To report laparoscopic resection of retroperitoneal paraganglioma close to the caudal vena cava in a dog.Case report.Twelve‐year‐old, neutered male Jack Russell terrier.The dog had undergone three previous cystotomies for bladder stones. On follow‐up ultrasonographic evaluation, a 14‐mm × 17‐mm tumour was incidentally detected in the dorsal midline of the caudal abdomen. The dog underwent computed tomography (CT) imaging and ultrasound‐guided fine needle aspiration of the tumour under general anaesthesia. CT imaging showed that the tumour was close to the caudal vena cava. There was no evidence of metastasis. Neuroendocrine tumour was suspected on cytologic examination. Based on these findings, laparoscopic tumour resection was performed using a vessel‐sealing device. The operation time was 136 minutes.The dog was stable after recovery from anaesthesia and discharged to home the next day. Histopathological diagnosis of the tumour was a paraganglioma. The dog remained without clinical evidence of recurrent tumour or metastasis for 670 days after the surgery.Retroperitoneal paraganglioma in dogs is uncommon, but it is one of the differential diagnoses of a retroperitoneal tumour. Laparoscopic resection of a retroperitoneal paraganglioma was successfully performed in the dog. Laparoscopic resection conferred the advantages over open surgery of being minimally invasive, providing better visualization of the surgical field through pneumoperitoneum and semisternal patient recumbency, and allowing for magnification of the operative field, which facilitated the ease and safety of the procedure. Laparoscopic resection of a retroperitoneal paraganglioma close to the caudal vena cava was successfully performed in a dog. The dog remained without clinical evidence of recurrent tumour or metastasis for 670 days after the surgery. Histopathological examination revealed primary hypoplasia of the portal vein. No clinical signs associated with primary hypoplasia of the portal vein had been observed.2.2Physical examination findings were unremarkable. Temperature, heart rate, and respiratory rate were 38.2°C, 102 bpm, 20 breaths per minute, respectively. On ultrasonographic examination, a 14‐mm × 17‐mm tumour was incidentally found near caudal vena cava (CVC) and abdominal aorta on the dorsal side of the abdomen.No other abnormal findings were detected. Based on the ultrasound findings, complete blood count, plasma blood chemistry panel, blood coagulation tests, abdominal ultrasonography, echocardiography, chest and abdominal radiography, and urinalysis were performed prior to anaesthesia. Plasma chemistry panel revealed mild increases in alanine aminotransferase concentration  and alkaline phosphatase concentration . Abdominal ultrasonography performed before general anaesthesia showed the retroperitoneal tumour, which was unchanged in size from 12 days earlier, as well as bladder stones. The results of other blood tests, echocardiography, radiography, and urinalysis were unremarkable.To further define the anatomical location of the tumour and to assess for metastasis, whole body computed tomography (CT) imaging was performed 12 days after the initial examination. Ultrasound‐guided fine needle aspiration (FNA) of the mass were also performed under general anaesthesia immediately after CT imaging.Prior to anaesthesia, a 22‐gauge peripheral intravenous catheter was placed in the cephalic vein. Anaesthesia for the CT and ultrasound‐guided FNA procedures began with midazolam (0.2 mg/kg) and butorphanol (0.1 mg/kg) administered intravenously as preanesthetic medications. Propofol 6 mg/kg was infused intravenously to effect to induce general anaesthesia, after which the dog was intubated with a 6‐mm diameter endotracheal tube. A surgical plane of anaesthesia was maintained with sevoflurane. CT  used a 16‐row, 32‐slice device.The dog was positioned in dorsal recumbency. Noncontrast CT of the head, chest, and abdomen was performed with a slice thickness of 1.25 mm. After the whole body was scanned, a contrast medium (iodine 300 mg/ml) was administered at 2 ml/kg intravenously over 15 s using a power injector . Immediately after iodine administration, images of the liver to the area around the tumour in the caudal abdomen were obtained in the arterial phase. The portal vein phase and equilibrium phase of the same area were obtained 40 and 120 s after the initiation of iodine administration, respectively. After completion of the CT scan, FNA of the retroperitoneal tumour was performed under ultrasound guidance using a 23‐gauge needle. Cytology slides were prepared and stained with Wright‐Giemsa stain.Based on CT images, the tumour (14 mm ×17 mm× 19 mm) was located on the midline caudal to the kidney and ventral to the abdominal aorta and caudal vena cava  was administered intravenously as a preanesthetic medication and meloxicam (0.2 mg/kg) was administered subcutaneously as an analgesic. Propofol (6 mg/kg) was infused intravenously to effect to induce anaesthesia. The dog was then intubated with a 6‐mm endotracheal tube and connected to a ventilator. Sevoflurane (2.0–3.0%) inhalation was used to maintain a surgical plane of anaesthesia. Intravenous constant rate infusion of fentanyl (5 μg/kg/hr) and ketamine (0.4 mg/kg/hr) were administered as analgesics during surgery. Ampicillin (25 mg/kg) was administered intravenously as a perioperative antibiotic.End‐tidal carbon dioxide was maintained at 35–45 mm Hg by adjusting the respiratory rate. The dog was placed in left lateral recumbency with the spine elevated to create a semisternal position by placing cushions and towels under the body so that the dog was tilted toward the side on which the surgeon was standing Figure  Mayhew .2  at 1.0 L/min, and the intra‐abdominal pressure was maintained at 8 mmHg. A 5‐mm telescope  was attached to the camera head, and the abdominal cavity was observed through the camera port. After confirming the presence of the tumour, a 5‐mm instrument port was placed both cranial and caudal to the camera port so that the three ports were aligned. The tumour was visualized close to the CVC and abdominal aorta, and the right ureter was identified near the CVC. A small incision was made between the tumour and the CVC using laparoscopic Metzenbaum scissors . The tumour was separated from the CVC by blunt dissection using Maryland‐type forceps , a dissector probe , and a vessel sealing device . Large vessels were elevated dorsally with a dissector probe to prevent iatrogenic injury. Bleeding from the tumour was controlled by compression and the vessel‐sealing device  was placed through the camera port using the modified Hasson technique. Pneumoperitoneum was induced by insufflation of COe Figure . The excAfter verifying the absence of bleeding, the abdominal cavity was de‐sufflated. The abdominal wall and subcutaneous tissue were closed with 3‐0 polydioxanone suture, and the skin was closed with 3‐0 nylon suture Figure . Bupivac2.4Histopathological examination of the resected tumour revealed alveolar clusters of polygonal cells Figure . On immu3Blood pressure and other vital signs were stable intraoperatively and after surgery. No complications occurred intra‐ or post‐operatively. The patient started eating several hours after the operation, did not seem to care about the surgical wound, and heart rate and respiratory rate were stable. Thus, no additional analgesics were required after surgery, and the dog was discharged home the following day. No post‐operative complications were observed. Follow‐up thoracic and abdominal radiograph and abdominal ultrasonography were performed at 1‐, 3‐, and 6‐month, 1‐year, and 1.8‐year post‐operative periods. No recurrence of the tumour or obvious metastatic findings were noted at the time of last follow‐up 670 days after surgery and the patient remined clinically well.4Retroperitoneal paragangliomas are considered rare as both primary and metastatic tumours and not often associated with biochemical changes, such as catecholamine release in dogs (Dorn et al., Surgical resection is the only treatment option for paragangliomas, and complete resection may be achieved depending on the tumour's location and size as well as the presence of metastasis in dogs (Carpenter & Childers, In humans, laparoscopic resection of retroperitoneal paragangliomas leads to less bleeding and shorter discharge time when compared to conventional open surgery (Ping et al., One report described the use of a semi‐sternal position for laparoscopic resection of a right adrenal tumour (Jiménez Peláez et al., Our laparoscopic approach may be an effective treatment option for resecting retroperitoneal tumours located near large blood vessels. One of the advantages of laparoscopic surgery is the magnified surgical field. Better visualization helps identify blood vessels so that they can be handled with greater caution, which minimizes bleeding from large blood vessels during tumour dissection (Naan et al., In previous reports, paragangliomas have caused sudden death in a dog (Obradovich et al., 5A paraganglioma should be considered as a differential diagnosis of a retroperitoneal tumour in dogs. As surgical treatment is recommended for paragangliomas with no evidence of local invasion or metastasis, laparoscopic resection could be reasonably considered based on a magnified surgical field and its minimally invasive nature. The recommended approach involves semi‐sternal position and insufflation to ensure a good laparoscopic field of view. Further research is necessary to investigate the criteria for defining resectability, the frequencies of functional or non‐functional tumours, how catecholamine secretion affects surgical resection, and the long‐term prognosis after surgery.The authors declare no conflicts of interest related to this report.Young Tae Park wrote the original draft and Tomomi Minamoto wrote the review and edited.https://publons.com/publon/10.1002/vms3.588.The peer review history for this article is available at SUPPORTING INFORMATIONClick here for additional data file.


