Vorticity, Rotation and Symmetry (V)—Global Results and Nonlocal Phenomena. It is the fifth conference in a series of international conferences on mathematical fluid mechanics held at Centre International de Rencontres Mathématiques (CIRM) in Luminy . This conference was scheduled for April 6–10, 2020, for about 70 participants but had to be cancelled due to the COVID-19 pandemic. We were very glad to have the chance to organize a hybrid conference in the last week of October 26–30, 2020, with about 30 speakers, either on site in Luminy or connected by video conference to a larger international community.This special issue covers the proceedings of a conference entitled The conference series deals with a lot of recent topics in mathematical fluid mechanics such as the classical questions of existence, regularity and uniqueness as well as asymptotic structure and decay of solutions in time and space of the Euler and Navier–Stokes systems. Further topics are active scalar equations related to fluid mechanics, complex fluids, mixtures, flocking and aggregation, dynamical systems approaches, stabilizing and destabilizing effects, influence by nonlocal terms, free surface flows, and problems of fluid-structure interaction.x, t)-depending directions of the velocity gradient and of the vorticity. A generalized Stokes operator with a bounded measurable matrix-valued coefficient of viscosity and its semigroup is estimated by P. Tolksdorf in local N well-separated holes with solutions of a related Stokes-Brinkman system when the radius of the holes ranges from 1/N to the third root of 1/N. Recent endpoint regularity estimates in This volume contains 15 selected research articles and survey papers of participants of this conference and their coauthors. Classical questions on Navier–Stokes equations are discussed in the papers by M. Beneš, P. Kučera, P. Vacková focussing on various types of boundary conditions in the two-dimensional instationary case, whereas E. Miller wrote a survey of geometric constraints on the possible blowup of three-dimensional solutions in exploiting components in arbitrary Reinhard Farwig (Darmstadt) Šárka Nečasová (Prague) Jiří Neustupa (Prague)


A novel concept of polymer railway sleeper is proposed in this study that has the potential to meet static performance requirements within the cost of hardwood timber. The existing challenges of composite sleepers, such as low performance or high cost, can be overcome using this innovative concept. Such a proclamation is proven through limit state design criteria and a series of experimentations. Results show that polyurethane foam as an infill material can provide sufficient strength and stiffness properties to the sleeper, but the inadequate screw holding capacity could be a problem. This limitation, however, can be overcome using a particulate filled resin system. The findings of this study will help the railway industry to develop a timber replacement sleeper. The global railway industry is looking for an alternative material railway sleeper that can replace traditional timber sleeper which is suffering from premature deterioration ,2. To taFerdous et al.  investigThe commonly used polymeric resins for structural applications are epoxy, polyester, vinyl ester, phenolic, polyurethane foam and some others. Several classes of synthetic resins manufactured by the esterification of organic compounds are available. The advantage and disadvantage of different resins are summarised in This concept of sleeper is based on a glass fiber reinforced polymer (GFRP) rectangular hollow pultruded section filled with polyurethane (PU) foam as shown in The increase of sectional modulus and moment capacity with the increase of tube thickness from 1 mm to 10 mm are summarised in A solid component made with polyurethane (PU) based resin was used as a core material of sleeper a. The tyThe PU core was prepared in a closed mould before applying laminates. The PU core was wrapped with GFRP laminates using hand lamination process. The wrapping was done by two steps to avoid sagging due to gravity (a) applied on two sides and the top surface and (b) flipped over and repeat the same process . The fibres were overlapped at two sides of the sleeper for full depth.3), NOROX CHM-50 hardener (density 1.06 g/cm3) and fly ash with a mixing ratio of Resin: Hardener: Fly ash = 2000:40:200 g. The resin and hardener were mixed before the fly ash added to the mix. The gel time (working time) of the resin mix was around 1 h. The samples were post cured at 80 °C for 3 h. The overall casting process are shown in A particulate filled resin mix was prepared using polyester resin  and able to overcome the limitation of heavy weight for concrete sleepers (2000 kg/m3). Moreover, the density is also slightly lower than the recycled plastic sleepers (850–1150 kg/m3) and close to FFU (fibre-reinforced foamed urethane) synthetic sleeper (740 kg/m3) [The dimensions of the PU foam core sleeper were 243 mm (W) × 120 mm (D) × 2130 mm (L). The overall weight of the sleeper was 39 kg that provided an equivalent density of 640 kg/m0 kg/m3) . The lowBending modulus of elasticity is an important property on which the deflection characteristics of sleeper are dependent. The MOE is the function of the initial slope of the load-displacement curve obtained from midspan bending test. Therefore, a non-destructive test a up to 5a, L, I and ΔP/Δδ are shear span, span, effective moment of inertia of the sleeper, and slope of the load–displacement curve, respectively. The MOE for softwood timber sleeper can be as low as 7.4 GPa. Moreover, the MOE for recycled plastic sleeper varies between 1.5 GPa and 1.8 GPa while it is 8.1 GPa for FFU synthetic sleeper [MOE for polymer composite sleeper should be at least 1.17 GPa [ sleeper . The Ame1.17 GPa . The incThe compression modulus of elasticity of sleeper indicates the deformation characteristics when the sleeper is being compressed. A non-destructive rail-seat compression test was conducted to determine these properties. The compression load a was appD, A and ΔP/Δδ are the depth of sleeper, effective compression area and maximum slope of the load-displacement curve. The compressive modulus of elasticity was obtained 450 MPa which is higher than the same property for recycled plastic sleepers (176–269 MPa) but lower than timber sleepers (650 MPa) [The compression load-displacement behaviour is plotted in sleepers 6–269 MPaMOR of 94 MPa according to Equation (3) where M is the average ultimate moment and c is the distance of outer fibre from the neutral axis. The MOR obtained for this sleeper is significantly higher than the MOR of softwood timber (22–34 MPa) and hardwood timber sleepers (55 MPa) [Modulus of rupture measures the load carrying capacity of sleeper in bending. The test was conducted following the procedure described in the AS 1085.22 . Two spePull-out resistance measures the screw holding capacity of sleeper. This is an important property that ensures rail alignment in correct gauge. The pull-out resistance is determined based on the force required to pull-out the screw. Three standard 16 × 105 rail screws  supplied by Cold Forge Pty Ltd  were inserted into the sleeper. The test setup is shown in It is obvious that the alternative sleeper has to perform satisfactorily as per standard requirements. The performance of the sleeper Concept-1 obtained from the extensive testing program is tabulated in The low strength capacity of PU foam core in sleeper Concept-1 is the main issue identified for low screw holding capacity as no crack was observed on the FRP. Therefore, the strength properties of core material need to be improved. One approach to improve the core properties is to use the particulate filled resin (PFR) system. Ferdous et al. and Khotbehsara et al. ,21,22 deThe 5.2 mm thick FRP tubes used in Concept-2 were fabricated by pultrusion process and were made of E-glass fibre and vinyl-ester resin with a fibre volume fraction of 60%. The hollow pultruded section is a preferred choice over the laminates overlapping method discussed in Concept-1 as the latter one was failed due to skin buckling a. The PFSix standard screws were inserted in the PFR core FRP sleeper (Concept-2) as shown in Since the primary component of drillable PFR is resin, it is more expensive than normal OPC concrete which is non-drillable. Therefore, an optimal use of PFR only at rail-seat region where the screws are drilled could significantly minimise the cost of sleepers. The other parts should be filled with normal OPC concrete. In such a case, several bond areas between PFR and OPC concrete are obvious which could create weak zones due to the variation of their elastic modulus properties. To address this concern, one of the two FRP tubes was filled with PFR and the other one was filled with a combination of normal OPC concrete and PFR by equal volume a,b. Cent2) was tested to understand whether the concept is suitable for manufacturing sleeper. In this study, 2.15 times larger  deflection of the beam than the deflection of the actual size sleeper is expected as the dimensions of the actual sleeper are 2.3 times wider  and 1.15 times deeper .There still remains the question of how the full-scale sleeper will behave in rail-track. To understand the in-track deflection behaviour of full-scale sleeper, a five-point bending test was conducted. The justification of the five-point bending test setup for railway sleeper is discussed in . The loaSleeper Concept-2 was introduced to overcome the challenge of low screw holding capacity in sleeper Concept-1. Replacing PU foam core by PFR can significantly improve and meet the requirements of pull-out resistance for polymer sleepers. Since PFR is an expensive material, its optimal use is important to minimise the cost of sleepers. Therefore, the rail-seats region of the FRP tube need to be filled with drillable PFR while other parts can be filled with non-drillable OPC concrete. It was found that the joint between PFR and OPC concrete does not affect the overall behaviour. Filled FRP tube is a promising concept for developing polymer railway sleepers. To meet the strength and stiffness requirements, a minimum tube thickness of 4 mm is necessary.Polyurethane foam as an infill material can provide sufficient bending and compression properties. However, it cannot provide sufficient resistance to hold screws.Particulate filled resin (PFR) system as an infill material can overcome the limitation of low screw holding capacity that was observed in polyurethane foam.The expensive and drillable infill PFR material can be replaced by the inexpensive and non-drillable OPC concrete except rail-seat locations.The joint between PFR and OPC concrete does not affect the overall performance of the sleeper as the behaviour of the sleeper is governed by an external FRP tube and the type of in-fill materials have only minimal impact due to the confinement effect.The proposed design of sleeper only requires 50% volume of PFR as infill material that lead to manufacture a high performance and cost effective railway sleeper technology.This study proposed a new concept for manufacturing fibre reinforced polymer railway sleeper. The systematic design approach and experimental program identified new materials and method for developing composite sleeper concept. The following conclusions are made:Railway sleepers are often subjected to impact and fatigue loading due to flat wheel and repeated movements of the train. Therefore, an in-depth understanding of the impact and fatigue behaviour of the polymer sleeper will ensure its suitability to replace existing timber sleepers. To understand the fatigue behaviour of a polymer sleeper, the author attempted to investigate the fatigue behaviour of GFRP laminates ,25.


