the lymphatic system. The current evidence base is reviewed for the use of OMT to assist in restoring or augmenting lymphatic system function to help patients recover from illness and maintain health and wellness. An overview is provided on how osteopathic principles and philosophy relative to the immune system are applied in practice. A literature search was conducted using databases such as Medline, PubMed, Ostmed-DR, and Scopus, focusing on osteopathic approaches to the lymphatic system. Keywords used included osteopathic manipulative medicine, OMT, and lymphatic manual treatment or therapy. Current osteopathic textbook information was also surveyed. There is support for the application of osteopathic principles and OMT for certain conditions that involve the lymphatic system. More prospective research is needed. Osteopathic physicians (DOs) have long recognized how important the lymphatic system is for the maintenance of health. Andrew Taylor Still, the founder of the osteopathic profession, saw the lymphatic system as an absolutely indispensable component of the body and urged physicians not to neglect the system in their treatment of patients. For example, he stated, “We lay much stress on the uses of blood and the powers of the nerves, but have we any evidence that they are of more vital importance than the lymphatics? ... the system of the lymphatics is complete and universal in the whole body” . He furtDefinitionThe lymphatic system functions as a part of the circulatory system maintaining fluids in the body at balanced levels and as a part of the immune system by playing a role in the body’s defense system against infections. The lymphatic system is comprised of lymphoid organs, lymph tissues, lymph ducts, lymph capillaries, and lymphatic vessels that transport lymph and miscellaneous materials throughout the body . With thAnatomy of the lymphatic systemThe lymphatic system is comprised of lymphatic vessels, lymphoid organs, and lymphatic fluid. DOs utilize certain OMT procedures to stimulate the lymphoid organs, such as the liver and spleen, to enhance the body’s immune system functions to help the patient recover from certain disease states -7. OtherLymphatic VesselsThe lymphatic vessels consist of the following structures: lymphatic capillaries, pre-collecting vessels, collecting vessels, lymphatic trunks, and lymphatic ducts.Lymphatic CapillariesThe lymphatic capillaries’ structure ensures that they can permit the flow of interstitial fluid to absorb proteins and other large molecules Figure  14]. Th. Th14]. Pre-Collecting VesselsAfter the fluid from the interstitium travels through the lymph capillaries or "initial lymphatics," the fluid then travels to the pre-collecting vessels . Like thLymph CollectorsIn the same way that the pre-collecting vessels are seen as comparable to capillaries, lymph collectors are similar to the veins Figure  18]..18].The lymphatic pathways have their own intrinsic active capacity to transport the lymph. Actin filaments have been found in the lymphatic vessels (into smooth muscle cells), which are able to contract and create their own tone. Actin exhibits a transient but constant depolarization due to calcium-dependent chloride currents. This contraction may depend on the sensitivity of actin to feel flow variations (shear stress); the vessels have nerve endings, particularly sympathetic endings, which regulate the contraction of actin filaments .The major differences are that the lymph collectors have less space between their valves and the lymph collectors have slimmer walls than the walls found in veins. The direction of the fluid flow is determined by the passive movement through their one-way valves which prevent backflow of fluid and maintain transport to regional lymph nodes proximally . The lymThe lymphangion is the name given to the portion of the collector between a distal and a proximal valve. It is the smallest functional unit of the lymph collector . The lymLymphatic Trunks and DuctsThe lymphatic trunks and ducts are the largest of all the lymphatic vessels. While the trunks collect excess fluid from the organs and structures in their localized quadrants, the ducts serve as the vessels that return the lymph back into venous circulation Figure  19,22].,22.19,22There are a total of five lymphatic trunks in the lymphatic system. The five lymphatic trunks are as follows: (1) Jugular lymph trunks, which drain lymph from the cervical lymph nodes of the neck; (2) subclavian lymph trunks, which drain lymph from the apical lymph nodes in the armpit; (3) bronchomediastinal lymph trunks, which drain lymph from the regions around the chest including the lungs, heart, and mammary glands; (4) lumbar lymph trunks, which drain lymph from the legs, pelvic region, and kidneys; (5) intestinal lymph trunk which receives chyle from the intestines .There are two lymphatic ducts in the body known as the right lymphatic duct and the thoracic duct . The thoThe glymphatic systemHistorically, scientists were not able to identify a lymphatic system in the brain. Intuitively, however, it seemed implausible that the brain would not have a system for the removal of waste products. Finally, in 2012, just such a system was discovered. The name “glymphatic” was given to describe this system. The term is a combination of the words “lymphatic” referring to the function of the system, and “glial,” referring to the type of cells that surround the perivascular spaces through which the cerebrospinal fluid (CSF) flows .CSF enters the Virchow-Robin space (VRS) which surrounds the walls of paravascular arteries that serve as the blood supply for the brain parenchyma. The exchange of interstitial fluid (ISF) is facilitated by cerebral arterial pulsations and aquaporin-4 (AQP4) water channel proteins . CSF floThis knowledge of the glymphatic system is of great interest to osteopathic physicians (DOs). DOs have commonly used OMT to remove impediments, or augment lymph flow, and stimulate the body’s immune system defenses . KnowledLymphatic fluidLymph, also called lymphatic fluid, is normally a clear or yellowish liquid that is slightly less viscous than blood. As the blood circulates, some fluid is lost at the arterial capillary end and becomes interstitial fluid. This interstitial fluid is absorbed into, and removed by, the initial lymphatic capillaries. It continues to pass through a series of increasingly larger collecting vessels, lymph nodes, and ducts and is eventually returned to the blood. Once the interstitial fluid makes its way into the collecting duct, it is referred to as lymph .Lymph consists of water and other substances. These other substances include colloids and minerals, proteins, lipids , cells , waste products, and other foreign substances. Still, other substances may include (in varying amounts) damaged cells, nutrients, cancer cells, viruses, and bacteria ,31.Lymphoid organsThe following structures are considered lymphoid organs Figure  32]: th: th32]: Physiologic functions of the lymphatic systemThere are three major functions attributed to the lymphatic system. The first major function of the system is the reabsorption and return of excess interstitial fluids into the circulatory system . The lymPhysiologic circulation of the lymphatic fluid (lymph flow)The circulation of the lymphatic fluid begins with the lymphatic capillaries or "initial lymphatics" located within the interstitial fluid space (IFS). The reabsorbed excess fluid, lymph, is then transported to the lymphatic collecting vessels, which contain one-way valves to prevent backflow of lymph. Simultaneously, the lacteals absorb fats and fat-soluble vitamins in the small intestine. Along the way, the lymph passes through one or more groups of regional lymph nodes, which filter the lymph. The next stop will be the lymphatic trunks, and then the left and right thoracic collecting ducts, and finally it enters the