Previous studies have used magnetic resonance imaging (MRI) to quantify the fluid in the stomach and small intestine of children, and the stomach, small intestine and colon of adults. This is the first study to quantify fluid volumes and distribution using MRI in the paediatric colon. MRI datasets from 28 fasted (aged 0–15 years) and 18 fluid-fed (aged 10–16 years) paediatric participants were acquired during routine clinical care. A series of 2D- and 3D-based software protocols were used to measure colonic fluid volume and localisation. The paediatric colon contained a mean volume of 22.5 mL ± 41.3 mL fluid,  in 15.5 ± 17.5 discreet fluid pockets (median 12). The proportion of the fluid pockets larger than 1 mL was 9.6%, which contributed to 94.5% of the total fluid volume observed. No correlation was detected between all-ages and colonic fluid volume, nor was a difference in colonic fluid volumes observed based on sex, fed state or age group based on ICH-classifications. This study quantified fluid volumes within the paediatric colon, and these data will aid and accelerate the development of biorelevant tools to progress paediatric drug development for colon-targeting formulations. Oral drugs are the most common form of drug administration due to both patient convenience and the favourable cost:benefit ratio ,2. DespiColon-targeting formulations are often designed to exploit local intestinal environment characteristics for drug release, such as pH or the presence of bacterial-derived metabolising enzymes ,12. AlthData on intestinal fluid  are necessary for in vitro and in silico models in drug development ,14,25,26As XR-formulations are associated with high inter-individual variability in vivo ,34, it iThere is a recognised knowledge gap about the amount and distribution of free fluid in the GIT of the paediatric population ,37,38, rThe free fluid in the GIT can be quantified by magnetic resonance imaging (MRI), a non-invasive tool that permits undisturbed visualisation of the GIT . In contTherefore, the aim of this study was to use MRI to locate and quantify the fluid volumes and number of pockets in the colon of a paediatric population.Our research group has access to an MRI databank showing the abdomen of 49 paediatric participants, which was used previously to quantify the volume of free fluid in the stomach and small intestine. This observational, retrospective study used the same MRI datasets to quantify the volume of free fluid in the colon ). Following exclusion criteria were applied to ensure the cohort was as healthy as possible: patients with acute abdomen (appendicitis or perforated viscus), malignant bowel disease, surgery , bowel wall thickening/stricture/fistula/abscess. The datasets originated from two sites: Birmingham Children’s Hospital (BCH) and University Hospitals Coventry and Warwickshire NHS Trust (UHCW). All participants were fasted overnight and in addition, the UHCW site required the children to ingest 500 mL of Oral Klean Prep (a macrogol-based osmotic laxative) 60 min before MRI acquisition. This enabled the study to include a fasted and a fluid-fed population. The MRI acquisition parameters are listed in Only datasets with good resolution and correct T2-weighted MRI sequence were included in the study; this was a total of 46 MRI acquisitions. A 2D- and 3D-model were used to quantify the fluid within the MRI datasets.Horos  to identImageJ to calculate the area of the marked fluid regions.Two software packages were used for the 2D-fluid volume determination :Horos [4Horos is an open-source code software program distributed free of charge under the LGPL license at Horosproject.org and sponsored by Nimble Co LLC d/b/a Purview in Annapolis, MD, USA. Fluid identification within the MRI slices was based on the intensity of cerebrospinal fluid (CSF) which is routine in interpretation of these images ,31,42,45Horos to identify and highlight the fluid pockets in the MRI dataset;Blender to remove artefacts and isolate and compute the fluid pockets.Two software packages were used to calculate the number of fluid pockets in addition to quantifying the free fluid volume via a 3D-model :Horos toIn order to measure individual fluid pockets within the colon, the MRI dataset after the CSF-based thresholding was converted into a 3D-model. Initially, the 3D-volume rendering feature within Horos was used A, which p-value (p) was smaller than 0.05. To investigate correlations, Pearson’s correlation tests were used. To compare between two groups (such as investigating feed state), independent sample t-tests were used. Where multiple groups were compared, a one-way ANOVA with Bonferroni correction was used. Where multiple measurements on the same participants were investigated (such as a difference in volume in the three colon segments), an ANOVA of repeated measures with a Greenhouse–Geisser correction and post hoc Bonferroni correction was used.SPSS  was usedThe participants MRI datasets were stratified into age groups according to the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) classifications ,52 Tabl with theThe paediatric colon contained an average of 22.5 (±41.3 standard deviation) mL of fluid in 15.5 (±17.5 standard deviation) discreet fluid pockets; the data were not normally distributed, and the median volume was 0.80 mL with a median number of 12 pockets .The majority of the fluid pockets were small ; a total of 90.4% of the pockets (557/616) were smaller than 1 mL, which accounted for 5.5% of the total fluid volume (51.5 mL/933.2 mL) in the database. The other 9.6% of the pockets (59/616) were larger than 1 mL and contributed 94.5% to the total fluid volume (881.7 mL/933.2 mL) in the database. The paediatric colon contained on average 3.6 ± 2.9 pockets larger than 1 mL .There was a large variation observed in the datasets, both in fluid volume and number and size of pockets. No colonic fluid was found in nine participants, whereas eight datapoints with a total fluid volume higher than 60 mL were classified as statistical outliers, however these were not excluded from the subsequent analysis . ANOVA of repeated measures with a Greenhouse–Geisser correction and post hoc Bonferroni correction showed that the ascending colon contained the most fluid (p < 0.001). No statistically significant difference was detected between the fluid volumes in the two other segments. However, in four MRI datasets  either the transverse or descending colon contained all the fluid (and thus none was present in the ascending colon).There was a trend for decreased fluid volume in the segments towards the distal colon B. The flp < 0.001); no statistically significant difference was found in the number of fluid pockets between the transverse and descending colon. Only the ascending colon contained fluid pockets larger than 1 mL, apart from one dataset where a pocket of 249.6 mL was observed that spanned the entire colon. A graphical representation of the distribution of the individual pockets in the entire colon with a volume of up to 25 mL is shown in The ascending colon contained the most fluid pockets compared with the other two segments , age was positively correlated (Pearson’s coefficient >0.65) to total and ascending colonic fluid volume  is considerably lower than that observed in adults  of healthy children aged 14–18 years is reported to be 227 mL (interquartile range 180 mL–263 mL) , whereasAs the generic ICH-classification is based on days/months/years after birth, stratification on a parameter that considers the physicochemical properties of GIT anatomy and contents might have been more appropriate, such as height, body surface area (BSA) or body mass index (BMI) as these are typically used for dose adjustments. However, such data were not collected from the participants. A limitation in this study is that data were only available from 49 patients, extracted from two tertiary centres with differing MRI acquisition protocols. Paediatric MRI small bowel is a scarce resource offered mainly in tertiary centres (UHCW and BCH). It is a specialist modality only performed on selected cases, following discussion at a small bowel multidisciplinary meeting and requires interpretation by specialists. Therefore, the number of paediatric small bowel available for scientific study is small when compared with their adult counterparts. Together, these factors led to a small sample size and even smaller sub-sample size, when comparisons were made based on fed-status, sex or age; in addition to this, the numbers of participants in these sub-groups were not well balanced that may introduce bias into the statistical analysis and interpretation from these groups. Although the sample size is statistically small, it is sizeable considering population type and complexity of the modality; as such, these data should be viewed as preliminary where a larger, well defined cohort is warranted to provide robust data that will lead to more conclusive claims.n = 12) were all 20–22 years old and healthy, to minimise variability in the dataset. The paediatric participants have potential underlying morbidities, resulting in variability which may have introduced bias into the data. The large variability in colon fluid volumes was also observed in studies on healthy adults via MRI quantification [As the influence of age was investigated by comparing four paediatric age groups, the sample size per age group was similar to sample sizes used in the previous studies on healthy adult volunteers. However, the demographics of the participants in the adult studies were closely controlled; for example, those in Murray et al.  (n = 12)fication ,42,58,59fication ,60.There were on average 3.6 ± 2.9 fluid pockets larger than 1 mL, mainly in the ascending colon. Similarly to adults, the majority of the fluid pockets were very small [n = 12), was age significantly positively correlated to fluid in the total and the ascending colon , which could be a result of several factors. Firstly, it was unlikely that Oral Klean Prep had an impact on the colon fluid volumes in the timeframe of this study, although an effect could not be excluded a priori, as literature is conflicting about when an ingested solution reaches the colon since this is subjected to inter- and intra-variability . In addiThe ascending colon typically contained the highest portion of fluid . CorrelaThere is substantial variation in the choice of software used to interrogate MRI datasets in order to calculate fluid volumes and pockets ,42,45,58This pioneering study quantified the free fluid in the paediatric colon, which is of great interest to the pharmaceutical industry  for paedThis study successfully quantified the fluid volumes within the paediatric colon. Two methods were employed to quantify fluid in MRI datasets and their robustness was demonstrated via cross-analysis between operators and methods.The small overall sample size and even smaller sub-population sizes meant that these data are preliminary, and a fuller cohort study is required to verify the findings presented here. The median fluid volume for the total paediatric colon (0.80 mL) is comparable to literature data for the adult colon, although the mean paediatric volume (22.48 ± 41.30 mL) is nearly double than the adult value standardly used, i.e., 13 mL. The high variability was also observed in adults. No overall correlation was detected between colonic fluid volumes and age, similar to results in the paediatric SI. Fed status, sex or age across the whole population did not influence the colon fluid volumes. The fact that a significant correlation was observed in the 2–5 years group  between age and fluid volumes in the entire and ascending colon warrants further investigation. No such correlation was observed for other age groups. Furthermore, the ascending colon contained the most fluid compared with the transverse and descending colon. This study demonstrates the feasibility of obtaining real-world data from MRI to inform physiologically based models, which minimises the burden to special populations.The novel output from this study will improve the physiological understanding of the paediatric colon, aid biopredictive in silico simulations and establishing novel, more accurate in vitro assays and thus support paediatric drug development that targets the colon, resulting in more age-appropriate medicines for the paediatric population ,72.


