Preoperative evaluation of nipple-areola complex (NAC) tumour involvement is crucial to select patients candidates for nipple-sparing mastectomy. Our aim was to validate a previously developed automated method able to compute the three-dimensional (3D) tumour-to-NAC distance (the most predictive parameter of nipple involvement), using magnetic resonance imaging (MRI) datasets acquired with a scanner and protocol different from those of the development phase.We performed a retrospective analysis of 77 patients submitted to total mastectomy and preoperatively studied with MRI. The new method consisted of automated segmentation of both NAC and tumour and subsequent computation of the 3D distance between them; standard manual two-dimensional segmentation was independently performed. Paraffin-embedded section examination of the removed NAC was performed to identify the neoplastic involvement. The ability of both methods to discriminate between patients with and without NAC involvement was compared using receiver operating characteristic (ROC) analysis.p = 0.431).The 3D tumour-to-NAC distance was correctly computed for 72/77 patients (93.5%); tumour and NAC segmentation method failed in two and three cases, respectively. The diagnostic performance of the 3D automated method at best cut-off values was consistently better than that of the 2D manual method , even if the difference did not reach statistical significance (The introduction of the 3D automated method in a clinical setting could improve the diagnostic performance in the preoperative assessment of NAC tumour involvement. The distance between tumour and nipple-areola complex (NAC) on magnetic resonance imaging (MRI) is the best predictor of occult nipple involvement, essential for planning nipple-sparing mastectomy.A novel three-dimensional (3D) automated method for computing tumour-to-NAC distance on MRI datasets was developed.This new method was compared with two-dimensional (2D) manual measurement on an external dataset, i.e. different from that used in the development phase.In the external validation, the 3D automated method showed a sensitivity and a specificity higher than those of 2D manual measurement, even though not reaching the statistical significance.Since its introduction, nipple-sparing mastectomy (NSM) has become a frequent surgical option for both prophylactic and therapeutic indications.The shift towards more conservative types of mastectomy, which began in the early 1990s with the introduction of the skin-sparing mastectomy , combineIn patients who might be offered NSM, the preoperative assessment of NAC tumour involvement is crucial to optimise the surgical planning. Some predictive models have been developed , 11 to pDespite the available evidence, this data is often difficult to obtain, primarily because the analysis of MRI requires interpretation by experienced breast radiologists and secondly because it is subject to operator-dependent variability, thus resulting in a difficult standardisation of the parameter . FurtherTo overcome these issues, recently Giannini et al.  developeThe aim of the current study is to assess the automated three-dimensional (3D) tumour-to-NAC distance calculation with the algorithm proposed by Giannini et al.  for the Patients who received total or SSM for breast cancer between January 2010 and May 2016 at the Academic Breast Unit of the Umberto I Hospital, University of Turin, were considered eligible for the present study.Data from 403 mastectomy specimens were retrieved. Patients who had the nipple preserved were excluded, based on the lack of definitive data on histological examination of the terminal ducts. In addition, patients who had not been studied with preoperative breast MRI, including T2-weighted images and dynamic study, were excluded. Metastatic disease and age at surgery were not considered as exclusion criteria. Of the 124 cases initially selected, 103 had undergone MRI examination at our institution and MRI data were therefore effectively accessible. Among these, it was possible to retrieve in the archives of the Department of Radiology 81 full MRI studies. Of these, four did not contain all the image sequences and were excluded. Overall, 77 cases were available for the analysis. A study flow diagram is provided in Fig. Written informed consent to the anonymous use of their clinical and instrumental data for scientific purposes is signed by all the patients treated at our institution.@ Bayer) at a flow rate of 2 mL/s, followed by saline flushing of 20 mL at a flow rate of 2 mL/s. Table Breast MRI was performed using a 1.5-T scanner  and a dedicated phased-array 16-channel coil, with the patient in the prone position, following the recommended requirements . In partThe tumour-to-NAC distance was defined as the minimum distance between the base of the NAC  and the nearest margin of tumour focus. As described in a previous study , the tumThe automated calculation of 3D tumour-to-NAC distance was performed using the previously described algorithm  which coAverage execution time was 8 min to segment the tumour lesions, 2 min to segment the nipples, and 1 min to compute the 3D tumour-to-NAC distance. The execution time was measured on a computer equipped with a CPU Intel Core i7 940 Quad Core @#2.93 GHz architecture and 8 GB RAM. Figures The pathological sections were used as the reference standard for the diagnostic performance of the two methods of calculation of tumour-to-NAC distance on MRI. Final histological reports were reviewed by a pathologist with more than 10 years of experience in breast pathology. NAC involvement was defined by the presence of invasive ductal or lobular carcinoma and/or ductal carcinoma in situ (DIN1c–DIN3) within the retro-areolar margin (2–3 mm below the areola-nipple junction). Lobular carcinoma in situ/lobular intraepithelial neoplasia (LIN1–LIN3) was not considered as malignant lesions.Diagnostic performances of both automated and manual methods to compute tumour-to-NAC distance were recorded: sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and overall diagnostic accuracy.True positives were defined as those cases in which the automated or manual method provided a tumour-to-NAC distance lower than the chosen cut-off and the pathologic report confirmed tumour NAC involvement. False positives were defined as those cases in which the automated or manual method provided a tumour-to-NAC distance lower than the chosen cut-off, but the pathologic report showed the absence of tumour NAC involvement. True negatives were defined as those cases in which the automated or manual method provided a tumour-to-NAC distance equal or higher than the chosen cut-off and the pathologic report confirmed the absence of tumour NAC involvement. False negatives were defined as those cases in which the automated or manual method provided a tumour-to-NAC distance equal or higher than the chosen cut-off and the pathologic report showed tumour NAC involvement.When 95% confidence intervals were provided, they were calculated according to the binomial distribution.p value < 0.05 was considered as statistically significant.Receiver operating characteristic (ROC) curves were obtained, and the best cut-off value was searched for optimal balance between sensitivity and specificity for both methods. Different cut-off values were also considered, according to those used in the literature . A Statistical analysis was performed using SPSS, version 24 for Windows , and MedCalc, version 13.2.2.0 . All calculations were performed on both softwares.From the raw series of 77 patients with 77 tumour lesions, 2 were excluded because the tumour lesion was not correctly detected by the algorithm , 2 because of an error in the segmentation or malposition of the nipple, and 1 because the possible NAC involvement at the final pathology examination was not specified in the report. The two false-negative tumour lesions were one invasive ductal carcinoma, with a maximum diameter of 7 mm, and one ductal carcinoma in situ, with a maximum diameter of 23 mm, both of them with a poor contrast enhancement on the axial DCE sequence.Thus, we tested the algorithm on both nipples for each of the 72 patients, and the nipples properly segmented were 141/154 . Of the 72 patients with evaluated tumour-to-NAC distance, 23 (32%) showed tumour infiltration of the nipple at final pathology.p = 0.431).The ROC curves obtained for both 2D manual and 3D automated tumour-to-NAC distances are shown in Fig. Sensitivity, specificity, PPV, NPV, and accuracy calculated at different cut-off values for the two methods are shown in Table p < 0.001). However, the manual method showed a lower specificity than the automated method (p < 0.001). At the bottom line, the difference between the two methods as regards PPV (p = 0.471) and PNV (p = 0.164) was not significant.Considering the 20 mm cut-off, which is currently the most widely used in clinical practice, the sensitivity of the automated method was significantly higher than that of the manual method  demonstrated that this 3D automated method could represent a reliable method to preoperatively compute tumour-to-NAC distance, improving the management of patients candidate for NSM. The algorithm had a failure rate of only 5% because of the failure of the nipple (two cases) or tumour (two cases) segmentation.z-/y-axes is lost, with the consequent chance that the lesion and the nipple appear closer, as lying on the same x-/y-axis. Actually, the lesion and nipple are often more distant, as they are seated in different slices of MRI volume. In fact, the distance calculated by the automated algorithm was greater than that manually calculated in 55/72 cases (76%). In a recent study [The automated system presented in this study may show many advantages. First, the 3D tumour-to-NAC measurements were more reliable than the 2D measurements calculated using MIP images. In fact, when a measurement is carried out on the axial/sagittal projection, the information along the nt study , the issnt study  did not Interestingly, no fully automated methods for the nipple segmentation on MRI are available yet. The nipples often differ in form and intensity of the signal in different patients; in addition, the nipple is not always perfectly located at the centre of the T2-weighted image and, when inverted, cannot always be distinguished from glandular tissue. In our experience, 92% of the nipples were properly segmented by the algorithm.Taken for granted that the tumour-to-NAC distance is up to date the most useful parameter for the preoperative assessment when NSM is under consideration, the main issue is to define the best cut-off value capable of predicting NAC involvement. In this regard, the literature is inhomogeneous. Some authors propose 10 mm as the ideal cut-off , while oIn the present series, the 3D automated method improved the diagnostic performance when compared to 2D manual measurement, even though not significantly. In particular, the best compromise between sensitivity and specificity for each method was reached using the cut-off of 30 mm for the automated method and of 21 mm for the manual method. This difference is consistent with the increase by 11.5 mm in the average tumour-to-NAC distance when processed by the automated method versus the manual one and with the previously mentioned greater distance in 76% of cases as compared to the manual measurement.As shown in Table Since the aim of the assessment before surgery is to propose NSM to all patients who may potentially preserve the nipple (i.e. patients without NAC tumour involvement at pathology), specificity and PPV are the most useful preliminary parameters to know. As a high specificity is related to a low sensitivity, many patients with NAC involvement (i.e. patients with NAC tumour involvement at pathology) will still be candidates for NSM.By choosing the cut-off at ≤ 10 mm, the high specificity (100%) allows to exclude all the patients who certainly will not be able to keep the nipple, while the low sensitivity (26%) causes the inclusion in the selection for NSM of most patients (74%) with NAC involvement. However, the current protocol for the NSM mandates the intraoperative histological examination of the retroareolar tissue, which shows a good negative predictive ability. In such cases, the surgery may be converted to SSM at the same surgical time.The data obtained from our study are similar to those we have previously obtained with the same 3D methods on images obtained on a different equipment, as shown in Table Limitations of our study are mainly related to its retrospective design. Our series covers a period of almost 6 years, during which there have been technological advances that, although modest, could have influenced the signal to noise ratio and the image quality, changing the segmentation capabilities  of the automated algorithm, as shown in Fig. In conclusion, our study suggests that breast MRI is a promising method for the preoperative assessment of patients candidate for NSM by predicting the occult involvement of the NAC. Our novel 3D automated method seems to improve the results obtained with the 2D manual distance measurement also being validated on an independent external dataset. A cut-off value of ≤ 10 mm provided great accuracy as all the patients with a tumour-to-NAC distance ≤ 10 mm require the removal of the nipple. If integrated into clinical practice, this method could be useful to reduce the variability in selecting patients who may have the nipple preserved.


