A-type crystalline structure. Their short-range ordered structure and lamellar structure had no significant difference. However, the rhizome starch had a significantly bigger granule size and lower amylose content than the bulbil starch. The swelling power and water solubility were significantly lower in the rhizome starch than in the bulbil starch. The onset and peak gelatinization temperatures were significantly higher in the rhizome starch than in the bulbil starch. The rhizome starch had a significantly higher breakdown viscosity and a lower setback viscosity than the bulbil starch. The thermal stability was lower in the rhizome starch than in the bulbil starch. The rhizome starch had a significantly lower resistance to hydrolysis and in vitro digestion than the bulbil starch. The above results provide important information for the utilization of rhizome and bulbil starches of Chinese yam.Chinese yam is an important edible starch plant and widely cultivated in China. Its rhizome and bulbil are starch storage tissues below and above ground, respectively. In this paper, starches were isolated from the rhizome and bulbil of Chinese yam, and their structural and functional properties were compared. Both starches had an oval shape with an eccentric hilum and a C Starch consists mainly of linear amylose and highly branched amylopectin and is stored as semicrystalline granules . Starch Dioscorea of the monocot family Dioscoreaceae. World production of yams reached over 65.9 million metric tons in 2016, and the top three producers are Nigeria, Côte d’Ivoire, and Ghana with production quantities over 31.5, 4.9, and 4.5 million metric tons, respectively, every year from 1997 to 2016 [Dioscorea opposite Thunb.) is a rhizome crop mainly cultivated in China, Japan, and Korea [Yam is a generic name for the plants of twining climbers that form tubers or rhizomes in the genus  to 2016 . Chinesend Korea . The rhind Korea ,15,16,17nd Korea . The bulnd Korea ,19. The nd Korea ,20. The nd Korea ,20. HoweIn this study, starches were isolated from rhizomes and bulbils of Chinese yam cultivar Shuishanyao. Their morphology, granule size, amylose content, crystallinity, short-range ordered structure, lamellar structure, swelling power, water solubility, thermal and pasting properties, thermal stability, and hydrolysis and digestion properties were investigated using many physicochemical measuring methods and spectral analysis machines. Our objective was to compare the structural and functional properties of rhizome and bulbil starches and provide some information as to how these starches can be used.Lilium lancifolium [The soluble sugar and starch contents in dry rhizome and bulbil are presented in cifolium . The higLilium lancifolium [The isolated starch granules were observed under normal and polarized light. The starches from rhizome and bulbil were similar and exhibited an oval shape with the hilum at one end of the granule . The precifolium . Starch cifolium .The rhizome and bulbil starch contained 35.2% and 38.3% amylose content, respectively , which wA-(closer to A-type), CC-, and CB-(closer to B-type) type according to the proportion of A- and B-type crystallinity. The CA-type starch has a shoulder peak at 18° 2θ, and CB-type starch has two shoulder peaks at 22° and 24° 2θ [A-type starch. CA-, CC-, and CB-type starches have been reported in bulbils of different Chinese yam cultivars [Starches from different plant sources can be divided into A-, B-, and C-type according to their X-ray diffraction (XRD) patterns. Usually, normal cereal seeds have A-type starch, most tubers contain B-type starch, and some rhizomes and leguminous seeds have C-type starch . The XRDd 24° 2θ . Accordi−1 are associated with ordered/crystalline and amorphous regions in starch, respectively. The ratio of absorbance, 1045/1022 cm−1, can be used to quantify the ordered degree, and that of 1022/995 cm−1 is used to measure the proportion of amorphous to ordered carbohydrate structure in the starch [−1 are shown in Lilium lancifolium also present similar IR ratios at 1045/1022 and 1022/995 cm−1 [−1 [The short-range ordered structure of starches, defined as the double-helical order, can be detected using a Fourier transform infrared (FTIR) spectrometer. The bands at 1045 and 1022 cme starch . The FTI995 cm−1 , but stacm−1 [−1 . The difThe lamellar structure of starch can be analyzed using small-angle X-ray scattering (SAXS) spectra. The SAXS spectra of the rhizome and bulbil starches are shown in The swelling power and water solubility of both rhizome and bulbil starches are shown in The thermal properties of both rhizome starch and the bulbil starch were analyzed by a differential scanning calorimeter (DSC), and their thermograms and thermal parameters are given in The pasting properties of starch were determined by a rapid visco analyzer (RVA), and their RVA patterns and parameters are given in The thermogravimetric analysis (TGA) curves of the rhizome and bulbil starches are presented in The rhizome and bulbil starches were hydrolyzed with α-amylase from porcine pancreatic (PPA) by measuring the released soluble carbohydrate A. The hyAspergillus niger (AAG)  B. DuringDioscorea opposita Thunb. cultivar Shuishanyao) were obtained from a local natural food market  in December 2016.The fresh rhizome and bulbil of Chinese yam  equipped with a charge-coupled device (CCD) camera  as previously described . The graThe amylose content was determined using the iodine colorimetric method as previously described .The crystalline structure of starch was analyzed using an X-ray powder diffractometer  at 200 mA and 40 kV as previously described . The scaThe short-range ordered structure of starch was analyzed using a Varian 7000 FTIR spectrometer with a DTGS detector equipped with an attenuated total reflectance single-reflectance cell containing a germanium crystal (45° incidence angle)  as previously described .The swelling power and water solubility of starch were determined by heating starch-water slurries at temperatures from 50 to 95 °C in 5 °C intervals as previously described .The thermal properties of starch were measured by a DSC  as previously described .The pasting properties of starch (8% solids) were analyzed using an RVA  following the temperature program: holding at 50 °C for 1 min, heating to 95 °C at 12 °C/min, maintaining at 95 °C for 2.5 min, cooling to 50 °C at 12 °C/min, and holding at 50 °C for 1.4 min.Ten milligrams of starch were placed on a platinum pan and heated from room temperature to 800 °C using a Pyris 1 TGA system  at a heating rate of 10 °C/min. Nitrogen was used as a purge gas at a flow rate of 20 mL/min. Change in sample weight against temperature was measured.g for 5 min. The supernatant was used for the measurement of the solubilized carbohydrates to quantify the degree of hydrolysis using the anthrone-H2SO4 method.The starch was hydrolyzed by PPA  for different times as previously described . After hThe in vitro digestion of starch was determined using both PPA (Sigma A3176) and AAG  as previously described . The relThe mean value and significant difference were analyzed using SPSS software .A-type crystallinity and showed a similar short-range ordered structure and lamellar structure. The rhizome starch had lower swelling power, water solubility, and setback viscosity and higher onset and peak gelatinization temperatures and breakdown viscosity than the bulbil starch. The bulbil starch had higher thermal stability, resistance to hydrolysis, and in vitro digestion than the rhizome starch.The bulbil and the rhizome of Chinese yam had 66.9% and 84.6% starch content, respectively. Their starches were oval in shape and had eccentric hila. The volume- and surface-weighted mean diameters were 16.4 and 8.5 μm for the bulbil starch, respectively, and 17.7 and 9.4 μm for the rhizome starch, respectively. The bulbil starch and the rhizome starch had 38.3% and 35.2% amylose content, respectively. Both rhizome and bulbil starches had C


Adzuki beans are used to prepare foods with glutinous and non-glutinous rice in Japan, and adzuki bean pigments are able to color rice starch a purplish red. This study deals with the adzuki bean extract-dependent suppression of starch digestion of non-glutinous rice flour (joshinko in Japanese), which was gelatinized in boiling water and then cooled to 37 °C. Accompanying the treatment of joshinko with pancreatin, amylose and amylopectin were released from the joshinko particles, and the released amylose and amylopectin were further digested. The adzuki extract suppressed the release and digestion by binding to amylose and amylopectin, which were present in the particles and at the surfaces of the particles. Fatty acids and flavonoids in the adzuki extract contributed to the suppression. In addition, the starch digestion in the joshinko particles appeared to be suppressed if the amylose/fatty acid complexes and amylose/flavonoid and amylopectin/flavonoid complexes, which are poor substrates of α-amylase, surrounded the particles. It is discussed that the suppression was due to the prevention of α-amylase access to the particles. Adzuki bean is a normal food in Japan and is used in cooking with glutinous and non-glutinous rice to prepare sekihan and adzuki-meshi, respectively. The adzuki bean is also used to prepare the paste for sweets. In addition, flour of non-glutinous rice (joshinko in Japanese) is normally used to prepare various Japanese foods, including dumplings, uiroh (sweet jelly or pudding), kashiwa-mochi (rice cake wrapped with an oak leaf), kusa-mochi (rice cake mixed with boiled leaves of mugwort), etc. The latter two types of cakes are filled with adzuki bean paste. It has been reported that adzuki contains health-promoting antioxidants, such as flavonoids and tocopherols ,2,3, andThis study deals with (i) the adzuki bean extract-dependent suppression of starch digestion in retrograding joshinko, which was catalyzed by α-amylase in pancreatin; (ii) the binding of adzuki components to amylose and amylopectin in retrograding joshinko; and (iii) the elucidation of the adzuki bean components that suppressed starch digestion. Taking the above results and the structures of retrograding joshinko particles into consideration, it is discussed that amylose/fatty acid complexes and amylose/flavonoid and amylopectin/flavonoid complexes can suppress starch digestion in the particles by surrounding them. The elucidation of the mechanism may provide some procedures to prepare foods using joshinko, the digestion of which is slow. Vigna angularis (Willd.) Ohwi & H. Ohashi cv. erimo) were supplied by the Japan Pulse Foundation  and the Tokachi Agricultural Experiment Station . Flour of non-glutinous rice (joshinko) was obtained from Hinode-Seifun Co. . (+)-Catechin, taxifolin, rutin, and quercetin were from Sigma-Aldrich Japan . Oleic acid, linoleic acid, linolenic acid, and pancreatin from pig pancreas were from Wako Pure Chemical Ind. Co. . Quercetin 7-O-glucoside and vignacyanidin were prepared from adzuki bean as reported previously [Seeds of adzuki  was added to the soaked adzuki bean after removing the water not absorbed by the bean. The adzuki bean in methanol was kept at 25 °C in the dark for 1 h with gentle shaking to extract components contained in the seed coat. This extraction was repeated