left and right subclavian veins, respectively .While in many ways the lymphatic system parallels the cardiovascular system, the lymphatic system does not have its own pump like the heart in the cardiovascular system. Movement in the lymphatic system requires that lymph is moved through the lymphatic system by two types of forces: extrinsic and intrinsic. Extrinsic forces include respiration, the contraction of skeletal muscles that surround the lymphatic vessels, and muscles that act as extrinsic pumps, namely the thoracoabdominal and pelvic diaphragms. Intrinsic forces include spontaneous, pacemaker-like contractions of the smooth muscle walls of larger lymphatic vessels, and stimulation of lymphatic vessel contractility through sympathetic innervation ,42.SD and its effects on the lymphatic systemThe crucial task attributed to the lymphatic system is the transport of lymphatic fluid from the interstitium to the central circulatory system. The energy for this process comes by way of pumping mechanisms that propel the lymph along with the lymphatic network of vessels. As mentioned above, these pumping mechanisms are extrinsic and intrinsic. The extrinsic pump produces cyclical compression and expansion of lymphatic vessels by such actions as joint movements, muscular contractions, myofascial flexibility, mechanical respiration, and postural changes. The intrinsic pump relies on the rhythmic, spontaneous contractions of lymphatic muscle, and neural modulation via the autonomic nervous system .SD is defined as “impaired or altered function of related components of the somatic (body framework) system: skeletal, arthrodial and myofascial structures, and their related vascular, lymphatic, and neural elements” . SDs thaIndicationsThe indications for use of OMT to influence the lymphatic system include, but are not limited to, diseases or conditions that demonstrate some measure of edema, tissue congestion, or lymphatic or venous stasis. Some specific indications are acute SDs, sprains/strains, pregnancy, infection, inflammation, and pathologies with significant venous and/or lymphatic congestion ,44.ContraindicationsClinical judgment must be used in employing lymphatic techniques, paying attention to the patient’s diagnosis, clinical condition, and current medical therapy. These factors influence the choice of the appropriate technique, dose, duration, and frequency of treatment.Absolute contraindications include anuresis if not on dialysis, due to potential risk of pulmonary edema, necrotizing fasciitis (in the area involved), and lack of patient consent and/or cooperation.Relative contraindications entail the inability of congestive heart failure (CHF) patients to tolerate excessive preload, chronic obstructive pulmonary disease (COPD) due to increased residual volume post-treatment, acute asthma exacerbation due to the risk of increasing reflexive bronchospasm, unstable cardiac conditions, and cancer (immune system activation vs. lymphatic spread). As of today, there are currently no risks associated with lymphatic treatment for patients with cancer, but this area is still under study . Most auOther contraindications include osseous fracture or crushed tissue (in the area involved), bacterial infections with risk of dissemination, chronic infections with risk of reactivation , diseased organ , pregnancy , circulatory disorders , untreated coagulopathies or patients on anticoagulants (modify the technique to be less vigorous), and finally acute herniated nucleus pulposus ,44.Safety and efficacyThe safety and efficacy of osteopathic lymphatic treatments have been demonstrated in many published studies without any reports of significant complications ,44. It iDiagnostic principles for SD affecting the lymphatic systemTo determine the presence of SD and any effects upon the lymphatic system, a systematic approach is helpful. This approach includes the following elements ,44.History and Physical ExaminationAs in any medical or surgical situation, a thorough history and physical examination are essential. The physical examination should give special attention to any evidence of areas of swelling or puffiness, infection, inflammation, organ dysfunction or disease, shortness of breath, and tissue trauma.Terminal Lymphatic Drainage SitesPalpable tension, tenderness or ticklishness, or full, boggy tissue texture changes in the regional terminal lymphatic drainage sites are particularly useful in determining if regional tissue congestion exists Figure .Central myofascial pathways palpate anatomical transition zones for the presence of fascial restrictions which may inhibit lymphatic flow.Fluid pumps determine whether the physiologic motion of any of these muscular, fascial, and/or membranous regions is restricted. The cranial dura membranes, especially the tentorium cerebelli , the supraclavicular fossa region (often referred to by DOs as the “thoracic inlet”), the thoracoabdominal diaphragm, and the pelvic diaphragm.Spinal InvolvementSDs of the spinal vertebrae, rib cage, and sacrum result in reduced biomechanical efficiency, and increased tension in their associated muscular and connective tissue structures, all of which may contribute to less optimum lymphatic flow.Peripheral PathwaysAfter evaluating terminal lymphatic drainage sites, palpate other regional and local tissues to evaluate the presence of congestion and excess fluid in the interstitial tissues. This helps identify any regions that might benefit from local fluid techniques.Treatment principles for SD affecting the lymphatic systemEfficient and effective treatment for SD that hinders optimum lymphatic system function must be based on clear treatment goals. Such treatment goals will guide the physician to the most appropriate therapeutic interventions for any given patient, including the use of OMT to help ensure optimum lymphatic system function. Treatment protocols based on clear therapeutic goals should aim to reduce impaired lymphatic fluid movement; enhance the mechanical process of respiration, by reducing the work of breathing and increasing the efficiency of air and fluid flow influenced by this respiratory pump; enhance the movement of lymph and lymphocytes from local, regional, and peripheral tissues that need decongestion .In theory, all OMT techniques can help to optimize lymphatic system functioning through changes in muscle tone, alleviation of myofascial restrictions, modulation of neural reflexes, maintenance of balanced autonomic tone, and beneficial effects on respiration .To establish a rational therapeutic osteopathic approach, DOs use a clinical reasoning process known as the Five Models of Osteopathic Patient Care. The specific models are biomechanical, neurological, respiratory-circulatory metabolic-nutritional, and behavioral-biopsychosocial . Using tThe use of a treatment protocolSome DOs prefer to use a more protocol-based approach to the use of OMT to treat SDs that affect the lymphatic system. This method allows for a structured approach to determining which components of the lymphatic system need to be restored to more optimum function, and in what order. In general, treatment addresses the more central components of the lymphatic system and then progresses to the more regional and peripheral components. The principles involved in this approach are : treat tFrom the time of its founding, the osteopathic profession recognized the important role of the lymphatic system. The diagnosis and treatment of the lymphatic system were vital for the maintenance of health and the treatment of disease. This paper has presented a more in-depth exploration of these concepts, demonstrating that the application of osteopathic principles and practices to help restore and enhance immune system functions and lymphatic fluid flow can be an important factor in disease prevention, recovery from illness, and the maintenance of health and wellness.