To evaluate the use of preoperative virtual Magnetic Resonance Elastography (vMRE) for patients undergoing transsphenoidal resection of pituitary adenomas (PA).Ten patients  were prospectively examined with the vMRE-method prior to transsphenoidal surgery. vMRE-images, reflecting tissue stiffness were reconstructed. From these images, histograms as well as the mean stiffness values over the tumor body were extracted. Finally, vMRE-data was compared with the PA consistency at surgery blinded to vMRE.In all patients, successful vMRE-examination was performed enabling evaluation of even small PAs. For tumors with homogenous tissue, the mean stiffness value increased with surgical consistency grading. For heterogenous tumors, however, the mean stiffness value did not consistently reflect the grading at surgery. On the other hand, the vMRE-images and histograms were found to be able to characterize the tumor heterogeneity and display focal regions of high stiffness that were found to affect the surgery outcome in these PAs. The vMRE-images and histograms showed great promise in characterizing the consistency at surgery for these PAs.Evaluation of PA consistency in preparation for surgery seems to be feasible using the vMRE-method. Our findings also address the need for high resolution diagnostic methods that can non-invasively display focal regions of increased stiffness, as such regions may increase the difficulty of transsphenoidal PA-resection.The online version of this article (10.1007/s11102-021-01129-4) contains supplementary material, which is available to authorized users. Pituitary adenomas (PA) account for 10–15% of all intracranial tumors . The majTo characterize the consistency of PAs, various magnetic resonance imaging (MRI) methods based on standardized T1- and T2-weighted imaging, as well as contrast media enhanced MRI imaging and standardized diffusion weighted imaging have been investigated –10. Up tIn this exploratory study, a novel method is proposed. Besides being a non-invasive method that relies on a clinically available MRI sequences with high speed and image resolution, this so called virtual elastography (vMRE) method is attractive for evaluation of the PA consistency due to an intrinsically high sensitivity to the viscoelastic property of the tissue. To our knowledge, however, the method has not previously been used for tumor evaluation.Therefore, the aim of this study was to evaluate the potential usefulness of the vMRE method for preoperative evaluation of tumor consistency in patients undergoing transsphenoidal resection of PA.The study was conducted according to the Declaration of Helsinki. Ethical approval was given by The Regional Ethics Review Board and oral and written informed consent was obtained from all participants. The data was derived from the Gothenburg Pituitary Tumor (GoPT) study, which is a prospective study that is enrolling patients scheduled for pituitary surgery at Sahlgrenska University Hospital, the sole provider of neurosurgical services for 1.8 million people in the western region of Sweden .The patients were included among patients referred to the radiology department from 17/10/2018 to 31/03/2020, where patients with macroadenoma (≥ 1 cm) who underwent an MRI the day before surgery for neuro-navigation, was included in the study. The indications for surgery were (1) tumor causing chiasmal compression and/or (2) hormonal hypersecretion and/or (3) sequential imaging showing growth of the tumor.The tumor type was verified by histology.All patients were examined on a 3T MRI scanner . A standardized preoperative examination protocol, including morphological imaging in the sagittal view and standardized angiography in the transversal view Table  was usedb = 200 and 1000 s/mm2) were used. With the use of higher b-values, the normally high sensitivity to isotropic gaussian diffusion was shifted toward non-isotropic non-gaussian diffusion and thereby, the diffusion weighted imaging sequence was made sensitive to fibrotic tissue  and one for the higher b-value (b1000 images). These images were then used to quantitate vMRE stiffness values. The quantification is summarized below and described in detail by le Bihan et al. [Sb200 and Sb1000 are the signal values in the b200 and b1000 images and the scaling factor of − 9.8 and the shift factor of 14 were previously determined in a calibration step by le Bihan et al. [.After scanning, two diffusion weighted stacks of images were automatically generated on the scanner; one for the lower n et al. . In shorn et al. .Then, the tumor region was segmented on all vMRE stiffness images that included the tumor body. The border of the segmentation was located approximately one pixel away from the visible edge of the tumor, ensuring that no background tissue was included in the tumor region Fig. . Also, pFrom the segmented tumor regions, the mean vMRE stiffness and the standard deviation of the mean were automatically extracted within 1 sec. Also, histograms were automatically extracted to visualize the distribution in vMRE stiffness values within the tumor region.The quantification of the vMRE stiffness was performed off-line using MATLAB R2016a .All surgeries were performed by one of two surgeons (D.N. or T.S.). The surgeons, blinded to the vMRE stiffness images, gave a single classification of tumor stiffness in each case as very soft, soft, moderate, hard or very hard (I–V). The system for classification of the consistency was jointly developed by the surgeons and has systematically been performed as a part of the prospective GOPT-study since 2015. In addition, the surgeons also made separate notes about the consistency of the tumor in free text.Categorical variables were expressed by numbers and continuous variables by mean ± SD.The vMRE stiffness images and histograms, as well as the quantitative parameters (mean vMRE stiffness and the standard deviation of the mean) were compared with surgical findings as well as between individuals.p = 0.05 significance level.The Chi-squared test was conducted to compare vMRE-values and ADC-values between surgical gradings at The inter-observer agreement was determined using intra-class correlation coefficients (ICC) with 95% confidence intervals, where the two-sided random ICC model was used to determine the consistency of the agreement.The statistical analyses were performed using the MatLab software.Ten patients  underwent preoperative MRI examination before endoscopic transsphenoidal resection of PA. The patient characteristics and results on PA consistency at surgery are shown in Table The diffusion weighted imaging was successfully performed in all patients, resulting in vMRE stiffness images with high image quality.Overall, the mean vMRE stiffness did not differ between groups of patients with different grading of PA consistency at surgery Table . HoweverBetween but also within individuals, the PAs displayed a large variation in vMRE stiffness values. Some PAs displayed focal regions of increased tissue stiffness, depicted as clusters of pixels with higher values in the vMRE stiffness images. The histograms characterized such heterogeneity as right-skewed distributions while streaky tissue was characterized in histograms as a large distribution of stiffness values. For example, ID 6 that was re-operated showed a region of higher stiffness values around 8.5 in the lower part of the tumor (close to the point of entry of the surgery) while deeper into the body, the tumor appeared soft with low stiffness values around 4 Fig. . The corr = 1.00 and p < 0.001) with near identical mean vMRE stiffness values in all three cases .The ICC analysis showed excellent agreement  but the results were encouraging, suggesting that stiffness have the possibility to predict PA surgery outcome. The considerably higher resolution, but also the shorter scan time and availability of the proposed method makes the proposed method an attractive alternative to the standardized magnetic resonance elastography method for evaluation of PA consistency. Also, the present method does not rely on additional hardware for induction of measurable shear wave vibrations.A recently published study has utilized standardized magnetic resonance elastography to determine PA consistency at surgery, where a correlation between the stiffness values and consistency grading was reported . As in pIn similar to the standardized magnetic resonance elastography method, the vMRE method seemed to offer mean stiffness values that increased with surgical grading for homogenous tumors. The high-resolution vMRE method also offered detailed information about the tumor tissue in terms of color vMRE stiffness images and histograms, where our findings showed the importance of characterizing tumor tissue heterogeneity in detail. Focal regions of tissue with higher stiffness may alter the surgical approach and have an impact on the surgical outcome. For example, one of the PA patients (ID 6) was re-operated and as a result displayed a region of very stiff values due to scarring, clearly displayed in the vMRE stiffness images at the point of surgical entry. This might have limited the removal of the tumor. Preoperative knowledge of the position of the focal region might have altered the course of the surgery in this subject. Future studies are encouraged to confirm the diagnostic value of the vMRE method and the clinical usefulness of this novel stiffness contrast.Since the proposed method has not previously been used for patients undergoing transsphenoidal resection of PAs, there is a lack of knowledge regarding the appearance of the PA tumors in the vMRE stiffness images and histograms, associated to the surgeon’s experience at surgery. For homogenous PAs, there seems to be a consistency between the mean vMRE stiffness value and the surgery grading. For heterogenous tumors, however, deeper knowledge seems to be needed, as exemplified by ID 4. While the consistency of most PAs was found to be successfully characterized with the present method, this softer PA displayed a mean vMRE stiffness value in similar to PAs with high consistency grading. No focal regions of higher stiffness values were seen in the vMRE stiffness images. However, the histogram displayed a wide distribution of stiffness values, representing a patchy heterogenous texture pattern. Such texture pattern might give a soft experience at surgery.Being a feasibility study, a small number of patients were included in the evaluation and, as such, this limits the strength of the conclusion. Future investigations on larger study cohorts and different types of adenomas are therefore encouraged to confirm our findings and evaluate the predictive value of the vMRE method for surgical PA outcome.b-values were used. Even though present findings show great promise, we cannot neglect the fact that the vMRE-method might have benefitted from even higher b-values, however, at the expense of increased spatial resolution or extended scan time.The proposed vMRE-method utilized a diffusion weighted imaging sequence that was sensitized to enhance tissue textures with dense diffusion barriers, such as fibrotic tissue. For that purpose, high . [In similar to the work by Hughes et al. , the stuThe vMRE stiffness calculation used in this study is straightforward, where the mean tumor stiffness can be estimated directly from the mean signals of the b200 and b1000 images. Future MR scanner releases may implement the proposed method and offer automatic evaluation. Today, however, an off-line software is needed to calculate the vMRE stiffness on a pixel by pixel basis and reconstruct vMRE stiffness images and histograms for PA heterogeneity evaluation. This may limit the evaluation in centers with no technical/programming competence.Present study showed high feasibility of the vMRE method for preoperative evaluation of the PA consistency. The vMRE-method produced artifactual-free images of high quality and resolution, enabling evaluation of tumor heterogeneity. Also, the mean vMRE stiffness value was found to increase with surgical stiffness grading for solid tumors with homogenous texture. Further studies are encouraged to confirm our findings and determine whether the vMRE method provides useful information in the preoperative planning of PAs.Figure S1. The vMRE stiffness image (above left) and histogram (above right) for PA with ID 7 displayed a small focal region with higher stiffness values around 10 point of entry of the surgery. (PNG 66 kb)Figure S2. The vMRE stiffness image (above left) and histogram (above right) for PA with ID 9 displayed in consensus with surgent consistency grading, the PA consistence as homogenously soft with low variation in stiffness values over the tumor body. (PNG 84 kb)Figure S3. The vMRE stiffness image (above left) and histogram (above right) for PA with ID 10 displayed a homogenously stiff tumor. At surgery, this tumor was graded as soft but left residual tumor tissue, seen on the 6 months follow-up MRI. (PNG 77 kb)Figure S4. The vMRE stiffness image (above left) and histogram (above right) for PA with ID 5 displayed a heterogenous PA with low stiffness and a variation in stiffness from left to right. At surgery, this tumor was graded as very soft. (PNG 67 kb)Below is the link to the electronic supplementary material.