twice. After combining the two methanol extracts, methanol was removed in vacuo and the pH of the remaining aqueous solution (about 700 mL) was adjusted to 3 by HCl to extract with 200 mL of ethyl acetate three times. Water not absorbed by the adzuki bean (about 500 mL) was extracted with 200 mL of ethyl acetate once after the pH was decreased to 3. The ethyl acetate extracts were combined and washed with 100 mL of 1% HCl five times. After removing the water of the combined ethyl acetate extracts with anhydrous sodium sulfate, ethyl acetate was evaporated in vacuo. The residue was dissolved in 2 mL of ethanol with 0.1% HCl and kept at −20 °C. Flavonoids in the extract were identified from their retention times, UV-visible absorption spectra, and mass spectra ; their a−1) and then incubated for 5 min at 37 °C. The reason that the volumes were selected was that 10 μL of adzuki extract, which was equivalent to 5 g of adzuki bean, significantly suppressed pancreatin-induced starch digestion under the conditions of this study. Retrograding joshinko with and without adzuki extract was centrifuged at about 1500× g for 2 min. The supernatants and sediments were reddish purple and purple, respectively. The purple sediments were washed once with 1 mL of the above buffer solution and then suspended in 0.9 mL of the buffer solution. In the following experiments, the samples used were (1) retrograding joshinko, (2) the sediment, and (3) the supernatant. Starch digestion was initiated by adding 5 or 10 μg of pancreatin mL−1 at 37 °C. Joshinko (0.5 g) was suspended in 50 mL of 0.1 M sodium phosphate (pH 7.0) with 0.15 M NaCl, and gently heated for 5–6 min to start boiling. After gentle boiling for 2 min, the gelatinized joshinko was cooled to 37 °C in a water bath. The cooled joshinko was used for experiments as retrograding joshinko. Adzuki extract (0–10 μL) was added to 1 mL of the retrograding joshinko (0.01 g mLv/v) just before the experiments [2, and 0.5 M NaOH. Samples (1)–(3) were incubated for defined periods with pancreatin, and 20 μL of the samples was added to the mixture of the two solutions and kept in boiling water for 6 min. After cooling in iced water and centrifuging at 11,000× g for 2 min, absorbance of the supernatant was measured at 410 nm using a spectrophotometer . The path length of the measuring beam was 4 mm.Starch digestion was estimated by the reducing sugar formation and starch-iodine complex formation. The solution to quantify reducing sugars was prepared by mixing solutions I and II , 30 μL of the reaction mixture was withdrawn and added to the mixture of 10 mM iodine in 1.0 mL of 0.1 M sodium phosphate (pH 7.0) with 0.15 M NaCl. As large joshinko particles were precipitated rapidly after the addition of the reaction mixtures prepared from samples (1) and (2), i.e., joshinko and the sediment, the absorption spectra of starch-iodine complex were recorded 1 min after the addition of the reaction mixtures using a UV-2450 spectrophotometer combined with an integrating sphere (MPS-2000) (Shimadzu). The path length of the measuring beam was 4 mm. Data are presented as the difference in absorbance between the starch-iodine complexes and the iodine solution at 550 and 700 nm.Iodine solution (100 mM) was prepared as described previously . After ig for 5 min, and then the absorbance of the supernatant was measured at 490 nm. The concentrations of total sugar were estimated from the standard curve prepared using glucose, and the total sugar concentrations were expressed equivalent to mg or μmol of glucose. Total sugar was estimated by the phenol-sulfuric acid method . An aliq−1) incubated with 10 μL of adzuki extract for 5 min at 37 °C was centrifuged at about 1500× g for 2 min. The sediment was washed once with 1 mL of 0.1 M sodium phosphate with 0.15 M NaCl (pH 7), and then the sediment was extracted with 10 mL of 0.1% HCl in methanol. The supernatants obtained by the first and the second centrifugation were combined and then extracted with 10 mL of 0.1% HCl in methanol. The methanol extracts were dried in vacuo using a rotary evaporator. Each residue was dissolved in 0.5 mL of 1% HCl in methanol, and an aliquot (50 μL) of each solution was applied to a HPLC column  combined with a pump (CL-20AD) and a photodiode array detector (SPD-M20A) . The mobile phase was a mixture of methanol and 0.2% formic acid and the concentration of methanol increased stepwise as previously reported [−1. Each flavonoid was quantified from the area under the peak.One milliliter of retrograding joshinko (0.01 g mLv/v), and the flow rate was 1 mL min−1. Fatty acids separated were detected at 210 nm and quantified from the area under the peak.Fatty acids in adzuki extract were also estimated using the above HPLC system. The mobile phase was a mixture of methanol and 0.2% formic acid  at 1500× g for 5 min. The supernatant, which might have contained starch fragments, was incubated with the adzuki extract, fatty acid, and flavonoid for 5 min at 37 °C. The supernatant became turbid by the incubation. Then, the turbid mixtures were centrifuged at 11,000× g for 5 min to obtain the precipitates. The precipitates were suspended in 0.8 mL of 0.1 M sodium phosphate (pH 7.0) with 0.15 M NaCl and mixed with the sediment obtained by the centrifugation of retrograding joshinko at 1500× g to study starch digestion induced by 10 μg of pancreatin mL−1. The digestion was estimated using iodine as described above.About 0.8 mL of the supernatant was obtained by the centrifugation of 1 mL of joshinko  when the supernatant was prepared in the presence of 0, 5, and 10 μL of adzuki extract, respectively. The results in The pancreatin-induced reducing sugar formation in retrograding joshinko was suppressed by 5 and 10 μL of adzuki extract by about 50% and more than 95%, respectively A. Rates A spectrum of trace a minus KI–I2 in Traces a in The adzuki extract suppressed the amylose–iodine complex formation, which had an absorption maximum around 700 nm, in retrograding joshinko A. Accordn = 3), respectively. Here, the concentration of oleic acid was much higher than that of linoleic acid, but it has been reported that the content of linoleic acid is greater than that of oleic acid in the adzuki bean [As the components to suppress the starch-iodine complex formation in retrograding joshinko, fatty acids were considered first as adzuki beans contain fatty acids , which cuki bean ,24. TherIn addition to fatty acids, flavonoids can suppress starch iodine complex formation ,25,26 beFrom the effects of the adzuki extract on starch-iodine complex formation, it could be postulated B that . This result suggests the release of not only amylopectin but also amylose from the joshinko particles. The adzuki extract suppressed the absorbance increase around 700 nm more effectively than that around 550 nm (traces b–d). This was confirmed by the time courses of the initial increase in the starch concentrations  by incubation in the presence of 0, 5, and 10 μL of adzuki extract, respectively. The above results coincided with the observation that adzuki extract could suppress the release of starch fragments from joshinko particles and the digestion of the released fragments. The suppression of the released starch digestion by 6 μL of adzuki extract seemed to continue for 4 h, and the release of amylopectin in the presence of 10 μL of adzuki extract also seemed to continue for 4 h C,D. JoshAccording to the illustration that starch fragments surround joshinko particles in A spectrum of 3 min minus 0 min of incubation in the absence of adzuki extract showed a much greater absorbance increase around 550 nm than around 700 nm (trace a), suggesting that amylopectin was mainly released from the sediment. The adzuki extract suppressed the release, but the suppressive effects were smaller than those in The pancreatin-induced release of starch fragments from the sediment is shown in A spectra of 40 min minus 20 min of incubation  of the initial volumes in the presence of 0, 3, 6, and 10 μL of the adzuki extract, respectively. The above data support the postulation that the amylose and amylopectin combined with adzuki components had important functions to suppress the initial release of amylopectin from joshinko particles. The ∆cubation B indicatcubation C,D, the A spectra of pancreatin-induced digestion , respectively, and the amounts of (+)-catechin, taxifolin, quercetin 7-O-glucoside, rutin, quercetin, and vignacyanidins in the blue precipitate prepared by adding 5 μL of adzuki extract were 2.20 ± 0.20, 1.59 ± 0.19, 1.61 ± 0.24, 0.27 ± 0.05, 18.7 ± 2.23, and 5.53 ± 0.67 (n = 3) nmol, respectively. The above data suggest that amylose and amylopectin in the supernatant became precipitable by combining with the components in the adzuki bean.To exclude the above possibility that the suppression of starch digestion was due to the inhibition of α-amylase in pancreatin, the precipitate, which contained amylose and amylopectin combined with adzuki components, was prepared from the supernatant of retrograding joshinko as described in n = 3), respectively, at 550 nm, and 13.7 ± 10.3, 22.1 ± 6.5, and 62.1 ± 15.1% (n = 3), respectively, at 700 nm. The suppressions by 0.6 mM quercetin plus 0.2 mM vignacyanidin were about 31 and 16% (average of two experiments) at 550 and 700 nm, respectively. To clarify the components that contributed to the suppression of starch digestion, the effects of fatty acids, quercetin, and vignacyanidin on pancreatin-induced starch digestion were studied using retrograding joshinko. There have been reports on the suppression of amylose digestion by fatty acids ,28,29 ann = 3), respectively, at 550 nm and 31.3 ± 12.5, 38.0 ± 11.6, and 63.3 ± 15.3% (n = 3), respectively, at 700 nm . The release of starch fragments from the joshinko particles and the digestion of the released starch were further suppressed by 0.6 mM quercetin, 0.2 mM vignacyanidin, and 0.6 mM quercetin plus 0.2 mM vignacyanidin. The above results suggest that (i) fatty acids can effectively suppress the release of amylose, resulting in the preferential digestion of the released amylopectin; (ii) the greater suppression of released starch digestion by flavonoids may be due to the release of amylose and amylopectin, which combined with the flavonoids from the joshinko particles and/or the combining of the released starch fragments with free flavonoids present in the reaction mixtures; and (iii) flavonoids could suppress the starch digestion cooperating with fatty acids.Starch precipitates prepared using the adzuki extract suppressed starch digestion of the sediment A,B. ThenThe present study indicated that fatty acids, quercetin, and vignacyanidin in the adzuki bean extract could suppress starch digestion in retrograding joshinko by binding to amylose and amylopectin in joshinko particles and that these compounds could also suppress the digestion of fragmented amylose and amylopectin present in the retrograding joshinko by forming complexes with them. It was found that starch fragments combined with fatty acids and flavonoids, which are poor substrates of α-amylase, could suppress the digestion of the starch in joshinko particles and starch fragments released from the particles by surrounding them. According to the findings, it is possible to prepare starch-containing foods using fatty acids and hydrophobic flavonoids, the digestion of which is slow. Although the structure of amylose/fatty acid complexes has been proposed , the str