Klebsiellae are Gram-negative pathogens responsible for serious nosocomial and community-acquired infections. Carbapenem resistance, both intrinsic and acquired, complicates therapy. Taniborbactam  is a bicyclic boronate β-lactamase inhibitor (BLI) that inhibits all four Ambler classes of β-lactamase enzymes, both serine- and metallo-, with the notable exception of class B IMP β-lactamases. Taniborbactam is currently undergoing phase 3 clinical trials in combination with cefepime  as part of the β-lactam-BLI (BL-BLI) combination FEP-taniborbactam (FTB).Figure 1. Structures of taniborbactam and cefepime. The β-lactamase inhibitor is in red and the β-lactam antibiotic is in black.Klebsiellae (CRK) strains collected as part of the Antibiotic Resistance Leadership Group (ARLG) Consortium on Resistance against Carbapenems in Klebsiella (CRACKLE) study. Among these strains, 193 expressed class A KPCs, one expressed a class B NDM, and six expressed class D OXA-48 or variants. Broth microdilution minimum inhibitory concentrations (MIC)s were determined using the ThermoFisher Sensititre system with custom assay panels. American Type Culture Collection strains were used for quality control. The susceptible-dose-dependent breakpoint for FEP was provisionally used for FTB, where taniborbactam was fixed at 4 µg/mL.We determined the activity of FTB against 200 carbapenem-resistant Klebsiella strains tested, susceptibility for β-lactams alone ranged from 1% for ceftazidime (CAZ), 2.5% for meropenem, and 13.5% for FEP (Table 1). The addition of BLIs increased % susceptibility compared to BL alone to: 98% for CAZ-avibactam (CZA); 95.5% for MEM-vaborbactam (MVB); and 99.0% for FTB. MIC50 and MIC90 were in the susceptible and provisionally susceptible range for CZA and MVB, and in the provisionally susceptible range for FTB. Analyzing the CZA and MVB non-susceptible strains, 7 of 9 MVB non-susceptible strains and 2 of 4 CZA-resistant strains were provisionally susceptible to FTB.Among the 200 Table 1. MIC50 and MIC90 values (μg/mL) and percent susceptibility for Klebsiella pneumoniae strains (n=200). AMK, amikacin; CST, colistin; CAZ, ceftazidime; CZA, ceftazidime-avibactam; FEP, cefepime; FTB, cefepime-taniborbactam; MEM, meropenem; MVB, meropenem-vaborbactam; TGC, tigecycline. * The breakpoint for CST is intermediate, as no susceptible breakpoint is available. ** The susceptible-dose-dependent breakpoint for FEP alone was provisionally applied to FTB, where taniborbactam was fixed at 4 μg/mL. Breakpoints from CLSI M100, 31st ed, 2021.The addition of taniborbactam restored susceptibility to FEP in 99.0% of CRACKLE isolates studied, comparable to CZA and MVB. Taniborbactam also restored FEP activity against some MVB- and CZA-resistant strains. FTB may provide a promising therapy for CRK infections.Robin Patel, MD, 1928 Diagnostics (Consultant)BioFire Diagnostics (Grant/Research Support)ContraFect Corporation (Grant/Research Support)Curetis (Consultant)Hylomorph AG (Grant/Research Support)IDSA Infectious Diseases Board Review Course Mammoth Biosciences (Consultant)NBME Netflix (Consultant)Next Gen Diagnostics (Consultant)PathoQuest (Consultant)PhAST (Consultant)Qvella (Consultant)Samsung Selux Diagnostics (Consultant)Shionogi & Co., Ltd. (Grant/Research Support)Specific Technologies (Consultant)TenNor Therapeutics Limited (Grant/Research Support)Torus Biosystems (Consultant)Up-to-Date  Robin Patel, MD, BioFire  Involved: Self): Grant/Research Support; Contrafect  Involved: Self): Grant/Research Support; IDSA  Involved: Self): Editor’s stipend; NBME, Up-to-Date and the Infectious Diseases Board Review Course  Involved: Self): Honoraria; Netflix  Involved: Self): Consultant; TenNor Therapeutics Limited  Involved: Self): Grant/Research Support; to Curetis, Specific Technologies, Next Gen Diagnostics, PathoQuest, Selux Diagnostics, 1928 Diagnostics, PhAST, Torus Biosystems, Mammoth Biosciences and Qvella  Involved: Self): Consultant David van Duin, MD, PhD, Entasis (Advisor or Review Panel member)genentech (Advisor or Review Panel member)Karius (Advisor or Review Panel member)Merck Pfizer Qpex (Advisor or Review Panel member)Shionogi Utility (Advisor or Review Panel member) Vance G. Fowler, Jr., MD, MHS, Achaogen (Consultant)Advanced Liquid Logics (Grant/Research Support)Affinergy Affinium (Consultant)Akagera (Consultant)Allergan (Grant/Research Support)Amphliphi Biosciences (Consultant)Aridis (Consultant)Armata (Consultant)Basilea Bayer (Consultant)C3J (Consultant)Cerexa Contrafect Debiopharm Destiny (Consultant)Durata Genentech Green Cross Integrated Biotherapeutics (Consultant)Janssen Karius (Grant/Research Support)Locus (Grant/Research Support)Medical Biosurfaces (Grant/Research Support)Medicines Co. (Consultant)MedImmune Merck (Grant/Research Support)NIH (Grant/Research Support)Novadigm (Consultant)Novartis Pfizer (Grant/Research Support)Regeneron sepsis diagnostics Tetraphase (Consultant)Theravance Trius (Consultant)UpToDate Valanbio xBiotech (Consultant) Daniel D. Rhoads, MD, Becton, Dickinson and Company (Grant/Research Support) Michael Jacobs, MBBS, Venatorx Pharmaceuticals, Inc. (Grant/Research Support) Focco van den Akker, PhD, Venatorx Pharmaceuticals, Inc. (Grant/Research Support) David A. Six, PhD, Venatorx Pharmaceuticals, Inc. (Employee) Greg Moeck, PhD, Venatorx Pharmaceuticals, Inc. (Employee) Krisztina M. Papp-Wallace, Ph.D., Merck & Co., Inc. (Grant/Research Support)Spero Therapeutics, Inc. (Grant/Research Support)Venatorx Pharmaceuticals, Inc. (Grant/Research Support)Wockhardt Ltd.  Robert A. Bonomo, MD, entasis (Research Grant or Support)Merck (Grant/Research Support)NIH (Grant/Research Support)VA Merit Award (Grant/Research Support)VenatoRx (Grant/Research Support)


Although Pinterest has become a popular platform for distributing influential information that shapes users’ behaviors, the role of recipes pinned on Pinterest in these behaviors is not well understood.This study aims to explore the patterns of food ingredients and the nutritional content of recipes posted on Pinterest and to examine the factors associated with recipes that engage more users.Data were collected from Pinterest between June 28 and July 12, 2020 (207 recipes and 2818 comments). All samples were collected via 2 new user accounts with no search history. A codebook was developed with a raw agreement rate of 0.97 across all variables. Content analysis and natural language processing sentiment analysis techniques were employed.taste more heavily than complexity  and health .Recipes using seafood or vegetables as the main ingredient had, on average, fewer calories and less sodium, sugar, and cholesterol than meat- or poultry-based recipes. For recipes using meat as the main ingredient, more than half of the energy was obtained from fat . Although the most followed pinners tended to post recipes containing more poultry or seafood and less meat, recipes with higher fat content or providing more calories per serving were more popular, having more shared photos or videos and comments. The natural language processing–based sentiment analysis suggested that Pinterest users weighted Although popular pinners tended to post recipes with more seafood or poultry or vegetables and less meat, recipes with higher fat and sugar content were more user-engaging, with more photo or video shares and comments. Data on Pinterest behaviors can inform the development and implementation of nutrition health interventions to promote healthy recipe sharing on social media platforms. Healthy eating patterns and their effect on disease prevention have been demonstrated robustly across the scientific