Background and Objectives. The gut microbiota has been shown to be involved in the development and severity of type 2 diabetes (T2D). The aim of the present study was to test the effect of potential functional food ingredients, alone or in combination, on the gut microbiota composition in diabetic rats in a pilot study of 1 week of feeding. Methods. In a pilot study to modulate the composition of the gut microbiota, (i) native taro starch, (ii) modified taro starch, (iii) beet juice, (iv) psicose, (v) the probiotic L. plantarum IS-10506, (vi) native starch combined with beet juice, (vii) native starch to which beet juice was adsorbed, (viii) modified starch combined with beet juice, and (ix) modified starch to which beet juice was adsorbed were fed to rats in which T2D was induced with streptozotocin (STZ). After one week, the composition of the gut microbiota was evaluated by sequencing the PCR-amplified V3-V4 region of the 16S rRNA gene. Results and Conclusions. The next-generation sequencing showed that 13 microbial taxa of the gut microbiota were significantly different between groups, depending on the treatment. The results of this pilot study will be used to design a 4-week intervention study in STZ-induced T2D rats to determine the best functional food for counteracting T2D, including their effects on satiety hormones. This should ultimately lead to the development of functional foods for prediabetic and diabetic individuals. The global incidence of T2D is predicted to reach 578 million cases by the year 2030 and 700 million cases by 2045, a predicted increase of 51% from the 463 million cases in 2019[In Indonesia, taro (Cocoyam) is cultivated and traditionally used as food crop by several ethnic communities in Borneo , and itsAnother functional dietary component that has been shown to affect T2D is D-psicose , a rare monosaccharide. In an attempt to produce functional foods aimed at low calorie and less sugar intake for T2D, D-psicose is considered as a substitute for sugar with proven antihyperglycemic, antihyperlipidemic, and anti-inflammatory effects.L. plantarum IS-10506 [Also probiotics have been shown to be beneficial in treatment of T2D . ProbiotIS-10506 , 21.ß-cells, modulation of hepatic glucose output, activation of insulin receptors, or modulation of glucose uptake in insulin-sensitive cells [A last functional class of dietary components considered to be beneficial in T2D is polyphenols. Polyphenols interact heavily with the gut microbiota, either because they have been shown to inhibit certain species or on the other side because they stimulate other taxa and are converted in other bioactive phenolic components . Polypheve cells . Moreoveve cells . Beetroove cells , and beeve cells . PolypheL. plantarum IS-10506, and phytonutrients of beet juice, alone or in combination, on the gut microbiota composition in streptozotocin-induced T2D rats in a pilot study of 1 week of feeding.The aim of the present study was to find out the effect of (modified) taro starch, psicose, the probiotic Streptozotocin (STZ) was purchased from Enzo Life Sciences . Purified Rodent Diet AIN-93M, a modified AIN-76A standard diet (American Institute of Nutrition) , was usead libitum access to water and commercially available rat normal pellet diet (NPD) purchased from local market, prior to the dietary manipulation.All animal procedures undertaken were approved by Ethics Committee of the Faculty of Medicine, University of Indonesia (Ref:1196/UN2.F1/ETIK/2018). A total of 40 Male Sprague Dawley rats were purchased from Animal Experimental Laboratory, National Agency of Drug and Food Control , at 6 weeks of age and were allowed to adapt for 14 days. They were housed in individual cages and maintained at 21–23°C and 55% ± 5% humidity with 12-hour light/dark cycle. During the 14 days' acclimatization period, all rodents were given −1 nicotinamide in 0.9% NaCl, followed after 15 minutes by STZ in citrate buffer pH 4.4 (70 mg kg−1), and, four days after the STZ injection, fasting and postprandial blood glucose levels were measured using a freestyle glucose meter (Easy Touch GCU 3 in 1) from tip of the tail. The rats with a fasting glucose of >100 mg dl−1 and/or postprandial blood glucose levels of ≥140 mg dl−1 were considered as type 2 diabetic, and those rats which had not yet developed T2D within these 4 days were injected for the second time with 120 mg kg−1 nicotinamide and STZ in citrate buffer pH 4.4 (70 mg kg−1). After confirmation of T2D, the eight-week-old 190–220 g rats were divided into 10 groups of n = 4 each, namely, (i) AIN-93M (control), (ii) AIN-93M with two times in a day 3 ml psicose by gavage, (iii) AIN-93M with two times in a day 3 ml beetroot juice by gavage, (iv) native taro starch, (v) modified taro starch, (vi) native taro starch with beetroot juice adsorbed and then dried, (vii) modified taro starch with beetroot juice adsorbed and then dried, (viii) native taro starch combined with two times in a day 3 ml beetroot juice by gavage, (ix) modified taro starch combined with two times in a day 3 ml beetroot juice by gavage, and (x) L. plantarum IS-10506. The native or modified taro starch was replacing the corn starch in AIN-93M.Four rats in each group were allocated to the dietary treatments, AIN-93M (AIN), modified AIN-93M by replacing corn starch with taro starch, sucrose and cellulose were replaced with maltodextrin, respectively, for 3 days, with 75, 50, and 25% commercial rat pellet diet and 25, 50, and 75% dietary intervention formulation, respectively. Then the rats were injected intraperitoneally (i.p.) with 120 mg kgThe feed and water intake of the animals were measured. The rats were allowed to continue to feed on their respective diets until the end of the study. Food intake and bodyweight were monitored every day and at the end of one week, respectively, during the study, and the average intake per rat was calculated. Fecal pellets were collected at baseline and at the end of one week of feeding.Taro starch “HASILBUMIKU” was purchased from a local supplier in Bantul, Yogyakarta (Indonesia). Modified taro starch was manufactured by autoclave-cooling according to a modification of the method of Zhao and Lin . In brieBeet juice was adsorbed to both native and modified taro starch by absorbing beetroot juice, at a ratio of 1 : 1, and then drying in an oven at 40°C for 16 hours. These were prepared and fed according to the dose of beet juice of 6 ml/day but in adsorbed beet juice form.10 colony forming units (CFU)/day.The probiotic was given by gavage at 10DNA of feces samples was extracted using the Quick-DNA™ Fecal/Soil Microbe Miniprep Kit (Zymo Research) according to manufacturer's instructions, using the Precellys 24 tissue homogenizer , applying 3 cycles of 30 seconds each, with 5-minute cooling on ice in between. DNA concentration and purity were checked by measuring absorbance at 260 and 280 nm.μl using the 341F (5′-CCTACGGGNGGCWGCAG-3′) and the 785R (5′-GACTACHVGGGTATCTAATCC-3′) primers appended with Illumina adaptor sequences . PCR products were purified and the sizes of the PCR products were checked on Fragment analyzer  and quantified by fluorometric analysis. Purified PCR products were used for the 2nd PCR in combination with sample-specific barcoded primers . Subsequently, PCR products were purified, checked on a Fragment analyzer , and quantified, followed by multiplexing, clustering, and sequencing on an Illumina MiSeq with the paired-end (2x) 300 bp protocol and indexing.PCR-amplifying the V3-V4 region of the 16S rRNA gene and next-generation sequencing Illumina 16S rRNA gene amplicon libraries were generated and sequenced at BaseClear . In short, barcoded amplicons from the V3-V4 region of 16S rRNA genes were generated using a 2-step PCR. 10–25 ng isolated genomic DNA was used as template for the first PCR with a total volume of 50 α- and ß-diversity and visualization of the (un)weighted UniFRac principal coordinate analyses were also done using QIIME. The software package R (3.5.1) [q-values  were considered significantly different at <0.05. Correlation between OTUs and continuous values are indicated by the rho-value. Permutational multivariate analysis of variance  differences. Hierarchical clustering was performed in R using the function “hclust,” based on the relative abundances of the samples. Dissimilarity values were calculated on the basis of Euclidean distance. Clustering was performed using Ward's minimum variance method.The sequencing run was analyzed with the Illumina CASAVA pipeline (v1.8.3) with demultiplexing based on sample-specific barcodes. The raw sequencing data produced was processed removing the sequence reads of too low quality (only “passing filter” reads were selected) and discarding reads containing adaptor sequences or PhiX control with an in-house filtering protocol. A quality assessment on the remaining reads was performed using the FASTQC quality control tool version 0.10.0. Subsequently, the sequences were further analyzed using the Quantitative Insights Into Microbial Ecology (QIIME) software pipeline, version 1.9.1 . Operati (3.5.1)  was usedRMANOVA; ) was perß-diversity in α-diversity did not differ between groups . Kruskal-Wallis comparison was used to indicate which operational taxonomic units (OTUs) were significantly different between baseline and week 1 . These are listed in Compared to baseline (when confirmed T2D was established by STZ injection) the microbiota of the rats was different at week 1 after the interventions with the potential functional food ingredients (displayed as unweighted UniFRac q < 0.05) different between groups  is thought to decrease  pathogenic bacteria [Leuconostoc, another lactic acid producing genus, was increased by similar interventions as for Lactococcus, although in addition modified starch showed some increase as well. Beet juice exclusively increased Eubacterium, a genus known to produce butyrate, a microbial metabolite considered to be beneficial for the host, as it has been shown to be an important energy source for the colonocytes and has anti-inflammatory effects [L. plantarum IS-10506. Anaerovibrio was increased by several of the starch interventions. RA of Bacillus was reduced by most treatments compared to control, while RA of Phascolarctobacterium was increased by all interventions compared to control. The RA of members of the Christensenellaceae was increased by treatment with all taro starch containing interventions. Christensenellaceae has been correlated with leanness, as they have been shown to be enriched in individuals with low body mass index [Bifidobacterium seemed to be reduced by all treatments except native starch, while Bifidobacteriaceae were increased by more treatments but particularly by native starch with beet juice (either in combination or adsorbed).After 1 week of dietary intervention, the different interventions did not lead to a clear separation between the groups of rats .  Figuren groups .  Figurebacteria , althoug effects . The RA ss index . The genq < 0.05), the correlations were rather weak (A few OTUs were correlated to body weight (BW) and one OTU was correlated to fasting glucose. Although these were statistically significant even after correction for multiple comparisons (Being an initial pilot and screening experiment, the study has some inherent limitation. The first is the length of the time of intervention. To allow for an initial quick screening, we chose to feed the functional ingredients for a single week. In the meantime, based on these screening results, we have carried out a study for a 4-week period in rats. Also, based on the results of this study, a clinical trial in prediabetic individuals has been started. In an animal trial, the separate individual animals, coming from the same breeding colony, more or less have the same microbiota composition (although In conclusion, given the correlations of different OTUs to the different treatments  in this