Shingles  and disseminated zoster are caused by the reactivation of latent varicella zoster virus (VZV). Reactivation of VZV is related to impaired cell-mediated immunity. Extensive burns affecting a patient result in burn-related immunosuppression and cytokine storm. Despite immunosuppression in burn patients, the reactivation of VZV is extremely rare, whereas eczema herpeticum, caused by reactivation of latent herpes simplex virus (HSV), is common. We have found only 1 published case of VZV reactivation during burn treatment in the literature.Pseudomonas aeruginosa (MDRP) was detected from a blood specimen, and the patient developed multiple organ failure (MOF). On day 31, compact aggregations of small vesicles appeared on the intact skin of his left knee and left buttock. The vesicles were located within the 4th lumbar (L4) spinal dermatome. From day 32 to day 34, similar new vesicles arose on his intact skin and epithelializing skin-graft donor sites. We diagnosed disseminated zoster, based on the patient’s age, the characteristic occurrence of the initial vesicles within a limited area of intact skin in the left L4 dermatome, and a positive Tzank smear. Serologic testing on day 36 showed a high level of anti-VZV immunoglobulin (Ig)G with low levels of anti-VZV IgM, anti-HSV IgG, and anti-HSV IgM. The patient was isolated in a negative-pressure room to avoid air-borne spread of VZV. On day 52, the patient died.A 51-year-old man was burned in a fire, which affected 60% of his total body surface area (TBSA), and also received inhalation injury (day 0). Despite fluid resuscitation, he showed persistent renal failure. Continuous hemodialysis and filtration (CHDF) combined with polymyxin B-immobilized fiber column direct hemoperfusion (PMX-DHP) therapy was used for cytokine modulation. Autologous and allogeneic skin grafting was performed. On day 15, multiple-drug-resistant To the best of our knowledge, our patient is the second case of reactivation of VZV during burn treatment. It is unclear why reactivation of VZV is rare in patients with burn-related immunosuppression, whereas HSV reactivation is common. Cytokine modulation throughout the treatment period using CHDF combined with PMX-DHP might have been related to the rare reactivation of VZV in our patient. Our case provides an additional information on the relationship between the immune status of a patient with extensive burns and reactivation of latent VZV or HSV. Disseminated zoster, caused by the reactivation of latent varicella zoster virus (VZV), is sometimes seen in patients with immunocompromised status, such as post-organ transplantation, lymphoma, and acquired immunodeficiency syndrome (AIDS) . ExtensiA 51-year-old unemployed Asian man sustained a thermal burn affecting 60% of his total body surface area (TBSA) in a gasoline fire. He arrived at our hospital 4 h after injury (day 0). Upon initial examination, he was in shock and found to have inhalation injury. He had third degree burns on his face, head, neck, chest, back, bilateral upper extremities, abdomen, and bilateral thighs. Fluid resuscitation and artificial ventilation with tracheal intubation were started. According to his family, he did not have any obvious comorbidities. The patient’s history of VZV vaccination and VZV infection were unknown. Despite fluid resuscitation and catecholamine support, he had persistent oliguria and hypotension. Continuous hemodialysis and filtration (CHDF) combined with polymyxin B-immobilized fiber column direct hemoperfusion (PMX-DHP) was started on day 1 to treat his renal failure, reduce cytokine storm, and remove endotoxins in an attempt to prevent development of multiple organ failure (MOF). Multiple eschar debridements and autologous and allogeneic skin grafting were performed. Almost complete escharectomy was achieved by day 13. However, throughout the treatment period, the patient’s general condition remained critical. He could not be weaned from artificial ventilation and CHDF, and he continuously needed catecholamine circulatory support. On day 10, he developed cardiac arrest that was treated by cardiac massage and intravenous adrenaline, with return of spontaneous circulation (ROSC).Pseudomonas aeruginosa (MDRP) was detected in specimens from his burn wound and blood. His burn sepsis was refractory to treatment, and he developed multiple organ failure (MOF). On day 31, compact aggregations of small vesicles appeared on the intact skin of his left knee and left buttock. The vesicles were located within the 4th lumbar (L4) spinal dermatome  family, which consists of 8 known types of viruses . VaricelPatients with extensive burns have burn-related immunosuppression , 8. HoweHSV, a member of the HHV family, is also associated with the occurrence of herpetic lesions in burn patients. HSV includes HSV types 1 (HSV-1) and 2 (HSV-2) and is transmitted by direct contact. Primary infection of HSV or reactivation of latent HSV can result in herpetic skin or mucous membrane lesions. Typically, oral lesions are due to HSV-1 and genital lesions to HSV-2. In burn patients, herpetic lesions associated with HSV are likely to occur in second degree burns and split-thickness skin graft (STSG) harvested sites during the healing process –18. DissSometimes differentiating between KVEs, varicella, and disseminated zoster is difficult. In our case, a dermatologist examined the patient and diagnosed disseminated zoster 3 days after the onset of the eruptions. The diagnosis of disseminated zoster was made based on the age of the patient, the primary onset of exanthema on the skin of the left L4 dermatome, aggregated pattern of vesicular eruptions, vesicles appearing on normal skin rather than involving healing burn lesions or STSG donor sites, asynchronous generalized eruption of vesicles, and concomitant existence of vesicles, pustules, and crusts. Those findings all support the diagnosis of disseminated zoster instead of varicella or KVE. Serologic testing performed 5 days after the onset of vesicular eruptions also supported the diagnosis of disseminated zoster. However, completely denying the possibility of HSV involvement in our case is difficult, because the simultaneous involvement of HSV and VZV has been reported. Giehl et al. reported that in 1.8% of patients in whom the clinically suspected pathogen was VZV, an additional HSV infection was identified by polymerase chain reaction (PCR) . Althoug2 of creatinine clearance [The first-line treatment of disseminated zoster is acyclovir, which is also true for shingles, varicella, and HSV infection. VZV infection of immunocompromised patients is life-threatening, and early high-dose acyclovir has been reported . In our learance . A conselearance , 10. VZVlearance .The molecular trigger of the reactivation of latent VZV is unknown. However, it is clear that deficient cell-mediated immunity in elderly and immunocompromised patients is associated with the reactivation of latent VZV . ImmunosFever itself is one of the stimulations that promote HSV reactivation. Warren et al. reported that in 46% (190 of 411) of patients undergone artificially induced fever, oral-labial herpes whose cause is thought to be reactivation of HSV occurred, whereas no herpes zoster by reactivation of VZV were observed . Fever cNerve growth factor (NGF) and epidermal growth factor (EGF) inhibits HSV reactivation in neuron in vitro , 27. BotIn severely burned patient, the number of CD8+ T cells decreases . CytolytIn contrast to the reduction of the number of T cells in severely burn patient, the number of B cells do not alter after burn injury . B cellsThe rate of cytomegalovirus (CMV) infection is considerably high in severely burned patients, that 71% in CMV seropositive burn patients and 13% in CMV seronegative patients . SuperinVZV infects not only neurons but also many other cell types. In severe systemic cases of VZV infection, infected cells can be found in almost every organ, including the spleen, liver, lungs, thymus, digestive tract, kidneys, pancreas, aorta, heart, and brain , 37–42. CHDF blood purification combined with PMX-DHP might have been associated with the occurrence of disseminated zoster in our case. CHDF plus PMX-DHP was initiated on day 1 and continued until the day the patient died (day 52). CHDF plus PMX-DHP was used not only to treat renal failure, but also to modulate inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1, and IL-6 . ExtensiThe prevention of airborne transmission of VZV to other patients and health care workers is clinically important. The reported incidence rate of disseminated zoster is lower than the incidence rate of KVE in burn patients. However, there is a possibility that disseminated zoster in burn patients is overlooked, because disseminated zoster has been rarely known to occur as a complication in burn patients. Varicella outbreaks among pediatric burn patients and other ICU patients have been reported , 12. AnuIn conclusion, we treated a severely burned patient with suspected disseminated zoster basing on his clinical findings and results of serology. Immediate isolation of the patient in a negative-pressure room was implemented to prevent airborne infections of VZV. Burn-related immunosuppression and long-term cytokine modulation by CHDF plus PMX-DHP might lead to rare, fatal reactivation of latent VZV infection.