literature -9. The USocial media has become a new and efficient way to distribute and consume influential information that shapes people’s dietary behaviors ,14. Sevepins) and upload it to the board (a collection of pins from different users) [Pinterest, launched in 2010, is a unique social media platform where users can save images (t users) . It has t users) , and ovet users) . PintereTo the best of our knowledge, no prior study has evaluated the recipe content on Pinterest. This study provides a first glimpse of this domain to advance the understanding of the relationship between social media use and dietary behavior. We aim to achieve the following 2 goals. First, we aim to examine the patterns of food ingredients and nutrients prescribed by recipes posted on Pinterest. Second, by employing both traditional content analysis and a natural language processing (NLP) technique, we sought to understand the factors that distinguish the most popular recipes among users.recipe, breakfast, lunch, and dinner were used to identify samples on Pinterest. Pins with recipes containing all the required information were selected by scrolling down the search results page for each keyword. This approach was developed based on previous studies on Pinterest content [Data were collected between June 28 and July 12, 2020. Although there is no “rule of thumb” on how long the data collection should persist, we adapted a proper time frame based on previous literature that specifically focused on Pinterest ,22. All  content ,22. PinsFor the content analysis, following the 2015-2020 United States Department of Agriculture Dietary Guidelines, food ingredients were classified as dark green vegetables, red and orange vegetables, legumes (beans and peas), starchy vegetables, other vegetables, fruits, seafood, meats, poultry, eggs, nuts or seeds or soy products, dairy, oil, and butter . The rechealth, healthy, calorie, and cholesterol), taste , and the complexity of the recipe . Keyword searching was applied to assess sentiment of comments posted by Pinterest users.For the comment analysis, 3 keyword dictionaries were created with keywords related to health , whereas the number of comments indicates medium engagement (users created and shared such content) [Descriptive analyses were performed for each type of food ingredient and their corresponding nutrient content. In addition, the popularity of recipe ingredients was assessed by the number of recoded followers (presented in tertiles). The level of engagement for each recipe was also evaluated by categorizing comments and shared photos or videos into tertiles, with regard to the fat, sugar, and fiber content of the recipes. Comments and shared photos or videos were chosen as indicators of engagement based on prior literature that suggested that, in the context of Pinterest, the number of content) . High encontent) . To proccontent) . VADER rcontent) . VADER hThe bar charts in U shape. Although not statistically significant, this pattern indicated an upward trend in the number of shared photos or videos and comments received (between the first and second tertiles) when a recipe contained high sugar content, but the trend turned down when the recipe’s sugar level reached the third tertile. In terms of sugar, our data suggested that the number of comments was similar regardless of the fiber content. However, although not statistically significant, recipes containing the highest amount of fiber (third tertile) were less popular, having fewer shared photos and videos compared with those in the first or second tertile.taste related, 25.9% (141/544) were positive, significantly higher than negative  and neutral . The complexity of a recipe and its health attributes were commented on less frequently: less than 8% (225/2818) of the comments contained text related to complexity, and less than 3% (84/2828) of the comments contained text related to health. We found that taste and complexity were the most important factors in shaping Pinterest users’ sentiments.In this study, recipes posted on Pinterest were collected, analyzed, and compared for ingredients and nutrients. We found that, in most cases, recipes using seafood or vegetables as the main ingredient, for example, tuna salad , had, on average, fewer calories and less sodium, sugar, and cholesterol than meat- or poultry-based recipes, for example, crispy chicken wraps  and Mongolian beef . Recipes using meat as the main ingredient, for example, creamy herbed pork chops , provided more energy by fat. Although the most followed pinners tended to post recipes containing more poultry or seafood and less meat, recipes serving higher fat or providing more calories per serving were more popular, having more shared photos or videos and comments. Sentiment analysis based on text mining showed that Pinterest users, in general, valued taste more than health qualities when making comments or sharing photos or videos.With the sharp increase in the number of social media users, platforms such as Pinterest have become influential mechanisms to transform knowledge sharing and acquisition, including dietary choice . Accordisocial elites do not eat meat at all; they only eat vegetables or seafood. It is not surprising that celebrities or social influencers, such as those with many followers on Pinterest, embrace this elite social norm, considering that most Pinterest users are from high-income households [From the perspective of content providers, we found that the most popular pinners, by sharing recipes containing more seafood and poultry  with lesuseholds , which ruseholds . In theouseholds . Popularuseholds . Red meauseholds  and is auseholds ,36. Suchuseholds ,37. Pinnlike on a particular pin [tasty recipes will be disseminated quickly on social media through users’ social networks  and make their way into regular recipe rotations for more people [From the users’ perspective, they are often learners in pursuit of inspirational recipes . We founular pin . The socular pin . Our senular pin . Previouular pin ; therefoe people . Our finThere appears to be a discrepancy between what pinners posted and how users consumed information, leading to an opportunity for future health interventions via Pinterest. Previous studies have shown that social media interventions can have a positive effect on nutritional outcomes . StrategThis study had some limitations. First, because of the restrictions imposed by Pinterest, the content scrolling process is not automated. The manual data collection resulted in a relatively small sample size and a large margin of error. To address the issues related to the small sample size, we applied a machine learning technique to mine text from the comments. A total of 100 comments were randomly selected to assess sentiment error rates. We found that the error rate was 18%, which is better than the acceptable level used in previous studies by convention . Second,In this study, we used both content analysis and NLP techniques to analyze recipes posted on Pinterest. Seafood-based recipes and vegetarian recipes had fewer calories and less sodium, sugar, and cholesterol than meat-based recipes. Although the most popular pinners tended to exhibit more health consciousness by posting recipes with more seafood, poultry, and vegetables and less meat, recipes with higher fat and sugar content had higher user engagement, as demonstrated by the higher numbers of photo or video shares and comments. Population health could be improved with targeted interventions to address this disparity through efforts to enhance interest in and adoption of healthy recipes by Pinterest users.