After this article  was publPLOS ONE’s Editorial Board advised in the post-publication discussions that they did not have concerns about the study design or analyses. However, PLOS ONE concluded that the following updates and clarifications are needed.PLOS followed up on the concerns and conducted a post-publication editorial assessment to evaluate the potential impacts of undisclosed competing interests on the article’s peer review. The outcome of this assessment supported the conclusion that the disclosure issues did not unduly impact the peer review process; a member of PLOS ONE Editors concluded that based on available information about BLOOM’s activities, the affiliations of FLM and CN and other previously undisclosed associations to BLOOM and the MSC need to be declared as potential competing interests. Accordingly, the article’s Competing Interests statement is updated to:FLM and CN listed their affiliations with BLOOM in the article’s “About the Authors” section but the published Competing Interest statement says, “The authors have declared that no competing interests exist.” After careful review, the As noted in the article’s “About the Authors” section, FLM and CN are affiliated with BLOOM. CN is Founder and Honorary President of BLOOM. As such, she has never received any salary from BLOOM from its inception in 2005 to 2020. FLM is the organization’s Scientific Director. CJ worked for BLOOM from 2014–2016, and JJ was an unpaid member of BLOOM’s Board of Directors from 2010–2012. FLM was a member of the public chamber of the MSC Stakeholder Council in 2016–2017 and received travel funds from the MSC to attend a meeting for this Council.The Data Availability Statement is updated to:http://dx.doi.org/10.17632/gpynbmn7f9.1. The authors did not obtain specific permissions to publish or make publicly available the primary images, which are available through the Mendeley repository. These images are all screenshots from public reports and social media, and are not covered by the PLOS ONE article’s CC-BY 4.0 license.All data and processing scripts are available from Sentences 3 and 4 in the Abstract are revised to cite sources and identify assertions as hypotheses:Original: “While the MSC is increasingly recognized by decision-makers as an indicator for fishery success, it is also criticized for weak standards and overly-lenient third-party certifiers. This gap between the standard’s reputation and its actual implementation could be a result of how the MSC markets and promotes its brand.”Revised version: “While the MSC is increasingly recognized by decision-makers as an indicator for fishery success, it has also been criticized for having weak standards and lenient third-party certifiers . We hypothesized that these disparate views of the MSC’s reputation and program implementation could be a result of how the MSC markets and promotes its brand.”In the Introduction, the study aim statement is revised to clearly and accurately describe the reported study.Original: “As with agricultural production, there is a widespread perception that ‘small is beautiful’ [37–40], which led us to question whether the MSC was accurately representing its certified fisheries in its promotional materials.”Revised version: “As with agricultural production, there is a widespread perception among consumers that ‘small is beautiful’ [37–40]. In this study, we assessed images used in MSC promotional materials and compared the relative proportion of different fishery types shown in these materials to the relative abundance of different fishery types certified by the MSC.”The following paragraphs are added to the article’s Discussion to address some of the study’s limitations:The study analyzed images only, it did not assess the materials’ overall communications, including text discussion, about different types of fisheries, their representation in the overall portfolio of MSC-certified fisheries.Conclusions cannot be drawn from this study’s results as to the MSC’s objectives in selecting images to show in their materials, or about why there are differences between the relative abundance of different fishery types shown in marketing materials versus certified by the MSC. Images used in promotional materials may be selected for various reasons, such as the company’s marketing objectives, the intended audience of the materials, and/or to represent aspects of the message(s) relayed in accompanying text.Readers should note that the following text in the Discussion represents the authors’ opinions and is not intended to relay conclusions following from the data reported in the article.“…it appears that the MSC strongly appeals to the idealization of fisheries by consumers and policy-makers by promoting fisheries involving small-scale gears and passive gears in much higher proportions than in reality, as is the case in other sectors such as agriculture [37–39]. The MSC may be trying to appeal to the needs and desires of its consumers, providing them with the symbolic satisfaction of not having harmed the environment [58].”“there is a credible risk of misunderstanding for those who scroll quickly through their documents and websites, which may explain to some extent the positive public image of the MSC.”“We hypothesize that this discrepancy between MSC-certified fisheries and what the MSC advertises aims to ‘green’ its image with consumers. We further posit that this discrepancy might be the reason behind a perceived gap between its ‘supporters’—including policy-makers—and other stakeholders that have gradually disengaged or become critical of the MSC .”*Except for Reference #1, the citations included in this notice refer to the published article’s References list.


Analysis of genome variation provides insights into mechanisms in genome evolution. This is increasingly appreciated with the rapid growth of genomic data. Mitochondrial genomes (mitogenomes) are well known to vary substantially in many genomic aspects, such as genome size, sequence context, nucleotide base composition and substitution rate. Such substantial variation makes mitogenomes an excellent model system to study the mechanisms dictating mitogenome variation. Recent sequencing efforts have not only covered a rich number of yeast species but also generated genomes from abundant strains within the same species. The rich yeast genomic data have enabled detailed investigation from genome variation into molecular mechanisms in genome evolution. This mini-review highlights some recent progresses in yeast mitogenome studies. The greymbiosis . The excymbiosis . These eSaccharomyces cerevisiae is among the best-studied model organisms, an abundant number of mitogenomes have been sequenced for S. cerevisiae   of related yeast strains is well known to show hypervariability in sequence [reviewed in –) . Petite isolates . The forisolates , which pisolates . Some deisolates , which aisolates   in exon regions approaching intron boundaries . Althougvar1 gene  is another well-studied example of unidirectional inheritance in S. cerevisiae  in the coding region (S. cerevisiae (Magnusiomyces capitatus (in the Dipodascaceae family), GC-clusters inserted in protein-coding regions are transcribed in mRNAs, but the GC-cluster region in mRNA gets bypassed (or ignored) during translation , Non-allelic mtDNA recombination can lead to deletion and insertion of sequences. Allelic mtDNA recombination, on the other hand, can prevent mtDNA deletion and repair mitogenome damage . MitochoNakaseomyces  and one ribosomal protein gene var1, SSU and LSU rRNAs, and 22–24 tRNAs. This is in contrast with a recent report that “no gene is universally conserved in fungal mitogenomes” (S. cerevisiae mitogenomes (accessions: CM002421 and CP046458) reportedly missing atp6 by S. cerevisiae mitogenomes  . The losgenomes” . Althouggenomes” , the varogenomes . The Saca uvarum . This isludwigii , a relatNakaseomyces bacillisporus at 107 kb contains no intron, while mitogenomes in Candida glabrata at around 20 kb contain 3–4 introns in at least four intron-distribution patterns [ have compact mitogenomes. Unfortunately, many yeast species either lack intraspecific genomic data or suffer insufficient sequence diversity, making it impossible to estimate the degree of genetic drift. To address this issue, extended efforts are needed to sample and sequence an abundant number of intraspecific strains in a broad range of yeast species.The question on whether genome size is under selective constraint has been a subject of debate . The mut species  and has  species , yet manHanseniaspora uvarum mitogenome is 29.3%  used in the MIP1) linked to reduced fidelity of mtDNA replication  to slow-evolving mitogenomes inflated by large introns, repeats and non-coding sequences (similar to plant mitogenomes). The fast-growing yeast mitogenome data have allowed us to begin to identify mechanisms driving genome diversity. Specific efforts are needed to sequence and study an abundant number of intraspecific strains from closely related species for contrast genomic features. The future of yeast mitogenome studies is bright, and the generated knowledge will no doubt benefit our understanding of mitogenomes much beyond the fungal kingdom.WH wrote the manuscript.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Pseudoaneurysm of ascending aorta is a rare but serious complication of cardiovascular surgeries and it infrequently occurs in the normal prosthetic graft materials. We share our experience with an unusual case of ascending aorta Dacron graft pseudoaneurysm caused by a fractured sternal wire.A 34-year-old man, known case of Marfan syndrome, with history of two prior aortic surgeries for aneurysm of ascending aorta, arch and thoracoabdominal aorta, presented with hemoptysis. The hemoptysis originated from an aortobronchial fistula secondary to a huge ascending aorta Dacron graft pseudoaneurysm. The graft erosion and subsequent pseudoaneurysm was caused by a fractured sternal wire. Surgical repair of the pseudoaneurysm was performed successfully and a Gore-tex patch was placed behind the sternum over the graft to prevent further direct contact of the wire and the graft.Sternal wires can damage the adjacent vascular grafts and lead to fatal complications such as pseudoaneurysm formation. Thus, preventive measures such as using sternal bands and placing a covering layer between the sternal wires and aortic grafts are recommended in patients with dilated or replaced ascending aorta. Pseudoaneurysm of ascending aorta is a rare but serious complication of cardiovascular surgeries. It is typically associated with aortotomy, aortic cannulation sites, anastomotic suture lines, and needle puncture sites, but it infrequently occurs in the normal prosthetic graft materials , 2. ThisA 34-year-old man, known case of Marfan syndrome, presented with three episodes of non-massive hemoptysis. Seven years earlier, he had undergone Bentall operation and arch replacement with E-vita graft using frozen elephant trunk (FET) technique with a stent-graft in the descending aorta due to acute aortic dissection and aortic arch aneurysm. Six years later, he had returned with a thoraco-abdominal aortic aneurysm and a huge pseudoaneurysm around the distal part of the descending aorta. An extensive surgery with thoracoabdominal incision replacing all the thoracic and abdominal aorta by Dacron graft had been done, and the proximal part of the graft had been sutured to the distal end of the stent-graft.Finding out the cause of hemoptysis, he was admitted and a comprehensive evaluation including computed tomography angiography (CTA), bronchoscopy, and transesophageal echocardiography set out. The CTA showed a new collection, most probably a pseudoaneurysm sized 4 × 6 × 9 cm around the ascending aorta extending to the posterior border of the manubrium at the maximum diameter Fig. B, C. TheThe episodes of hemoptysis decreased to only one in his admission course, but the patient progressed with a fever. Work-ups ruled out COVID19 infection but revealed a positive Enterobacter blood culture for which the patient received wide spectrum antibiotics for two weeks. Even though later cultures were negative and transesophageal echo did not indicate endocarditis, a low-grade fever was