Automation is one of the trending terminologies in the field of engineering to achieve various sensors and actuators such as the hydraulic system. Smart fluid is also one of the hot topics for researchers to develop a type of actuator in many control systems since the fluid’s rheological characteristics can be controlled or tuned by the intensity of the external stimuli. In this work, a new smart fluid of magnetorheological (MR) fluid is proposed and its field-dependent rheological characteristics are experimentally identified. An MR fluid using the carrier fluid as the blend of three different fluids, namely silicon oil, honey, and organic oil is prepared. In addition, two types of natural oils are used, sunflower oil and cottonseed oil. The samples are prepared using the blend as the carrier fluid, electrolytic iron powder coated with guar gum as the dispersed phase, and oleic acid as an additive. The quantity of oleic acid is optimized for 30% by weight of electrolytic iron powder. Two samples based on sunflower and cottonseed oil are synthesized and characterized for shear viscosity and shear stress with respect to shear rate subjected to a variable magnetic field. The blend-based MR fluid shows about 10% improvement over the sedimentation rate of silicon oil-based MR fluid as compared to that to conventional MR fluid. The cottonseed oil blend-based MR fluid performs better than sunflower-based fluid in terms of the viscosity and structure. Magnetorheological (MR) fluid is one of the kinds of smart fluids where the behavior of fluid can be shifted from Newtonian fluid to a semi-solid material (Bingham fluids) by being subjected to a magnetic field. The rheological properties of MR fluids are the function of an applied variable magnetic field; hence, to obtain precise control over rheological properties a variable magnetic field is desirable. Additionally, the yield stress in MR fluid which is directly proportional to the viscosity and intern to the applied variable magnetic field, can be formulated as .(1)τ=τy2, and identified improved compatibility of non-polar silicone oil with coated particles. Kumar [Synthesis of fluid dates back to 1949 by Jacob Rabinow, further the development of fluid was carried out in various aspects and stages. Sarkar and Hirani  presentes. Kumar  studied s. Kumar .In this study, the basic and critical constituent of MR fluid, that is the, carrier fluid is the center of importance which is the blend of organic oils. For the study, two organic oils are considered, namely, sunflower oil and cottonseed oil. The blends are prepared using fluid silicon oil (25%), honey (25%), and organic oil (50%) weight ratio. An electrolytic iron powder (EC10TR) is employed as magnetizable particles, these particles are coated with guar gum to enhance dispersibility and finally the oleic acid which forms a bond between the guar gum film and a carrier fluid is added. In this study, the carrier fluid used employs organic fluid (75% by weight), which ensures the eco-friendly nature of the MR fluid. The obtained MR fluid makes it a right candidate for various MR applications such as brake, clutches, damper, etc.The manufacturing process of MR fluids follows an exclusive process involving the coating of magnetic particles with guar gum and also obtaining a carrier fluid by blending various fluids to enhance the physical properties of MR fluid ,9.The physical and rheological characteristics of a magnetorheological fluid are the combined effort of each constituent put together in the process of synthesis. The optimization of these parameters and constituents for various applications is one of the biggest challenging issues for research. In this paper, an effort was made to optimize the additive in synthesis process. There are various parameters and factors which influence the overall performance of MR fluid, out of these the critical aspects are discussed below.−1 maintained at 40 °C [In order to obtain a high performing MR fluid, a researcher has to consider off-state viscosity of MR fluid as a critical parameter. The field-independent or off-state viscosity  and is one of the challenging aspects for automobile industries ,13. As aAs the application of MR fluid-based systems in the automobile sector is of importance and the operating temperature of these automobiles can range from sub-zero to prolonged operating time resulting from temperature rise. Commercially available MR fluid is reported to withstand temperature of 80 °C.The carrier fluid plays a critical role in deciding the characteristics of the MR fluid as it forms the major essentials of fluid (50%–80% by volume) . CommercCommonly the particle size of magnetizable particles dispersed in MR fluid varies from 1 to 10 µm ,16. The The stabilizers are employed in order to keep the magnetizable particles suspended in the carrier fluid, whereas surfactants serve in enhancing polarization by adhering to the surface of magnetic particles under the applied magnetic field. In this study, guar gum is used as a coating to magnetic particles that provides antifriction behavior to the application system. Hence, this enhances sedimentation stability ,9. FurthThe preparation of MR fluid starts with the preparation of carrier fluid, that is, the organic oil blend and the coating of magnetic particles using guar gum. Two types of organic oil blends are prepared (1) silicon oil (25%) + honey (25%) + sunflower oil (50%), called sunflower-based blend, and (2) silicon oil (25%) + honey (25%) + cottonseed oil (50%), called cottonseed-based blend. The electrolytic iron powder particles are used as magnetic particles in the study, the coating of electrolytic iron powder is done using guar gum, the irregular shape and surface texture of EI particles promotes proper surface interaction for coating.The coated magnetic particles are mixed with oleic acid to obtain proper bonding between oleic acid and the guar gum present on the surface of EI particles. The weight proportion of EI powder in prepared samples is kept constant to 30-weight percentage, and the proportion of oleic acid is varied to investigate the best suitable amount of oleic acid for EI composition. After the mixture is ready, it is supplemented with the prepared organic oil-based blend carrier fluid in parts of four for every half hour, which helps the thorough mixing and dispersion of the particles in carrier fluid, followed by two hours of stirring. The prepared MR fluid is subjected to sonication for two hours to confirm uniform dispersion of particles in the fluid to obtain a stable MR fluid. The pictorial representation of the whole process is shown in The scanning electron microscope (SEM) is used to study the morphological characteristics of the sample, and hence the quality of particles is tested. The details on the structure of particles and the size distribution carried out using SEM is presented in The study was carried out for two batches of MR fluid samples and in each sample series, five different samples for each batch were synthesized by varying the weights of the oleic acid (OA) ranging from zero to 0.35% (wt). The optimal weight percentage of OA in the MR fluid composition is identified by plotting graphs between the shear rate and shear stress of the samples, and is presented in −1 as a function of shear stress are plotted in The results for shear rate varied from 100 to 1000 s−1. The sample exhibits a non-Newtonian behavior, even under off-state conditions which is due to the high concentration of magnetic particles in the samples. The off-state viscosity of cottonseed oil-based MR fluid is slightly higher than sunflower oil-based fluid because of its inherent high viscosity.With the optimized quantity of additive OA, the final samples were prepared with blends of cottonseed oil and sunflower oil. It is noted that the rheological characterization of the MR fluid for the samples is carried out under a static magnetic field and constant static shear stress. The plot in The viscoelastic characteristics of MR fluid under magnetic field are studied using an oscillatory test on a magneto-rheometer under different magnetic fields: 0, 45, 81, 136 kA/m. The loss modulus amplitude measurement plot is shown in The G′ value increases with an increase in magnetic strength. In addition, the loss modulus data develops two zones, first zone loss modulus with increase up to the peak implying the development of a micro-size crack on particles which occurs in semi solid-state (G′ > G″). Once the plot reaches the second zone implying growth of bigger cracks, at the end it yields to fluidic state (G′ < G″). The plots show that both sunflower and cottonseed oil blend-based MR fluid with 0.5 weight % of oleic acid respond similarly to the externally applied magnetic field with an increase in shear stress corresponding to the magnetic field, which is also observed in MR fluid characterization ,21. The The investigation on the dispersibility of particles in carrier fluid for both the samples is done using Turbiscan stability index. The particle suspension samples are transferred into a cylindrical cell as shown in The guar gum coating over particles and OA facilitates suspension with the variation in viscosity of blends which are used as carrier fluid. This is considered as one of the influencing factors among the samples. Sedimentation in the MR fluid needs to be controlled to the lowest possible value, as the rate of sedimentation has a direct effect on the reduction in yield strength value. The sedimentation of Lord MRF-132DG shows a sedimentation ratio of 2.3 for a period of 72 h , while MThis work focused on the carrier fluid in the manufacturing of new MR fluid. The carrier fluid was blended with three different fluids; silicon oil (25%), honey (25%) and organic oil (50%). Two types of organic oil are employed for the comparison purpose; sunflower oil and cottonseed oil. The reason for the choice of such natural organic oils is closely related to the eco-friendly environment which has not been considered in existing MR fluids, including commercially available MR fluid. The MR fluid characterization was studied for various compositions of oleic acid to optimize the value which was identified by 0.25% and hence this was used to synthesize the final two samples for the rheological and sedimentation study. The results obtained in this experimental work demonstrate that the prepared samples show solid-like behavior under an external magnetic field. In addition, the coating of guar gum to the particles yields better sedimentation stability. It was also identified that the cottonseed oil-based MR fluid shows better sedimentation resistance compared to sunflower-based MR fluid. It is finally remarked that MR fluids proposed in this work are eco-friendly due to the use of edible organic oils. More specifically, considering the environmental aspect, the synthesized MR fluid consists of more than 75% organic ingredients which do not cause any harm to the environment. This environmental issue has not been reported yet in the development of MR fluids. In addition, it should be remarked that the proposed MR fluids can provide comparable field-dependent rheological properties such as the storage modulus with cost-effectiveness. In the future, an optimal recipe to make eco-friendly MR fluids will be further explored with certain applications such as MR brake and MR damper.It is finally remarked that the field-dependent rheological properties of the synthesized MR fluids need to be identified at a wide temperature range and a comparative work on various properties (such as the yield stress and wear) between commercial MR fluids and proposed MR fluids should be undertaken to demonstrate advantages and disadvantages of the proposed MR fluids. These studies will be performed as a second phase of this work.