Ultrasound revealed a right liver tumour, segment VI, measuring at 56 × 53 mm. According to images of ultrasound and MRI, the diagnosis was mesenchymal hepatic sarcoma. The paediatric patient had surgery to remove the entire liver segment containing the tumour. Micropathological examination showed that the tumour was a MHL. The serum AFP level fell rapidly to near normal following the surgery. The MHL benign liver tumour with an atypical presentation caused a very high AFP level. This was a rare clinical case, and it was difficult to diagnose.Mesenchymal hamartoma of the liver (MHL) is a benign tumour that most commonly occurs in children. In most cases of MHL, the α fetoprotein (AFP) level is within the normal limits, only in a few cases, increased AFP has been described which usually causes misdiagnosis of hepatoblastoma. We report a case of a 3-month-old paediatric patient who was incidentally detected with a very high level of AFP, at 6388.4 ng ml This benign tumour usually occurs in children, from birth to 2 years.3The oldest MHL cases were reported in 1959 and 1966. Additional cases have been reported until 2012. In total, less than 200 cases have been reported in the English literature.4MHL is a benign developmental malformation composed of a mixture of loose mesenchymal tissue, bile ducts, connective tissue and hepatocytes with cysts formed either from degenerative areas of mesenchymal or from dilated bile ducts and lymphatics. It is not a true tumour.5The tumour is usually in the form of a large cyst, with macrolobulated margin, multiple septae or a dense tumour containing of small cysts.−1 have been reported in the medical literature.3In most MHL cases, α fetoprotein (AFP) levels are within the normal limits. Only in a few cases with increased AFP, it is easily confused with hepatoblastoma. Very few reports of AFP level above 6000 ng mlWe report a special clinical case of a MHL of a 3-month-old paediatric patient with very high AFP level.A 3-month-old male paediatric patient was with a normal birth to full term. He was breastfed by his mother and weighted at 7.2 kg. His mother was healthy and there was no known liver disease in his family.−1. AFP-L3 and PIVKA (DCP) were within the normal limits, (AFP-L3 <0.5% and DCP was at 15 mAU/mL)  .At that time, an abdominal ultrasound revealed a right hepatic lesion, located in the segment VI, measuring 56 × 54 mm. The mixed echo cystoid tumour was unclearly delineated, multi curvilinear, with a solid content, hyper-echogenicity, echogenic vasculature and liquid with many echogenic septa inside. The solid content had low stiffness around 6.6 kPa in real-time 2D shearwave elastography. .Contrast-enhanced MRI detected a right hepatic lesion in the segment VI, measuring 56 × 53 mm. The lesion had heterogeneous high signal intensity to the liver parenchyma in T2W, heterogeneous low signal intensity in T1W, heterogeneous gadolinium contrast opacification, peripheral enhancement of the arterial phase, progressive centripetally in the venous phase and late phase without filling the whole lesion. Delayed phase was taken through dynamic T1WI vibe pulse with contrast agent, which was recorded after 5–7 min from beginning of artery phase. .−1. AFP L3 and DCP remained within normal limits .5 MHL is rare in adults. Commonly, males are more affected in children while females are more affected in adults.1,6MHL is a rare disease. This benign tumour usually occurs in children, between birth and 2 years of age,3The oldest MHL cases were reported in 1959 and 1966. Additional cases have been reported until 2012. In total, less than 200 cases have been reported in the English literature.5These tumours often have many cysts, ranging in size from a few millimetres to more than 10 cm, and are heterogeneous filled with a thick fluid. However, they may also be solid or of mixed forms. Ultrasound, CT and MRI showed images of a large cystic liver tumour, with a scalloped border, multiple septa or a solid mass of small cysts. The solid tissue composition of the tumour was revealed after the contrast agent injection.7,8Radiographically, MHL is easily misdiagnosed as mesenchymal hepatic sarcoma. However, mesenchymal hepatic sarcoma is very rare, with clinical manifestations of abdominal pain, weakness, loss of appetite and weight loss. Mesenchymal hepatic sarcoma often shows signs of vascular invasion and biliary obstruction.−1 in 256 normal infants. AFP levels decreased by 50% after the first 5 days, then continued to decrease to 16–1995 ng ml−1 (average 178 ng ml−1) by 2 months of age and to 6–1045 ng ml−1 (average 80 ng ml−1) at 3 months of age. From 6 months to 2 years, AFP ranged from 0.8 to 87 ng ml−1 (average 8 ng ml−1).9AFP is a plasma glycoprotein that has a high level in fetal blood, is relatively high in newborns and gradually decreases to normal. M. E. C. Blohm et. al. analysed 524 samples who were aged from infants to 2 years at Düsseldorf University Hospital (Germany). At birth, the mean serum AFP level was 41687 ng ml−1 have been reported in the medical literature.3,10In most MHL cases, the AFP is within the normal range. Only a few cases demonstrated increased AFP, so it was easily confused with hepatoblastoma. Very few reports of an increase in AFP above 6000 ng ml11,12A.Habador et. al reported a case of mesenchymal hepatic hamartoma, which was diagnosed as hepatoblastoma before surgery. The author believes that mesenchymal hamartoma and hepatoblastoma are common liver tumours in children. They have similar geometrical and pathological features. If the serum AFP is measured in a patient, an accurate diagnosis can be made before surgery, since AFP is largely elevated in hepatoblastoma. When in doubt, serum AFP is a good guide for differentiating hepatoblastoma from mesenchymal hamartoma.−1. After 1 month, there was a decrease, but it was still at a high level at 2176.4 ng ml−1. AFP decreased rapidly in 1 month after the surgery removed the whole segment containing the tumour, at 74 ng ml−1. Two months after the surgery, AFP continued to decrease to near the normal limit at 44.4 ng ml−1. It showed that the increase in AFP in this patient was related to the benign MHL tumour. AFP L3 and PIVKA II (DCP) were in normal range before and after the surgery.In this case study, the AFP was very high. Before surgery, AFP level was 6388.4 ng ml13Increased AFP and AFP-L3 are indicators for more accurate diagnose of malignant liver tumours for the newborn.Mesenchymal hepatic hamartoma is a rare benign tumour that sometimes increases AFP level in the blood, causing a misdiagnosis of hepatoblastoma.MHL usually presents as a large cystic, multilobulated, multi-septae or solid tumour with small cysts inside. The solid tissues of the tumour were revealed after enhancement by a contrast agent injection.On imaging diagnose, MHL was misdiagnosed as mesenchymal hepatic sarcoma. To diagnose more accurately, we need to look for signs of vascular and biliary tract invasion in order to differentiate between MHL and mesenchymal hepatic sarcoma.It is very difficult to diagnose liver tumours in children. The diagnosis should rely not only on imaging results but also many other factors.In the event of a mismatch between the imaging results and the tumour markers, a liver biopsy should be performed.Before performing surgery, the patient should be monitored to ensure the malignant diagnosis is correct. If the tumour markers decrease, a diagnosis of benign tumour should prevail.


Spin—an intrinsic angular momentum carried by elementary particles—has long been thought to be exclusive to quantum mechanical systems. The spin of photons, for example, is associated with the vectorial nature of electromagnetic waves . In clasIt comes down to the question of whether the scalar theoretical framework is a complete description of acoustics in some scenarios. Fundamentally speaking, vector field theory is required for the accurate depiction of hydrodynamics, and some information about the degree of freedom of microscopic fluid motion might be lost when using the scalar-pressure acoustic equations. Indeed, most of the time, the vector potential in Stokes flows is regarded as an arbitrary quantity that does not participate in the sound emission and momentum transfer. Recently, researchers from the University of California, Berkeley and Tongji University observed the spinning motion of fluids in sound propagation for the first time , with brZitterbewegung, ‘trembling motion’ in German). Furthermore, it has been demonstrated that the concept of Bohm quantum potential in the Madelung fluid model is an equivalent way of showing the existence of spin and Zitterbewegung of particles, thus providing a physical interpretation of the non-classical potential. For a given fluid flow at position x,X and V ≡ Scientists have long been seeking to establish a connection between quantum mechanical wave functions and fluid dynamics. For example, a set of well-established Madelung equations that illustrate the link between spin and orbital angular momentums  can be ootential . Note thotential :(2)\docs reflects the fluid's intrinsic property and is origin-independent; this is in contrast to the orbital angular momentum, which generally requires a reference point to define: r is the position vector and the total angular momentum admits the following decomposition:One would immediately recognize that the second-to-last term is analogous to the Bohm potential in quantum mechanics, which reflects the non-classical spinning nature of the fluid motion, while the classical center-of-mass momentum mentclass2pt{minimThis implies that acoustic vortex beams are eigenmodes of the total angular-momentum operator instead of et al. offered new physical insights into the spinning-vector potential by evaluating the rotation angle on a coiled space meta-atom serving as an acoustic probe [In summary, Shi ic probe . In addiic probe . It alsoConflict of interest statement. None declared.