In the resting state, heartbeats evoke cortical responses called heartbeat-evoked responses (HERs), which reflect cortical cardiac interoceptive processing. While previous studies have reported that the heartbeat evokes cortical responses at a regional level, whether the heartbeat induces synchronization between regions to form a network structure remains unknown. Using resting-state MEG data from 85 human subjects of both genders, we first showed that heartbeat increases the phase synchronization between cortical regions in the theta frequency but not in other frequency bands. This increase in synchronization between cortical regions formed a network structure called the heartbeat-induced network (HIN), which did not reflect artificial phase synchronization. In the HIN, the left inferior temporal gyrus and parahippocampal gyrus played a central role as hubs. Furthermore, the HIN was modularized, containing five subnetworks called modules. In particular, module 1 played a central role in between-module interactions in the HIN. Furthermore, synchronization within module 1 had a positive association with the mood of an individual. In this study, we show the existence of the HIN and its network properties, advancing the current understanding of cortical heartbeat processing and its relationship with mood, which was previously confined to region level. Complex brain processing usually occurs at a network level, which requires an interaction between brain regions. However, despite its importance in homeostasis and affective processing, a network level processing of cardiac interoception has not been investigated. Here, we first provided an evidence that the heartbeat induces phase synchronizations between cortical regions those comprise a heartbeat-induced network (HIN) with control analyses excluding the possibility of an artificial synchronization. Furthermore, by applying graph-theoretical analysis, we find hubs of the HIN and found out that it is a modularized network with five modules. Finally, we also showed the relationship between the participants’ mood and the HIN. These results provide the first evidence of network-level heartbeat processing and its relevance with emotion.One of the important purposes of the brain is to maintain homeostasis by continuously sensing the homeostatic state , which is termed interoception, and the brain regulates the bodily condition using this homeostatic information even in the resting state . RecentlBecause the brain processes interoceptive information, there exist brain responses related to visceral signal processing. For example, the heartbeat signal evokes cortical activity defined as heartbeat-evoked responses HERs; . HERs arWhile previous studies have shown that the heartbeat evokes region-level or sensor-level cortical responses and is related to a variety of psychological states and functions, whether the heartbeat also induces interactions between cortical regions remains unknown. However, previous fMRI studies have shown that network-level fluctuations vary with visceral signal-related measures such as heart rate variability HRV; , indicatSCR_008749). All subjects were young (22–35 years of age) and healthy. MEG recordings were collected on a whole-head Magnes 3600 scanner (4D Neuroimaging) with 248 magnetometer channels at a sampling rate of 2034.51 Hz. Recordings were performed in three sessions, and each session lasted 6 min. HERs were extracted from the preprocessed version of the MEG dataset, which is publicly available at Connectome DB (Resting-state MEG data from 89 subjects collected from the Human Connectome Project (HCP) S1200 data release were used in this study (ctome DB . The prectome DB  was applctome DB  source tSCR_004849). First, preprocessed HCP MEG data, which were initially segmented into 2-s epochs, were concatenated into one continuous time series such that each segment was realigned to its original location in the raw MEG recording (preprocessed data contained information about the locations of each segment in the raw MEG recording). Because bad segments were removed, the concatenated continuous time series data had empty spaces where the bad segments existed. These empty spaces were replaced by NaN. Next, the R-peak was detected in the ECG recordings using the Pan-Tomkins algorithm  to perform a region of interest (ROI)-based connectivity analysis. Among the total of 246 brain regions, because an HER is known to mainly reflect cortical heartbeat processing (All sensor HER data were source-reconstructed using the linearly constrained minimum variance (LCMV) beamformer methods  providedocessing  and the ocessing , we exclocessing  were ave10.1523/ENEURO.0200-19.2019.t1-1Extended Data Table 1-1Table 1-1, DOCX file.List of 195 cortical regions used in the analysis and the graph theoretical properties of each region Download The debiased estimator for weighted phase lag index wPLI-D;  was usedWe compared the wPLI-Ds between the baseline period, which was defined as a time window 300–100 ms before the R-peak onset, and a time window 200–600 ms after the R-peak onset, which is the time window in which the effects of HERs were reported in most previous HER studies , to deteSCR_002454) analysis, which is a statistical method that controls multiple comparisons at the network level. This analysis enabled us to identify a network composed of significantly increased wPLI-Ds between cortical regions in the induced period compared to the baseline period at the group level. First, baseline and induced wPLI-D matrices were computed by averaging wPLI-Ds from each time window for every subject, which resulted in one baseline wPLI-D matrix and one induced wPLI-D matrix for each subject. Second, multiple paired t tests comparing wPLI-Ds from the induced period and the baseline wPLI-D were performed for every pair of cortical regions, which resulted in one matrix of t values from these paired t tests. Then, a threshold t value of 2.51 was applied to the matrix of t values, and a t value < 2.51 was therefore set to 0. The network statistic was computed by adding the t values of all the connected components in the thresholded matrix (a connected component refers to any two nodes within a component are connected by a path of edges). Next, a null distribution of the network statistic was created from 5000 permutations by randomly permuting an element of the induced wPLI-D matrices and the baseline wPLI-D matrices within each subject. Finally, network-level familywise-error (FWE)-corrected p values of the network were obtained using the original network statistic and null distribution. Next, we constructed a heartbeat-induced synchronization (HIS) matrix whose elements corresponded to the increase in the wPLI-D in the induced period compared to that in the baseline period, and each element was significant in the NBS results. Therefore, the HIS matrix was composed of elements with significantly increased wPLI-Ds in the group-level NBS analysis and represents the structure of the HIN.We then performed a group-level NBS  compared to that in the baseline period to test this hypothesis. We calculated the wPLI-Ds between ECG signals and 195 cortical regions in the theta band for every subject, which resulted in two 195 by one vector of ECG-brain region wPLI-Ds from the baseline and induced periods for every subject. Then, we performed 195 group-level paired We next tested whether phase synchronization in the HIN reflected artificial synchronization due to evoked responses within distributed regions, which are phase-locked to the heartbeat , using tAfter confirming that the HIN does not reflect artificial synchronization induced by either CFAs or evoked responses, we identified the following characteristics of the HIN. First, we identified the hubs of the HIN, which play an important role in connecting regions within the HIN. Then, we identified whether the HIN is one homogeneous network or can be divided into subnetworks called modules, indicating that the HIN is modularized. Additionally, we also investigated how synchronization within the HIN changes over time by summing the wPLI-D of every HIN edge at every time point from –300 to 600 ms.SCR_004841).One of the important features of a network is the hub of the network, which is defined as a node that plays an important role within a network, such as connecting nodes . To idenBecause we expected that the HIN would have a modular structure, we applied a community detection algorithm to the HIS matrix to determine how the HIN is partitioned into different subnetworks. However, to identify whether the HIN is modularized, one should examine the extent of modularity compared to random networks. Therefore, we compared the “modularity index” of the HIN with 100 random networks . OptimalIn this equation, tition S  using thtition S . The contition S , is brieus study . Fourth,z score  > 0.3  > 0.3 .5C2 pairs of modules was computed by the sum of the edge weights between each module, which resulted in a 5 × 5 between-module synchronization matrix. By applying graph-theoretical analysis to this between-module synchronization matrix, we identified which module plays a central role within the HIN using module-level nodal strength and betweenness centrality.We next investigated synchronization patterns between modules. Between-module synchronization of To identify relationships between the HIN and participants’ emotional states, we used the emotional statuses included in the HCP data. The HCP data included a measure for six negative emotional affective states, including anger-affect, anger-hostility, anger-aggression, fear-affect, fear-somatic, and sadness, and positive affect surveys were retrieved from the NIH Toolbox . To redup < 0.001), revealing the existence of the HIN. The density of the network was 9.2%, indicating that among the total of 195C2 pairs of regions, 9.2% of the region pairs showed significantly increased phase synchronization after the heartbeat. Additionally, no HIN formed in the α-frequency or β-frequency bands.The NBS analysis showed a network displaying a significant increase in phase synchronization in the induced period compared to that in the baseline period  comparing responses between the induced period (200–600 ms after the R-peak) and the baseline period did not reveal a significant increase or decrease in wPLI-Ds between ECG signals and cortical regions in the theta band -corrected] with t(86) = 2.97 in the “right postcentral gyrus A2”; The paired 10.1523/ENEURO.0200-19.2019.f1-1Extended Data Figure 1-1Figure 1-1, PDF file.wPLI-D in the induced and baseline period for real and surrogate data. This figure shows the wPLI-D in the induced (blue) and baseline (orange) period separately. R represents the wPLI-D of the real data and others represents wPLI-D of 20 surrogate data. One can notice that the synchronization within the HIN is much stronger in the real data for both induced and baseline period. Note that, an increase of induced synchronization was also strongest in the real data compared to the surrogate data B. Downlop < 0.05; We compared the synchronization within the HIN in 20 surrogate datasets and in the real data. The results showed that the HIS within