Electrocardiogram signal analysis is based on detecting a fiducial point consisting of the onset, offset, and peak of each waveform. The accurate diagnosis of arrhythmias depends on the accuracy of fiducial point detection. Detecting the onset and offset fiducial points is ambiguous because the feature values are similar to those of the surrounding sample. To improve the accuracy of this paper’s fiducial point detection, the signal is represented by a small number of vertices through a curvature-based vertex selection technique using polygonal approximation. The proposed method minimizes the number of candidate samples for fiducial point detection and emphasizes these sample’s feature values to enable reliable detection. It is also sensitive to the morphological changes of various QRS complexes by generating an accumulated signal of the amplitude change rate between vertices as an auxiliary signal. To verify the superiority of the proposed algorithm, error distribution is measured through comparison with the QT-DB annotation provided by Physionet. The mean and standard deviation of the onset and the offset were stable as  Electrocardiogram (ECG) signals are electronically converted signals from the depolarization and repolarization of the atria and ventricle . GeneralThe fiducial point detection step identifies the most important fiducial points of the ECG signal, which are represented by the onset, the peak, and the offset of the P-wave, the QRS complex, and the T-wave ,21,22,23The R-peak, which is the peak of the QRS complex, is the easiest to detect because it has the largest amplitude value. It is used not only for measuring heart rate, but also for detecting other fiducial points. Typical R-peak detection methods are based on the Pan’s method, which is approximately 99% accurate. However, accurate detection methods of the fiducial points of the QRS complex other than the R-peak have been not clearly determined. Their low detection rate and inaccuracy are problematic due to signal deformation caused by various arrhythmias. These difficulties stem from the ambiguity of the reference regions that serve as a starting point.Among various signal compression techniques, a method using polygonal approximation compresses a signal through a small number of vertices. In this case, since the onset and offset of the waveform represent the boundary between the waveform region and the baseline region, they are well-preserved as vertices.As shown in On the other hand, since the polygonal approximation uses the atomic vertices, there is a large difference in the feature values of the vertices. As shown in In this paper, we propose a curvature-based vertex selection technique to solve the ambiguity of the fiducial points in the QRS complex. Our approach is roughly divided into three stages. The first stage consists of initial vertex selection using curvature-based polygonal approximation. Since the curvature value of the fiducial point is large, most of the fiducial points are represented as vertices. The second stage is an incremental vertex selection using repetitive sequential polygonal approximation, and the third stage performs an additional vertex optimization step using dynamic programming. These steps are applied for a missing case due to ambiguous curvature value.After applying the polygonal approximation, unwanted variations of the QRS complex—such as the presence of Q-waves, S-waves, and the polarity of the waveform—remain a major problem. To mitigate the side effects of this problem, we have prepared an auxiliary signal by accumulating data from the polygonal approximation signal. This cumulative signal monotonically increases by accumulating the absolute value of the rate of change of the amplitude between the vertices of the polygonal signal. Thus, by effectively expressing and emphasizing the feature of the fiducial point, we could effectively represent changes in the QRS complex’s shape and polarity. From the cumulative signal, we analyze feature values for each vertex, such as the amplitude difference between R-peak and vertex, the time difference between reference point and vertex and the angles with neighbor vertices. Then, we determine the vertex with the largest sum of these feature values as a fiducial point.This paper is organized as follows. The ECG signal, which consists of the P-wave, QRS complex, and T-wave, includes the corresponding onset, offset, and peak points, which are referred as fiducial points. Power line interference: various high frequency noise according to country.Baseline wander: a low-frequency noise (0.15 up to 0.3 Hz). This noise results from the patient breathing and leads to a baseline shift in the signals.Electrode contract noise, electrode motion artifacts, muscle contractions, electrosurgical noise, instrumentation noise, and so on.As shown in 1 and 2 are typical high- and low-frequency ECG signal noises, respectively. Since noise complicates the baseline, it is difficult to estimate baseline and thus detect the boundary with an ambiguous waveform. Most of study use the bandpass filter for suppress noises and in some case, it uses a notch filter  for aimiPeaks, such as the R-peak, can be easily detected because the amplitude has a local maximum or minimum and the rate of amplitude change is large enough. In contrast, the boundary between the baseline and the waveform is still ambiguous, even when filtering is applied. This is because the amplitude change occurs slowly, and the features are similar to the surrounding samples. In this paper, we propose an effective fiducial point detection technique by emphasizing ambiguous fiducial points based on polygonal approximation.Signal approximation techniques have been widely studied by using polygonal approximation, such as sequential polygonal and cyclic polygonal approximation, and polynomial approximation, such as B-spline. However, these techniques are problematic, since they select too many vertices and errors are not minimized.In the curvature-based vertex selection technique , curvatuS) is summarized as follows. Separate the R-R section of the input signal. In this paper, we detect the R-peak by Pan’s method.After calculating the curvature for the separated R-R section, the curvature-based polygonal approximation technique is applied to select the initial vertices. Equation  represeni-th initial vertex and the We apply the sequential polygonal approximation method to the interval between each initial vertex to select additional vertices. Equation  represenDynamic programming is applied to the additional vertices to optimize their position. Equation  is a setRepeat steps 2–4 to proceed with polygonal approximation for the entire input signal. Equation VD={With the absolute value of the amplitude difference obtained using Equation , that vaThis simplifies the signal as monotonically increased, even if the QRS complex appears as a downward wave or includes Q-peaks and S-peaks that appear as downward or upward waves. The vertex corresponding to the fiducial point also maintains the feature of dividing the baseline and waveform regions.As shown in The feature value of each vertex of the accumulated signal is analyzed to determine the fiducial point. In this paper, we propose three types of features for each vertex to determine the fiducial point: the amplitude difference between R-peak and vertex, the time difference between the reference point and vertex, and the angles with neighbor vertices.The onset and offset of the QRS complex are the boundary points between it and the baseline region. The amplitude difference between the R-peak and the vertex in the accumulated signal is close to the maximum value near the fiducial point. Therefore, the Q-peak with a largest amplitude difference in Equation  represenIn this paper, we suggest the time difference as a second feature value for excluding the case of detecting the onset of the P-wave. Generally, the normal width of the QRS complex is about 0.08 to 0.12 s. We use a reference point based on the point 0.3 s away from the R-peak for estimating the time difference and consider that the larger the time difference is, the more likely it is to be the fiducial point.If the ventricular arrhythmia occurred and the width of the QRS complex is increased, fiducial point may have lower time difference feature value when there is a vertex at notch in QRS complex. However, this problem can be easily solved from the amplitude difference between previous R-peak and vertex, since the amplitude of notch and fiducial point is similar to previous R-peak and baseline, respectively.Equation  represenF denotes a sampling frequency and Since the fiducial point is boundary between the waveform region and the baseline region, most significant feature of vertices of fiducial point is angle between the horizontal line and straight line connecting the neighbor vertex.i-th vertex, respectively.In the case of the onset, the angle with the left vertex is close to 0 degrees, and the angle with the right vertex is close to 90 degrees. In the case of the offset, it is reversed. Based on the feature values of the fiducial point, we can calculate the feature value of each vertex in the searching interval. Our approach provides a method to select the point with the highest probabilities of being the fiducial point by summarizing all feature values. Equations  and 10)10) repreIn the case of the baseline direction angle, the square root is added to consider that the vertex corresponding to the fiducial point may have a high value of about 30 to 40 degrees due to noise or signal distortion. On the other hand, since the waveform angle has a low possibility of distortion because of the large amplitude change, a square is