still running. The mediastinal hematoma and perhaps its connection to the respiratory system was presumed to be the main reason for the fever. Endovascular approach and stent grafting was excluded because of the possibility of the presence of an infection. So, our team decided on surgical treatment.Since the lower half of the sternum had severe inward deviation due to pectus excavatum and the suspected graft perforation was at the level of the manubrium, minimally invasive upper reverse J sternotomy was selected. The femoral vessels were exposed and cannulated. The innominate artery was also exposed and controlled above the sternal notch in case there was any need for selective cerebral perfusion. Cardiopulmonary bypass was started, and the sternum was opened at a temperature of 32°. The blood outburst from the aorta was recovered by sump suction back into the circulation, and the bypass flow rate was decreased to minimize the bleeding. An approximately 5 × 8 mm hole was identified on the Dacron graft, finger controlled and then directly repaired by reinforced 3-0 Prolene suture (Fig. Sternal wire fractures which occur after the median sternotomy are not rare radiologic findings. Although the strength induced by force under normal conditions is far below the ultimate tensile strength of the wire, the fracture of steel wires might occur after routine procedures. The mechanism is attributed to mechanical or chemical cracking secondary to bending and twisting or exposure to body fluids highly concentrated in chloride ions. Upper body activities and repetitive thorax movements such as respiration and coughing might also precipitate wire failures. Imperfect manufacturing and sterilization processes are other factors influencing the structure of wires , 4.In our case, the incident of wire fracture early after the second surgery despite no infection or sternal dehiscence made us contemplate whether the force caused by the thoracoabdominal incision and rib retraction during the second surgery had affected the normal constitution and strength of the sternal wire.Different types of intrathoracic injuries caused by the fracture and migration of wires have been reported such as right ventricle to aorta fistula , intrava


Dimocarpus longan Lour.) produced annually are processed into many products, one of which is black longan, from which the dried, dark-brown meat has been used medicinally in traditional medicine, while the starch-containing seeds are discarded. In this study, starch samples (BLGSs) were isolated from seeds of black longan fruits prepared using varied conditions. The in vitro digestibility was determined in comparison with those extracted from fresh (FLGS) and dried (DLGS) seeds. Scanning electron microscopy (SEM), X-ray diffraction (XRD), differential scanning calorimetry (DSC), and Fourier transform infrared (FTIR) spectroscopy were employed to evaluate the starch properties. The results showed that the yields of FLGS, DLGS, and BLGSs were 20%, 23%, and 16–22% w/w, respectively. SEM images showed starch granules of mixed shapes, with sizes up to 15 µm in all samples. XRD patterns confirmed an A-type crystallinity for FLGS and DLGS, with strong refraction peaks at 2θ = 15°, 17°, 18°, and 23°, while BLGSs also showed detectable peaks at 2θ = 10° and 21°, which suggested V-type structures. Thermal properties corroborated the changes by showing increases in peak gelatinization temperature (Tp) and enthalpy energy (ΔH) in BLGSs. The paste viscosity of BLGSs (5% w/w) decreased by 20–58% from that of FLGS. The FTIR peak ratio at 1045/1022 and 1022/995 cm−1 also indicated an increase in ordered structure in BLGSs compared to FLGS. The significant increase in the amounts of slowly digestible starch (SDS) and resistant starch (RS) in BLGSs compared to FLGS, especially at a prolonged incubation time of 20 (4.2×) and 30 days (4.1×), was proposed to be due to the heat-induced formation of starch inclusion with other components inside the seed during the black longan production process. Thus, black longan seed could be a new source of starch, with increased RS content, for potential use in the food and related industries.A large quantity of longan fruits ( Dimocarpus longan Lour.) is a popular tropical fruit grown in many parts of Asia. In addition to fresh consumption, longan fruits were processed into many products, ranging from canned longan in syrup to oven-dried longan, all of which subsequently yielded seeds as the by-product. Although longan seeds were widely researched and reported to contain phenolic compounds with many biological activities [w/w starch, with high amylose contents (26.6–30.1%), while a study by Guo et al. [w/w starch, with a 25.1% amylose content. In the production process of dried longan, the whole fruits were placed in a drying chamber of an oven, with controlled temperature and hot air circulation. While the moisture was gradually taken out from the flesh (aril), the seed inside the fruit was subjected to heat via the air and the moisture in the flesh. This created an environment of the heat-moisture treatment (HMT), which is known to alter the thermal and digestibility properties via changes in starch granular arrangement [Longan  are shown in p values compared to that of FLGS. The thermal transition ranges were broader in samples incubated for 5 and 10 days at all three temperatures, while narrower ranges were recorded in samples incubated for 20 and 30 days. ΔH values of BLGS samples were higher than that of FLGS, with the exception of the samples incubated at each temperature for 30 days in which the ΔH values were significantly lower.The peak gelatinization temperature (Tp), the thermal transition range (To–Tc), and the gelatinization enthalpy (ΔH) of FLGS registered respectively at 76.9 °C, 71.7–85 °C, and 14.9 J/g . BLGS saw/w, FLGS paste yielded the highest viscosity (0.56 ± 0.01 Pa.s), while the DLGS viscosity was 7% less. BLGS samples showed 20–58% decreases in the viscosity, with BLGS-60-20 being the least viscous paste (0.24 ± 0.02 Pa.s).The rheological profiles of FLGS, DLGS, and four representative BLGSs are shown in The deconvoluted ATR-FTIR spectra of FLGS, DLGS, and four representative BLGSs are shown in Starch-isolated fresh longan seeds (FLGS) contained mainly RDS (65%), 15% SDS, and relatively low RS (7.6%) . The staStarch-isolated fresh longan seeds (FLGS) contained mainly RDS (65%), 15% SDS, and relatively low RS (7.6%) . The sta−1 obtained from the deconvoluted ATR-FTIR was known as an effective means to characterize the degree of order in starch granules [−1 was suggested to correlate with the formation of a double helix in starch molecules [The starch yields 16–23%), calculated from the actual starch obtained in each sample, were significantly lower than those reported by previous studies, i.e., 44.9–49.5% %, calcul. No signgranules , while tolecules . The higolecules , althougLongan fruits, grade B, were obtained from a local supplier . The seeds of fresh fruits were acquired by manual peeling, washed, and air-dried before subjecting to starch extraction. The Resistant Starch Assay Kit (AOAC Method 2002.02) was a product of Megazyme . All organic solvents and chemicals used were reagent grade, unless indicated otherwise.Whole longan fruits were dried in a hot-air oven using a proprietary drying condition of Honeyqueen Co., Ltd. . For quality control purpose, the pulp of the resulting dried whole longan fruits was analyzed to contain 14–17% moisture content and 0.5–0.6 water activity (aw) according to the Thai Agricultural Standard (TAS 10-2006) . Seeds wSubsequently, twelve (12) sets of 500 g of oven-dried, whole longan fruits (control sample) were incubated at 60, 70, and 80 °C, in desiccators previously equilibrated at approximately 75% relative humidity using saturated NaCl solution, for 5, 10, 20, and 30 days. At the end, the seeds of incubated “black” longan samples were separated from the pulp/meat, washed, and collected for starch extraction.All seed samples were subjected to starch extraction using a method described previously for Jackfruit seed starch , with thSEM experiments to analyze the granule surface, shape, and size were conducted using a JEOL instrument, model JSM-5410LV , equipped with a large field detector. The acceleration voltage was 15 kV under low-vacuum mode (0.7–0.8 torr). The sample was placed on a copper stub covered with adhesive tape and coated with gold under vacuum. The images were taken at 1000X magnification.XRD patterns were recorded in the reflection mode on a Siemens D-500 X-ray diffractometer . Diffractograms were registered at a Bragg angle (2θ) range of 5–40° at a scan rate of 2.5°/min and step size of 0.02°.−1. Spectra were baseline-corrected using Omnic ver.6.2. The region at 1200–900 cm−1 was deconvoluted and the absorbance values at 1045, 1022, and 995 cm−1 were determined. The peak ratios of 1045/1022 and 1022/995 were calculated for each sample.FTIR spectra were recorded on a Nicolet Nexus 470 FTIR equipped with a DTGS detector  using an attenuated total reflectance (ATR) mode. For each spectrum, 64 scans were recorded at a resolution of 4 cmw/w) starch–water mixture sample. An empty pan was used as a reference. The temperatures of the characteristic transitions, including onset (To), peak (Tp), and conclusion (Tc) temperatures, were recorded. Enthalpy change of gelatinization (ΔH) was calculated and expressed as J/g of dry starch.Thermal properties were assessed using a Perkin Elmer DSC-7  differential scanning calorimeter. The analysis was carried out at a temperature range of 30–120 °C, at 10 °C/min, on a 1:3  held at 100 s−1 for 1 min, and (3) a decrease of the shear rate from 100 to 0 s−1 in 1 min. All measurements were performed in triplicate, at a controlled temperature of 25 ± 1 °C. The data were analyzed with Brookfield Rheo 2000 software. The apparent viscosity for all samples in this study was measured at a shear rate of 100 s−1 and was expressed in Pa s.Apparent viscosity of longan starch paste was determined using a Brookfield R/S-CPS rheometer . The samples were prepared by dispersing 2.5 g of starch sample in 50 mL distilled water, mixed thoroughly, and stir-heated at 90 °C for 15 min. The measuring system was CC48 DIN. The mode used was CSR (controlled shear rate). The measured parameters consisted of three steps: (1) an increase of the shear rate from 0 to 100 sThe digestibility of starches obtained from fresh, dry, and black longan seeds was analyzed using a Megazyme Resistant Starch Assay Kit (AOAC Method 2002.02) according to a procedure previously described . In briep < 0.05). Significant differences among mean values were determined by Duncan’s multiple range test.All tests were performed at least in triplicate. The statistical significance tests were performed using analysis of variance (ANOVA) at the 95% confidence level (The enormous amount of longan seeds from the fresh fruits and the dried longan industry, together with previous reports on starch isolation, suggested that this biowaste could potentially be a new source of commercial starch. The starches extracted from black longan seeds showed similar granule size and structure but exhibited different in vitro digestibility profiles compared to those extracted from seeds of fresh or oven-dried fruits. The amounts of resistant starch found in BLGS samples were 4× higher than that of FLGS. Additional peaks in XRD, transition parameters in DSC, and decreased paste viscosity of BLGS suggested that the increases in resistant starch content were possibly due to the starch inclusion complexes, which were formed through interaction with other components in the seeds during the temperature/time-controlled incubation. The optimum temperatures and time for incubation of dried longan fruits to obtain black longan seeds with high RS content were 60–70 °C for 20–30 days. The applied heat altered starch properties in a way similar to that caused by dry heat treatment. Black longan seed starch could be a new source of resistant starch for applications in the food, pharmaceutical, and related industries.