the HIN in the real data were significantly stronger than that in all 20 surrogate datasets (Monte Carlo t(84) = 5.22, p < 0.001 in a paired t test comparing induced synchronization between the right and left hemispheres; Within the HIN, left inferior temporal regions including the temporal pole and parahippocampal gyrus had high strength and betweenness centrality . Figure p < 0.01), indicating that the HIN is modularized rather than one homogeneous network. Importantly, among the five modules, the synchronization within module 1 was the strongest  = 8.95, p = 0.004, R2 = 0.10, β of the within-module synchronization of module 1 = 0.57, t = 3.00, p = 0.004; p < 0.05).The first PC explained 47% of the variance of the emotion survey data and had positive loading on the positive affect score and negative loading on the other negative affect scores. Therefore, we surmised that this PC reflected the moods of the participants. Stepwise linear regression analysis resulted in a model that only contained the within-module synchronization of module 1 in the HIN, which explains ∼10% of the variance of the mood data  than with other brain regions , and theA limitation of our study is that although interoceptive processing typically includes subcortical regions, such as the amygdala , we usedIn conclusion, we first showed the existence of HIN interactions with hubs in inferior temporal regions. The HIN was modularized and contained five modules, with module 1 as the center of module interactions. The synchronization within module 1 of the HIN had a positive association with the mood scores of the participants. Considering recent theories on abnormal interoceptive processing in mood disorder patients , investi


A 41-year-old female presented with complaints of right arm claudication, weakness, and pain associated with serous drainage from a previous incision site to the right anterior chest. At age 16, this patient was involved in a motor vehicle accident, which resulted in a right innominate artery and brachiocephalic vein avulsion. The two vessels were immediately ligated and oversewn. The perfusion to her right arm was supplied by cerebral collateral circulation down the right vertebral to the right subclavian artery. It is quite uncommon to encounter difficult cases of innominate artery repair via an aorto-subclavian graft. We are reporting a complicated case of innominate artery avulsion (secondary to motor vehicle accident) and repair via an aortic to subclavian arterial Gore-Tex  bypass graft with subsequent occlusion and re-grafting, after 25 years of initial grafting.A 41-year-old female presented with complaints of right arm claudication, weakness, and pain associated with serous drainage from a previous incision site to the right anterior chest. At age 16, this patient was involved in a motor vehicle accident, which resulted in a right innominate artery and brachiocephalic vein avulsion. The two vessels were immediately ligated and oversewn. The perfusion to her right arm was supplied by cerebral collateral circulation down the right vertebral to the right subclavian artery.The patient was stable until the age of 35 years when symptoms of ischemia set in and she developed right arm pain. This led to her readjusting her right arm frequently and an inability to sleep secondary to ischemic pain. It required surgical intervention done at the University of Chicago with the placement of a right aortic to axillary bypass graft.Six years passed before the patient began developing her recent symptoms of right arm claudication and weakness. Upon the patient’s recent presentation, a computed tomography (CT) angiogram of this graft showed occlusion of the aortic-axillary bypass graft due to thrombosis. Interventional radiology decided to thrombolyse the clot and re-canalize the graft. However, the patient developed an infection of the graft with serous discharge, bacteremia, and persistent symptoms of right upper extremity arterial occlusion Figure .The persistence of these symptoms and the infection of the graft led to the decision to perform a two-staged procedure: 1) removal of the infected/occluded aorto-subclavian bypass graft with the exception of the aortic stump; this was performed without sternotomy. The patient was treated with antibiotics and once the infection was cleared, the next stage involved redo sternotomy with 2) removal of the old aorto-axillary graft stump and creation of a new aorta to right subclavian artery stump with a direct path using an open harvest of a piece of the left greater saphenous vein.ProcedureThe previous incision over the right anterior chest was reopened. The sternum was not opened at this time. The graft was controlled and the subclavian artery was exposed. A large number of adhesions was noted, requiring extreme care to liberate and identify the right subclavian artery and vein. The patient was heparinized using 8000 units. The left greater saphenous vein was harvested open. Once the subclavian artery was isolated, all graft material was removed up to the incorporated graft material just medial to the aorta. A remnant of the graft was left attached to the lateral position of the aorta, as it appeared to be incorporated into the surrounding tissue. The laterally surrounding tissue was noted to have induration but no gross pus present. However, once the lateral graft was incised, pus spilled out. All material was sent for culture. Proximal and distal control of the subclavian artery was obtained and the defect of the arterial wall was closed using a saphenous venous patch and 6-0 Prolene sutures. The distal flow was noted following repair and clamp removal. Hemostasis was obtained. Due to active infection, it was deemed unsafe to remove the stump and re-implant another graft. The incision was left open and a wound vacuum was placed medially. The incision over the axillary artery was loosely closed and a drain was placed.Post-operative care and follow-up procedureThe drain was removed at three days post-procedure. Home wound vacuum and intravenous antibiotics were initially started with vancomycin for two weeks followed by Bactrim DS twice daily for two weeks. The infected graft and pus were tested for aerobic, anaerobic, and fungal cultures. The patient did not show signs of infection after the antibiotics Table .On follow-up after the first staged procedure, the patient denied fever, chills, weight loss, or any other constitutional symptoms. There was excellent granulation tissue, and the incision was approximately 50% closed with no surrounding erythema, odor, drainage, or fluctuance. The incision subsequently healed and the wound vacuum was removed.Two months after the initial procedure to remove the graft, the patient underwent redo sternotomy to remove the remainder of the old graft. During this same procedure, a right subclavian to aorta graft was placed using an 8 mm Gore-Tex graft . This procedure resulted in total relief of the symptoms. The patient continues to do well.The first successful repair of an innominate artery injury after blunt trauma was reported by Binet in 1962 . Motor vVarious surgical options are available to deal with such an innominate artery blunt trauma. If the laceration is close to the origin, bypass grafting can be done from the ascending aorta to the distal part of the axillary artery. If the injury is to the mid-region, interposition grafting can be done. If there is no extensive tissue loss, a primary repair can be done . A cardiIn our patient, aorto-subclavian bypass grafting was initially performed for innominate artery avulsion after a motor vehicle accident occurring 26 years prior in 1992. The initial graft was patent for 20 years prior to occlusion (2011). The patient developed occlusive symptoms rather early, for which interventional radiology attempted clot thrombolysis, but the procedure was unsuccessful. This resulted in persistent occlusion symptoms and was later complicated with a chronic serous infection of the graft. Removal of the graft, repair of the subclavian artery, and re-grafting was performed, which led to the resolution of symptoms.Graft stenosis can be asymptomatic or have symptoms similar to those in patients with subclavian or innominate artery stenosis due to atherosclerosis. These patients may present with symptoms of vertebrobasilar ischemia, including episodes of dizziness, diplopia, ataxia, vertigo, limb claudication, paresthesia, and steal syndrome. Physical examination may reveal diminished pulse and decreased blood pressure (>20 mmHg reduction as compared to the opposing extremity) in the affected limb.In this case, the graft was occluded and infected, which required the removal of the infected graft and subsequent re-grafting. Re-grafting posed more challenges due to multiple adhesions from previous procedures. Exposing the subclavian artery is associated with risks of possible complications, such as injuries to the phrenic nerve, the recurrent laryngeal nerve, the vagus nerve, the cervical sympathetic chain, and the thoracic duct, which are located in the vicinity of the subclavian artery.An aorto-subclavian bypass graft is an uncommon, but effective, way of repairing an innominate artery injury subsequent to blunt trauma resulting in avulsion. Long-term complications may include graft occlusion requiring re-grafting. All measures should be taken to avoid infection postoperatively.


Journal of Cellular and Molecular Medicine on 14 September 2016 in Wiley Online Library (wileyonlinelibrary.com), and in Volume 19, pp. 2136‐2142, has been retracted by agreement between the authors, the journal Editor in Chief, Stefan Constantinescu, and John Wiley & Sons Ltd. The retraction has been agreed due to unattributed overlap of the language used in the “Materials and method” and “Discussion” sections of this study and the following article published in Lung Cancer: “CYP2E1 Rsa I/Pst I polymorphism is associated with lung cancer risk among Asians” by Ping Zhan, Jing Wang, Yu Zhang, Li‐Xin Qiu, Su‐feng Zhao, Qian Qian, Shu‐Zhen Wei, Li‐Ke Yu and Yong Song, Volume 69, 2010, pages 19‐25.The above article, published in the REFERENCEhttps://doi.org/10.1111/jcmm.12579Shen Z‐T, Wu X‐H, Li B, Shen J‐S, Wang Z, Li J, Zhu X‐X. CYP2E1 Rsa Ι/Pst Ι polymorphism and lung cancer susceptibility: a meta‐analysis involving 10,947 subjects. J Cell Mol Med. 2015;19:2136‐2142.  Journal of Cellular and Molecular Medicine on 14 September 2016 in Wiley Online Library (wileyonlinelibrary.com), and in Volume 19, pp. 2136‐2142, has been retracted by agreement between the authors, the journal Editor in Chief, Stefan Constantinescu, and John Wiley & Sons Ltd. The retraction has been agreed due to unattributed overlap of the language used in the “Materials and method” and “Discussion” sections of this study and the following article published in Lung Cancer: “CYP2E1 Rsa I/Pst I polymorphism is associated with lung cancer risk among Asians” by Ping Zhan, Jing Wang, Yu Zhang, Li‐Xin Qiu, Su‐feng Zhao, Qian Qian, Shu‐Zhen Wei, Li‐Ke Yu and Yong Song, Volume 69, 2010, pages 19‐25.The above article, published in the REFERENCEhttps://doi.org/10.1111/jcmm.12579Shen Z‐T, Wu X‐H, Li B, Shen J‐S, Wang Z, Li J, Zhu X‐X. CYP2E1 Rsa Ι/Pst Ι polymorphism and lung cancer susceptibility: a meta‐analysis involving 10,947 subjects. J Cell Mol Med. 2015;19:2136‐2142.