added to have a low feature value for the vertices except the fiducial point.In the preprocessing step, noise suppression and R-peak detection are performed. Breathing and muscle movements cause the low-frequency noise, and power noise of 30 Hz or 60 Hz causes the high-frequency noise. In this paper, these noises are suppressed using a Butterworth bandpass filter of 1–25 Hz and the most widely known Pan’s method is applied to R-peak detection. The QT-DB and MIT-BIH ADB provided by Physionet are used to evaluate the performance of the proposed algorithm.The QT-DB contains a total of 105 fifteen-minute excerpts of two-channel ECGs, which were carefully selected to avoid significant baseline wander or other artifacts. Within each record, around 30 numbers of beats were manually annotated by cardiologists, who can identify the onset, peak, and offset of the QRS complex. The proposed algorithm works on a single-channel signal, while cardiologist manually recorded one annotation considering both channel of signal simultaneously. Therefore, to compare our approach with the manual annotations on the QT-DB for each of the two single-channels, it is reasonable to choose the annotation result of the channel with less error [ng party , toleranWe summarized the experimental results by our proposed method in Most of the data is regarded as having satisfied the tolerance or shown on the detection result similar to the tolerance. Especially in the case of MIT-BIH normal sinus rhythm DB (represented as a red marker), it can be confirmed that the standard deviation of the total error satisfies tolerance. On the other hand, MIT-BIH arrhythmia DB and sudden-death patient DB, represented by a black marker, contains various arrhythmia heartbeats and caused large errors compared to another DB.As with other algorithms, the standard deviation of error of the onset is out of tolerance. On the other hand, the offset satisfies the tolerance and confirmed a lower error than other algorithms.To analyze experimental results in detail, we tried to cluster the experimental results for each data set. The experimental results of some data according to the types of QRS complex are shown in Clustering was performed around the R-peak, and the position of the R-peak was shown as the origin. As shown in Since MIT-BIH ADB only annotate the arrhythmia type of each heartbeat, statistical analysis for fiducial point detection is hard. We use the mean length of QR section, which start from the onset to R-peak, and RS section, which start from R-peak to the offset, as reference for onset and offset detection, respectively. According to the type of heartbeat, we separate into normal heartbeats and abnormal heartbeats, and calculate the mean and standard deviation of each type of heartbeats, respectively.However, in the case of arrhythmia, there are various types of abnormal heartbeats. Most occur with premature atrial contraction  or premature ventricular contraction , with the remainder very rare. For example, supraventricular premature or ectopic beat  only appears twice in record 208 over the total 107,265 heartbeats in MIT-BIH ADB. Thus, the test for abnormal heartbeat only proceeded for one of the most typical abnormal heartbeat types for records with more than 30 PACs or PVCs.Generally, since the PAC does not affect the QRS complex, the stability of the fiducial point detection is high as shown in The proposed algorithm detected the fiducial points regardless of whether Q- or S-waves are present or not in the normal heartbeat. In addition, we could reliably detect the fiducial points for the various types of QRS complex with arrhythmia.From this experiment, we can confirm that the proposed polygonal approximation method can effectively detect fiducial points compared with other algorithms. In addition, it obtains stable results for various types of QRS including arrhythmia. In some cases, detection error is high due to the acquisition of signals such as the V2 and V5 channel, rather than signal acquisition by the modified lead II (MLII) channel, which emphasizes the QRS complex. The distortion in the preprocessing process causes an error. Therefore, we consider that the signal acquisition clearly reflecting the shape of the QRS complex—such as the MLII channel—or the stable noise suppression technique can improve the performance of the proposed algorithm.In this paper, the proposed algorithm focuses on mitigating the ambiguity of the fiducial point by polygonal approximation to extract the feature points of ECG signals, motivated from the characteristic that the polygonal approximation preserves the fiducial point as a vertex. Therefore, our approach resulted in better performance compared to other signal compression techniques. In addition, we propose an effective auxiliary signal for stable detection results in various types of QRS complex using these features. Experimental results show that the proposed auxiliary signal-based technique enables stable detection for various applications in real ECG databases equipped with QT-DB and MIT-BIH ADB.In future research, the proposed method is required not only to expand the detection of P- and T-waves, but also to apply post-processing to improve the existing technique. This method can also be extended to study adaptive signal compression technique according to importance of intervals by reapplying fiducial point detection results. The trade-off between the accuracy and energy consumption could be achieved by adjusting the fiducial points compression ratio, so the extremely long-time ECG monitoring services are capable in the battery-operated wearable systems.


We characterize the class of nondeterministic


LMW) regress dependent on T cells. Yet, TAMS not T cells express FGF receptors. Bone marrow derived-macrophages from Fgf2LMW−/− mice co-injected with cancer cells reduce tumour growth and express more inflammatory cytokines. FGF2 is induced in the tumour microenvironment following fractionated radiation in murine tumours consistent with clinical reports. Combination treatment of in vivo tumours with fractionated radiation and a blocking antibody to FGF2 prolongs tumour growth delay, increases long-term survival and leads to a higher iNOS+/CD206+ TAM ratio compared to irradiation alone. These studies show for the first time that FGF2 affects macrophage programming and is a critical regulator of immunity in the tumour microenvironment.Regulation of the programming of tumour-associated macrophages (TAMs) controls tumour growth and anti-tumour immunity. We examined the role of FGF2 in that regulation. Tumours in mice genetically deficient in low-molecular weight FGF2 (FGF2 Macrophages contribute to tumour progression and response to therapy. Here, the authors show that absence of FGF2 in the tumour microenvironment reduces tumour growth and enhances the anti-tumour immune response by altering macrophage polarization. As a result, disruption of this macrophage programming by anti-FGF2 blocking antibodies enhances the outcome from radiotherapy.  Whilst some clinical data indicates an association with reduced patient survival3, others suggest specific subsets of macrophages can improve patient outcomes5. These disparities may be due to macrophages having both pro- and anti-tumourigenic effects. Macrophage nomenclature has evolved over recent years to take account of these differing phenotypes, and although some debate still remains, it is largely accepted that most TAMs lie on a spectrum that can encompass features of both classically activated ‘M1’ macrophages, exhibiting immune-stimulatory functions, and alternatively activated ‘M2’ immune-suppressive macrophages9. Experimental evidence demonstrates that TAMs are often skewed towards a pro-tumourigenic phenotype. Tumour microenvironmental stimuli, including tumour-derived cytokines, can polarise macrophages towards an M2 phenotype11, promoting tumour growth through mechanisms including angiogenesis, proliferation, local invasion, metastatic potential and immune escape12. Macrophages also mediate resistance to therapy including radiotherapy, and irradiation can augment angiogenic and pro-tumourigenic macrophage phenotypes15. Recent data suggest a key role for macrophages in modulating the host adaptive immune response, primarily via suppressive effects on CD8+ T lymphocytes16.Tumour-associated macrophages (TAMs) are highly abundant in a range of solid tumours20. Of these receptors, FGF2 has the highest affinity for FGFR1, 2 and 3b21. FGF2 has many isoforms, generated by the use of alternative translational start sites22. The lowest molecular weight isoform, FGF2LMW, is the only secreted form24, and is a highly potent angiogenic molecule25. FGF2LMW also affects the response to injury in skin, nerve and cartilage27. The higher molecular weight forms of FGF2 are often nuclear in location and can regulate transcription independently of FGFRs. In previous work, we found that FGF2LMW was highly expressed by myeloid cells in colorectal cancer (CRC) liver metastases. Blocking FGF2 with an antibody delayed growth of murine CRC liver metastases through vascular renormalisation25.In this study, we demonstrate that fibroblast growth factor 2 (FGF2) plays a pivotal role in shifting TAMs towards a pro-tumourigenic, M2-like phenotype in the tumour microenvironment. FGF2 is one member of a family of related proteins (FGFs) that exert mitogenic activity by binding to FGF receptors (FGFR)LMW(Fgf2LMW−/−) while shifting TAM polarisation. Cancer cells failed to develop substantial liver metastases or subcutaneous tumours in Fgf2LMW−/− mice. Fgf2LMW−/− TAMs were more inflammatory, (M1-like) than in C57Bl6 wildtype (WT) mice, and subcutaneous tumour regression was T cell-dependent. In