Tumour hypoxia is significantly correlated with patient survival and treatment outcomes. At the molecular level, hypoxia is a major driving factor for tumour progression and aggressiveness. Despite the accumulative scientific and clinical efforts to target hypoxia, there is still a need to find specific treatments for tumour hypoxia. In this review, we discuss a variety of approaches to alter the low oxygen tumour microenvironment or hypoxia pathways including carbogen breathing, hyperthermia, hypoxia-activated prodrugs, tumour metabolism and hypoxia-inducible factor (HIF) inhibitors. The recent advances in technology and biological understanding reveal the importance of revisiting old therapeutic regimens and repurposing their uses clinically. The presence of low oxygen concentrations, known as hypoxia, is a prevalent characteristic of the microenvironment of solid tumours . Tumour For over a century there have been consistent efforts to target tumour hypoxia to promote radiation and chemotherapy responses. However, due to technical challenges and/or the lack of understanding of tumour biology, most trials failed in the clinical setting. Therefore, there is still a strong demand to solve these unmet needs to alleviate tumour hypoxia. In this review, we focus on the historical approaches to targeting hypoxia and how these can be better applied in the current times, in combination with new advances in techniques and molecular targeted treatments. A summary of the approaches discussed in this review is depicted in The most relevant approach to overcome tumour hypoxia is to promote oxygen delivery. Hyperbaric oxygen (HBO) treatment consists of breathing 100% oxygen at 2–4 times the normal atmospheric pressure, which results in saturated haemoglobin and increased oxygen in the circulation ,6,7. WhiNicotinamide is a vitamin B3-derived molecule and its initial radiosensitising effect was thought to be mediated by the inhibition of the DNA damage repair (DDR) pathway specifically by blocking poly(ADP-ribose) polymerase 1 (PARP1), which is responsible for nucleotide excision repair (BER) . The comHyperthermia (HT) is a treatment based on applying heat (40 to 45°C) to tissues to above physiological temperatures. HT can be directly applied to treat tumours, but due to the technical difficulties in achieving cytotoxic temperatures, it is mainly used as an adjuvant therapy in combination with radiation or chemotherapy ,22,23,24Although HT has been used since ancient times to treat a variety of diseases. Its clinical use for cancer patients was introduced in the 1900s, which was accelerated by the development of the devices to heat tumours ,28. In pThe biggest clinical challenge when delivering HT is to achieve cytotoxic temperatures >43 °C) and to overcome thermotolerance ,43. It i °C and tMultiple studies suggest that tumour oxygenation is caused by increased blood flow, perfusion, and decreased oxygen consumption, specifically at mild HT temperatures (39–43 °C) ,51,52,53The effect of HT in DDR is another aspect that can be exploited for radiosensitisation. HT was reported to induce increases in ATM and γH2AX pathways as well as p53 expression ,63,64,65The modulation of the immune system by HT also suggests the potential of using HT as a systemic treatment. The increased augmentation of the major histocompatibility complex (MHC) class I, and the enhanced infiltration of NK cells, CD8+ T cells, and B cells were observed after HT ,68,69,70Taken together, mild HT has a strong potential as an adjuvant treatment to overcome tumour hypoxia. To obtain clearer insights into the relationship between HT treatment and tumour oxygenation at the molecular level, further investigation is required. Furthermore, the underlying mechanisms of heat resistance  need to be considered to identify therapeutic targets or to better stratify patient groups into benefitting from HT ,72,73,74In the 1980s, several studies proposed targeting hypoxia by developing hypoxia-specific cytotoxins, which are now known as hypoxia-activated prodrugs (HAPs) ,76. HAPsSince the 1960s, the discovery of a direct relation between the electron affinity of the sensitising agent and the degree of radiosensitisation led to the study of nitroaromatic compounds as potential hypoxia modifiers . These tThe first nitroaromatic drugs, which were tested as radiosensitisers, were metronidazole and misonidazole. These nitroaromatic molecules showed a high effectiveness both in vitro and in preclinical models ,81. HoweIn contrast to etanidazole and pimonidazole, nimorazole showed promising clinical results. It improved the radiotherapeutic effect in supraglottic and pharynx tumours without side effects in a Danish head and neck cancer study (DAHANCA) . As a reFollowing the development of the initial set of oxygen-mimetic HAPs, new agents targeting both hypoxia and DNA were developed. Several compounds have been reported to reduce tumour hypoxia in vitro and were studied in the clinic, including mitomycin C, PR-104, evofosfamide (TH-302), A4QN, and tirapazamine (TPZ), as extensively reviewed elsewhere ,92. In t2−) and the activity of NQO1 [2−.; O2− can then be converted into hydrogen peroxide or hydroxyl peroxide, which can damage normal tissues [TPZ or 3-amino-1,2,4-benzotriazine1,4-dioxide is a HAP which can be reduced to its cytotoxic form by one electron reductases . This type of reduction leads to the formation of a TPZ radical and the release of hydroxide radical (OH·), then the TPZ radical is further reduced to TPZ 1-oxide by acquiring another electron ,94. TPZ  of NQO1 . NQO1 ca tissues . While t tissues ,98,99.TPZ was found to have an additive cell killing effect with radiation and cisplatin tumour cell survival ,101,102.Phase II trials of TPZ also showed effectiveness when combined with cisplatin, radiation, or gemcitabine in HNSCC, non-small cell lung cancer (NSCLC), advanced or recurrent cervical cancer, and metastatic melanoma ,115,116.Phase III clinical trials of cervical carcinoma did not find that TPZ and cisplatin to improve overall survival or progression free survival compared to cisplatin alone in stage IB2, IIA, IIB, IIIB, and IVA patients . The advInterestingly, in paediatric cancers (refractory solid tumours and rhabdomyosarcoma), TPZ did not further increase toxicity when combined with cyclophosphamide or cyclophosphamide/doxorubicin ,124. HowAlthough TPZ did not progress past the clinical trials due to the limited success in Phase III, recent studies suggest that TPZ might still be efficacious. Second generation TPZ analogues might protect cardiomycites from doxorubicin-induced toxicity . RecentlAnother possible usage of HAPs is to help deliver other anticancer agents. For instance, 2-nitroimidazole prodrugs were used to deliver the PARP1 inhibitor, 5-bromoisoquinolon . Using mTPZ was also found to be capable of suppressing osteosarcoma cells under hypoxia in part through SLC7A11-mediated ferroptosis, which is a recently discovered type of cell death . InteresAt the molecular level, hypoxia stabilises the hypoxia-inducible transcription factor (HIF), which regulates more than 100 genes involved in tumour angiogenesis, apoptosis, invasion, metastasis and metabolism ,136. HIFHIF pathways are highly upregulated in most tumour types and their expression is correlated with poor patient outcomes ,142,143.Multiple attempts have been made to target HIF to inhibit hypoxia-driven cancer progression. However, only those indirectly acting on HIF are currently used in clinical trials, including camptothecin and bortezomib ,150. OthAn emerging alternative approach to alleviate tumour hypoxia in order to increase radiosensitivity is to reprogramme the cellular metabolism into lower oxygen consumption rates (OCR). Oxygen is mostly used by mitochondria through oxidative phosphorylation (OXPHOS) and the electron transport chain (ETC). Therefore, reducing cellular OCR by inhibiting OXPHOS in well-perfused and peri-hypoxic areas of tumours increases oxygen availability and facilitates the diffusion of the non-utilised oxygen into hypoxic areas, reducing tumour hypoxia and increasing radiation responses ,165.There are several advantages to reducing tumour hypoxia by lowering OCR in tumours. Computational analysis indicated that inhibiting oxygen consumption inadequately perfused tumour regions, with a decrease as little as 30%, was more effective at reducing tumour hypoxia than increasing blood flow or elevating the oxygen concentration in blood . AnotherVarious treatments that aim to target the ETC to impair OXPHOS and decrease oxygen consumption are currently being tested. One of these treatments is the commonly prescribed, anti-diabetic biguanide, metformin. Metformin was first highlighted as a potential cancer treatment after the observation that diabetic patients treated with metformin had a lower cancer incidence and better outcome than patients with or without diabetes taking other medications ,169,170.One of the controversies around using metformin to reduce tumour hypoxia is that the OCR reduction reported was considerably low (<20%). Hence, a more potent biguanide, phenformin, was proposed as an alternative. Phenformin was reported to decrease OCR levels and reduce radioresistance in a xenograft model of colorectal cancer . SimilarArsenic trioxide is a well-known inhibitor of the complex IV in the ETC; it is an FDA-approved treatment for acute promyelocytic leukaemia (APL) and is involved in numerous clinical trials in other cancer types. It was shown that arsenic trioxide reduced tumour hypoxia in Lewis lung carcinoma (LLC) and transplantable mouse liver (TLT) with a consequent increase in radiotherapy response . ArsenicPneumocistis carinii pneumonia in children with leukaemia [Atovaquone is an FDA-approved anti-malarial agent that also showed OCR inhibition and its subsequent tumour hypoxia reduction in vitro and in vivo. This ubiquinone analogue acts as a complex III inhibitor reducing cellular oxygen consumption without inducing cell death. Atovaquone was reported to reduce the OCR by more than 80% in several types of cancer in vitro, as well as a significant tumour growth reduction in xenograft models . The fireukaemia . Other aeukaemia .Overall, targeting OXPHOS to reduce tumour hypoxia and increase radiosensitivity represents a shift away from focusing on hypoxic regions and overcomes the need to supply drugs into diffusion-limited hypoxic areas. Although at an early stage, several ETC inhibitors have been developed and are being tested clinically with promising results and will hopefully lead to new therapies to alleviate tumour hypoxia.For over a century, extensive efforts have been made to target tumour hypoxia in both experimental and clinical settings. It is critical to develop treatments that aim to reduce low oxygen environments within tumours and inhibit tumour progression and treatment resistance. To overcome hypoxia, multiple attempts were made to increase oxygen delivery, decreasetumour oxygen consumption, specifically target hypoxic cells, and inhibit HIF pathways. Although most of the trials did not accomplish a successful translation into the clinic, these studies still support the importance of hypoxia targeting to promote tumour control and patient survival. Technological advances and a better comprehension of tumour biology revived some of these historical treatments. Here, we reviewed several examples of these advances, such as the development of real time MRI-guided thermotherapy (MR thermometry). MR thermometry allows for the monitoring and control of tumour tissue temperature during HT, hence improving the application of HT. We also reviewed several second-generation molecules with improved pharmacokinetics and a higher potency, such as nimorazole or the HIF-2 inhibitor, PT2977, which showed promising results. Furthermore, we discussed the alternative usage of treatments that failed in clinical trials but are now being tested in combination with other inhibitors, such as TPZ in combination with PARP1 inhibitors. Lastly, we highlighted examples of drugs that could be repurposed, such as the anti-diabetic metformin and the anti-malarial atovaquone. These compounds were shown to reduce tumour hypoxia and are currently being tested as radiosensitisers. Therefore, we hope that combining efforts to find old and new remedies to target hypoxia will soon translate into more effective therapeutic regimens.


It is estimated that there are 1.59 million cases of chronic hepatitis B virus (HBV) infection (CHB) in the United States. HBV infection is highest among men and non-Hispanic Asian adults. CHB can lead to liver damage, cirrhosis, hepatocellular carcinoma, or death. However, the population that is most likely to develop severe outcomes is not as well-defined.We evaluated electronic health record data from Kaiser Permanente Southern California adult members from 2008-2019 with at least 1 year of continuous membership, and with 2 successive, positive HBV lab results  at least 6 months apart (indicative of CHB). Severe outcomes included incident hepatic decompensation, hepatocellular carcinoma (HCC), liver transplant and death, and prevalent and incident liver cirrhosis. For each outcome, we estimated the distribution of characteristics including age, sex, race/ethnicity, and lab values .Our final study population included 5,427 CHB-diagnosed patients with 411 (7.6%) cases of liver cirrhosis, 123 (2.3%) of hepatic decompensation, 65 (1.2%) of HCC, 8 (0.1%) of liver transplant, and 164 (3.0%) deaths. Compared to the total cohort, those who developed severe outcomes were older . Among those with severe outcomes, the majority were male ( >56%) and Asian. Diabetes was more prevalent in patients with hepatic decompensation, HCC, and death versus the entire cohort , and twice as prevalent among those with cirrhosis. All severe outcomes were associated with >2 x upper limit of normal ALT levels. The characteristics of those with severe outcomes were consistent with those of overall CHB, although there was a 2-3 times higher prevalence of diabetes in those with severe outcomes. Identifying characteristics that are more prevalent in those with severe outcomes can help inform screening and management of CHB.Ana Florea, PhD MPH, Gilead Inc. (Grant/Research Support) Prabhu Gounder, MD, Gilead Inc. (Grant/Research Support) Amandeep Sahota, MD, MS, Gilead Inc (Grant/Research Support) Katherine J. Pak, MS, Gilead (Grant/Research Support) Vennis Hong, MPH, Gilead Inc. (Research Grant or Support) Theresa M. Im, MPH, Gilead Inc. (Grant/Research Support) Sara Tartof, PhD, Gilead


Dear Editor,Atherosclerosis, as the leading cause of coronary artery disease is one of the major contributors of death globally.We reanalyzed scRNA‐seq data of COVID‐19 patient blood samples and found SARS‐CoV‐2 infection may affect CD36 and MSR1 expressions which mediate lipid uptake of macrophages contributing to atherosclerosis progression , and a grant from the Department of Science and Technology, Zhejiang Province (Grant no. LGF19H020011), a Grant from the Department of Science and Technology, Zhejiang Province (Grant no. Z16H020001) and the Natural Science Foundation of China (Grant no. 81873484), People's Republic of China.Z.L.R., Z.J., and W.S. conceived the research and participated in the study design. C.M., H.J., Y.X., and H.X.T. assisted in collecting the data. Z.W.T., W.Z., and C.W.J. analyzed the data and drafted the manuscript. All authors read and approved the final manuscript.Supporting InformationClick here for additional data file.