Incorrect anterior-posterior orientation of the Descemet endothelial complex (DEC) is one of the causes of failure of Descemet membrane endothelial keratoplasty (DMEK). We evaluated a new marking technique to avoid such a misorientation.  A new marking technique of the DEC was evaluated in patients requiring primary DMEK. A Braille-“R”-letter was applied dot by dot onto the stromal surface of the DEC after lifting it by injecting an air-bubble into the interface between the endothelial surface of the partially stripped graft. The positioning of the graft was intraoperatively controlled by an orientation of the Braille-“R”-letter. Laboratory tests were conducted to test the impact of the marking technique on endothelial cell count. 2, respectively. Only one patient (3%) needed re-bubbling.  We included prospectively 37 eyes of 30 patients. Four eyes were phakic and 33 pseudophakic. Five grafts (14%) presented an undifferentiated rolling tendency in the anterior chamber, and evaluation of their positioning was possible due to orientation of the mark alone. In case of an upside-down orientation, grafts were flipped immediately. A correct orientation of the graft was achieved in all cases at the end of the surgery. The endothelial cell loss due to the mark was estimated to be less than 0.3%. At 3- and 6-month follow-ups, the mean best-corrected visual acuity was 0.21 ± 0.15 and 0.15 ± 0.11 logMAR, respectively, and endothelial cell density was 1661 ± 349 and 1618 ± 396 cells/mm To rely on the natural rolling tendency of the graft alone does not assure its correct positioning in all cases. Creation of the mark with 4 dots punctuated on the air-lifted stromal side of the DEC is a simple and endothelial cell saving marking method to ensure correct orientation of the graft during DMEK. In many countries, Descemet membrane endothelial keratoplasty (DMEK) became the standard treatment of hereditary or acquired corneal endothelial pathologies in recent years . Quick vIncorrect anterior-posterior orientation of the Descemet endothelial complex (DEC) is one of the reasons for graft detachment and failure in DMEK . Many suWe developed an alternative cost and endothelial cell saving marking technique in order to avoid the upside-down positioning of the lamella and to preserve the maximum of endothelial cells and investigated the results of this technique in a prospective study.This observational clinical study was carried out in accordance with the Declaration of Helsinki. Ethics approval was obtained from the Ethics Committee of Rhineland-Palatinate, Germany. All patients were evaluated at the Department of Ophthalmology University Medical Center of the Johannes Gutenberg University Mainz. Written informed consent was obtained from all participants. Donor tissue has been obtained from the Eye Bank of Rhineland-Palatinate, Mainz, Germany, and its use has been consented for transplantation and research. At the eye bank, the tissue has been stored at 34°C in medium 1  and at least 24 hours before use in medium 2 . Medium 2 is supplemented with 60 g Dextran 500 per 1000 ml as opposed to medium 1. Both media 1 and 2 were supplemented with gamma-irradiated fetal calf serum 10% . The inclusion criteria were male or female; >18 years of age; phakic or pseudophakic; and diagnosis of Fuchs endothelial dystrophy or corneal endothelial failure. The exclusion criteria were previous intraocular surgery except for uncomplicated cataract surgery.All graft preparations and DMEK surgeries were performed by the same experienced surgeon (AG). The graft preparation was performed immediately before the surgery using the stripping technique. A cornea was centered on the punch base using suction . A superficial cut of the DEC was performed using a 9.5 mm donor trephine. The DEC was detached circularly from the peripheral corneal tissue using a Sinskey hook  and peeled longitudinally in order to avoid stress lines towards the center of the cornea using straight pointed tweezers (Geuder)   on the dried area . AfterwaLaboratory tests were conducted to test the impact of the marking technique on endothelial cell count.Graft preparation including the marking technique was conducted on sclerocorneal buttons of nonoptical quality obtained from the Eye Bank of Rhineland-Palatinate. After preparation, the graft was placed back into medium 2 and stored at 34°C for 24 hours.μm (0.637 mm2) were made in the area where the mark was set and in an area of corresponding localization of the opposite side of the same graft. Next, the endothelial cell number was counted and the number of endothelial cells per mm2 calculated.On the following day, the graft was transferred into a Petri dish and stained as described previously . After aThe DMEK procedure was performed under topical anesthesia (Tetracain 1% eye drops) in 35 cases and under general anesthesia in 2 cases. The descemetorhexis was performed within an area of 9.0–10.0 mm using an irrigation Descemet hook  and an irrigation Descemet scraper . The single-use DMEK-cartridge  was used to insert the DEC into the anterior chamber. The graft was unfolded by gentle tapping of the cornea. The positioning of the graft was checked by the orientation of the Braille-“R”-letter, which was visible in all cases. An air/10% sulfur hexafluoride (SF6) bubble was injected underneath the graft as soon as the proper orientation of the Braille-“R”-letter (to the right looking from the limbus towards the center of the cornea) was achieved. In case of an upside-down positioning (orientation of the Braille-“R”-letter to the left looking from the limbus towards the center of the cornea), the graft was flipped over with a flush of balanced salt solution and the position was checked again Figures .The results of the marking technique were evaluated in a prospective clinical trial. The follow-up examinations were performed three months and six months postoperatively. The outcome measures were best-corrected visual acuity (BCVA) in logMAR, endothelial cell density (ECD), and re-bubbling rate. Furthermore, rolling tendencies of the graft in the anterior chamber and preparation- and surgery-time durations were documented.2 was reduced by 22% compared to the corresponding areas of the opposite graft side  and posterior polymorphous endothelial dystrophy (n = 1). Four eyes were phakic and 33 pseudophakic.We included 37 eyes of 30 consecutive patients  requiring primary DMEK due to Fuchs endothelial dystrophy (The mean duration of the graft preparation was 20.9 ± 8.8 minutes (range 8–53 minutes). The marking procedure took about 2-3 minutes. The mean duration of the DMEK procedure was 24.0 ± 8.7 minutes (range 12–45 minutes). The DMEK procedure was uneventful in all cases. Intraoperatively, five grafts (14%) presented an undifferentiated rolling tendency in the anterior chamber meaning the grafts rolled with the direction of the fluid flush, and evaluation of their positioning was possible due to orientation of the mark alone. The Braille-“R”-letter mark was visible in all cases at the end of surgery. The correct orientation of the graft was achieved in all cases at the end of surgery. In the following 6 months, only one patient (3%) required re-bubbling.The baseline BCVA was 0.44 ± 0.23 logMAR. At the 3- and 6-month follow-up visits, the mean BCVA was 0.21 ± 0.15 and 0.15 ± 0.11 logMAR, respectively.2. At the 3- and 6-month follow-up, ECD was 1661 ± 349 cells/mm2 and 1618 ± 396 cells/mm2, respectively. The endothelial cell loss was 33.6% at the 3-month follow-up and 35.3% at the 6-month follow-up. Moreover, no graft decompensation and no immunological rejection were observed during follow-up.The mean ECD of the donor corneas was 2500 ± 179 cells/mmThe new air-bubble-Braille-“R”-letter marking technique is an effective supplementation of the stripping technique used for the DEC preparation. It is a simple and cost-saving procedure. In contrast to other methods determining the donor graft orientation, it does not require any specific instruments like dermatological biopsy punch or intraoperative OCT –12.Many surgeons rely on the natural endothelium-out orientation of the graft roll in the anterior chamber alone. However, the orientation of the DEC may vary depending on the donor's age, endothelial cell density, or its elastic properties . In the 2, and the diameter of the graft is 8.0–8.25 mm.The need for establishing marking techniques enabling reliable recognition of the endothelial surface of the DEC has been addressed since the beginning of the DMEK era. Matsuzawa et al. proposed two pairs of asymmetrical semicircular marks of 1.0 and 1.5 mm in diameter placed on the edge of the 7.5 mm donor graft using dermatological biopsy punches . BachmanAs the endothelial cell loss after DMEK ranges from 33.9% to 56% in the first 6 months anyway, some surgeons neglect the ECD loss associated with the punch technique , 19. HowThe marking techniques associated with S or F letters stamped through the stromal hole require a dermatological biopsy punch , 10. In In case the graft is asymmetrically trephined and the periphery cut of, the orientation of the Braille-“R”-letter can often still be identified . We expeThe stripping preparation technique with a superficial trephine cut was shown to be accompanied by the lowest cell death rate in comparison to other stripping and bubble techniques. However, it is the most time-consuming and expensive preparation method . The marIn summary, the air-bubble-Braille-“R”-letter marking technique is a simple and in some cases cost-saving supplementation of the DEC preparation to ensure correct orientation of the DEC graft intraoperatively during the DMEK procedure. The estimated loss of 0.3% of the endothelial cells is low in comparison to other marking techniques.