Dengue fever is a mosquito-borne viral disease endemic in 128 countries. An unmet clinical need remains for an effective vaccine that can be used more broadly than the vaccine presently available. A clinical development program has evaluated the long-term safety, immunogenicity, and vaccine efficacy (VE) of TAK-003, a live attenuated tetravalent dengue vaccine with a DENV-2 backbone engineered to elicit immune responses to all 4 dengue serotypes. nd dose, and for another 25 months after a booster dose. Data up to 3 years after the second vaccination are currently available.18 clinical trials in 13 countries have involved 28,175 seropositive/seronegative participants aged from 1.5-60 years from endemic/non-endemic regions. In the ongoing pivotal phase III study, 4–16-year-old healthy children  were randomized 2:1 to receive two doses of TAK-003 or placebo, 3 months apart for an evaluation of VE and safety over a multi-year period stratified pre-vaccination dengue serostatus. Active surveillance throughout the trial detected symptomatic dengue. The trial will continue up to 4–4.5 years post 2nd dose. At 18 months, VE was 66.2% (95% CI: 49.1–77.5) in dengue-naive and 76.1% (95% CI: 68.5–81.9) in dengue pre-exposed participants, with VE of 90.4% (95% CI: 82.6–94.7) and 85.9% (95% CI: 31.9–97.1) for prevention of hospitalized VCD and dengue hemorrhagic fever, respectively. Cumulative VE against VCD from first dose to 3 years post 2nd dose was 62.0% (95% CI: 56.6–66.7) and 83.6% (95% CI: 76.8–88.4) in prevention of hospitalized VCD. Some decline in VE was observed over time mainly driven by outpatient dengue. Two doses of TAK-003 3 months apart were well-tolerated with no important safety risks identified up to 3 years after completion of the vaccination schedule. Safety and immunogenicity data from Phase I/II studies established the final formulation and dosing schedule. Overall VE in the pivotal phase III study was 80.2% [95% CI: 73.3–85.3] against virologically confirmed dengue (VCD) at 12 months post 2TAK-003 is immunogenic against all 4 dengue serotypes and continues to be efficacious, well-tolerated, and with no evidence of disease enhancement in seronegative population up to 3 years post-vaccination.Vianney Tricou, D Phil, Takeda Pharmaceuticals International (Employee) Shibadas Biswal, MD, Takeda Vaccines, Inc (Employee) Sanjay S. Patel, PhD, Takeda Pharmaceuticals International AG (Employee) Olaf Zent, MD, Takeda Pharmaceuticals International AG (Employee) Martina Rauscher, PhD, Takeda Pharmaceuticals International AG (Employee) Gonzalo Perez, MD, Takeda group companies (Employee) Walid Kandeil, MD, Takeda Pharmaceuticals International AG (Employee) Nicolas Folschweiller, PhD, Takeda (Employee)


Globally, millions of patients are affected by myocardial infarction or lower limb gangrene/amputation due to atherosclerosis. Available surgical treatment based on vein and synthetic grafts provides sub-optimal benefits. We engineered a highly flexible and mechanically robust nanotextile-based vascular graft (NanoGraft) by interweaving nanofibrous threads of poly-L-lactic acid to address the unmet need. The NanoGrafts were rendered impervious with selective fibrin deposition in the micropores by pre-clotting. The pre-clotted NanoGrafts (4 mm diameter) and ePTFE were implanted in a porcine carotid artery replacement model. The fibrin-laden porous milieu facilitated rapid endothelization by the transmural angiogenesis in the NanoGraft. In-vivo patency of NanoGrafts was 100% at 2- and 4-weeks, with no changes over time in lumen size, flow velocities, and minimal foreign-body inflammatory reaction. However, the patency of ePTFE at 2-week was 66% and showed marked infiltration, neointimal thickening, and poor host tissue integration. The study demonstrates the in-vivo feasibility and safety of a thin-layered vascular prosthesis, viz., NanoGraft, and its potential superiority over the commercial ePTFE.The online version contains supplementary material available at 10.1186/s12951-022-01268-1. Worldwide > 220 million patients are at risk of stroke, myocardial infarction, lower limb gangrene, and amputation due to severe atherosclerosis, a disease mainly affecting arteries with < 6 mm diameter . The estTo address these limitations, our group at Amrita (India) developed a novel concept of biodegradable, flexible vascular graft using bundles of nanofibrous threads called yarns , which cThe full version of Materials and Methods is shown in the Additional file 2).Polymeric yarns were produced from poly(L-lactic acid) (PLLA) previously subjected to electrospinning. These were then utilized to make the NanoGraft using a weaving system designed and fabricated in-house , 18. TheAs the woven NanoGraft was intended for arterial applications, it was subjected to a pre-clotting protocol as described by Sauvage et al. . to prevThe physical properties of the nanotextile conduit were evaluated at the Bristol Heart Institute under arterial pulsatile flow conditions using a bioreactor  primed with fresh heparinized porcine blood , fixed with 4% paraformaldehyde (PFA), washed in PBS, dehydrated in gradient ethanol, and imaged using SEM. We then undertook cell cultures on the whole NanoGraft to assess cytotoxicity and amount of engrafted cells. First, HUVECs (105 cells) were seeded in the NanoGraft at static conditions by placing them in a petri dish for 24 h. Next, the NanoGraft was mounted in the bioreactor primed with culture media and run at physiological conditions  for additional 24 and 48 h. Then, the NanoGraft was cut into 5 mm segments. Cell presence and viability were tested using the Alamar blue assay, as reported in the Supplemental file. The number of viable cells remaining adherent in each segment of the NanoGraft was obtained from the standard curve. The percentage of adherent cells was calculated by comparing it with static control.In-vitro biocompatibility was tested by seeding human umbilical cord vein endothelial cells (HUVECs) described in the Supplemental file. Briefly, we undertook static cell culture over 72 h on small nanotextiles samples aseptically transferred to 96-well plate and seeded with 2 × 10Upon establishing the in-vitro mechanical safety of the NanoGraft in the dynamic bioreactor study, we undertook the in-vivo feasibility trial in an advanced porcine carotid artery replacement model. The in-vivo feasibility trial was conducted at the Translational Biomedical Research Centre (TBRC) for the large animal at the University of Bristol, Bristol, UK.The animal procedures were in line with the U.K. Home Office regulations  and were undertaken under a Project Licenses (PPL 30/3064 and PPL: 30/2854) granted by the Home Office after formal review and approval by the University of Bristol Animal Welfare and Ethics Review Body (AWERB). We used female Yorkshire pigs (approx. 60 kg) receiving daily aspirin (300 mg) with food. The procedure was in line with established approaches by our group . GeneralIn-vivo vascular ultrasound doppler (USD) was acquired before the implant, 5 min after implant, 2-week for three Nanografts and three ePTFE grafts, and 4-week for three additional NanoGrafts. The USD was acquired under general anesthesia and continuous hemodynamic and surface electrocardiographic monitoring. Lumen size and blood flow velocity were measured and compared within groups and across groups. The inner diameter and intimal thickening of the proximal and distal anastomosis and mid graft portion were determined by 2D Doppler . Blood flow through the implanted graft was determined both qualitatively and quantitatively by using the color Doppler mode.All the carotid arteries inclusive of all the grafts were explanted at 2 and 4 weeks under general anesthesia at termination and placed in 0.9% saline solution. Next, the lumen area of the excised grafts was assessed using OCT , with an ex-vivo pullback being performed throughout the length of each specimen.More detailed methods are presented in the Additional file Samples were stained with Hematoxylin  for 2 min, then rinsed in distilled water for 3 min. Sections were stained using 0.5% Eosin , washed for 3 min in tap water, dehydrated in 100% ethanol, rinsed in xylene for 5 min, and imaged using Leica compound microscope .Sections were stained with Verhoeff’s solution  as detailed in the Supplementary file and checked microscopically for elastin fibers. The slides were treated with 5% sodium thiosulfate and washed in running tap water prior to counterstaining with Gieson’s solution for 3–5 min. The specimens were dehydrated quickly with alcohol and mounted with coverslips using DPX mountant .The slides were prepared as described in the Additional file The slides were stained in a 1% alcian blue solution with 3% acetic acid  for 30 min, washed in running tap water for 2 min, and counterstained in the nuclear fast red solution for 5 min.Samples were fixed in 10% NBF, washed twice with PBS for the 30 s, and tissue exposed to 200 µl of permeabilization buffer  for 10–15 min. Next, additional PBS washing, incubation with 1 mg/ml Bovine Serum Albumin  for 1 h and exposed to antibody solutions in sequence  as described in Additional file Flexible and kink-resistant nanotextile conduits of 4 mm diameter were developed from polymeric nanofibrous yarns of electrospun PLLA using a modified weaving system as described earlier , see FigOne key aspect of the in-vitro validation of the Nanograft before in-vivo testing was to assess its biocompatibility under arterial dynamic conditions. Specifically, it was important to assess the endothelialisation