WT mice, TAMs were the major source of FGF2, and were the only immune cell to abundantly express FGFR1 and 2. FGF2 has been reported to be induced in irradiated human tumours31 raising the possibility that FGF2 might be a useful therapeutic target for patients who have received radiotherapy. Accordingly, we examined the effect of FGF2 on the irradiation response, and found that a blocking antibody to FGF2 in combination with radiotherapy reduced or even eliminated tumour regrowth in association with an increase in the TAM iNOS+/CD206+ ratio .Here, we show that tumours regressed in mice genetically deficient in FGFFgf2LMW−/− mice as they were by blocking anti-FGF2 antibody32, we examined the growth of liver metastases in Fgf2LMW−/− mice. After intrasplenic injection of the murine CRC cell line, MC38 or the pancreatic cancer cell line, KPC into WT or Fgf2LMW−/− mice, Fgf2LMW−/− mice had significantly less liver tumour burden  and KPC (2 × 105 cells) tumours reached endpoint by day 21 in WT mice, yet only 1 of 10 (10%) MC38 and 5 of 8 (63%) KPC tumours in Fgf2LMW−/− mice reached endpoint by day 120 , which extended into the clusters of tumour cells  with over a 100-fold increase in Fgfr2 , while fewer MDSCs or granulocytes expressed FGF2, 9.7% and 29.9% respectively. In contrast, FGF2 was expressed in <4% of T cells Fig. . TAMS exAMs Fig. . It shousis Fig. . Macrophion Fig. . These d41. Therefore, we assessed TAM polarisation in Fgf2LMW−/− mice by flow cytometry and RNA analysis   Fig. . TAMs isYm2 Fig. . These dFgf2LMW−/− or WT mice to polarisation stimuli, LPS (M1) or IL4 (M2). Without stimulation, Fgf2LMW−/− BMDM had higher levels of the pro-inflammatory cytokines, Cxcl1, Il1β, Il6 and Tnfα, and decreased Ym2, a M2 marker compared with WT BMDM  did not reduce cellular viability, proliferation or clonogenic survival after radiation  ratio of TAMs decreased with irradiation in the SW1222 tumours, but was unchanged in the other two models  have the capacity for both pro- and anti-tumourigenic functionsFgf2LMW−/− mice with tumour regression dependent on T cells. In these tumours TAMs were the predominant immune cells that expressed FGF2 and its receptors. Although the proportion of macrophages infiltrating the tumours was equivalent in WT and Fgf2LMW−/− mice, the nature of these TAMs was substantially different. Macrophage polarisation or activation can lead to a complex variation of expression patterns and functions47. These range from inflammatory, anti-tumourigenic phenotypes (M1) to pro-tumourigenic and immunosuppressive functionality (M2). In general, TAMs have been described as more M2 in nature50. TAMs from Fgf2LMW−/− mice expressed many characteristic pro-inflammatory cytokines and markers compared with WT TAMs, and Fgf2LMW−/− BMDM suppressed tumour growth after co-injection. This suppressive effect was associated with enhanced infiltration of T cells in immunocompetent mice, but was also evident in immunosuppressed mice. This enforces the concept that FGF2 mediates tumour growth through alteration of TAMs, both dependent and independent of adaptive immunity. In addition, other cell types in the tumour microenvironment, such as fibroblasts are also likely to be affected by the Fgf2LMW−/− genotype and the altered phenotype of TAMs almost certainly includes indirect effects.In our experiments, cancer cells including MC38 at lower innocula failed to generate continued tumour growth in 51. As a result, in some glioblastoma models, inhibition of CSF1 led to a shift in TAM polarisation away from pro-tumourigenic phenotypes resulting in tumour regression independent of adaptive immunity51. In other models, inhibition of CSF1 resulted in an enhanced immune response52. Furthermore, inhibition of PI3Kγ blocked M2 polarisation of macrophages, promoted T cell recruitment, and enhanced anti-tumour immunity41. Here we showed that co-injection of BMDM conditioned by tumour cells had a tumour growth promoting effect. We also found that BMDM deficient in Fgf2LMW−/− mice have an anti-tumour effect. These actions were both adaptive immune dependent and independent.Macrophages have multiple effects on tumour growth, some of which can involve tumour immunity, with or without an adaptive immune response. For example, CSF1 which acts as both a chemotactic and survival factor for macrophages also promotes M2-like polarisation55, but nonetheless an effective anti-tumour response often fails to occur even after administration of immune checkpoint therapy56. Macrophage recruitment and polarisation after irradiation of tumours is one factor contributing to immunosuppression, which can counteract the immune stimulatory effect of irradiation57. It should be noted that alteration of macrophage recruitment and polarization has been shown to radiosensitize tumours. Agents such as anti-CSFR1 antibody and SDF1 inhibition have been used in this way58. The dose of radiation makes a difference59. Klug et al. showed that at relatively low doses, up to 2 Gy, radiation led to improved T cell infiltration into tumours due to induction of NOS2 expressing macrophages37. Some of these effects were due directly to irradiation of the macrophages. In our hands at higher doses a similar macrophage phenotype also was influenced by the absence or blocking of FGF2. We observed an increase in FGF2 following irradiation of tumours in mice similar to those described in several clinical reports44. Blocking FGF2 with a specific antibody led to diminished tumour regrowth after irradiation that was associated with an increase in M1 polarisation of TAMs. It should be noted that there are at least 22 human FGFs. Blocking only FGF2 is likely to have very different effects than blocking less specifically the FGF-FGFR signalling pathways. Treatment with the blocking anti-FGF2 antibody did not affect tumour growth or macrophage polarisation unless irradiation had been applied, similar to the tumours formed from high innocula in Fgf2LMW−/−mice. We failed to find evidence that FGF2 was acting as an autocrine growth factor for these cancer cells. Macrophages influence tumour angiogenesis40. FGF2 certainly might affect angiogenesis directly in these models, but the effect could also be indirect through reprogramming of macrophages. In our hands blocking FGF2 or its absence did not have a generalisable effect on tumour vascular density before or after radiation. Inhibition of tumour growth after irradiation occurred in both immunocompetent and immunosuppressed mice. The extent of the additional effect of the anti-FGF2 antibody correlated with the extent of the tumour response to radiation alone. These data are consistent with previous reports showing inhibition or depletion of macrophages after irradiation leads to prolonged tumour growth delay or in some cases augments immune checkpoint therapy61.Radiotherapy is known to stimulate adaptive immunity to tumoursHere, we directly associate TAMs with increased tumour growth based on experiments showing enhanced tumour growth after co-injection of cancer cells with pre-conditioned BMDMs. Macrophages were the only immune cell population in tumours to abundantly expression FGFR1 or 2, and in tissue culture, we showed that tumour cells can induce the expression of FGFR1 and 2 in BMDM. Furthermore, FGF2 levels are increased in the tumour microenvironment following irradiation in vivo. Collectively, these findings suggest that TAMs in the tumour microenvironment are ready to engage with FGF2 in the irradiated tumour microenvironment leading to reprogramming to generate a more pro-tumourigenic milieu. These TAMs can then contribute to tumour growth in both an immune dependent and independent fashion.Fgf2LMW−/− mice. It also suggests that FGF2 in the tumour microenvironment is an important regulator of macrophage differentiation, particularly during radiotherapy. Further recognition of factors, which contribute to TAM polarisation could lead to novel therapeutic strategies. In non-cancer models, effects on macrophages by FGF2 have not been directly described. However, enhanced leucocyte recruitment directed by FGF262, and promotion of autoimmune arthritis61 are both suggestive of potential macrophage involvement. FGF2 deficiency leads to enhanced colitis in murine models63, demonstrating the capacity of FGF2 to mediate inflammation. Furthermore, FGF2 supports wound healing, a process associated with a rapid influx of myeloid cells64, and depletion of FGF2 delays wound healing65. Even though effects on macrophages have not been described directly in these settings, our work introduces these possibilities and raises additional questions about effects on fibroblasts, other cell types and other more complex indirect actions. Overall our work defines FGF2 as a key modulator of macrophage polarisation in tumours and demonstrates that FGF2, as well as other factors which regulate macrophage activity, can be targeted during radiotherapy.Our work suggests that macrophage polarisation could be a component driving delayed tumour growth and tumour regression in All methods are available in the Further information on research design is available in the Supplementary InformationPeer Review FileReporting Summary