Cardiovascular disease (CVD) has emerged as one of the leading causes of death worldwide. Elevated blood cholesterol and low‐density lipoprotein levels are crucial risk factors that contribute to the development of CVD and other metabolic diseases. Dietary fat is believed to be the key factor in modulating circulating cholesterol levels. Thus, reducing dietary intake of fat appears to be an effective strategy to reduce the risk of heart disease. Also, excessive intake of fat and high‐calorie foods is also related to the development of obesity, which contributes to the development of CVD. Therefore, the consumption of low‐fat low‐calorie foods is part of a healthier dietary pattern. However, simply removing fat from foods may lead to compromised overall quality and reduced acceptance of the food products. Thus, fat replacers have emerged as ideal alternatives to dietary fat, which can not only reduce the overall fat and calorie content of the foods but also mimic the physiochemical properties of dietary fat. Starch‐based fat replacers are one kind of fat mimetic that can be produced either chemically as modified starch or enzymatically as maltodextrins. Both modified starch and maltodextrins have been demonstrated to have a promising ability to improve the overall quality of reduced‐fat foods. Modified starch granules act directly as fat globules in modulating the structure and sensory characteristics of the foods, whereas maltodextrins can form thermoreversible gels. Both modified starch granules and maltodextrins can create a fat‐like mouthfeel and therefore are potential fat replacers. This review article aims to discuss the following topics: (a) the effect of carbohydrates and fat on human cardiovascular health and other disease risks, (b) the functionality of starch‐based fat replacers in foods, (c) the applications of starch‐based fat replacers in various foods, and (d) the current and future market value of starch‐based fat replacers. This review article discusses the following objectives: (a) effect of carbohydrate and fat on human cardiovascular health and other disease risks, (b) functionality of starch‐based fat replacers in foods, (c) applications of starch‐based fat replacers in various foods, and (d) current and future market value of starch‐based fat replacers. According to Health Canada, approximately 2.4 million Canadians aged 20 years and older have heart disease  to compare the effects of carbohydrates and fat on human cardiovascular health and other disease risks, (b) to discuss the functionality of starch‐based fat replacers in foods, (c) to discuss the applications of starch‐based fat replacers in various foods, and (d) to briefly review the current and future market value of starch‐based fat replacers.2The link between dietary fatty acids, circulating cholesterol levels, and heart disease has been extensively studied in recent decades. Modulating dietary fat intake appears to be one of the most effective approaches to control the prevalence of heart disease. A typical type of dietary fat, trans fat, was found to significantly increase TC/LDL‐C levels, contributing to the pathogenesis of atherosclerosis  maltodextrins are ideal fat replacers (Chavan et al., 4The first commercial maltodextrin‐based fat replacer called N‐Oil was invented by the National Starch & Corporation in 1984, and later, a modified starch‐based fat replacer was developed by American Maize Products Company in the late 1980s Jones, . Using sIn addition, starch‐based fat replacers have been largely implicated in cheese and yogurt processing. In 1993, Mcmahoh and colleagues compared the effects of different types of fat replacers, including Stella made from modified corn starch and protein‐based fat replacers, on low‐fat Mozzarella properties (McMahon, Alleyne, Fife, & Oberg, However, it should be noted that the abovementioned food products are classified either as concentrated or as semisolid food systems. In food systems, fat globules act as barriers between protein particles and prevent cross‐link interactions between proteins, producing a softer and tenderer texture (Peng & Yao, Maltodextrins are starch derivatives that are hydrolyzed from starch and lose the physical properties of starch granules. However, maltodextrin has a strong water holding capacity and is able to form thermoreversible gels with water, creating a similar mouthfeel as fats do. Since the functionalities of maltodextrins as fat replacers are slightly different than those of native or modified starch granules, which require water, solid food systems such as breads and muffins are not ideal applications. Nevertheless, maltodextrins can be used in several food products as fat replacers and can reduce the overall energy content of the products. For instance, confectionery filling contains a substantial amount of dietary fat and is usually considered a high‐fat food. Hadnadev et al.  reported5Several authorities, such as the AHA and Health Canada, recommend limiting overall fat and energy intake, and a study conducted in the UK reported that fat intake has decreased significantly over the past 30 years (Pot et al., According to a recent press release article, the market for fat replacers is anticipated to reach an annual growth rate of 6.5% by 2023, whereas the growth rate may reach 7.2% in societies in Asia–Pacific (Research and, 6Reducing total fat and energy intake by consuming low‐fat low‐calorie foods has emerged as one of the strategies to prevent CVD and obesity. While several health organizations have recommended that dietary fat intake be strictly controlled, food industries have also been extensively developing and utilizing fat replacers in low‐fat foods. Starch‐based fat replacers, including native starch, modified starch, and maltodextrins, play crucial roles in the low‐fat food industry. These fat replacers mimic the physiochemical and sensory properties as fat globules. Starch‐based fat replacers have been widely used in various food products, such as dairy products, baked goods, salad dressings, and mayonnaise. Through decades of research, starch‐based fat replacers have exhibited promising efficacy in substituting fats without safety issues and are generally considered safe. The market size of fat replacers is also continually growing. However, there is very limited evidence of how starch‐based fat replacers can reduce the risk of CVD and obesity compared to fat in clinical studies. The lack of human intervention trials leads to a poor understanding of other possible side effects that starch‐based fat replacers have while reducing calorie and fat intake in humans, and if there are other side effects, the extent of these effects remains unknown. Therefore, future research should also focus on the physiological efficacy and safety of fat replacers based on human intervention studies while continuing to explore novel fat replacers.All authors declare no conflict of interest.The present work does not involve any animal and human subjects.


Despite accumulating evidence on the role of glial cells and their associated chemicals in mechanisms of pain, few studies have addressed the potential role of chemokines in the descending facilitation of chronic pain. We aimed to study the hypothesis that CXCL1/CXCR2 axis in the periaqueductal gray (PAG), a co-restructure of the descending nociceptive system, is involved in descending pain facilitation.Nfkb, Cxcl1, and Cxcr2 and their protein expression in the ventrolateral PAG (vlPAG). Immunohistochemical co-staining with NeuN, GFAP, and CD11 were used to examine the cellular location of pNFκB, CXCL1, and CXCR2. The effects of NFκB and CXCR2 antagonists and CXCL1 neutralizing antibody on pain hypersensitivity were evaluated by behavioral testing.Intramedullary injection of Walker 256 mammary gland carcinoma cells of adult female Sprague Dawley rats was used to establish a bone cancer pain (BCP) model. RT-PCR, Western blot, and immunohistochemistry were performed to detect pBCP induced cortical bone damage and persistent mechanical allodynia and increased the expression of pNFκB, CXCL1, and CXCR2 in vlPAG. The induced phosphorylation of NFκB was co-localized with GFAP and NeuN, but not with CD11. Micro-injection of BAY11-7082 attenuated BCP and reduced CXCL1 increase in the spinal cord. The expression level of CXCL1 in vlPAG showed co-localization with GFAP, but not with CD11 and NeuN. Micro-administration of CXCL1 neutralizing antibody from 6 to 9 days after inoculation attenuated mechanical allodynia. Furthermore, vlPAG application of CXCL1 elicited pain hypersensitivity in normal rats. Interestingly, CXCR2 was upregulated in vlPAG neurons (not with CD11 and GFAP) after BCP. CXCR2 antagonist SB225002 completely blocked the CXCL1-induced mechanical allodynia and attenuated BCP-induced pain hypersensitivity.The NFκB-dependent CXCL1-CXCR2 signaling cascade played a role in glial-neuron interactions and in descending facilitation of BCP.The online version of this article (10.1186/s12974-018-1391-2) contains supplementary material, which is available to authorized users. Bone cancer pain (BCP) is a severe and chronic pain that has a negative impact on the quality of life of cancer patients. Advancements in the mechanisms and pharmacotherapy of BCP have achieved limited success, and commonly-used analgesics have resulted in little or no response. Therefore, novel and more efficacious therapies are urgently needed for improving the patients’ quality of life.Recent studies have demonstrated that hyperalgesia in animal models with persistent pain is closely associated with the activation of “top-down” modulatory circuits involving descending facilitation or descending inhibition , 2. The Accumulating evidence demonstrated an important role of neuroimmune interactions in chronic pain . Glial hNuclear factor kappa B (NFκB) is a transcription factor that transduces extracellular signals to affect gene expression . NFκB isHence, the present study was aimed to investigate the hypothesis that CXCL1/CXCR2 signaling in vlPAG is involved in descending the pain facilitation. We examined the expression and distribution of NFκB and CXCL1 in vlPAG after inoculation of Walker 256 cells. We also evaluated the role of NFκB in CXCL1 production at vlPAG and pain hypersensitivity after BCP. We further investigated the expression and distribution of CXCR2 in vlPAG and the antinociceptive effect of CXCR2 antagonist. The release of CXCL1 from astrocytes mediated by NFκB and the subsequent activation of its CXCR2 receptor on neurons may present astroglial-neuronal interaction that is involved in the descending facilitation of BCP-like behavior in rats.Female Sprague Dawley rats (180–200 g) were housed in plastic cages and maintained on a 12-h light/dark cycle at 22–25 °C with food and water available. All efforts were made to minimize the animal suffering and the number of animals used. All experimental procedures received prior approval from the Animal Use and Care Committee for Research and Education of the Jiaxing University  and the ethical guidelines to investigate experimental pain in conscious animals .n = 8). PWT of the ipsilateral hind paw was observed 1 day before and 6, 12, and 18 days after surgery. Additional 8 rats were then randomly divided into two groups: sham and 12 days after surgery (n = 4). These rats were then radiographed . Sixteen rats were randomly divided into BCP and sham  groups (n = 8). After each time point, rats were anesthetized and then perfused with fixative or decapitated for immunohistochemistry (n = 4) or Western blot/RT-PCR analysis (n = 4). Tissue sections from additional four rats of day-12 surgery animals were labeled with a fluorescent marker for CXCL1 and co-labeled with CD11 , GFAP (astrocytic marker), or NeuN  . Fifty-six rats were randomly