potential of this nanotextile conduit. The Alamar blue assay demonstrated the presence of a large number of viable endothelial cells adherent to the NanoGraft and distributed uniformly at 24 and 48 h throughout its length, with the exception at both ends where the cannula was inserted to secure the graft to the flow circuit Fig. g–h. NotiHaving validated in-vitro the physical properties and biocompatibility of the NanoGraft, we then undertook an in-vivo feasibility trial of pre-clotted NanoGrafts, using clinical-grade ePTFE grafts as control for the 2-week time point. The range of internal diameter of the NanoGrafts was 3.6–3.9 mm. Both NanoGraft and ePTFE grafts exhibited good surgical suturability and handling properties during anastomosis and had a similar vascular cross-clamping time of 12–15 min Fig. : Video 3USD results showed that the synthetic grafts had an average wall thickness of 400–500 µm, similar to the native carotid artery. All the grafts were patent soon after the surgical implantation. Late graft patency, lumen size, and blood flow velocity are shown in Fig. Preliminary histology showed no signs of gross thrombi on the luminal surface of the NanoGrafts Fig. a in cont. This finding was confirmed by the en face staining of the endothelial surface of NanoGrafts, as shown in Fig. Finally, the intimal thickening observed in the NanoGraft was minimal, with uniform and complete endothelial coverage which can be attributed to its transluminal capillary in-growth as illustrated in Fig. A significant transmural neo-capillary formation was observed in NanoGrafts at 4-weeks after surgery, via hematoxylin & eosin staining Fig. a, i–iv. Biodegradable grafts have attracted recent interest due to their inherent regenerative potential and ability to resorb over time, eliminating life-long foreign-body inflammatory activation –28. ManuIn-vivo feasibility studies were conducted at clinical standards, as this is of paramount importance to ascertain the safety and pre-clinical efficacy of the device to enhance its translation potential. Biodegradable grafts must withstand the pressures exerted by pulsatile flow without bursting or experiencing permanent and dangerous deformation through aneurysm or tearing. At the same time, colonization of the graft by inflammatory cells may trigger structural weaknesses. Noticeably, thrombus formation within the grafts can be mitigated if alterations in blood flow velocity across the graft length are minimal, and the luminal graft surface characteristics are non-thrombogenic . In addivasa-vasorum, which was absent in ePTFE grafts. The formation of these transmural neo-capillaries may be attributed to the porous nature of the NanoGraft. Similar observations have been made by others related to graft porosity [The excellent in-vivo biocompatibility of NanoGraft is an essential factor that may facilitate longevity, which was not observed in ePTFE grafts at the same time point. This observation might be associated with the uniform transmural neo-capillary formation within the walls of the NanoGraft, resembling porosity , 40. Addporosity , supportporosity , 40, 42.porosity , which fporosity .Synthetics grafts like ePTFE and Dacron® are incapable of spontaneous endothelialisation on the luminal surface of the graft , 44. ThiThe deposition of extracellular matrix components such as elastin, collagen and mucopolysaccharides would also provide mechanical integrity during the resorption of NanoGraft. Elastin is expected to increase only after the loss of mechanical integrity of the synthetic graft, observed typically in long-term in-vivo studies. Collagen improves the mechanical resilience , while eThe study confirmed the in-vivo feasibility and safety of the single-layered vascular prosthesis, viz., NanoGraft. Its use appears to be associated with increased transmural in-growth of host cells and neo-capillaries, complete neo-endothelial coverage, patency, and reduced neointimal thickening when compared to ePTFE, although more data is needed to confirm this. Suturability and surgical handling were as good as ePTFE grafts. Physical properties of the NanoGraft were in concurrence with ISO standards, with no dilatation/aneurysm formation observed at 4-week. A pivotal porcine long-term preclinical study is warranted to confirm the safety/efficacy of the proposed graft.Additional file 1: Materials and Methods, Figure S1. Mechanical properties of NanoGrafts a) High radial stiffness to resist distortion and compression b) Superior suture retention that demonstrates resistance to wear and tear edges c) Adequate tensile strength to prevent graft rupture. Figure S2. ePTFE grafts showed significant suture line oozing after the restoration of arterial blood flow following anastomosis with 7-0 sutures. Figure S3. The histopathological finding of the midsection of grafted ePTFE at 2 weeks a) total thrombotic occlusion at the luminal region b and c) higher magnification showing intact fibrin-clot at the graft interface. Figure S4. The histopathological finding of the midsection of grafted ePTFE at 2 weeks showing dense infiltration of immune cells in the abluminal section characterised by presence of lymphocytes (white arrows), macrophages (green arrow) and neutrophils (yellow arrows). Figure S5. En face immunofluorescence staining of NanoGraft at 2 weeks showed tight endothelial junctions .Additional file 2:Video 1. Physiological properties of the NanoGraft were evaluated under arterial pulsatile flow conditions using a bioreactor  primed with heparinized porcine blood.Additional file 3:Video 2. Nanotextile based graft showed excellent suturability and lack of fraying at the edge.Additional file 4:Video 3. ePTFE grafts showed significant suture line oozing after the restoration of arterial blood flow through the graft wall.Additional file 5:Video 4. Lack of postoperative suture line bleeding and transmural blood leakage of NanoGraft.Additional file 6:Video 5. Percutaneous ultrasound shows the luminal patency and pulsatile blood flow through the Nanograft at 2 weeks.Additional file 7:Video 6. Percutaneous ultrasound shows the luminal patency and pulsatile blood flow through the ePTFE graft at 2 weeks.Additional file 8:Video 7. Percutaneous ultrasound shows the total occlusion of ePTFE graft at 2 weeks.Additional file 9:Video 8. Ex-vivo OCT post-termination confirmed the patency of Nanograft at 2 weeks.Additional file 10:Video 9. En face staining of NanoGrafts confirms complete endothelial coverage at two weeks of post-implantation.


Function by Roy et al.“Choose your parents wisely” is a popular adage coined by the British Philosopher Bertrand Russell, and often used when discussing issues such as socioeconomic position and genetic contributions to disease risk. When it comes to vascular function, a predictor of cardiovascular disease risk, the recent study in HCR) and vice versa (HCR-mtLCR). Specifically, HCR female offspring were backcrossed with male LCR  via inbreeding, and this backcross procedure was repeated over several generations to generate the LCR-mtHCR and HCR-mtLCR. The investigators performed echocardiography to assess cardiac function and left ventricular mass, wire, and pressure myography to assess arterial vasodilatory function (with and without PVAT) and mechanics, macroscopic tissue imaging of PVAT, and bioenergetic assays with vascular smooth muscle cells (VSMCs).The investigators studied rats artificially selected (within-family) for intrinsic aerobic endurance running capacity to generate LCR and HCR male rats.HCR) but decreased left ventricular mass and several indices of cardiac performance in HCR-mtLCR relative to HCR.Compared to HCR, LCR rats had higher body mass, epididymal fat mass, and blood pressure. Regarding cardiac measures, HCR rats presented higher left ventricular mass than LCR rats see . CompareHCR), however, it did not reduce vascular function in HCR-mtLCR. Using mesenteric arterioles and plotting internal lumen and external diameters with intraluminal pressure curves, the authors revealed that the arterioles from LCR rats exhibited hypotrophic remodeling . Moreover, mitochondrial swap did not have an effect on either the LCR or HCR phenotype.The key findings were that compared to HCR, LCR rats presented with lower endothelium-dependent (acetylcholine) and endothelium-independent (sodium nitroprusside) vasodilation in mesenteric resistance arteries. Mitochondrial swap rescued endothelium-dependent and endothelium-independent vasodilation in LCR  to what extent intrinsic exercise capacity and mitochondrial DNA influence vascular function in female and older rodents, (2) if the reduced endothelium-dependent vasodilation in LCR rats is mediated by reduced NO synthesis and/or bioavailability or a combination of NO and/or other vasodilators  or prostacyclin), (3) the role of mitochondrial ROS and other vascular sources of ROS  in contributing to the vascular phenotypes associated with HCR and LCR, and (4) determining whether exercise, similar to mitochondrial swapped conplastic strains, can also rescue endothelial dysfunction in LRC rodents .,HCR on LCR rodents demonstrates that mitochondrial DNA is an essential oddment to the protective phenotype conferred by high CRF. These findings will hopefully pave the way for exciting future research to determine mitochondrial-based targets to help prevent and treat cardiovascular disease.At the population level, we cannot do much to improve an individual’s intrinsic CRF. Indeed, the HERITAGE family study established that intrinsic capacity and trainability are genetically heritable in humans.