Esophagectomy is the cornerstone in curative treatment for esophageal and gastroesophageal junctional cancer. Esophageal resection is an advanced procedure with many complications, whereof anastomotic leak is the most dreaded. This study aimed to monitor the microcirculation with microdialysis analysis of local lactate levels in real-time on both sides of the esophagogastric anastomosis in totally minimally invasive Ivor-Lewis esophagectomy.® Probe, along with the CliniSenz® Analyser  was utilized for measurements. Lactate levels from both sides of the anastomosis were analysed in real time, on site, by a transportable analyser device. Measurements were made every 30 min during the first 24 h, and thereafter every 2 hours for up to 4 days.Twenty-five patients planned for esophageal resection with gastric conduit reconstruction and intrathoracic anastomosis were recruited. A sampling device, the OnZurfAll probes could be positioned as planned and on the third postoperative day 19/25 and 15/25 of the esophageal and gastric probes, respectively, continued to deliver measurements. In total, 89.6% (1539/1718) and 72.4% (1098/1516) of the measurements were deemed successful. The average lactate level on the esophageal side of the anastomosis and the gastric conduit ranged between 1.1–11.5 and 0.8–7.0 mM, respectively. Two anastomotic leaks occurred, one of which had persisting high lactate levels on the gastric side of the anastomosis.® analyser system, in combination with the OnZurf® Probe was feasible and safe. Continuous monitoring of analytes from the perianastomotic area has the potential to improve care after esophageal resection.Application and use of the novel CliniSenz Esophageal cancer and cancer of the gastroesophageal junction (Siewert I and II) is the eight most common type of cancer globally and the incidence rate is rising , 2. The Anastomotic leak can be the result of technical error in anastomotic construction or caused by impaired healing, the latter considered to be a result of compromised perfusion of the anastomosis. Several attempts to assess the perfusion in the gastric conduit have been made using pulse oximetry , doppler® probe , has been developed. The principle of using surface dialysate has been validated on the small bowel [. The present study aimed to monitor the microcirculation in real-time on both sides of the esophagogastric anastomosis in totally minimally invasive Ivor-Lewis esophagectomy.Microdialysis is a method for measuring analytes in target organs and has been widely used in neurosurgery  as well ll bowel , liver [ll bowel  and the ll bowel  in experAll patients planned for esophageal resection with gastric conduit reconstruction and intrathoracic anastomosis at a tertiary referral centre were invited to participate in this pilot study.® Probe, a 150-cm-long microdialysis catheter equipped with a 15 mm dialyse membrane with a pore size of 10 kDa, along with the CliniSenz® Analyser  was utilized for measurements. The analyser featured an enzyme-based heat flow detection device incorporated on a microfluidic chip, which aided in avoiding substance interferences. The system enabled continuous and fast sampling of small volumes of either lactate, glucose or pyruvate, the first of which was analysed in the present study.A sampling device, the OnZurfAll operations were performed as minimally invasive two-stage procedures. A standard gastric mobilization and abdominal lymphadenectomy was performed in the supine position and the gastric conduit was constructed 4–5 cm wide by multiple firings of linear staplers from the incisura angularis to the top of the fundic dome. The right gastro-epiploic arcade and the right gastric artery were preserved. The patient was then placed in the prone position. After mobilization of the esophagus and completion of the thoracic lymphadenectomy, the specimen was resected and the gastric conduit pulled up to the level of the anastomosis. A side-to-side esophagogastric anastomosis was constructed with a 45-mm linear staple cartridge and hand-sewn closure of the remaining defect and subsequently wrapped with omentum. Finally, one 28-Fr active chest drain was introduced.® probes were introduced through an existing trocar and one of the probes was secured on the proximal side of the anastomosis between the esophagus and pleura. A second probe was secured on the serosal surface of the tip of the gastric conduit under the omental wrap. The probes were tested for functionality before and after placement in the thoracic cavity. In the first 8 cases, the probes were externalized along with, and taped to, the chest drains and in the subsequent 17 cases, they were passed through a separate trocar incision. Post procedure, all patients were extubated in theatre and upon arrival to the high-dependency unit, the two probes were connected to one CliniSenz® Analyser instrument each , also equipped with battery back-up. Staff were instructed to remove the probes from the measuring devices in case of an emergency. Following a routine chest CT with intravenous and peroral contrast on day 5, the probes were removed. CT-scans were scrutinized for anastomotic leak, pneumonia, pneumothorax , and subcutaneous emphysema .Systemic lactate levels were recorded during care at the high dependency unit. Lactate levels in the dialysates were analysed in real-time on site by the transportable analyser device. Measurements were made every 30 min during the first 24 h, after which the patients were transferred from the high dependency unit to ward level care. Following re-calibration of the CliniSenzAll lactate measurements were blinded to clinical personnel, and thus not used in the clinical management of patients. The postoperative lactate microdialysate measurements were considered successful if being within the prespecified range of 0.5–12 mM.No sample size calculation was performed due to the exploratory nature of the study. Data regarding clinical characteristics are presented as median with range. Average and standard error of lactate levels are presented as well as percentage of successful measurements.Ethical statement: The study was conducted according to the guidelines of the declaration of Helsinki and approved by the Regional Ethics committee in Uppsala (Dnr 2016/562).