Perioperative fluid management – including the type, dose, and timing of administration –directly affects patient outcome after major surgery. The objective of fluid administration is to optimize intravascular fluid status to maintain adequate tissue perfusion. There is continuing controversy around the perioperative use of crystalloid versus colloid fluids. Unfortunately, the importance of fluid volume, which significantly influences the benefit-to-risk ratio of each chosen solution, has often been overlooked in this debate.The volume of fluid administered during the perioperative period can influence the incidence and severity of postoperative complications. Regrettably, there is still huge variability in fluid administration practices, both intra-and inter-individual, among clinicians. Goal-directed fluid therapy (GDFT), aimed at optimizing flow-related variables, has been demonstrated to have some clinical benefit and has been recommended by multiple professional societies. However, this approach has failed to achieve widespread adoption. A closed-loop fluid administration system designed to assist anesthesia providers in consistently applying GDFT strategies has recently been developed and tested. Such an approach may change the crystalloid versus colloid debate. Because colloid solutions have a more profound effect on intravascular volume and longer plasma persistence, their use in this more “controlled” context could be associated with a lower fluid balance, and potentially improved patient outcome. Additionally, most studies that have assessed the impact of a GDFT strategy on the outcome of high-risk surgical patients have used hydroxyethyl starch (HES) solutions in their protocols. Some of these studies have demonstrated beneficial effects, while none of them has reported severe complications.The type and volume of fluid used for perioperative management need to be individualized according to the patient’s hemodynamic status and clinical condition. The amount of fluid given should be guided by well-defined physiologic targets. Compliance with a predefined hemodynamic protocol may be optimized by using a computerized system. The type of fluid should also be individualized, as should any drug therapy, with careful consideration of timing and dose. It is our perspective that HES solutions remain a valid option for fluid therapy in the perioperative context because of their effects on blood volume and their reasonable benefit/risk profile. Perioperative fluid therapy is a routine aspect of daily clinical practice for most anesthesiologists but remains a therapeutic challenge. One of the most complex aspects of perioperative fluid therapy is determining how much fluid to give each patient. Numerous observational studies have reported a strong association between both excessive and insufficient perioperative fluid administration and an increased risk of postoperative complications –6. As itThe choice of intravenous fluid type has also been a subject of passionate debate, recently refueled by the publication of several large, prospective, randomized studies in different patient populations. These studies have demonstrated the impact of intravenous fluid solutions on patient outcomes, particularly in critically ill patients in the intensive care setting . AlthougYES.P = 0.04)”. Another point of interest is that use of a GDFT protocol has never been associated with a deleterious effect for the patient. It is therefore not surprising that GDFT has been included in the national expert recommendations of several countries, including the United Kingdom and France  in trauma patients and PHOENICS trial [N = 2280] in elective abdominal surgical patients) are on the way. They should clearly help to precise the benefit to risk ratio associated with the use of HES in the two specific contexts. In the final debate, the price should be taken into account as HES solutions are much more expensive than crystalloid solutions, but significantly cheaper than albumin solutions in most European countries.Not surprisingly, these four studies confirm what physiology tells us: to achieve a predefined hemodynamic target, a smaller volume of colloid solution, namely HES 130/0.4, is required, compared to crystalloid solution, resulting in a less positive intraoperative fluid balance. Of note, several studies have reported an association between positive perioperative fluid balance and worse outcome –80. HoweIn the perioperative setting, not only the type of fluid but also the volume administered can impact a patient’s outcome following major surgery. Fluid volume should be guided by predefined physiologic targets, using an individualized hemodynamic algorithm. Clinician compliance with such protocols can be improved by using automated closed-loop systems that enable automation of some of the simple but restrictive therapeutic tasks. It is of prime importance that intravenous fluids be administered with the same care as any other drug, with strict indications and contraindications and precautions with regard to potential adverse effects. Finally, perioperative colloid solutions, and in particular HES solutions, may have a place in optimizing a patient’s hemodynamic status while limiting the total volume of fluid administered. Large multicenter studies that assess the impact of crystalloids vs colloids and that apply a strict approach that ensures high protocol compliance, for example with a closed-loop system, are needed.


Mitochondria are vital regulators of plant growth and development, constitute the predominant source of ATP, and participate in multiple anabolic and catabolic metabolic pathways. But the mechanism by which dysfunctional mitochondria affect plant growth remains unknown, and more mitochondria-defective mutants need to be identified.decreased vascular bundle 1 (dvb1) was isolated from rice mutant library mutagenized by EMS (ethylmethane sulfonate), which shows dwarfism, narrow leaves, short branches, few vascular bundles, and low fertility. Map-based cloning, genetic complementation, and phylogenetic analysis revealed that DVB1 encodes a structural protein classified in the Mic10 family and is required for the formation of cristae in mitochondria, and was primarily expressed in vascular bundles. The DVB1 protein is partially localized in the mitochondria and capable of forming dimers and polymers. Comparing with the wild type, disruption of amino acid metabolism and increased auxin synthesis were observed in dvb1 mutant which also showed increased sensitivity to the mitochondrial electron transport inhibitors.A mitochondria-defective mutant DVB1 belongs to Mic10 family and DVB1 is partially localized in the mitochondria. Further studies indicated that DVB1 is important for mitochondrial and plant development in rice.The online version contains supplementary material available at 10.1186/s12284-021-00454-3. Apart from energy, the roles of mitochondria in a variety of processes, such as amino acid metabolism, hormone biosynthesis, CaMitochondria and auxin are both critical regulators of plant growth and development, and more and more evidence suggest that mitochondrial function and auxin are interconnected. Mitochondrial dysfunction regulates auxin signaling, which in turn which can regulate mitochondrial metabolic and energy pathways to adjust plant growth  complex, and was partially localized in the mitochondria. In the dvb1 mutant, the mitochondria exhibited an abnormal structure, amino acid metabolism was disrupted, and the auxin content was increased. The results demonstrated that DVB1 is indispensable in mitochondrial and plant development.In our study, we identified a rice mutant dvb1 mutant were relatively indistinct, but the differences became more obvious with the passage of time. At seedling stages, there are no apparent difference in root between the wild-type and the dvb1  and observed that the cells in the dvb1 mutant appeared disordered and shorter than those of the wild type , which indicates that the mutant phenotypes were controlled by a single recessive nuclear gene. The DVB1 locus was mapped to a 52 kb region flanked by the markers W3–100 and InD3–2 on chromosome 3. Sequencing analysis revealed a single-nucleotide substitution from G to A in exon 2 within LOC_Os03g62420 in the dvb1 mutant, causing an amino acid conversion from Gly-90 to Glu-90 . DVB1 transcripts were abundant in the root, culm, leaf, sheath, and spikelet at each stage, and thus showed a non-specific spatiotemporal expression pattern. DVB1 transcripts showed higher abundance in mature leaves at the tillering stage  and the 2 × 35S::DVB1-GFP fusion protein were transiently expressed in rice protoplasts. Green fluorescence from expression of GFP alone was detected uniformly throughout the protoplasts except in the vacuole, whereas the signal from the 2 × 35S::DVB1-GFP fusion protein exhibited a dispersed dot-like and large polymeric cluster-like subcellular distribution  to examine the mitochondrial ultrastructure of the wild type and tae Fig. a, b. Theons Fig. c. Mitochant Fig. d, e, whiG90E could interact with themselves and with each other , which did not contain the mutation site of the DVB1 gene, was unable to interact with itself in a yeast two-hybrid assay , and could bend the inner membrane of mitochondria to form cristae  assays to evaluate differential gene expression between the .05 Fig. a. Kyoto ism Fig. b.Fig. 7dvb1 mutant phenotypes, namely dwarfism, narrow leaves, short branches, fewer vascular bundles, and abortive pollen, all these suggests that the dvb1 mutant was affected by auxin deficiency  was identified in rice, showing dwarfism, narrow leaves, short branches, few vascular bundles, and low fertility. DVB1 encodes a structural protein classified in the Mic10 family and is required for the formation of cristae in mitochondria, and is primarily expressed in vascular bundles. The DVB1 protein is partially localized in the mitochondria. Both DVB1 and DVB1G90E are capable to forming dimers and polymers. Disruption of amino acid metabolism and increased auxin synthesis were observed in the dvb1 mutant. Compared with the wild type, the dvb1 mutant showed increased sensitivity to the mitochondrial electron transport inhibitors. Results from this work suggest that DVB1 