divided into seven groups: naïve, BCP  group, and sham  groups (n = 8). After each time point, rats were anesthetized and then perfused with fixative or decapitated for immunohistochemistry (n = 4) or Western blot/RT-PCR analysis (n = 4). Tissue sections from additional four rats of day-12 surgery animals were labeled with a fluorescent marker for p-NFκB and co-labeled with CD11, GFAP, or NeuN . Fifty-six rats were randomly divided into seven groups: naïve, BCP  group, sham  groups (n = 8), BCP + serum group (n = 8), BCP + CXCL1 neutralizing antibody 8 μg group (n = 8), BCP + CXCL1 neutralizing antibody 4 μg group (n = 8) groups, sham + CXCL1 neutralizing antibody 8 μg group (n = 8), sham + veh (PBS) group (n = 8), BCP + veh group (n = 8), BCP + BAY 10 μg group (n = 8), BCP + BAY 1 μg group (n = 8) groups, and sham + BAY 10 μg group (n = 8). The rats continuously received a vlPAG microinjection of above drugs once daily for four consecutive days (from day 6 to 9 after BCP), and we then detected the analgesic effect of CXCL1 and pNFκB inhibition 1 h later after micro-injection on postoperative days (POD) 6, 7, 8, 9, 10, 11, and 12, respectively . Eighty rats were randomly divided into ten groups: sham + serum group (n = 8), BCP + veh (n = 8), and BCP + LAA (n = 8). After repeated drug administration from days 6 to 9 after BCP (once daily for four consecutive days), rats were decapitated for CXCL1 Western blot analysis of the vlPAG (n = 4) and perfused with fixative for immunohistochemistry for GFAP (n = 4). Additional 12 rats were then randomly divided into three groups: sham + veh (PBS) group (n = 4), BCP + veh group (n = 4), and BCP + BAY 10 μg group (n = 4). We blocked the NFκB pathway with BAY (once daily from day 6 to 9 after BCP) and then detected CXCL1 protein level after the last injection . Twenty-four rats were randomly divided into three groups: sham + veh  (n = 8), CXCL1 100 ng (n = 8), and CXCL1 10 ng (n = 8). The naive rats received a vlPAG microinjection of above drugs, and we then detected the analgesic effect of drugs 0.5, 1, 2, 3, 4 and 5 h later after micro-injection respectively. Additional 40 rats were then randomly divided into five groups: PBS + SB (SB225002) 20 μg group (n = 8), 5%DMSO + CXCL1 100 ng group (n = 8), SB 20 μg + CXCL1 100 ng group (n = 8), SB 5 μg + CXCL1 100 ng group, and PBS + 5%DMSO group (n = 8). The naive rats received a vlPAG microinjection of above drugs, and we then detected the analgesic effect of drugs 0.5, 1, 2, 3, 4 and 5 h later after micro-injection respectively . Twenty-four rats were randomly divided into three groups: PBS (n = 8). After each time point, rats were anesthetized and then perfused with fixative or decapitated for immunohistochemistry (n = 4) or Western blot/RT-PCR analysis (n = 4). Another 40 rats were randomly divided into five groups: BCP + veh (5%DMSO) group (n = 8), BCP + SB 20 μg group (n = 8), BCP + SB 5 μg group (n = 8), sham + veh group (n = 8), and sham + SB 20 μg group (n = 8). The rats continuously received a vlPAG microinjection of above drugs once daily for four consecutive days (from day 6 to 9 after BCP), and we then detected the analgesic effect of CXCR2 inhibition 1 h later after micro-injection on postoperative days (POD) 6, 7, 8, 9, 10, 11, and 12, respectively. Tissue sections from additional four rats of day-12 surgery animals were labeled with a fluorescent marker for CXCR2 and co-labeled with CD11, GFAP, or NeuN . Fifty-six rats were randomly divided into seven groups: naïve, BCP  group, and sham  groups :Rats were anesthetized with isoflurane  in a gas mixture of 25% O2/75% nitrogen and then placed in a stereotaxic instrument . A 7-mm stainless steel guide cannula (23-gauge) was implanted into the unilateral vlPAG  [The CXCL1 neutralizing antibody and control serum were from Sigma  and diluted in sterile artificial cerebrospinal fluid (ACSF). The potent and selective antagonist of CXCR2 SB225002 was from Sigma  and dissolved in 5% dimethylsulfoxide (DMSO). BAY11-7082, an NFκB inhibitor, was from Abcam  and dissolved in phosphate buffer saline (PBS). Exogenous CXCL1 was purchased from R&D Systems  and dissolved in phosphate buffer saline (PBS). The astrocytic cytotoxin, L-α- aminoadipic acid (LAA), was from Sigma  and dissolved in 0.9% saline. The injection cannula remained in place for an additional 2 min to prevent the backflow of the drug after completion of drug infusion. The microinjection of the drug was administered at 8 a.m. daily, which was 30 min before the behavioral test. The doses of CXCL1 neutralizing antibody , SB225002 , BAY11-7082 , CXCL1 , and LAA (100 nmol) used in this experiment were based on our preliminary results and previous use in relevant studies , 17, 18.5 cells/ml) or heat-killed cells (sham group) were cautiously injected into the bone medullary canal. The cells were allowed to fill the bone cavity for 2 min. After that, the syringe was withdrawn and the injection site was immediately closed with bone wax.The BCP model has been established as previously described . In brieTo confirm cancer development in the tibia, rats were radiographed after 12 following implantation. After the rats were anesthetized with pentobarbital sodium, the right hind limbs were placed on X-ray film  and exposed to X-ray for 1/20 s at 40 kVp. The images were taken from sham rats and BCP rats.Mechanical allodynia of the right hind paw was measured with a von Frey monofilaments  as described in our previous methods . AnimalsImmunofluorescence staining and double/ triple immunostaining were performed as previously described . Briefly− ΔΔCT).Total RNA was extracted from the vlPAG with Trizol reagent . One microgram of total RNA was converted into cDNA using PrimeScriptRT reagent kit . The cDNA was amplified using the following primers: CXCL1 forward,5′-GGCAGGGATTCACTTCAAGA-3′;CXCL1 reverse, 5′-ATCTTGAGCTCGGCAGTGTT -3′;pNF-кBforward,5′-TCTGCTTCCAGGTGACAGTG-3′;pNF-кB reverse, 5′-ATCTTGAGCTCGGCAGTGTT-3′;CXCR2 forward, 5′-CGTTCTGGTGACTTTGCTGA-3′;CXCR2 reverse,5′-ACAGAGCAGGTGCTTCGATT-3′; GAPDH forward, 5′-TCTCTGCTCCTCCCTGTTC-3′;GAPDH reverse, 5′-ACACCGACCTTCACCATCT-3′. The PCR reactions were performed in a real-time detection system  by SYBR PremixEx Taq™ II kit (Takara). The reaction conditions consisted ofan initial denaturation step at 95 °C for 10 min, followed by 45 cycles at 95 °C for 5 s, 58 °C for 30 s, and 72 °C for 30 s. The data were analyzed using Rotor-Gene 6000 series software and evaluated using the Comparative CT method , 1 mM EDTA, 1% NP-40, 1 mM PMSF, protease inhibitor cocktail , and phosphatase inhibitor cocktail . Protein concentrations were determined by the BCA Protein assay . Equal amounts of protein (30 μg) were loaded for each lane, separated by SDS-PAGE gel (10%), and transferred onto the nitrocellulose blots. After the transfer, the blots were first saturated by incubation in 10% skim milk for 1 h and then incubated overnight at 4 °C with antibody against CXCL1 , CXCR2 , and p-NFkB p65 (Ser536) antibody . For loading control, the blots were probed with GAPDH antibody . All the blots were further incubated overnight at 4 °C with primary antibody. Bound primary antibodies were detected following incubation at RT for 1 h with appropriate horseradish peroxidase-conjugated anti-rabbit or anti-mouse secondary antibody . Immunoreactive bands were detected by using enhanced chemiluminescence (Thermo Scientific) and exposed to X-ray films. GAPDH served as an internal control.F test to test equality of variances in t test and Levene’s test of equality of error variances for ANOVA. p < 0.05 was considered to be statistically significant.SPSS version 20.0 was used to determine statistical significance. Results were expressed as mean ± SEM. Differences between the groups were compared using one-way ANOVA or two-way repeated measures or three-way repeated measures ANOVA followed by Newman-Keuls post hoc test. We used Shapiro-Wilk test to assess all the data distribution. As for the homogeneity analysis, we used the p > 0.05; Fig. F1,14 = 1315, ***p < 0.001 vs. sham group; n = 8, two-way repeated measures ANOVA, Fig. F1,14 = 1315, ***p < 0.001 vs. sham group; n = 8, two-way repeated measures ANOVA, Fig. To verify the validation of BCP model, radiological imaging of rat tibia to assess the bone destruction 12 days after cancer cell inoculation was performed. Ipsilateral proximal epiphysis was disrupted in the bone marrow cavity 12 days post-inoculation , CD11 (for microglia), and NeuN (for neurons). Confocal images showed that CXCL1-IR was co-localized with the GFAP  and DAPI was performed. Results showed that pNFκB was increased in vlPAG on day 12 after BCP Fig. a, f, k aF4,35 = 862.5, ***p < 0.001, **p < 0.01 vs. BCP + serum group; ###p < 0.001, ##p < 0.01 vs. BCP + CXCL1 neutralizing antibody 4 μg group; n = 8, two-way repeated measures ANOVA, Fig. To elucidate whether suppressing vlPAG CXCL1 could inhibit pain-related behaviors following BCP, we injected intra-vlPAG into a CXCL1 neutralizing antibody  or control serum into the BCP rats once daily for four consecutive days  and then detected the analgesic effect of CXCL1 inhibition 1 h later after micro-injection on postoperative days (POD) 6, 7, 8, 9, 10, 11, and 12, respectively. The injection of CXCL1 neutralizing antibody (8 μg/0.5 μl) into the vlPAG of sham rats showed no detectable behavioral changes relative to the baseline values. Compared with BCP + serum group, micro-administration of CXCL1 neutralizing antibody significantly elevated the PWT of the BCP rats in a dose-dependent manner . The injection of BAY11-7082 (10 μg/0.5 μl) into the vlPAG of sham rats showed no detectable behavioral changes relative to the baseline values. Repeated BAY11-7082 treatment with a dose of 1 μg demonstrated no effect on mechanical allodynia, but treatment with 10 μg of this compound significantly attenuated mechanical allodynia. The effect started on POD 6, i.e., on the day of treatment initiation  into the vlPAG of sham rats did not produce detectable behavioral changes relative to the baseline values (data is being published). In order to examine whether the production of CXCL1 is required for the activation of astrocytes, we micro-injected an astrocytic cytotoxin L-α-aminoadipate (LAA 100 nmol) into the BCP rats once daily for four consecutive days (from days 6 to 9 after BCP). Furthermore, the GFAP immunoreactivity and CXCL1 protein level after the last injection were detected. Compared with sham group rats, the immunofluorescence intensity of GFAP and protein expression level of CXCL1 in vlPAG were increased on day 9 in BCP rats. After repeated LAA administration, the BCP-induced GFAP immunofluorescence expression was abolished  into vlPAG for nocifensive behavior in naive rats. The results showed that CXCL1 produced mechanical allodynia in a dose-dependent manner ; F2,245 = 136.2, ###p < 0.001 versus 5%DMSO + CXCL1 (100 ng); @@@p < 0.001 @p < 0.05 versus SB225002 (5 μg) + CXCL1 (100 ng); n = 8, three-way repeated measures ANOVA, Fig. CXCR2 is a major receptor of CXCL1 , 30. To F3,12 = 86.13, ***p<0.001 vs. sham group; n = 4, one way ANOVA, Fig. F4,15 = 21.59, ***p<0.001 vs. naive group; n = 4, one way ANOVA, Fig. F6,21 = 39.48, ***p<0.001, **p<0.01 vs. naive group; n = 4, one way ANOVA, Fig. We further investigated CXCR2 expression and distribution in vlPAG in BCP. Immunostaining showed a low expression of CXCR2 in the sham group rats Fig. a and an F4,35 = 1039, ***p<0.001 vs. BCP + 5%DMSO group; ###p<0.001, ##p<0.01 vs. BCP + SB225002 5 μg group; n = 8, two-way repeated measures ANOVA, Fig. To investigate the role of CXCR2 in vlPAG after BCP, CXCR2 antagonist, SB225002  was micro-administered. The SB225002 at a dose of 5 μg gradually showed an ameliorative efficacy from days 7–9, but 20 μg significantly reduced