®  to the gastric conduit and esophagus were abandoned after the first patient due to technical issues and instead placement under the pleura and omental wrap, respectively, was employed.Twenty-five patients were included in the study, Table Patients stayed 18 14–24) hours in the high dependency unit. Systemic lactate levels were generally low, with a mean of 1.1 and a standard deviation of 0.5 mM. All, but two of the gastric probes (#9 and #25), started to deliver measurements when the system was activated. In retrospect, we believe that the two non-functioning probes were accidentally damaged when closing the incision site at the end of the procedure. In total, 89.8% (886/987) and 71.1% (604/850) of the half-hourly measurements were within the prespecified range from the esophageal and gastric probes, respectively. In individual patients, the average lactate level on the esophageal side of the anastomosis and the gastric conduit ranged between 1.1–8.0 and 0.8–5.3 mM, respectively. The measurements over time are demonstrated in Fig. 4 hours iSuccessful measurements at the ward were achieved during a period of 24–76 h. In 89.3% (653/731) and 74.2% (494/666) of these, the lactate levels were within the prespecified range, and ranging between 1.1–11.5 and 1.3–7.0 for the esophageal and gastric probe, respectively. On the morning of the third postoperative day (> 48 h), 76% (19/25) and 60% (15/25) of the esophageal and gastric probes, respectively, continued to deliver measurements. Figure p < 0.001).The rate of successful measurements was 82% overall, however, the functionality of the esophageal probe seemed superior . Lactate curves for these patients were plotted separately and one revealed persisting high surface lactate levels from the gastric conduit starting 4 h postoperatively . One patient had radiological signs of pneumonia.No emergencies requiring urgent removal of the probes from the measuring devices occurred. All probes could easily be removed as planned, without pain or any other immediate complications. The median length of stay was 10 days (range 8–55) and no 90-day mortality was observed. At follow-up, no local problems were noted at the externalization site of the probes.®-probe and direct analysis of lactate levels on both sides of the esophagogastric anastomosis was feasible and safe. In one out of two patients with anastomotic leak, high lactate levels, persisting throughout the measurements, were seen. Externalizing the probes through a separate incision seemed to reduce the risk for subcutaneous emphysema in absolute numbers.Application of the OnZurfThe clinical characteristics of the 25 studied patients  are in line with esophageal cancer care in Sweden . The opeTraditionally, microdialysis measurements have been performed in a more invasive manner, such as intracranial microdialysis in traumatic brain injury , 22. In The proximal probe was placed in a pocket in-between the mediastinal pleura and the esophageal wall and the gastric probe was placed under the omental wrap, without being fixated by sutures. The superior functionality of the esophageal probe might be a result of the snug fixation between the unopened mediastinal pleura and the untouched proximal esophagus.At the surgical ward, postoperative mobilization was not hindered by the probes to any greater extent. However, an increased catheter length would have been beneficial for increased mobility. The dedicated and specially constructed support walker worked adequately as well as the routine charging of the battery unit. However, a smaller battery-powered analysis unit would have facilitated the daily care of the patient at the surgical ward. Finally, the probes were not radiopaque enough to be visible on the postoperative routine CT-scans, a draw-back which should be corrected before launching the system into clinical practice.Although many authors describe the need for both lactate, glucose and pyruvate as well as various quotients of these variables, we could demonstrate high surface lactate values already during the first postoperative hours in one of the two patients having an anastomotic leak. Interestingly, the patient with high lactate levels in the gastric tube had an otherwise uneventful recovery the first postoperative days, consistent with the leak being a delayed result of early ischemia. Thus, this monitoring system has unique potential for detection, correction and follow-up of organ ischemia in the postoperative setting and might guide measures to improve oxygenation, rheology, end organ perfusion and other optimizable factors.® analyser in all patients. Also, the typical characteristics of the included patients, representing an average patient cohort with good postoperative outcome increase generalizability. A limitation is the sample size of the study, as there were not enough anastomotic leakages to fully evaluate the prognostic value of the system. Moreover, the choice not to suture the probes to the serosa of the studied organs could have increased the risk of catheter dislodgement, thus explaining some of the lacking measurements.Among the strengths of the present study is the direct collaboration with the manufacturer, being present in the operation theatre for the first procedures and setting up the CliniSenz® analyser system, in combination with the OnZurf® Probe, was feasible and resulted in firsthand clinical experience and several conceivable improvements. Most important, we are approaching the possibility of continuous evaluation of a high-risk intrathoracic anastomosis by an easy-to-use patient-monitoring system.In summary, the use of the novel CliniSenz


The XCTVTs harvested from NOD/SCID mice were then inoculated and grown in beagles and named mouse xenotransplantation of CTVTs (MCTVTs).Canine transmissible venereal tumours (CTVTs) can cross the major histocompatibility complex barrier to spread among dogs. In addition to the transmissibility within canids, CTVTs are also known as a suitable model for investigating the tumour–host immunity interaction because dogs live with humans and experience the same environmental risk factors for tumourigenesis. Moreover, outbred dogs are more appropriate than inbred mice models for simulating the diversity of human cancer development. This study built a new model of CTVTs, known as MCTVTs, to further probe the shaping effects of immune stress on tumour development. For xenotransplantation, CTVTs were first injected and developed in immunodeficient mice (NOD.CB17-PrkdcAfter the inoculation of CTVTs and MCTVTs into immune-competent beagle dogs separately, MCTVTs grew faster and metastasized more frequently than CTVTs did. Gene expression profiles in CTVTs and MCTVTs were analysed by cDNA microarray to reveal that MCTVTs expressed many tumour-promoting genes involved in chronic inflammation, chemotaxis, extracellular space modification, NF-kappa B pathways, and focal adhesion. Furthermore, several well-known tumour-associated biomarkers which could predict tumour progression were overexpressed