ASF1B is a member of the histone H3–H4 chaperone antisilencing feature 1 (ASF1). ASF1B reportedly acts as an oncogene in several cancers including, breast cancer and cervical cancer. To date, the role of ASF1B in lung adenocarcinoma (LUAD) is not elucidated. in vitro models, immunoprecipitation and immunofluorescence were utilized to investigate if BCAR1 interacted with ASF1B in LUAD. Further, transfection experiments were performed to validate the expression pattern of ASF1B in LUAD and examine its regulating role in tumor-associated processes including tumor cell proliferation and migration.  The TCGA database, containing data for 33 cancer types, was used to explore the dysregulation and prognostic value of the ASF1B gene in pan-cancer data. R software packages and public databases/webservers were applied for bioinformatics and statistical analyses. Using in vitro, it promoted the proliferation and migration of LUAD cells. ASF1B knockdown suppressed LUAD cell proliferation and migration and also diminished the expression of cell cycle, metastasis, and EMT signaling-associated proteins. BCAR1 was found positively correlated and interacting with ASF1B, and BCAR1 overexpression reversed the effects of ASF1B knockdown in LUAD cells.  ASF1B was found to be significantly elevated in LUAD and the majority of cancer types, except PCPG (pheochromocytoma and paraganglioma). The overexpression of ASF1B was associated with worse prognostic outcomes in most cancer types including LUAD. ASF1B was associated with lymph node metastasis, and  These findings indicated that ASF1B plays a significant role in the tumor progression of LUAD and BCAR1 mediates the tumor-promotive effects of ASF1B, acting as an intermediate protein. Therefore, the ASF1B/BCAR1 axis might be regarded as a putative therapeutic target for LUAD. Lung cancer, a highly prevalent malignant tumor, causes significant mortality and morbidity worldwide . Non-smaHistone H3–H4 chaperone antisilencing feature 1 (ASF1) is a key histone chaperone protein involved in chromatin-based cellular DNA replication, DNA damage repair, and transcription control –8. ASF1ABreast cancer antiestrogen resistance 1 BCAR1/p130Cas) is a scaffold protein, which is reportedly overexpressed, and serves to promote tumor proliferation and metastasis in multiple cancer types including lung cancer, breast cancer, and liver cancer 0Cas is a. In lungin vitro verification experiments.Therefore, the present study is aimed at exploring the mechanisms of ASF1B involvement in the pathogenesis of LUAD and also explored the putative role of ASF1B-BCAR1 interaction in this context by applying bioinformatic analysis and https://portal.gdc.cancer.gov/) and The Genotype-Tissue Expression (GTEx) cohort in TPM (transcript per million) format were downloaded from the University of California Santa Cruz (UCSC) Xena Browser (URL: https://xenabrowser.net) [2 transformed. A total of 15,776 samples were used for unpaired sample analysis and 10,534 samples were used for paired sample analysis. The Mann–Whitney U test (Mann–Whitney Wilcoxon test or the Wilcoxon rank-sum test) was used for comparing the ASF1B mRNA expression levels in healthy control and tumor groups. For paired sample analysis, if the data satisfied the Shapiro-Wilk normality test (P > 0.05), a paired sample t-test was used.RNAseq data from The Cancer Genome Atlas (TCGA) database (URL: ser.net) , 23 and https://portal.gdc.cancer.gov/) and GTEx cohort data from the UCSC Xena Browser (URL: https://xenabrowser.net) [2 transformed. The mRNA expression levels of ASF1B in LUAD were analyzed and visualized by using the “ggplot2” package (version 3.3.3) in R (version 3.6.3). Unpaired and paired sample analyses were both performed. For paired data satisfying the Shapiro-Wilk normality test (P > 0.05), a paired sample t-test was used. Unpaired data not satisfying the normality test (P < 0.05) were analyzed using the Mann–Whitney U test (Wilcoxon rank sum test). In addition, the dysregulation of the ASF1B gene reported in studies concerning LUAD was also examined by using the “Oncomine” web server (URL: https://www.oncomine.org/resource/login.html).Level 3 HT-seq data of LUAD patients in the FPKM format were downloaded from the TCGA database (URL: ser.net) , 23. Samx-axis abscissa represents the false-positive rate (FPR), and the y-axis represents the true-positive rate (TPR). ROC curves with an area under the ROC curve (AUC) > 0.9 indicates high diagnostic test accuracy, 0.7–0.9 indicates moderate accuracy, 0.5–0.7 indicates low accuracy, and 0.5 indicates a random result.ROC curve analysis of ASF1B gene expression data was conducted by using the “pROC” package (version 1.17.0.1) and visualized using the “ggplot2” package (version 3.3.3) in R (version 3.6.3). The clinical status  was used as the outcome parameter. The P < 0.05), the Wilcoxon rank sum test was used. Analysis was performed in R (version 3.6.3).Subsequent analysis was based on the TCGA-LUAD data obtained earlier. The expression levels of ASF1B mRNA, clinicopathological details, and general information pertaining to LUAD patients were obtained. Based on the median value of the ASF1B expression level, the LUAD samples were divided into two groups: a low-expression group of the ASF1B gene and a high-expression group of the ASF1B gene. The categorical variables analyzed included the TNM stage , pathologic stage, primary therapy outcome, gender, race, age, residual tumor, anatomic neoplasm subdivision divided by the left or right sites, anatomic neoplasm subdivision divided by central/peripheral lung, smoking history including the number of pack years smoked, OS event, DSS event, and PFI event. If all levels of a certain categorical variable satisfied the conditions of theoretical frequency > 5 and total sample size > 40, the chi-square test was used; else, Fisher's exact test was applied for testing the differences between high- and low-LUAD expression groups. If the data for a certain categorical variable were not normally distributed (P > 0.05), the Shapiro-Wilk normality test was used; otherwise, if the data of a certain clinical variable didn't satisfy the normality test (P < 0.05), the Kruskal-Wallis test was used. Such statistical analyses were performed and visualized by using the ggplot2 package in R program, based on the 329 LUAD tumor samples. In addition, the relationship between ASF1B expression and tumor characteristics was also investigated by applying a binary logistics model.ASF1B mRNA expression levels in multiple clinical variable categories were analyzed. If the data of a certain clinical variable were normally distributed (http://gepia2.cancer-pku.cn/#index) was used to plot a survival map for ASF1B expression in pan-cancer data. The KM plotter web server  was used to analyze the association between the ASF1B gene and the prognostic outcomes of LUAD tumor including overall survival (OS) and relapse-free survival (RFS).GEPIA  in R (version 3.6.3) was applied and the cox regression module was used. The outcome selected was overall survival. The tumor characteristics included the T stage, N stage, M stage, pathologic stage, primary therapy outcome, gender, race, smoking status, age, number of pack years smoked, anatomic neoplasm subdivision, anatomic neoplasm subdivision, and ASF1B gene expression group.P value) obtained from univariate and multivariate Cox regression analysis, two forest plots were plotted using the “ggplot2” package (version 3.3.3) in R (version 3.6.3). Each HR (hazard ratio) represents a relative risk of death that compares one level of a binary feature to the reference level. An HR > 1 indicates an increased risk of death, while an HR < 1 represents a decreased risk of death.Based on the results  in R (version 3.6.3). The Pearson correlation test to assess the linear relationship between the two groups was applied, and only protein coding genes were retained. The cor-pearson value or the Pearson correlation coefficient “The Cor-Pearson values of BCAR family genes  were obtained. A heat map was plotted to show the expression pattern of these BCAR family genes in LUAD tumor samples. In addition, a scatter plot was visualized to depict the correlation between ASF1B and BCAR1. The “ggplot2” (version 3.3.3) package in R (version 3.6.3) was applied for this purpose.P value < 0.05 were used for gene set enrichment analysis (GSEA). Significant differentially expressed genes (DEGs) between LUAD and healthy control samples were identified by using “DESeq2” (version 1.26.0) in R (version 3.6.3) and based on the TCGA dataset. The log2FC (fold change) values for ASF1B significantly correlated genes were obtained and used for the gene set enrichment analysis (GSEA). GSEA analysis was performed using the “clusterProfiler” package (version 3.14.3) in R (version 3.6.3). Pathways were obtained from three databases including the KEGG pathway database, WikiPathways (WP) database, and Reactome (REAC). The referenced gene set was c2.cp.v7.2.symbols.gmt (Curated) in the MSigDB Collections (URL: https://www.gsea-msigdb.org/gsea/msigdb/collections.jsp#C2). Functional terms satisfying a condition of P.adjust < 0.05, false discovery rate (FDR)  < 0.25, and ∣NES | >1 were considered as significantly enriched terms.Genes with an absolute value of correlation coefficient > 0.1 and http://org.Hs.eg.db” package (version 3.10.0) in R (version 3.6.3). Functional enrichment analysis was performed using the “clusterProfiler” package (version 3.14.3) in R (version 3.6.3). The species was selected as Homo sapiens and the Benjamini and Hochberg (BH) correction was applied to determine adjusted P values. GO terms including BP , CC (cellular component), and MF (molecular function) and KEGG pathways that were significantly enriched by the correlated genes were identified at a threshold of P.adj < 0.05 and q value < 0.2. If there were more than 30 terms, which were significantly enriched at this threshold setting, only the top 30 terms ranked in an ascending order of the adjusted P value were used to plot a bubble chart using “ggplot2” in R; otherwise, all of the terms were used.The top 100 genes ranked in a descending order of the cor_pearson value and the top 100 genes ranked in ascending order of the cor_pearson value were used for functional enrichment analysis to identify the significantly enriched functional terms among ASF1B-correlated genes. The gene names were converted to the Entrez ID by using the “2.Non-neoplastic lung epithelial cell lines (HBE) and six LUAD cell lines  were obtained from ATCC (American Type Culture Collection). Cells were cultured in RPMI-1640 medium  containing 10% fetal bovine serum  in an incubator at 37°C and 5% COsiRNA targeting ASF1B and negative control were obtained from RiboBio . Plasmids were purchased from Gene . siRNA and plasmid transfection was performed for ASF1B using Lipofectamine 3000 (Thermo Fisher Scientific