mechanical allodynia from day 6 to day 11 in a dose-dependent manner  and DAPI. Immunofluorescence triple staining showed that CXCR2 was increased in vlPAG from day 6 after BCP Fig. a, f, k aAmong the inflammatory chemokines, the role of CXCL1 in pain hypersensitivity has attracted considerable attention –18. HereCXCL1 is one of the major chemoattractants for neutrophils  and is kFurthermore, it was previously reported that CXCL1 is expressed intensely in spinal astrocytes, but not in the microglia and neurons in the pathological painstates , 17. NeuIt has been clearly proved that astrocytes in the central nervous system (CNS) play an essential role in several neuronal functioning aspects, including pain processing , 41, 42.Other chemokines, such as MCP1, are recently found to be involved in descending facilitation of neuropathic pain. Neutralizing CCL2 with an anti-CCL2 antibody into the rostral ventromedial medulla after spinal nerve ligation significantly attenuated the established thermal and mechanical hypersensitivity . In our NFκB, a transcription factor, is involved in the expression of genes related to the inflammatory process, including regulated CXCL1 expression . NFкB haThe PAG of the brainstem are considered as the essential structures of the endogenous pain modulatory system that alter the spinal cord processing of sensory input . DespiteTo check whether NFκB pathway in vlPAG was involved in BCP hypersensitivity, Western blot and RT-PCR results showed that the expression of pNFκB was gradually increased from 6 days to 18 days. We also showed that NFκB inhibitor reduced BCP and CXCL1 expression in vlPAG. These results indicated CXCL1 as an important downstream of NFκB activation in mediating the process of BCP and supported the role of NFκB in the maintenance of BCP via CXCL1 production in vlPAG.Our data further showed that pNFκB was predominantly expressed in astrocytes and neurons, but not in microglia of vlPAG. Consistently, pNFκB expression in astrocytes was found in the spinal cord following spinal nerve injury  and inocThe role of CXCL1 is dependent on its primary receptor CXCR2 , 55. AccCXCR2 was highly expressed by subsets of projection neurons in various regions of the spinal cord and brain , 59. TheOur present study suggests that inoculation of Walker 256 mammary gland carcinoma cells into the rat tibia induced bone destruction and pain hypersensitivity. In association with these changes, rats showed a prominent expression of CXCL1/CXCR2 in vlPAG, which in turn may activate astrocytes and neurons, respectively. Moreover, NFκB was involved in the production of CXCL1 in vlPAG astrocytes. Our data suggests that crosstalk between NFκB-dependent astrocytic CXCL1 and neuron CXCR2 plays a role in descending pain facilitation. Signal coupling between CXCL1 and CXCR2 receptors may lead to increased descending output from pain modulatory network, facilitating spinal nociceptive transmission and intensifying the perceived pain, and remains to be the focus of our future research.Additional file 1:Figure S1. A representative photomicrograph of a coronal slice at the level of the vlPAG stained with red fluorescent stain. Image shows the injection site in vlPAG. Coronal brainstem sections were stained with red fluorescent stain. Arrow indicates the cannulation track and circle shows the site of injection. Scale bars = 100 um; Aq, aqueduct. (TIF 5263 kb)


Which graft material is the optimal graft material for the treatment of aortic graft infections is still a matter of controversy. We used a branched xenopericardial roll graft to replace an infected aortic arch graft as a “rescue” operation. The patient is alive and well 37 months postoperatively without recurrence of the infection and any surgical complication. This procedure may have the possibility to serve as an option for the treatment of aortic arch graft infection. Orthotopic aortic reconstruction with an intraoperatively prepared xenopericardial branched roll graft was successfully performed as a rescue operation to treat a patient with a graft infection.1.1nd left posterolateral branch, 90%. All coronary artery bypass grafts were occluded. The origin of the left subclavian artery was ligated, and a 3rd graft branch was extended by a graft to the axillary artery via the left thoracic cavity. One month postoperatively, open‐chest drainage had been performed to treat left thoracic empyema. Subsequently, an incisional abscess was noted around the site of the left axillary anastomosis, and a graft‐branch‐to‐skin fistula was diagnosed. In 2013, at 64 years of age, the abscess was opened and the graft branch to the axillary artery was partially removed through a subaxillary skin incision. On admission to our hospital, the patient's serum C‐reactive protein level was elevated to 22.5 mg/dL, and a blood culture was positive for methicillin‐sensitive Staphylococcus aureus. The patient gradually became drowsy, and brain magnetic resonance imaging revealed multiple fresh cerebral infarctions and severe right internal carotid artery stenosis  for acute myocardial infarction at 55 years of age, and coronary artery bypass graft surgery x4 using left internal thoracic artery, radial artery, and saphenous vein grafts at 56 years of age. In 2012, at 63 years of age, a triplex 26 mm  had been used to perform a total aortic arch replacement procedure to treat a true aortic arch aneurysm at a nearby hospital. A reversed elephant trunk was inserted to facilitate the distal anastomosis using stepwise distal anastomosis technique. Preoperative coronary angiography showed the following stenosis: proximal left anterior descending artery, 75%; 1st diagonal branch, 75%; 1st left posterolateral branch, 75%; and 21.22 pericardial sheet  was simultaneously prepared intraoperatively from bovine pericardial sheets on a separate, sterile operating table. It takes 30‐40 min. to prepare a branched xenopericardial sheet. The method used to prepare the branched xenopericardial roll graft has been described in detail previouslyA cardiopulmonary bypass through a re‐resternotomy was established by means of two venous cannulas and an inflow cannula into the graft. The left subclavian abscess cavity communicated with the graft. Because of the dense adhesions, the left‐side of the heart was not dissected, and the coronary artery bypass graft was abandoned. Under deep hypothermic circulatory arrest with intermittent pressure‐augmented retrograde cerebral perfusionAs of 37 months postoperatively, the patient has been free of local recurrences of the graft infection, of pseudoaneurysms on the suture lines, and of other graft‐related complications, including of the branches  together with a xenopericardial sheet may be another option to achieve partial reconstruction of the aortic archIn conclusion, accumulation of clinical cases and confirmation of the long‐term durability of branched xenopericardial grafts may demonstrate their advantage as one of the options for the treatment for native aortic and aortic arch graft infection.Additional supporting information may be found online in the Supporting Information section at the end of the article.Supplementary VideoClick here for additional data file.


The elucidation of the molecular mechanisms of starch synthesis and mobilization in perennial woody tissues is of the utmost scientific and agricultural importance.Starch is the main carbohydrate reserve in plants and is fundamental in human nutrition and several industrial processes. In leaves, starch accumulated during the day is degraded throughout the night and the resulting sugars, glucose and maltose, are exported to the cytosol by the specialized transmembrane translocators pGT and MEX, respectively. Nevertheless, the degradation of the starch granule is a complex process not completely elucidated. While the mechanisms of starch mobilization during germination in the dead endosperm of cereal seeds are well described, the molecular and biochemical mechanisms involved in starch storage in the heterotrophic tissues of woody plants and its utilization in spring and winter are still puzzling. It is known that some biochemical steps of starch synthesis are conserved in heterotrophic tissues and in the leaves, but some aspects are particular to sink organs. From an agronomic standpoint, the knowledge on starch storage and mobilization in woody tissues is pivotal to understand (and to optimize) some common practices in the field that modify source–sink relationships, such as pruning and defoliation. Soluble sugars resulting from starch are also pivotal to cold adaptation, and in several fruits, such as banana and kiwifruit, starch may provide soluble sugars during ripening. In this review, we explore the recent advances on the molecular mechanisms and regulations involved in starch synthesis and mobilization, with a focus on perennial woody tissues. The transition from hunter-gathering to sedentary agriculture, also called the “Neolithic revolution”, occurred in different locations around the world. This revolution, that preceded the development of large and complex civilizations, was characterized by the cultivation of starchy staples, such as cereals . These starch excess (sex) potato mutant where a plastidic BAM is downregulated  and limit dextrinase (LDA), are also involved in starch degradation because isa3 and isa3/lda knock-out mutants display sex phenotypes. The role of α-amylases (AMY) in starch degradation in the leaves is still unclear and seems to vary between species. For instance, the Arabidopsis mutant amy3 does not display any sex phenotype  into the plastid to synthesize starch , found in roots, woody tissues, fruits, seeds, tubers, and pollen grains, relies on carbon obtained from long-distance sugar transport through the phloem Figs. . AlthougBrittle1 gene  is present in both source and sink tissues and it is exclusively localized inside the plastid in all tissues except in the cereal endosperm, which despite possessing the plastic form most of its activity is cytosolic  and translocates stachyose, most of the enzymes involved in raffinose synthesis are localized in the cytosol . Also, a pGT from rice is highly expressed in the seeds during development  for higher cluster/leaf ratios (Silva et al. As shown above, in woody plants, each growing season merges with the previous and following ones through the carbohydrate reserves accumulated in woody tissues, making this topic of the outmost agronomical and scientific importance. In grapevine, delaying pruning after bud burst could deplete reserve carbohydrates and the reiteration of this practice may cause carry-over effects on vine vigor and yield (Moran et al. Food scarcity due to an increase in the global population is a problem that is a challenge for scientists and policy makers, thus, unraveling the mechanisms of starch synthesis in both auto- and heterotrophic tissues is of the utmost scientific and societal importance. As shown in this review, several enzymes and plastidic proteins, such as MEX and pGT, modulate carbon allocation in plant tissues, but their role in starch mobilization in sink organs, such as woody tissues, deserves further investigation. Several experimental constrains, including the lack of model plants amenable for transformation and the absence of mutants, limit the study of these processes in woody tissues. Nevertheless, the advances that have been made in the understanding of starch metabolism in Arabidopsis and maize, together with the increasing availability of next-generation sequencing tools, will allow researchers to obtain a better picture of the molecular processes that occur in woody tissues.HG and HN conceived the idea of the review and prepared the first draft. All authors added new information to the manuscript, read and approved the final version.