Science paper ‘Increased cystine in leukocytes from individuals homozygous and heterozygous for cystinosis’ opened a new era of research towards understanding the pathogenesis and finding treatments for cystinosis patients. His tremendous contribution transformed cystinosis from a fatal disorder of childhood to a treatable chronic disease, with a new generation of cystinosis patients being now in their 40th and 50th years. Dr. Schneider wrote a fascinating ‘Personal History of Cystinosis’ highlighting the major milestones of cystinosis research. Unfortunately, he passed away before this manuscript could be published. Fifty-five years after his first paper on cystinosis, the ‘Personal History of Cystinosis’ by Dr. Schneider is a tribute to his pioneering discoveries in the field and an inspiration for young doctors and scientists who have taken over the torch of cystinosis research towards finding a cure for cystinosis.Cystinosis is a rare lysosomal storage disease that is tightly linked with the name of the American physician and scientist Dr. Jerry Schneider. Dr. Schneider (1937–2021) received his medical degree from Northwestern University, followed by a pediatrics residency at Johns Hopkins University and a fellowship in inherited disorders of metabolism. He started to work on cystinosis in J. Seegmiller’s laboratory at the National Institutes of Health (NIH) and subsequently moved to the UC San Diego School of Medicine, where he devoted his entire career to people suffering from this devastating lysosomal storage disorder. In 1967, Dr. Schneider’s seminal  When people write about the history of cystinosis, they usually start with Emil Abderhalden, who in 1903 described cystine crystals in the liver and spleen at the autopsy of a 21-month-old child . I w. I w6]. I also found that the solubility of cystine in human plasma at 37° and at physiological pH was about 50 times higher than the plasma cystine concentration in cystinosis patients. This proved to me that the cystine crystals must have formed intracellularly. I later found that a pathologist in Birmingham, England had come to the same conclusion by careful study of pathological sections from cystinosis patients.I isolated leukocytes in small volumes of blood from cystinosis patients and found that the amount of soluble cystine in these cells was 50–100 times higher than in leukocytes isolated from individuals who did not have cystinosis . We alsoMy two-year assignment in Seegmiller’s lab was coming to an end and I performed one more experiment that excited me. I wondered if there was an enzyme that reduced cystine to cysteine  that might be defective in cystinotic tissue. I decided to develop an essay for glutathione reductase and compare the activity of this enzyme in cystinotic compared to normal tissue. The idea was that glutathione reductase might indirectly reduce cystine to cysteine. In normal tissue, the cystine would be reduced to cysteine and be further metabolized, whereas in cystinotic tissue, the cystine would not be metabolized and would accumulate and form crystals. Sure enough, I found no glutathione reductase activity in the cystinotic tissue, but high amounts of glutathione reductase activity in the normal control tissue. I was very excited by this finding; fortunately, I decided to conduct control studies. I added cystine to normal tissue, and the activity of glutathione reductase disappeared. I dialyzed the cystinotic tissue for a long period with large volumes of solution to remove the cystine, and the activity of this enzyme returned to normal. It was obvious that cystine inhibited the activity of glutathione reductase. Some reading in the library informed me that cystine inhibits many common intracellular enzymes that are necessary for the normal life of cells. Just before leaving the NIH for further training in other areas of science, I conducted a brief experiment that demonstrated that cystine was somehow compartmentalized in the cell so that it did not inhibit the activity of important cellular enzymes.The next physician who studied cystinosis in Seegmiller’s laboratory was Joe Schulman. He found that the excess cystine in cystinotic tissue was located in the lysosomes of the cells. The Belgian scientist Christian de Duve shared the 1974 Nobel Prize in Physiology or Medicine for his discovery of lysosomes, an intracellular organelle that “digests” proteins and other cellular molecules . We now Seegmiller moved to the University of California San Diego (UCSD) in 1969, and Schulman took over his NIH laboratory. In 1970, Seegmiller recruited me to UCSD, where I was able to establish my own laboratory in the Department of Pediatrics. Schulman and I were friends and decided we would both continue to study cystinosis. I decided the major problem we had in these studies was how time consuming and difficult it was to assay cystine. Since each assay took 8 h, we could only perform three cystine assays a day on our one amino acid analyzer (I had to come to the laboratory each evening at midnight to start the third measurement.) Fortunately, I learned that a biochemist (Clement Furlong) at the University of California Riverside was isolating proteins from the bacteria E. coli that had the property of specifically binding small molecules and that there was a cystine-binding protein. I thought this might allow us to develop a sensitive assay for cystine. I called Furlong and asked if he could help me isolate this protein. He told me he was preparing some of these proteins later that week and he thought he knew where the cystine-binding protein came off the column he used to prepare these proteins. He invited me to visit and take some of this material. I had a new post-doctoral fellow (Robert Oshima) who had just arrived and we drove to Riverside later that week (90 min drive). We returned that evening with a beaker full of solution that we hoped contained the cystine-binding protein. I went home for dinner and sleep. Oshima was too excited to sleep and worked all night with the material we obtained and the next morning told me he was certain it would work. We submitted a paper to the Journal of Biological Chemistry two months later describing the use of this cystine-binding protein to measure small amounts of cystine . For manBefore I continue describing our laboratory studies, I must discuss some of the attempts to find a specific treatment for cystinosis. It is never easy to find a successful treatment for any disease, but in some cases it quickly becomes obvious that the treatment works. Of course, you have to be certain the treatment is not harmful, but at least you know if it works. For instance, if you develop an antibiotic for a certain bacterial infection, you soon learn if the antibiotic kills that bacteria. Cystinosis is much more difficult. The kidney damage we were attempting to arrest progresses very slowly, and our methods of measuring kidney function in children were not very exacting. We assumed that the cystine was causing the kidney damage, but we could not be certain. It was always possible that whatever was causing the storage of cystine was also causing the kidney damage, and that the two processes were not related. Fortunately, this was not to be the case, and measurement of leukocyte cystine proved to be an excellent marker of whether any particular drug was effective in treating cystinosis.Over the years many approaches were tried to specifically treat cystinosis and they all seemed to work! I will discuss two treatments that did not work and one that did. Horst Bickel directed the first treatment I will discuss. Dr. Bickel trained in pediatrics from 1947 to 1949 at the University Children’s Hospital in Zurich with Professor Fanconi. He then worked as a research fellow at the University Children’s Hospital in Birmingham, England from 1949 to 1954. While in Birmingham, Bickel developed the successful treatment for another inherited metabolic disease, phenylketonuria (PKU). This condition was treated with a specific diet very low in the amino acid phenylalanine . It was Science. I finally agreed to submit the paper being certain it would be promptly rejected and Dr. Kroll would learn to trust his mentor. To my astonishment, the paper was promptly accepted [Our group tried another treatment that did not work. A German post-doctoral fellow working in my lab (Wolfgang Kroll) found that adding fresh ascorbic acid (Vitamin C) to the culture media in which cystinotic fibroblasts were growing lowered the cystine content of the cells by over 50%. He was very eager to publish this finding. I thought we should try to understand why this occurred before submitting a paper for publication. Dr. Kroll was relentless in insisting we publish the finding even though we did not understand it. Furthermore, he wanted to submit it to the very prestigious journal, accepted . I was caccepted .John Crawhall, the scientist who helped me at the NIH, began the study that did work. He was then working in Montreal but came to UCSD for a sabbatical and decided to add a variety of compounds to the media in which cystinotic cells were growing to see if any had an effect on the cystine content of these cells. The only compound that seemed to have such an effect was cysteamine . By the Cysteamine occurs naturally in humans, but at very low concentrations . It was We joined with the NIH group of Joe Schulman, his post-doctoral fellow Bill Gahl and biostatistician James Schlesselman to start a large study. Many physicians contributed patients, including Michel Broyer in Paris and William van’t Hoff in London. In 1987, we reported the success of cysteamine in treating cystinosis . At firsMeanwhile, we continued to search for the metabolic defect in cystinosis. We all agreed that the most likely cause was a transport defect in the lysosomal membrane. As proteins are digested in the lysosomes, individual amino acids are released. Specific transporters exist in the lysosomal membrane that carry specific amino acids from the lysosome to the cytoplasm of the cell. Could the transporter for cystine be missing in cystinosis lysosomes? None of us knew how to test this idea. Eric Harms visited our laboratory from Germany and helped us isolate excellent preparations of lysosomes, but we still did not know how to study transport in the lysosomes. The breakthrough came when Frank Tietze, a biochemist at the NIH, saw a paper in the Journal of Biological Chemistry and pointed it out to Drs. Schulman and Gahl at the NIH. This paper described a method of loading lysosomes with a specific amino acid . Thus, wThe groups at the NIH and the University of Michigan  found that cysteamine works in the cystinotic lysosome by “attacking” the cystine molecule and forming a “mixed disulfide” of half cystine and cysteamine. This “mixed disulfide” resembles the amino acid lysine and exits the lysosome via the alternative transporting system that is normal in cystinotic lysosomes . FinallyIn the 1980s, we began to use cysteamine bitartrate to treat cystinosis. Although this drug is also difficult to synthesize, a company in Italy was producing it. The drug was stable as a solid and more acceptable for patients. In 1994, the United States Food and Drug Administration (FDA) approved cysteamine bitartrate for the treatment of cystinosis. Although this formulation of cysteamine was very effective in removing cystine from cystinotic cells, it still had several problems as a drug for cystinosis. It has a very bad smell and taste, and must be taken every 6 h to be completely effective. Elena Levtchenko, in Belgium, showed that if a patient went 8 h between doses, for instance while sleeping, the leukocyte cystine content went unacceptably high in the last two hours . For accIf started early in life, cysteamine bitartrate slows the progression of kidney glomerular but not kidney tubular damage. Studies from both the NIH and Hôpital Necker Enfants-Malades in Paris found that, even if started at an older age, cysteamine delays the progression of hypothyroidism, diabetes and neuromuscular disorders ,26. HoweScience 1967, the Special Issue of Cells is devoted to cystinosis and to the tremendous contribution of Dr. Schneider as a pioneer and founder of cystinosis research.Jerry Schneider devoted his whole career to unraveling the disease mechanisms and finding a curative therapy for cystinosis. His work transformed cystinosis from a fatal condition of childhood into a treatable disease with which patients can now reach advanced adult age. Dr. Schneider inspired several generations of clinicians and researchers to take over the torch of cystinosis research and to find cure for this devastating disorder. Fifty fine years after the first pioneering paper on cystinosis in