Inc.) according to the manufacturer's instructions. In order to examine the effects of potential ASF1B-BCAR1 interaction in LUAD cells, firstly, ASF1B and BCAR1 were each overexpressed. Next, ASF1B knockdown was performed, followed by BCAR1 overexpression.Total RNA was extracted using TRIzol RNA reagent . Reverse transcription was performed using the PrimeScript™ RT Reagent Kit with gDNA Eraser . qRT-PCR was conducted using SYBR Green™ Premix Ex Taq™ II  in an Applied Biosystems 7500 instrument . The primer sequence for each gene is described in Table α-tubulin, β-actin, ASF1B, BCAR1, Slug, CDK2/4, CCND1, P27 (CST), Snail, Slug, E-cadherin, and Vimentin (Abcam).Western blotting was performed according to a published protocol . In brieIP assay was performed to examine the possible binding between BCAR1 and ASF1B. IP was performed with the Pierce Co-Immunoprecipitation Kit (Thermo Scientific) according to the manufacturer's instructions. In brief, total proteins were extracted from cells and the amount of protein was quantified. After coupling the affinity-purified ASF1B antibody to amine- and carrier protein-free beads, proteins were incubated with the beads overnight at 4°C. The proteins were then pulled down with elution buffer, the samples were centrifuged, and the supernatant was collected. The samples were analyzed using mass spectrometry and Western blot. Anti-IgG (CST 2729S) was used as the negative control.Immunofluorescence was performed as previously described  to exami2. Colonies were washed thrice with PBS and stained with crystal violet for 15 min.Cells were plated into 96-well plates with 1000 cells per well. CCK-8 reagents  were added at every at 24 h for five days. The optical density was estimated at 450 nm wavelength to assess cell viability. For colony formation assay, the cells were plated in 6-well plates (500/well) and incubated for 2 weeks at 37°C, 5% CO6 cells were routinely seeded and inoculated in a 6-well plate. When the cell density reached 100%, scratching was performed with a 200 μl sterile tip, perpendicularly to the cell plane. Photos were obtained under a microscope at the time points of 0 h and 24 h.1 × 105 cells) were seeded with serum-free medium onto the top chamber and the bottom chamber was filled with 10% FBS medium. After 18 h, cells in all the chambers were collected and fixed with methanol for 30 min, followed by staining with 0.01% crystal violet for 15 min.Migration assays were performed using transwell chambers (Corning USA). For migration assays, cells .The “ESTIMATE” algorithm was applied, and calculated scores were downloaded from t-test was used, and when the SDs were different, the Student's t-test with Welch's correlation was used. If the variances were equivalent in multigroup sample statistics, one-way ANOVA was used; if not, Welch's ANOVA was used. Both were subjected to Bonferroni post hoc testing. The TCGA LUAD cohort samples were split into two categories based on median gene expression values. Statistically significant differences were denoted as ∗P < 0.05, ∗∗ denotes P < 0.01, and ∗∗∗ denotes P < 0.0001.For data processing, GraphPad Prism version 8.0 was used. Values were presented as mean and SD (standard deviation) unless otherwise specified. For independent survey contrasts between two groups, when the two groups' SDs were equal, the Student's (two-tailed) To assess the role of ASF1B in cancer, firstly, a pan-cancer analysis of ASFIB expression in the TCGA cancer cohort data was performed. The results depicted in Figures The prognostic value of ASF1B expression for pan-cancer data was assessed using the TCGA prognostic data Figures . First, To analyze the possible roles of ASF1B in LUAD pathogenesis, the relationship between the mRNA expression levels of ASF1B and the clinicopathological tumor parameters were examined. As ASF1B was earlier found to predict lymph node metastasis of LUAD, this finding was validated by conducting gene set enrichment analysis, based on the TCGA LUAD cohort. The GSEA results showed that several functional terms were significantly enriched by ASF1B-correlated genes including the formation of beta catenin TCF transactivating complex, G2_M_checkpoints, ESR-mediated signaling, signaling by Wnt, signaling by Notch, signaling by Rho_GTPASES, cell cycle, and MAPK signaling pathway . FunctioIn vitro, IP assay demonstrated an interaction between BCAR1 and ASF1B in A549 cells  is an adaptor protein that belongs to the CAS family of scaffold proteins. Studies have clarified that BCAR1 is involved in carcinogenesis, by promotion of cell survival, growth, invasion, and migration. BCAR1 has been identified as a new promising biomarker for lung cancer . MoreoveWhile the role of ASF1B has been reported in several cancers, the function of ASF1B in immune regulation in LUAD is poorly understood. Here, we focused on the putative relationship of the ASF1B expression level with immune scores and immune cell profiles in LUAD. ASF1B expression was found correlated with 44 types of noncancerous cells, including Th1/2 cells, which suggests its crucial role in regulating the tumor microenvironment. Overall, the present data contributes to the understanding of the function of ASF1B in tumor immunology and its use as a cancer biomarker in LUAD.Our research is the first to uncover the role of ASF1B in the development and pathogenesis of LUAD. In sum, our findings indicate that ASF1B can be considered a relevant drug target and a prognostic biomarker in LUAD. Furthermore, these results also depict the plausible value of developing highly selective and active ASF1B inhibitors with a goal to counteract chemotherapy resistance and improve lung cancer treatment outcomes. However, preclinical and clinical studies that explore the comprehensive molecular mechanisms and signaling pathways involved in mediating the effects of ASF1B in LUAD are warranted.in vivo experiments to validate the role of ASF1B in the proliferation and metastasis of LUAD. In addition, gene enrichment analysis predicted that ASF1B as involved in the regulation of many classic pathways including MAPK, which has been shown to be regulated by BCAR1 [The present study did not include by BCAR1 . Whether


The purpose of this study was to investigate the Staphylococcus aureus , methicillin-susceptible Staphylococcus epidermidis , other methicillin-susceptible coagulase-negative staphylococci , and vancomycin-susceptible Enterococcus faecalis (31). The isolates were collected primarily from pneumonia in hospitalized patients , urinary tract infections , and bloodstream infections . Organisms were tested using reference broth microdilution methods in a central laboratory. The susceptibility of 580 Gram-positive organisms were tested, including: methicillin-susceptible 90 value of 0.03 mg/L against MSSA and 0.015 mg/L against MSSE isolates. Ertapenem MIC90 values were 8-fold higher against MSSA  and 32-fold higher against MSSE . Tebipenem displayed an MIC90 value of 0.03 mg/L against MSCoNS species other than S. epidermidis. This result was 8- and 32-fold lower than those of meropenem  and ertapenem , respectively. Tebipenem inhibited all E. faecalis isolates at ≤1 mg/L , with an MIC90 value at least 2-fold lower than meropenem  and 16-fold lower than ertapenem .Tebipenem had an MICin vitro activity was greater than meropenem and ertapenem when tested against E. faecalis. These data indicate that tebipenem may be an option for treating urinary tract infections caused by these organisms or as an empiric option to provide broader coverage against Gram-negative and -positive organisms.Tebipenem displayed potent activity against methicillin susceptible staphylococci, including MSSA, MSSE, and other MSCoNS. Tebipenem S J Ryan Arends, PhD, AbbVie (formerly Allergan) (Research Grant or Support)GlaxoSmithKline, LLC (Research Grant or Support)Melinta Therapeutics, LLC (Research Grant or Support)Nabriva Therapeutics (Research Grant or Support)Spero Therapeutics (Research Grant or Support) Abby L. Klauer, n/a, Cidara Therapeutics, Inc. (Research Grant or Support)Spero Therapeutics (Research Grant or Support) Nicole Cotroneo, Spero Therapeutics  Ian A. Critchley, Ph.D., Spero Therapeutics  Rodrigo E. Mendes, PhD, AbbVie (Research Grant or Support)AbbVie (formerly Allergan) (Research Grant or Support)Cipla Therapeutics (Research Grant or Support)Cipla USA Inc. (Research Grant or Support)ContraFect Corporation (Research Grant or Support)GlaxoSmithKline, LLC (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, LLC (Research Grant or Support)Nabriva Therapeutics (Research Grant or Support)Pfizer, Inc. (Research Grant or Support)Shionogi (Research Grant or Support)Spero Therapeutics (Research Grant or Support)


The ePlex BCID Gram-Positive (GP) panel utilizes electrowetting technology to detect the most common causes of GP bacteremia (20 targets) and 4 antimicrobial resistance genes in positive blood culture bottles. Rapid detection of intrinsic vancomycin resistance and acquired resistance genes  Time of Flight mass spectrometry. Samples submitted for standard of care testing in Biomerieux BacT/Alert resin FA/FN blood culture bottles on the BacT/Alert VIRTUO automated blood culture system with GP bacteria on direct exam (n=100) were included. Bacillus, 1 Corynebacterium) were identified, and 7/7 coinfections with Gram negative (GN) bacteria were detected by the Pan GN target and identified by the BCID-GN panel. Discordant analyses revealed a positive percent agreement (PPA) of 96/97 (99%) with 1 false negative CNS and a negative percent agreement (NPA) of 92/97 (94.8%) with 5 false positives for either S. epidermidis or Corynebacterium. Detection of vanA yielded a PPA of 4/4 and NPA of 9/9. mecA gene detection exhibited a PPA of 14/14 and NPA of 14/14 for S. aureus and a PPA of 31/32 (97%) and NPA of 16/16 for coagulase negative staphylococci with 1 false negative methicillin resistant S. epidermidis. All GP bacteria were represented on the BCID-GP panel, most tests 97/100 (97%) yielded valid results, 53 common skin contaminants (50 coagulase negative staphylococci (CNS), 2 mecA gene detection in Staphylococcus species (n=30) represent opportunities for early optimization of antimicrobial therapy in 34/100 (34%) of samples. The BCID-GP panel provides rapid accurate detection of resistant isolates and common contaminants enabling high quality data driven optimization of antimicrobial therapy. Detection of acquired vancomycin resistance (n=4) and absence of Todd P. McCarty, MD, Cidara (Grant/Research Support)GenMark T2 Biosystems (Consultant) Sixto M. Leal, Jr., MD, PhD, Abnova (Grant/Research Support)AltImmune (Grant/Research Support)Amplyx Pharmaceuticals (Grant/Research Support)Astellas Pharmaceuticals (Grant/Research Support)CNINE Dx (Grant/Research Support)GenMark Diagnostics IHMA (Grant/Research Support)IMMY Dx (Grant/Research Support)JMI/Sentry (Grant/Research Support)mFluiDx Dx (Grant/Research Support)SpeeDx Dx (Grant/Research Support)Tetraphase Pharmaceuticals (Grant/Research Support)