in MCTVTs.This study demonstrated that defective host immunity can result in gene instability and enable transcriptome reprogramming within tumour cells. Fast tumour growth in beagle dogs and overexpression of tumour-associated biomarkers were found in a CTVT strain previously established in immunodeficient mice. In addition, dysregulated interaction of chronic inflammation, chemotaxis, and extracellular space modification were revealed to imply the possibly exacerbating mechanisms in the microenvironments of these tumours. In summary, this study offers a potential method to facilitate tumour progression and provide a niche for discovering tumour-associated biomarkers in cancer research.The online version contains supplementary material available at 10.1186/s12917-021-03093-4. Cancer is the leading cause of death in both humans and dogs, but current tumour treatments usually achieve undesirable therapeutic efficiency, especially in some knotty cancers. Syngeneic murine models or immunodeficient mice have allowed the engraftment of human xenotransplantation in cancer research for decades . HoweverCanine transmissible venereal tumours (CTVTs) are contagious round cell neoplasms that are frequently located in the external genitalia of both genders and are transmitted by the transplantation of cells during coitus . CTVTs cGenomic instability is a fundamentally important feature of almost all cancers, and the tumour microenvironment (TME) can also contribute to significant genetic changes in tumours . The gen8 CTVT cells in their backs for XCTVT preparation  mapping Fig. A, indicae < 0.05 . Total 1p < 0.05). KEGG pathway analysis revealed 6 significantly enriched KEGG pathways (p < 0.05). Most of the GO term and KEGG pathway analyses were related to “cell chemotaxis”, “chemokine activity”, “inflammatory response”, “NF-kappa B pathways”, “focal adhesion”, “pathways in cancer”, and other cancer-related functional pathways  [C5AR1, CXCL12, CCL19, and VCAM1, participated in numerous enriched functions, such as cell chemotaxis, regulation of the function of neutrophils, and NF-kappa B pathways, which potentially facilitated the growth of MCTVTs and establishment of the tumour-friendly microenvironment. The gene expressions of C5AR1, CXCL12, and CCL19  using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) for analysis. GO analysis of the DEGs identified 10 biological process (BP), 4 cellular component (CC), and 4 molecular function (MF) GO terms as significantly enriched (dex.jsp) , 18, theAPOC1 . Though the CTVT genome is highly conserved in canids [CTVTs are naturally occurring tumours that are transmitted during copulation as intact tumour cells and affect the external or internal genitalia in canids. CTVT is the oldest known somatic cell lineage, as it first affected a dog that lived about 11,000 years ago . As it hfor CTVT . Since Mn canids , we firsCCL19, CXCL12, and C5AR1, participated in the top 1 and 2  enriched biological processes, “cell chemotaxis ” and “positive regulation of neutrophil chemotaxis ”, as well as in several cancer-related KEGG pathways. Chemotaxis played a vital role in tumour formation and remodeling of the TME [CXCL12 maintained a 0.29 fold-change in MCTVTs, while loss of CXCL12 promotes tumour migration to the liver, bone marrow, and lung [CCL19 (which expressed a 2.6-fold-change in MCTVT) and C5AR1 (which expressed a 2.28-fold-change in MCTVT) is highly associated with the advanced tumour stage and promotes cell proliferation in several cancers, leading to poor patient outcomes [CCL19, CXCL12, and C5AR1, promote chemotaxis, tumour proliferation, and even metastasis, which are in line with previous studies and evidenced by the progressive MCTVT growth  patients, and a high level of APOC1 indicates a poor prognosis. In addition, APOC1 has been proven as a biomarker in CRC [MMP-1 contributes to invasion and metastasis in breast cancer, and it potentially serves as a diagnostic marker and target for breast cancer [KMO is a novel prognostic marker or oncogenic protein in human hepatocellular carcinoma [KMO is overexpressed in canine mammary tumours (CMT) and canine melanoma and plays an oncogenic role in tumour development [After surviving in the immunocompromised hosts, MCTVTs expressed numerous tumour-associated biomarkers Fig. , includir in CRC . Overexpt cancer . KMO is arcinoma  and triparcinoma , 40, sugelopment , 42. TheTumour biomarkers have become effective indicators of tumour categorization, malignancy, and prognosis . In addiCTVT is considered as one of the ideal animal models for cancer research, especially for investigating the interaction between host immunity and cancer progression. This study first demonstrates that host immune–tumour microenvironment interplay can remodel the transcriptomic profiles of CTVTs, thereby inducing the altered expressions of tumour-related biomarkers, which should serve as objective indicators for the prediction of tumour development or grading, and even be designed as therapeutic candidates for cancer immunotherapies. Though further evidence is needed, our data suggested that tumour-related biomarkers which were hidden by the cancers could be uncovered by just inoculating the cancer cells into immunocompromised mice. Collectively, the results provide a potential solution to discover tumour-related biomarkers for cancer research.8 freshly well-prepared tumour cells subcutaneously in their backs. The tumour volumes were measured with calipers twice per week and calculated as π × length × width × thickness / 4 (cm3). Tumours were classified into different stages based on the tumour growth curve. The P phase was defined as the tumour volume consistently increasing. As tumours achieved stability, they were regarded as the S phase. Last, the R phase was defined as tumours gradually shrinking in volume.Healthy 1- to 2-year-old beagles were purchased from Lasco Co., Ltd. (Taiwan) in this research. Dogs were fed with commercially available canine food  and offered water by automatic water suppliers ad libitum. All dogs were dewormed regularly and vaccinated against canine distemper, leptospirosis, parvovirus, and canine hepatitis. A spontaneous CTVT on the external genitalia of a male dog was used for the original transplantation. Tumours were processed following a protocol proposed by Hsiao et al. with modifications . Briefly8 viable CTVT cells on bilateral flanks. After sacrifice, the XCTVT tumours were harvested from these NOD/SCID mice and inoculated into others. In this study, mice were sacrificed due to ulcerations at the tumour site or tumour diameters exceeding 20 mm. After two serial passages in NOD/SCID mice, 