plays a crucial role in mitochondrial and plant development in rice.A mitochondria-defective mutant (Oryza sativa L. ssp. indica) with 1% ethylmethane sulfonate. In T1 generation, the dvb1 was found and was inherited stably after more than three years of observation. JH10, a restorer, was used as the wild-type (WT) in all experiments, all plants were bred in Southwest University, Chongqing, China, under natural conditions.We treaded the Jinhui 10 seeds (JH10) . The inner surface of the sheath was observed using a SU3500 scanning electron microscope  at an accelerating voltage of 5.0 kV at − 4 °C.The phenotypes of the dvb1 mutant plants, and fixed in 50% ethanol, 0.9 M acetic acid, and 3.7% formaldehyde overnight at 4 °C. The fixed samples were dehydrated with a graded series of ethanol , infiltrated with xylene, and embedded in paraffin . A rotary microtome RM2245  was used to cut 8-μm-thick sections, which were transferred onto poly-L-lysine coated glass slides, deparaffinized in xylene, dehydrated through an ethanol series, and stained with safranin-O and fast green. For cryosectioning, wild-type and dvb1 mutant leaves were cut into 0.5 cm pieces, placed in an embedding agent ENG50 , and rapidly frozen. The frozen samples were sliced to about 10 μm thickness, stained in hematoxylin and eosin solution, then dehydrated through a gradient ethanol series . The sections were observed using an Eclipse E600 light microscope .Leaves, panicle base, and culms were collected at the tillering stage from wild type and dvb1 mutant was crossed with rice ‘Xinong 1A’ (bred by the Southwest University Rice Research Institute) to generate the F1 population; the F2 population was derived by self-fertilization of the F1 population. The F2 individuals that exhibited the mutant phenotype were selected and used to map DVB1. Gene fine-mapping was conducted using simple sequence repeat markers obtained from the publicly available rice databases Gramene (http://www.gramene.org) and the Rice Genomic Research Program (http://rgp.dna.affrc.go.jp/E/publicdata/caps/index.html). Insertion/deletion markers were developed from comparison of the genomic sequences of ‘Xinong 1A’ and ‘Jinhui 10’.The http://Phytozome.jgi.doe.gov/pz/portal.html#!Search?show=BLAST). Vector NTI Advance 10 was used to align the amino acid sequences. The alignment was saved as a FASTA file and used for phylogenetic analysis with MEGA (v5.2). A phylogenetic tree was constructed using the neighbour-joining method based on the evolutionary model with the lowest Bayesian information criterion score. Bootstrap support values from 1000 replicates are presented for each node. The peptide sequences used for phylogenetic tree construction are listed in Supplementary Table SSequences of DVB1 homologous proteins from other plant species were acquired by conducting a BLAST search of the Phytozome portal (v12.1.6)  and 450 bp downstream sequences, was amplified from the wild-type genomic DNA and cloned into the binary vector pCAMBIA1301. The recombinant complementation plasmids were introduced into the dvb1 mutant using the Agrobacterium tumefaciens-mediated method. The primer sequences used for vector construction are listed in Supplementary Table STo complement the mutant phenotype of Ct method using ACTIN1 as an endogenous control  in accordance with the manufacturer’s instructions. The concentration, purity, and integrity of extracted RNA were determined using a NanoDrop 2000 spectrophotometer  and 1% agarose gel electrophoresis. All reverse transcriptions were performed using 1 μg total RNA with SuperScript® III Reverse Transcriptase  employing the oligo(dT) 18 primer in accordance with the manufacturer’s instructions. After cDNA synthesis, all samples were diluted tenfold with sterilized water, and qRT-PCR analysis was performed with the SYBR Supermix Kit  and the ABI 7500 Sequence Detection System  in accordance with the manufacturer’s instructions. Three replicates were performed and normalized relative expression levels were calculated by the ∆∆DVB1 was amplified and labelled using the DIG RNA Labelling Kit  in accordance with the manufacturer’s recommendations. Pretreatment of sections, hybridization, and immunological detection were performed as described previously  to generate the 2 × 35S::DVB1-GFP (green fluorescent protein) fusion protein. Both Pan580-DVB1-GFP and empty Pan580-GFP plasmids were introduced into rice protoplasts as described previously  for 30 min to stain the mitochondria  to generate the 2 × 35S:: DVB1G90E-GFP (green fluorescent protein) fusion protein. The transformation method is as above. The primer sequences used are listed in Supplementary Table SThe full-length coding region of Nicotiana benthamiana using an Agrobacterium (strain GV3101)-mediated infiltration. The pCAMBIA1300 vector was infiltrated as a control. After infiltration for 36–56 h, leaf cells were incubated with 0.5 mM MitoTracker™ Orange CMTMRos  for 30 min to stain the mitochondria  for at least 48 h, and then post-fixed in 1% osmium tetroxide for 2 h after washing with 0.1 mol L− 1 phosphate saline buffer. Tissues were stained with uranyl acetate, dehydrated in ethanol, and embedded in Spurr’s medium before thin sectioning. Samples were stained again and mitochondria examined using a H-7500 transmission electron microscope .Tissue fixation and preparation for TEM followed a previously described method with minor modifications  using an ACQUITY™ UPLC™ BEH-C18 column  and H-Class UPLC™ Tunable UV Detector .ATP extraction was performed as previously described  was cloned into the pGAD-T7 and pGBK-T7 vectors, and transformed into yeast cells. The DO-Leu-His-Ade and DO-Leu-Trp-His-Ade filter assays were performed in accordance with the manufacturer’s instructions to check self-activation of DVB1 BD and DVB1G90E BD and protein–protein interactions.The Matchmaker™ GAL4 Two-Hybrid System  was used to determine whether the DVB1 and DVB1G90E coding sequences were cloned into the expression vector pET-32a  using the Kpn1 and EcoR1 restriction enzymes, and introduced into E. coli strain BL 21 (DE3). Positive colonies were grown at 37 °C to an optical density at 600 nm (OD600). The fusion protein was induced by addition of isopropyl.To produce a His-tagged protein, the full-length DVB1 and DVB1β-D-thiogalactopyranoside to a final concentration of 1 mM, and then was purified with Ni-NTA Sefinose™ Resin  following the manufacturer’s protocol. Immunoblot analysis by SDS-PAGE and Native PAGE using a His-tag antibody  were performed as described previously . Total RNA was extracted from the samples using the TRIzol Reagent  in accordance with the manufacturer’s instructions. RNA sequencing was conducted on an Illumina HiSeq platform. The RNA-Seq clean reads were aligned to the TIGA rice genome using TopHat (v2.1.0). The RNA-Seq data were analysed using a previously described method. Differentially expressed genes were detected using the DESeq R package (v1.10.1) with a relative change threshold of two . Gene ontology (GO) categories were identified using the GOseq R package. KEGG pathways were assigned using the KEGG database (http://www.kegg.jp/) and were considered significant at P < 0.05.Leaves of wild-type and g for 10 min, the residue was diluted with 0.5 mL precooled sodium phosphate buffer for 2 h and then evaporated under N2 gas at 40 °C until the organic phase had disappeared. Petroleum ether (0.5 mL) was added to extract the residue three times, and the lower layer was evaporated to dryness under N2 gas at 40 °C. The residue was dissolved in 0.5 mL of the mobile phase  and then filtered. Free IAA in the extract was analysed using an Agilent 1100 HPLC  and Kromasil C18 reversed-phase column   sampled at the tillering stage was homogenized in liquid nitrogen, then 1 mL precooled sodium phosphate buffer was added and the homogenate was incubated at 4 °C overnight in the dark. After centrifugation at 8000 g at 4 °C for 5 min, the supernatant was diluted fivefold. An aliquot (100 μL) of diluted supernatant was added to 100 μl of 100 ppb double isotope internal standard  and vortexed for 30 s. After filtration, the supernatant was analysed by liquid chromatography–mass spectrometry (LC/MS) with regard to amino acid standards. An ACQUITY™ UPLC™ BEH C18 column  and electrospray positive ionization  source was used for LC/MS  sampled at the tillering stage was homogenized in liquid nitrogen and extracted with 1 mL precooled extraction mixture (1% formic acid–methanol) for 10 min at 25 °C. After centrifugation at 13,000 dvb1 mutant plants were treated with the inhibitor for about 4 days. The corresponding solution of water mixed with DMSO was used as the control. After treatment, the water, ATP and IAA contents were measured and analysed. The treated plants were photographed using a Canon 5DIII digital camera . All experiments were performed using at least six independent biological replicates.The antimycin A  and oligomycin  used were from acqueous dilutions of DMSO stock solutions  to 1.25 μM antimycin A and 0.5 μM oligomycin. Wild-type and OsDVB1 (LOC_03g62420), OsFIB (LOC_01g07500), OsTAR1 (LO05g07720), OsTDD1 (LOC_04g38950), OsCOW1 (LOC_03g06654).Sequence data from this article can be found in the GenBank/EMBL libraries under the following accession numbers: Additional file 1: Fig. S1. Root length of WT and dvb1. Fig. S2. Anthers and pollens in dvb1. Fig. S3. Narrowed leaves in dvb1. Fig. S4. Flower primordia and branches in dvb1. Fig. S5. Phylogenetic tree of DVB1. Fig. S6. Subcellular localization of DVB1 protein in Nicotiana benthamiana. Fig. S7. Subcellular localization of DVB1G90E. Table S1. Peptide sequences used for constructing the NJ tree. Table S2. Primer sequences used in this study.