Investments in the beginning was described as creating contact, showing the agenda and being adaptive, while explore the patient perspective included showing awareness, being explorative and creating a participating climate. Show empathy consisted of showing openness, being confirmative and creating acceptance, while showing a structured follow‐up plan, being open minded and invitational and creating motivation to build on were descriptions of invest in the end. Awareness of potential facilitators and barriers for patient‐centred communication during the beginning, middle and end of a continuous positive airway pressure treatment consultation can be used to improve contextual conditions and personal communication competences among practitioners working with continuous positive airway pressure treatment initiation.Adherence to continuous positive airway pressure treatment for obstructive sleep apnea tends to be poor. Communication influences adherence but has not previously been investigated from a practitioner perspective, although shared decision‐making is known to be of great importance. The aim was to describe how practitioners experience communication with patients with obstructive sleep apnea during the initial visit at a continuous positive airway pressure treatment clinic, with focus on facilitators and barriers related to the 4 Habits Model, a communication model comprised of four types of interrelated skills to make encounters more patient‐centred: investing in the beginning; exploring the patient perspective; showing empathy; and investing in the end. A descriptive design with qualitative content analysis was used. A deductive analysis was carried out based on interviews with 24 strategically selected practitioners from seven continuous positive airway pressure treatment clinics. The 4 Habits Model was used as a framework for identifying facilitators and barriers to communication.  The pilot tested interview guide was based on five questions, as follows. (a) Can you describe situations that affect communication between you and the patient during the initial visit? (b) Can you describe situations at the beginning of the visit that affect communication? (c) Can you describe situations that affect how you explore the patients’ situation? (d) Can you describe situations that affect how you create contact and show empathy during the conversation? (e) Can you describe situations that affect the ending of the conversation? Probing questions, such as: “Can you describe this in more detail?”, “How did it affect the conversation?”, “What did you say next?” and “What were you thinking?” were used. The duration of the audio‐recorded interviews ranged from 15 to 75 min.2.3Verbatim transcripts of all interviews were produced, resulting in 295 A4 pages of single‐spaced text in 12‐point Times font. The deductive analysis was performed by a multi‐professional team of clinicians and researchers . Credibility was based on the connection between the aim and the chosen method, i.e. qualitative content analysis (Graneheim & Lundman, 5Creating contact, showing the agenda and being adaptive were used in the beginning of conversation, while showing awareness, being explorative and creating a participating climate were used to explore the patient perspective. Showing empathy consisted of showing openness, being confirmative and creating acceptance, while showing a structured follow‐up plan, being open minded and invitational and creating motivation to build on were descriptions of investing in the end.None.


Fluid balance is an essential issue in critical care; however, the impact of early fluid balance on the long-term mortality in critically ill surgical patients remains unknown. This study aimed to address the impact of day 1–3 and day 4–7 fluid balance on the long-term mortality in critically ill surgical patients. We enrolled patients who were admitted to surgical intensive care units (ICUs) during 2015–2019 at a tertiary hospital in central Taiwan and retrieved date-of-death from the Taiwanese nationwide death registration profile. We used a Log-rank test and a multivariable Cox proportional hazards regression model to determine the independent mortality impact of early fluid balance. A total of 6978 patients were included for analyses . In-hospital mortality, 90-day mortality, 1-year and overall mortality was 10.3%, 15.8%, 23.8% and 31.7%, respectively. In a multivariable Cox proportional hazard regression model adjusted for relevant covariates, we found that positive cumulative day 4–7 fluid balance was independently associated with long-term mortality , and a similar trend was found on day 1–3 fluid balance, although to a lesser extent . In conclusion, the fluid balance in the first week of ICU stay, particularly day 4–7 fluid balance, may affect the long-term outcome in critically ill surgical patients.  Fluid balance is increasingly recognised as an essential issue in critically ill patients ,2; howevConservative initial fluid resuscitation and fluid balance is currently an essential issue in fluid management in patients with sepsis ,8, and oGiven that the data in the present study were anonymised prior to analyses, the informed consent was waived. This study was approved by the Institutional Review Board of the Taichung Veterans General Hospital (TCVGH: SE20249B) and conducted in accordance with the Declaration of Helsinki.This retrospective cohort study was conducted at TCVGH, a tertiary care referral hospital with 1530 beds and three surgical ICUs in central Taiwan. All adult patients who had been admitted to the ICU between 2015 and 2019 were included in this study, and we used the first ICU admission as the index of ICU admission.We used two databases in the present study: the critical care database of TCVGH and the death registration data of the Taiwanese National Health Insurance Research Database (NHIRD) . Data wiThe parameters in this study focusing on fluid status were daily fluid input, output and balance. Input fluid consisted of all intravenous fluid, the volume of drugs dissolved in solution, enteral fluid, parenteral fluids and transfusion of blood products. Output fluid was composed of urine output, haemodialysis and output from drains, orogastric and nasogastric tubes. We presented daily fluid status as daily and cumulative fluid input, output and balance in accordance with the format in previous studies, including our recently published studies ,16,17,18t-test for continuous variables as well as the Chi-square test for categorical variables. Kaplan-Meier analysis was applied to determine the association between mortality and day 1–3/4–7 cumulative fluid balance status. The Cox proportional hazards regression model was applied to identify independent predictors for long-term mortality, and the data were represented by the adjusted hazard ratio (adjHR) and the corresponding 95% confidence interval (CI). Statistical analyses were two-sided, and the level of significance was set at 0.05. Data analyses were conducted using R version 3.6.0.Continuous variables were presented as mean ± standard deviation, and data for categorical variables were shown as frequencies (percentages). The differences between the survivor and non-survivor groups were analysed by the Student’s p < 0.01), Charlson comorbidity index , APACHE II score  and a lower body mass index (BMI) . Non-survivors were also more likely to have shock , to receive mechanical ventilation for more than three days  and to receive renal replacement therapy during ICU stay  or end-stage renal disease (1.5% vs. 0.7%), whereas non-survivors were less likely to undergo a surgery during ICU stay   . p < 0.01) and a similar trend was found with regards to cumulative day 4–7 fluid balance .The fluid balance appeared to be generally positive on day one (1027.0 ± 1876.7 mL) as well as day two (119.3 ± 1040.8 mL) and then to be balanced or slightly negative on day three (−8.1 ± 923.5 mL), day four (−8.6 ± 827.9 mL) and the following days in the first week of ICU admission. Notably, we found a steady negative fluid balance between day four and day seven in the survivor group, whereas a slightly positive fluid balance was noted in the non-survivor group between days 4–7. Therefore, we divided fluid balance in the first week into day 1–3 fluid balance and day 4–7 fluid balance. Compared with the survivor group, a higher positive cumulative day 1–3 fluid balance was found in the non-survivor group  programme consists of the patient-centred, evidence-based, multidisciplinary team-developed pathways aiming to improve the outcome of surgical patients; however, an optimal peri-operative fluid strategy in critically ill surgical patients remains uncertain due to distinct surgical risks, varied hemodynamic monitors and complex patient-groups ,12,13. IGiven that the majority of the enrolled patients in this study were critically ill neurological patients, we hence further investigated the long-term mortality impact of early fluid balance in these patients. We validated a consistent trend that both day 1–3 and day 4–7 were independently associated with long-term mortality in critically ill neurosurgical patients, and the magnitude appeared to be slightly lower than those in the other surgical patients . We postThere are limitations to our study. First, given the observational nature of the present study, the decision of fluid and shock management was made by the individual attending physician and could potentially introduce a confounding effect. However, given the administration of intensivists in the reported ICUs, the aforementioned concern should at least partly be mitigated. Second, being a single-centre observational study, external validation is hence needed. However, the coding of fluid status and relevant data is in accordance with our previous published studies regarding fluid status in critically ill influenza and cancer patients ,18. ThirIn conclusion, we linked databases at TCVGH and Taiwanese NHIRD to investigate the impact of early fluid balance on long-term mortality in critically ill surgical patients. We found that days 1–3 slightly affected the long-term mortality in critically ill surgical patients with shock. Notably, we identified that day 4–7 fluid balance had a consistent impact on the long-term mortality in critically ill patients with and without shock, and the aforementioned impact was robust among those receiving surgery. These findings highlight the essential role of early fluid balance in critically ill surgical patients and support the implementation of restrictive or deresuscitative fluid strategy, particularly on days 4–7 after the stabilisation of the initial critical illness. More studies are warranted to prospectively confirm our findings and to explore the underlying mechanism.