1 × 108 viable XCTVT cells were grafted into beagles subcutaneously and then serially passaged in dogs as MCTVT. The transplantation methods were the same as those for CTVT described above. The MCTVT masses and cells were collected for further investigation. All beagles were maintained at the Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan, following the guidelines of the Institutional Animal Care and Use Committee (IACUC No. NTU98-EL-00002). Research was carried out in compliance with the ARRIVE guidelines.6- to 8-week-old NOD/SCID mice were obtained from the Laboratory Animal Center, College of Medicine, National Taiwan University, Taipei, Taiwan. All mice were kept in specific pathogen-free barrier facilities at the Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan, and subcutaneously inoculated with 1 × 10CTVT and MCTVT samples were surgically excised from tumour-bearing beagles, and XCTVT-bearing mice were euthanized with isoflurane and cervical dislocation to obtain their tumours. Single-cell suspensions were prepared from the tumour samples following the procedures described above. Tumour cell smears were stained with Diff-Quik stain  for subsequent cytological examination . For hisTotal RNA was extracted using TRIzol reagent  according to the manufacturer’s instructions. Briefly, tumour tissues were ground by mortar and pestle, resuspended in TRIzol reagent by vortex, and incubated at room temperature for 10 min. Following chloroform extraction, RNA was precipitated with isopropanol. The precipitated pellet was washed with 70% ethanol, dried in a vacuum chamber, and resuspended in diethylpyrocarbonate-treated water (DEPC-water). For Affymetrix analysis, total RNA samples were purified using RNeasy mini kits  following the manufacturer’s protocol. Briefly, total RNA was resuspended in 350 μl buffer RLT and 250 μl 99% ethanol was added to the homogenized lysate by pipetting. Six hundred microliter RNA suspension was placed into an RNeasy mini spin column installed with an RNeasy membrane sitting in a 2-ml collection tube and centrifuged at 10,000 rpm for 15 s. The column was washed with RPE with centrifugation. RNA adhered to the RNeasy membrane was eluted with 40 μl of RNase-free water at 10,000 rpm for 1 min and then analysed on 1% denaturing agarose gel.Canis familiaris probe sets for > 20,000 predicted genes. The GeneChip® was subsequently washed and developed according to the amplification staining protocol provided by Affymetrix and then scanned using an Affymetrix GeneChip® Scanner 3000. Affymetrix GeneChip® Operating Software, version 1.4, was used to quantify the expression levels of the genes represented on the GeneChip® Canine Genome 2.0 Array. Probe pairs were scored as positive or negative for the detection of a specific gene sequence by comparing signals from the perfect match and mismatched probe features. The number of probe pairs meeting the default discrimination threshold (= 0.015) was used to assign a call of absent (A), present (P), or marginal (M) for each assayed gene, and a P value was calculated. PCA reduced the dimensionality of the data  to three dimensions in order to visualize the similarity of the log-transformed expression ratios of the genes. For heatmap and hierarchical clustering, representative DEGs were chosen in gene expression profiles from triplicate CTVT and MCTVT samples.cDNA microarray was performed using GeneChip® Canine Genome 2.0 Array . For sample preparation, RNA was reverse-transcribed in the presence of a T7-(dT) 24 primer using a One-cycle cDNA Synthesis kit (Affymetrix). The cDNA product was then purified and transcribed in vitro with biotin-labelled ribonucleotides . The biotinylated DNA was then fragmented to 200 nucleotides or less, heated at 99 °C for 5 min, and hybridized for 16 h at 45 °C for the GeneChip® Canine Genome 2.0 Array, which contained 42,860 2 , 1 U/μL of DNaseI was added per 1 μg of RNA, and the mixture was incubated for 30 min at 37 °C. DNaseI activity was arrested by the addition of 1 μL of 25 mm EDTA , followed by incubation at 65 °C for 10 min. The RNA was denatured at 70 °C for 10 min and chilled on ice for 10 min, following which 4 μl of 5x first strand buffer , 1 μl of 10 mM dNTPs, 2 μl of 100 mM DTT , 1 μl of RNase-free water, and 2 μl of SuperScript II RT  were added to the RNA solution. The RT reaction was carried out at 42 °C for 2 h in a Mastercycler Personal . Quantitative PCR was performed (in triplicate) using SYBR Green PCR Master Mix  according to the manufacturer’s instructions and a qPCR machine . The primer sequences were designed using Primer 3 and are listed in additional file -(Ct of target gene – Ct of housekeeping gene), (2-DCT), in arbitrary units and analysed using IQ5 analysis software .RNA was digested with DNase I  to remove genomic DNA. DNaseI was dissolved in 10x reaction buffer  with MgClhttp://david.abcc.ncifcrf.gov/) to investigate the involvement of these genes in various BP, CC, and MF [https://www.gsea-msigdb.org/gsea/index.jsp) [https://software.broadinstitute.org/morpheus/) was illustrated to generate clustered heat map for comparison of the representative DEGs [http://string-db.org) was employed to evaluate the interactions among the identified DEGs [Identified DEGs were imported into DAVID  , 18 was ive DEGs , and theied DEGs . The resied DEGs . Moleculied DEGs . To obtaied DEGs .t-test (two-tailed) was used to determine the statistical significance of differences and a p value < 0.05 was regarded as a significant difference .All data are presented as mean ± SD. Student’s Additional file 1. The 136 up-regulated genes (≥ 2-fold) in MCTVTs in comparison with CTVTs.Additional file 2. The 37 down-regulated genes (≦ 0.5-fold) in MCTVTs in comparison with CTVTs.Additional file 3. Validation of Affymetrix data by quantitative PCR. Randomly selected genes were analyzed by quantitative PCR. The gene expression ratios (MCTVT to CTVT) of quantitative PCR and Affymetrix are consistent.Additional file 4. Sequences of primers used in quantitative PCR.


There was an error in the spelling of an author’s name; Juan M. Abofalia should be Juan M. Abolafia.The was also an error in one of the grant numbers in the funding statement for this article. The funding statement should read as follows:“This work was supported by EU H2020 Research and Innovation Programme under Grant Agreement No. 945539 (HBP SGA3), BFU2017-85048-R Spanish Ministry of Science to MVSV and by the project Async-Prop, Bournemouth University-IDIBAPS, RED11549, Partnering Project of the HBP SP3 (EU H2020 Research and Innovation Programme) to EB-B. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.”