Working from a dataset of 118 billion messages running from the start of 2009 to the end of 2019, we identify and explore the relative daily use of over 150 languages on Twitter. We find that eight languages comprise 80% of all tweets, with English, Japanese, Spanish, Arabic, and Portuguese being the most dominant. To quantify social spreading in each language over time, we compute the ‘contagion ratio’: The balance of retweets to organic messages. We find that for the most common languages on Twitter there is a growing tendency, though not universal, to retweet rather than share new content. By the end of 2019, the contagion ratios for half of the top 30 languages, including English and Spanish, had reached above 1—the naive contagion threshold. In 2019, the top 5 languages with the highest average daily ratios were, in order, Thai (7.3), Hindi, Tamil, Urdu, and Catalan, while the bottom 5 were Russian, Swedish, Esperanto, Cebuano, and Finnish (0.26). Further, we show that over time, the contagion ratios for most common languages are growing more strongly than those of rare languages. Users of social media are presented with a choice: post nothing at all; post something original; or re-post (“retweet” in the case of Twitter) an existing post. The simple amplifying mechanism of reposting encodes a unique digital and behavioral aspect of social contagion, with increasingly important ramifications as interactions and conversations on social media platforms such as Twitter tend to mirror the dynamics of major global and local events –4.Previous studies have explored the role of retweeting in the social contagion literature, though the vast majority of this research is limited to either a given language  or a short period , 2, 5, 6anything and the act of de novo generation .We describe distinct usage patterns of retweets for certain populations. For example, Thai, Korean, and Hindi have the highest contagion ratios, while Japanese, Russian, Swedish, and Finish lie at the other end of the spectrum. While there is a wide range of motives and practices associated with retweeting, our object of study is the simple differentiation of observed behavior between the act of replication of We acknowledge two important limitations from the start. First, while it may be tempting to naively view ideas spreading as infectious diseases, the analogy falls well short of capturing the full gamut of social contagion mechanisms –16, and Social contagion has been extensively studied across many disciplines including marketing –20, finaet al. [The body of research shows overwhelming evidence that retweeting is a key instrument of social contagion on Twitter , 51. Oneet al.  suggestset al. . The praet al. , 54. Staet al. . The useOther studies have quantified language usage on social media , 56, parWe structure our paper as follows. First, we discuss the state-of-the-art tools presently used for language detection of short and informal messages . We then describe our dataset and processing pipeline to answer some key questions regarding social contagion through the use of retweets. Based on our considerations, we deploy FastText-LID  to identet al. [For messages posted after 2013, we also analyze language labels provided by Twitter’s proprietary LID algorithm and justify using FastText-LID as an alternative LID tool to overcome the challenge of missing language labels in the historical feed from Twitter .We study the empirical dynamics of replication: The rate at which users choose to retweet instead of generating original content; and how that rate varies across languages temporally. We quantify the ratio of retweets to new messages (contagion ratio) in each language. In most common languages on Twitter, we show that this ratio reveals a growing tendency to retweet.http://compstorylab.org/storywrangler/papers/tlid/.Finally, we present a detailed comparison with the historical data feed in Appendix Twitter is a well-structured streaming source of sociotechnical data, allowing for the study of dynamical linguistics and cultural phenomena , 64. Of LID is often referred to as a solved problem in NLP research –100, espA key challenge of LID on Twitter data is the absence of a large, public, annotated corpus of tweets covering most languages for training and evaluation of LID algorithms. Although researchers have compiled a handful of manually labeled datasets of Twitter messages, the proposed datasets were notably small compared to the size of daily messages on Twitter and limited to a few common languages –83. TheyAs of early 2013, Twitter introduced language predictions classified by their internal algorithm in the historical data feed . Since tGiven access to additional information about the author of a tweet, the LID task would conceivably be much more accurate. For example, if the training data for prediction included any or all of the self-reported locations found in a user’s ‘bio’, the GPS coordinates of their most recent tweet, the language they prefer to read messages in, the language associated with individuals they follow or who follow them, and their collective tweet history, we expect the predictions would improve considerably. However, for the present investigation, we assume the only available predictors are found in the message itself. Our goal is to use the state-of-the-art language detection tools to get consistent language labels for messages in our data set to enable us to investigate broader questions about linguistic dynamics and the growth of retweets on the platform over time.Several studies have looked closely at language identification and detection for short-text –110, parMost studies share a strong consensus that language identification of tweets is an exceptionally difficult task for several reasons. First, language classification models are usually trained over formal and large corpora, while most messages shared on Twitter are informal and composed of 140 characters or fewer , 82 , hashtags , handles , html codes , emojis, and any redundant whitespaces.To avoid biasing our language classification process, we filter out Twitter-specific content prior to passing tweets through the FastText-LID model. This is a simple strategy originally proposed by Tromp et al.  to improet al. , 122. SpOnce we filter out all Twitter-specific content, we feed the remaining text through the FastText-LID neural network and select the predicted language with the highest confidence score as our ground-truth language label. If the confidence score of a given prediction is less than 25%, we label that tweet as Undefined (und). Similarly, if no language classification is made by the Twitter-LID model, Twitter flags the language of the message as undefined , 124. Wehttps://storywrangling.org/. See Alshaabi et al. [We subsequently extract day-scale time series and Zipf distributions for uni-, bi-, and tri-grams and make them available through an analytical instrument entitled Storywrangler. Our tool is publicly available online at: i et al.  for techet al. [We have collected a random 10% sample of all public tweets posted on the Twitter platform starting January 1, 2009. Using the steps described in Sect. et al. ).We find that our classification of tweets using FastText-LID notably improves the consistency of language labels when compared to the labels served with the historical streaming feed. In Fig. FastText-LID classifies up to 173 languages, some of which are rare, thus the occasional dropout of a language seen in this time series is expected. On the other hand, Twitter-LID captures up to 73 languages, some of which are experimental and no longer available in recent years. Nonetheless, Fig. We compute annual confusion matrices to examine the language labels classified by FastText-LID compared to those found in the historical data feed. Upon inspection of the computed confusion matrices, we find disagreement during the first few years of Twitter’s introduction of the LID feature to the platform. As anticipated, the predicted language for the majority of tweets harmonizes across both classifiers for recent years see Fig. . We notiFurthermore, we display a heatmap of the number of messages for each language as classified by FastText-LID in our data set see Fig. . We haveIn Fig. We take a closer look at the flow of information among different languages on the platform, specifically the use of the “retweet” feature as a way of spreading information. Encoding a behavioral feature initially invented by users, Twitter formalized the retweet feature in November 2009 . ChangesTo quantify the rate of usage of each language with respect to these different means by which people communicate on the platform, we categorize messages on Twitter into two types: “Organic Tweets” (OT), and “Retweets” (RT). The former category (OT) encompasses original messages that are explicitly authored by users, while the latter category (RT) captures messages that are shared (i.e. amplified) by users. We break each quote tweet into two separate messages: a comment and a retweet. We exclude retweets while including all added text (comments) found in quote tweets for the OT category.t and for each language ℓ, we calculate the raw frequency (count) of organic messages For each day To further investigate the growth of retweets, we use the ratio of retweeted messages to organic messages as an intuitive and interpretable analytical measure to track this social amplification phenomenon. We define the ‘contagion ratio’ as:In 2018, the contagion ratio exceeded 1, indicating a higher number of retweeted messages than organic messages Fig. . The ove6In 2020, Twitter’s developers redesigned their retweet mechanism, purposefully prompting users to write their own commentary using the Quote Tweet , along wFinally, we show weekly aggregation of the rate of usage entclass1pt{minimaUnderstanding how stories spread through and persist within populations has always been central to understanding social phenomena. In a time when information can flow instantly and freely online, the study of social contagion has only become more important.In the sphere of Twitter, the practice of retweeting is complicated from a social and psychological point of view. There is a diverse set of reasons for participants to retweet. For example, scientists and academics can use this elementary feature to share their findings and discoveries with their colleagues. Celebrities and political actors can benefit from other people retweeting their stories for self-promotion. Attackers can also take advantage of this natural feature of social contagion to pursue malicious intents, deploy social bots, and spread fake news.In this paper, we have analyzed over a hundred billion messages posted on Twitter throughout the last decade. We presented an alternative approach for obtaining language labels using FastText-LID in order to overcome the challenge of missing labels in the Decahose dataset, obtaining consistent language labels for 100+ languages. We acknowledge that shortcomings of language detection for short and informal text  are well known in the NLP literature. Using FastText-LID is not necessarily the best approach for language identification. Our analysis may be subject to implicit measurement biases and errors introduced by word embeddings used to train the language detection tool using FastText . We emphOur results comparing language usage over time suggest a systematic shift on Twitter. We found a recent tendency among most languages to increasingly retweet (spread information) rather than generate new content. Understanding the general rise of retweeted messages requires further investigation. Possible partial causes might lie in changes in the design of the platform, increases in bot activity, a fundamental shift in human information processing as social media becomes more familiar to users, and changes in the demographics of users .The metrics we have used to compute our contagion ratios are simple but rather limited. We primarily focused on tracking the rate of organic tweets and retweets to quantify social amplification of messages on the platform. While our approach of measuring the statistical properties of contagion ratios is important, it falls short of capturing how retweets propagate throughout the social networks of users. Future work may deploy a network-based approach to investigate the flow of retweets among users and followers. Whether or not the information is differentially propagated across languages, social groups, economic strata, or geographical regions is an important question for future research, and beyond the scope of our present work.Geolocation information for Twitter is also limited, and here we have only examined contagion ratios at the language level. Language, transcending borders as it does, can nevertheless be used differently across communities. For instance, characterizing the temporal dynamics of contagion ratios for English, which is used all around the globe, is very different from doing so for Thai—a language that is used within a geographically well-defined population. Different social and geographical communities have cultures of communication which will need to be explored in future work.vice versa. However, in order to conduct this line of research, it is necessary to have accurate geotagging of tweets, which is currently not the case except for a very small subsample [In particular, it is important to study the relationship between social contagion dynamics, geographical region, and language. It might be the case that contagion dynamics are more homogeneous across geographic regions even when each geographical region displays high language diversity, or ubsample . Future